On gut parasites and chronic fatigue

An interesting exchange on Twitter prompted this short post regarding a paper by Naess and colleagues* (full-text) on Giardia lamblia gastroenteritis and chronic fatigue syndrome (CFS). The tweets concerned another parasitic nasty called Toxomplasma gondii which has featured quite a bit on a sister blog with regards to its link to various behaviourally-defined conditions. I thought that T.gondii was a spine-tingling protozoa until someone posted about these other chaps and their brain-eating, behaviour-changing and belly exploding antics (pass the sauce, please).

Giardia lamblia is quite a special  protozoa in terms of its survival, persistence and ability to link into quite a few other health complaints particularly of the gastrointestinal variety and specifically links to lactose intolerance. The current observations by Naess et al are interesting in that based on an examination of over 1200 patients with laboratory-confirmed giardiasis following a large community outbreak in Bergen, Norway, approximately 5% of cases (58/1262) were diagnosed with CFS as classified by the CDC criteria (see here for a related post on the trials and tribulations of diagnosing CFS/ME).

Even assuming a CFS prevalence of 1% previously noted in children (not adults) in the UK, the 5% figure seems high bearing in mind correlation is not necessarily causation. What can perhaps be ascertained from this latest study is that it might be a good idea to screen for giardiasis where active functional bowel issues are present alongside fatigue-related conditions and further research on any mechanism of parasitic infection linked to long-term fatigue might be advisable.

* Naess H. et al. Chronic fatigue syndrome after Giardia enteritis: clinical characteristics, disability and long-term sickness absence. BMC Gastroenterology. February 2012.
DOI: 10.1186/1471-230X-12-13

Plasma amino acids and inflammatory bowel disease

My recent post on the application of metabolomics to food fingerprinting got me thinking about the examination of biological fluids and how our metabolome might have some interesting secrets to one day share. The 'promise' of the science of metabolomics is that one day, we should be able to look at various biological fluids across a range of conditions and based on the compounds excreted/detected determine diagnosis, disease progression and how well someone responds to intervention. I hasten to add that we are nowhere near that position yet.

In light of this, it was perhaps inevitable ('it is your destiny') that this paper by Hisamatsu and colleagues* (full-text) on possible plasma biomarkers for inflammatory bowel disease (IBD) would get some attention. I'm pretty sure that I don't have to explain this to viewers but the term inflammatory bowel disease covers quite a bit of diagnostic ground including Crohn's disease and ulcerative colitis. Diagnostic confirmation of these conditions is quite a complicated process normally involving a combination of peripheral measures (blood tests, stool analysis) coupled with more direct observation of the bowel.

Hisamatsu and colleagues reported that a previously trialed network analysis of plasma amino acid levels in patients diagnosed with IBD might provide novel, non-invasive and importantly, objective biomarkers potentially opening up some new research areas into the nature of IBDs.

The details:

• Fasting plasma 'aminograms' for a discovery group of 102 Japanese adult patients diagnosed with Crohn's disease (CD) and 102 patients with ulcerative colitis (UC) were initially compared against 102 healthy control participants. The majority of participants were male (70%) with disease duration ranging between an average of 7.8-11 years. A minority of participants were described as having 'active' disease (CD = 29/102; UC = 38/102).  Obtained aminogram results were also validated using a validation set of participants (CD: n=63; UC: n=120; controls: n=108).
• Serum albumin levels were reported as significantly lower in the IBD groups compared to controls.
• Various differences were noted between the groups in terms of amino acid levels. The authors seemed to have concentrated on histidine and tryptophan as primary examples with a view to disease activity and correlation with C-reactive protein levels (a marker for inflammation).
• Implementing their network analysis (MIAI), the authors came up with a formula based on 6 amino acids which discriminated CD and UC groups from controls with ROC values ranging from 0.894 to 0.955 depending on whether the discovery or validation group were used and comparing across the IBDs with controls. 
• Depending on whether disease was active or in remission, a formula incorporating data for 7 amino acids was suggested to have some power in discriminating disease activity with ROC values ranging from 0.894 (CD active vs. CD remission) and 0.849 (UC active vs. UC remission).

The data produced by this study is interesting. OK, it is not totally 'diagnostic' either in terms of classifying IBD from controls or looking at active vs. remissive symptom presentation but it's not a bad start at all. Indeed the use of discovery and training sets takes me back to a wonderful paper covered last year on schizophrenia, which did find a perfect classification based on a handful of serum and urinary markers. It makes me wonder if they were looking at serum and urine at the same time or maybe incorporating a few more well-known markers, whether those ROC values might further approach the magical number 1 (denoting a perfect classification).

The use of statistical models allied to biochemical data is also interesting. I note that similar linkages have been used in other areas of medicine, possibly even to recreate speech from brain activity...?

* Hisamatsu T. et al. 2012 Novel, objective, multivariate biomarkers composed of plasma amino acid profiles for the diagnosis and assessment of inflammatory bowel disease. PLoS ONE. January 2012
DOI: 10.1371/journal.pone.0031131

Newsflash: defining gluten-related disorders

In the style of one Homer J Simpson... can't stop... must finish for the day.. new guidance of what constitutes a gluten-related disorder just published by Sapone and colleagues*.

If there is one document that you absolutely have to look at which summarises where we are in relation to gluten-related conditions, not just coeliac disease, this is it.

It's full-text, has a myriad of gluten research names included on it (including Alessio Fasano, Marios Hadjivassiliou and David Sanders), so enjoy.

* Sapone A. et al. Spectrum of gluten-related disorders: consensus on new nomenclature and classification. BMC Medicine. February 2012.
DOI: 10.1186/1741-7015-10-13

Pesticides and vitamin D deficiency

Chemistry World, the public face of the UK Royal Society of Chemistry (RSC), carried an interesting report recently discussing research linking exposure to organochlorine pesticides (OCs) with vitamin D deficiency. The research in question is this paper by Jin-Hoon Yang and colleagues* (full-text) and before you ask, yes, it is a study of 'association', so we tread carefully.

Before wading into this study it is interesting to note that vitamin D is currently enjoying quite a trendy following in many areas. Important discussions are underway to determine whether a resurgence of conditions like childhood rickets means supplementation needs to be more closely inspected. 

I digress. A summary of the paper in question:

• An analysis of plasma levels of 7 OCs (including DDE and DDT) was undertaken for 2,337 people aged 12 and above recruited as part of the US National Health and Nutrition Examination Survey (NHANES). After exclusion of those where accompanying 25-hydroxyvitamin D (25(OH)D) were not available alongside other exclusions (e.g. pregnant women, being under 20 years old), the final participant group number was N=1275.
• The results: based across various grouping on concentrations of OCs, there were quite a few significant inverse relationships reported. That is, for the OCs -  p,p′-DDT, p,p′-DDE, and β-hexachlorocyclohexane - elevated plasma levels of these compounds individually and collectively were associated with lower vitamin D levels. Plasma levels of DDT in particular showed quite an enduring association with vitamin D levels. 
• There is a suggestion that the relationship may also be dose-dependent up to a certain point; that is up to a value of 200 ng/g lipid of DDT, vitamin D levels dropped, but increasing levels of DDT after that seemed to be linked with increasing vitamin D levels bearing in mind that most participants presented below this 200 ng/g lipid threshold.

As I said, this is a study based on association. Controlling for factors such as gender, age, race and vitamin D supplementation is an admirable quality of the study but association is normally only a guidepost to a possible relationship, not a dead cert. That and the fact that little information has been provided on why the results came out as they did, leaves the door open to further study in this area.

Noting that this blog is primarily concerned with the gut, I do wonder about these recent findings and how they may (or may not) fit into a past post on gut bacteria and organochlorine pesticides. It's a bit of a long shot but how about testing the suggestion that OCs affect gut bacteria which in turn affects other systems including vitamin D and its receptors?

* Yang J-H. et al. Associations between organochlorine pesticides and vitamin D deficiency in the U.S. population. PLoS ONE. January 2012. DOI:10.1371/journal.pone.0030093

Metabolomics and the food fingerprint

I've touched upon the beautiful science of metabolomics previously on a sister blog; that is, looking for 'chemical fingerprints' in various biological fluids to garner further information on underlying biological processes related to health. Already metabolomics has provided us with a few preliminary clues in relation to more behaviourally-led conditions such as autism and schizophrenia (AUC=1), but as the technology gets smarter and smaller (and cheaper), I foresee great things ahead for this flourishing branch of biochemistry similar to the genomic high-throughput screening technology carried out at places like the Sanger Institute.

When a story recently appeared on the BBC website discussing a food 'fingerprints' test, it was therefore always going to attract my attention. Indeed so much so that after a few quick emails to one of the people involved with the research, I was very satisfied to receive some of their peer-reviewed papers on the details behind the headlines. 

The BBC story discusses some collaborative research based on the notion that what we eat leads to the production of various chemical metabolites and that detecting those metabolites in fluids like urine means we can objectively find out what someone has consumed recently and if necessary promote any dietary changes. Food diaries are the normal way of collecting information on what people have eaten recently but recall is often not as accurate as you think.

My take on this work was slightly different in terms of the fact that the researchers are in effect, building up a large bank of information based on very controlled eating habits about what comes out when we eat certain foods. 

This paper by Favé and colleagues* (full-text) sums up the research aims and objectives; also providing a great overview of the various techniques used in metabolomics and importantly the statistical analyses required (normally PCA or some variant). The researchers have been very methodical in their approach to this issue. This paper again by Favé and colleagues** (full-text) details how they went about clearing a few potential interfering variables such as the fasting period and some information on the reproducibility of findings over various testing occasions. In short, they wanted to make sure that interesting collected metabolites were food derived and were consistent.

So what have they got so far?

Well, according to this paper by Lloyd and colleagues*** when compared to a standard breakfast made up of orange juice, tea with skimmed milk and sugar, butter croissant, and cornflakes with milk, an alternative dietary schedule made up of either salmon, broccoli, whole-grain wheat cereal or raspberries produced some notable differences in urinary metabolites. The differences were even more notable when the alternative dietary schedule was compared against fasting urine samples.

Looking more specifically at individual compounds formed as a result of dietary intake, a few interesting snippets were reported including:

• Smoked salmon intake was associated with increased levels of 1-methylhistidine and anserine (b-alanyl-L-methylhistidine), metabolites of the amino acid histidine, probably derived from fish skeletal muscle. Also alongside were findings of TMAO, a degradation product from carnitine (remember that?).
• Broccoli and raspberry were both associated with increased ascorbate and individually, xylonate/lyxonate and polyphenols.
• For each of these three dietary components, there were at least 7 putative biomarkers of intake reported although nothing was reported on for the wholegrain cereal consumption.

Another paper by Lloyd and colleagues**** reported that proline betaine was associated with citrus exposure, with detected levels using the same technology and methods explanatory of both acute and habitual exposure to citrus containing foods (makes it sound like an illicit drug or something).

Appreciating that this and related work is still an emerging area of investigation, subject to all manner of confounders that we as human beings do to ourselves every day, I am suitably impressed that there are groups out there looking at our food and how we metabolise it. I can envisage many applications for this kind of work, ranging from a non-invasive test of recent food consumption, to measuring adherence to certain kinds of dietary intervention, to examining whether there are any differences in the metabolism of certain foods as a function of say gut bacteria which might cast some light on the biology behind lots of conditions.

To finish how about a bit of artistic interpretation through the medium of dance from Kate Bush...

* Favé G. et al. Measurement of dietary exposure: a challenging problem which may be overcome thanks to metabolomics? Genes & Nutrition. 2009; 4: 135-141

** Favé G. et al. Development and validation of a standardized protocol to monitor human dietary exposure by metabolite fingerprinting of urine samples. Metabolomics. 2011; 7: 469-484

*** Lloyd AJ. et al. Use of mass spectrometry fingerprinting to identify urinary metabolites after consumption of specific foods. American Journal of Clinical Nutrition. 2011; 94: 981-91.

**** Lloyd AJ. et al. Proline betaine and its biotransformation products in fasting urine samples are potential biomarkers of habitual citrus fruit consumption. British Journal of Nutrition. 2011; 106: 812-824.

Greens, glutathione and the mucosal barrier

'Eat your greens' is a phrase most of us will have heard on several occasions when growing up. I have to admit to being a bit of a 'goody two-shoes' in this respect, having had a lasting affection for things like Brussels sprouts and broccoli. Indeed, sprouts still tend to be on my menu several times a week (boiled with a drop of Greek olive oil).

I say all this because an interesting article published a few years back has caught my eye recently. The paper by Hoensch and colleagues* (open-access) looked at the various factors potentially affecting the functioning of the gastrointestinal glutathione system with some interesting observations.

I will admit that glutathione (GSH) is something which I am becoming very interested in at the moment. Over on my autism research blog, I recently discussed the pretty remarkable findings emerging in some cases of autism with regards to plasma GSH which might (might!) eventually have some diagnostic usefulness in combination with other factors. The Hoensch paper talks about GSH levels in the upper gastrointestinal mucosa and how factors such as diet, some medications and even gender seemed to affect levels of GSH and accompanying enzyme activity (glutathione S-transferase, GST).

The particulars:

• Biopsy pinches taken from two sites (antral and duodenal mucosa) for 202 adults (104 males: 98 female) undergoing endoscopy were analysed for GSH and GST activities. Various background information was also taken from participants including details of family history, medication and current dietary habits via a food frequency questionnaire.
• The findings: different biopsy sites reflected different levels of GSH and enzyme activity. Female participants showed higher levels of GSH and GST activity in their antral mucosa samples than males. High intake of vegetables (more than 3 days a week) seemed to enhance aspects of GST activity.
• A subsequent author reply to a suggestion that Helicobacter pylori infection might also have had an effect suggested that indeed, one aspect of GST activity was negatively affected by H.pylori infection.

Bearing in mind that GSH and its related sub-systems represent an important part of our defences against those dastardly free radicals among other things, making sure that the system is in tip-top condition is probably quite important. I don't want to make sweeping gender generalisations but the fact that females seemed to be in a slightly more advantageous position compared with male participants leads me back to some interesting work on the fragile male. Having said that, let's not be too defeatist here; greater vegetable consumption seemed to have an enhancing effect as per other results so one could argue, guys in particular, eat more greens - especially more brassica vegetables to support your mucosal antioxidant defence system.

* Hoensch H. et al. Influence of clinical factors, diet, and drugs on the human upper gastrointestinal glutathione system. Gut. 2002; 50: 235-240.

Gut bacteria and heart health?

Happy (belated) New Year! Welcome back to Gutness Gracious Me in 2012. I start this year with a post on something pretty central to the ethos of this blog: gut bacteria.

I should perhaps first apologise to Alex Gazzola (#HealthJourno) who very kindly asked if I would be discussing the new Codex standards for labelling foods and gluten-free and the 20 parts per million threshold introduced. I did say I would have a look at this and I am still looking. I do however think that Alex has done a wonderful job of covering this topic on his own blog (see here) as per the reader response he has received.

Back to task. Despite being only 13 days into the New Year (Friday 13th...mmm?), already the research is coming thick and fast. Over at my Questioning Answers blog, the big news recently has been on the bacteria Sutterella and its detection via various methods in quite a few biopsy samples from children with autism and gastrointestinal (GI) issues. I say big news but in among the very detailed explanation of how that team came to find Sutterella, the main story for me is the suggestion that gut hyperpermeability in some cases of autism might be a route through which the immune system meets gut bacteria in places it really shouldn't and onwards formulates an antibody response. The template for this is Crohn's disease and leads into some interesting suggestions about whether gut bacteria itself might not necessarily be the bad guy but rather what happens when it is allowed to roam.

The other quite interesting news is from this paper by Vy Lam and colleagues* suggesting that different amounts and varieties of gut bacteria might influence the severity of and recovery from heart attacks in rats. I am well used to seeing papers talk about gut bacteria in relation to GI issues and conditions, even conditions like autism where functional and more systemic bowel disorders have been noted in some cases. This is however one of the first times that I have come across gut bacteria potentially so directly influencing the physical health of an organ like the heart.

So what did the researchers do?

• Three groups of rats were fed three different types of diet: a standard diet, a standard diet plus quite a powerful antimicrobial (vancomycin) and a standard diet plus a probiotic very aptly named GoodBelly. 
• The primary probiotic constituent of GoodBelly is Lactobacillus plantarum 299v quite commonly found in fermented foods like sauerkraut and subject to a few claims. One of those claims is that the bacteria might be able to reduce the production of leptin among other things.
• Both the vancomycin and GoodBelly supplemented groups showed a decrease in circulating leptin levels (38% and 41% respectively) alongside some changes to gut bacteria as one might expect.
• The vancomycin and GoodBelly supplemented groups also showed evidence of smaller heart attacks and improved recovery after heart attack compared with the standard diet group.

Whilst the authors describe this as a 'proof-of-concept' study, there are potentially many implications from this work not least that our gut bacteria might do far more than call us home and help digest our food. I have previously (jokingly) referred to gut bacteria as being our 'masters'. Assuming the results from this study are pertinent to humans as well as rats, I might well be offering a sacrifice to 'those who must be obeyed' in the near future.

Should we be surprised that the human body is interconnected? No probably not, although it might take a while and a few more bits of evidence of effect before cardio health care professionals start giving probiotics to help treat (prevent?) myocardial infarction (please note I am not giving any medical advice about this). Going back to the Sutterella post and the proposed link between bacterial translocation and gut permeability, I do wonder whether research should also be looking more closely at gut permeability and lots of other conditions just to see if findings such as these might provide a few more clues about health and illness.

*Lam V. et al. Intestinal microbiota determine severity of myocardial infarction in rats. The FASEB Journal. January 2012