Latest in Urinary Pyrrole Research

Latest in Urinary Pyrrole research – Applied Analytical Laboratories (AAL).

Below is the latest research from AAL, which although written for practitioners who send samples to the lab for testing from their clients, Brett from the lab has given me permission to share this valuable information with you. Whilst a lot of it may be quite technical, you will still be able to gather the important bits of information relevant to you. If not, feel free to pass this info on to your practitioner for further interpretation. Please pay particular attention to the details about collection of your sample.

The objective of this newsletter is to inform practitioners of the recent advances made to our urinary pyrrole test that further distinguish it from the others. Our test is in review for NATA ISO 15189 (medical) accreditation.

The information disclosed is to inform practitioners that the research we are carrying out is novel, and we have the scientific competencies to make leading discoveries in the field. We aim to separate ourselves from our national and international competitors in providing research that backs up our results.

There is a number of Ehrlich “positive” components found in urine that can be distinguished and it was brought to our attention by NATA during an assessment that the urinary pyrrole test was susceptible to significant interference by urobilinogen. We found this to be accurate following spiking studies where known quantities of urobilinogen were added to samples (the effect of which was measured).

Our spiking studies revealed data that enables us to now provide corrected results for the Urobilinogen, which is estimated to cause interference to the Ehrlich reading by approximately 40% (urobilinogen can cause an approximately 40% increase to the reading, but is different for all samples).

We found that small amounts of blood as well as supplementation with vitamin B6 do not interfere with results prior to testing.

However treatment strategies relying on uncorrected/false positive or false negative results, have given us cause for concern regarding general B6 toxicity. The role of Zinc and magnesium are of increasing importance.

Key Research
Pilot study research (involving 567 patients) to investigate the viability of diagnosis and treatment using “Pfeiffer/Walsh” regime;
ACNEM Journal; Vol 29 No.3 – 2010 “The Effectiveness of Targeted Nutrient Therapy in Treatment of Mental Illness – A pilot Study”; Stuckey, Walsh, & Lambert.
Study to verify pyrroles as a bio marker for schizo-effective psychosis (4 peer reviewed publications to-date, another in prep)
1 Biomarker Research, (2015) 3:3 DOI 10.1186/s40364-015-0028-1; “Biomarkers of a five-domain translational substrate for schizophrenia and schizoaffective psychosis” Fryar-Williams & Strobel.
2 Open Journal of Psychiatry, 2015, 51 78-112 dx.doi.org/10.4236/ojpsych.2015.51011; “Biomarker Symptom Profiles for Schizophrenia and Schizoaffective Psychosis”, Fryar-Williams & Strobel.
3 Frontiers in Psychiatry, 2016; doi 10.3389/fpsyt.2016.00048; “Biomarker Case-Detection and Prediction with Potential for Functional Psychosis Screening : Development and Validation of a Model Related to Biochemistry, Sensory Neural Timing & End Organ Performance.” Fryar-Williams & Strobel.
4 Frontiers in Psychiatry, 2016; doi 10.3389/fpsyt.2016.00172; “Fundamental Role of MTHFR 677C-T Genotype and Flavin Compounds in Biochemical Phenotypes for Schizophrenia and Schizoaffective Psychosis”, Fryar-Williams.
Study of treatment outcome based on Walsh protocols for child and adolescent violent offenders –
Journal of Child and Adolescent Psychopharmacology; doi 10.1089/cap.2016.0199 “Micronutrient Therapy for Violent and Aggressive Male Youth: An Open-Label Trial”; Hambly, Francis, Khan, Gibbons, Walsh, Lambert, Testa, & Haywood.
Study validating the aetiology of the elevated bio-marker and the result of treatment (in prep)
Interference studies – samples spiked with potential interferences and their effect determined on results (NATA requirement – AAL in-house development)
Characterisation of the Ehrlich and pyrrole chemistry (Combined Griffith University and AAL research – ongoing)

Treatment response studies
AAL will be involved in the bio-marker project phase III arm (with over 800 participants). This Precision Scientific research project will be collecting data on a national scale.

The key findings our research has led us to so far are as follows:

1 We have characterized the components of the fraction tested by “Pfeiffer” affiliated laboratories and found the fraction to comprise of urobilinogen and to lesser concentration, bilirubin. Therefore laboratories using the “Pfeiffer” method are measuring the interference components – not the analyte, ie the clinical significance of their results are questionable.

2 We have characterised the mechanism and products of the Ehrlich reaction by NMR and have gained profound insight into the chemistry occurring – casting doubts over some “currently accepted” chemical structures.

3 Urine urobilinogen concentration naturally peaks in the afternoon, which is why we have previously and still recommend 2nd or 3rd morning voids for testing (as urobilinogen interference is minimized at these times). We can also provide urobilinogen results.

4 Spectroscopic studies we have performed have resolved the difference between derivatives of urobilinogen, bilirubin and a ‘hydroxyhemepyrrole-like’ compound. Further elucidation is on-going.

5 We can now provide urobilinogen corrected results giving the most accurate urinary pyrrole results of any comparable laboratory in the world.

6 More accurate results have aided more accurate treatment strategies and we have found B6 toxicity to be of increasing concern.

Some changes we have made over the past few months.

AAL corrects for urobilinogen interference, although we still recommend collection of morning voids to keep the error margins to a minimum, and to provide the most accurate result possible. It is essential that the time of sample collection is provided to us on our request forms that accompany samples, so that we can distinguish between elevated pyrroles, peaks in urobilinogen concentration, and provide the most accurate reading for both.

Measurement of urobilinogen concentration is important and a low urobilinogen result can be just as significant as a high result. (ie low urobilinogen (and low pyrrole) with normal S.G., often with a trace of blood can indicate biliary obstruction and should be investigated further).

AAL provides this measurement and provides additional measurements (leukocytes, protein, nitrites, ketones, glucose, bilirubin, and blood (lysed and whole) using a Siemen Advantix urinalysis system) on samples that are outside the reference ranges (ie, both high and low).

Sample Collection
The compounds of interest in the sample are unstable and reactive with natural and artificial UV light and x-rays. Thus, it is pivotal that whatever sample pots are provided, they must be covered with foil to protect and preserve the analyte. The opaque pots provided by other laboratories (without foiling) are inadequate. It is also essential the sample must be immediately frozen and must remain frozen until testing starts.

Reference Ranges –
We have made changes to the reference ranges for the results we provide, based on real population and study data (validation of the testing protocol).

Paediatric (<14yrs)
[HPL] < 10 ug/dL Normal
10 ug/dL < [uHPL] < 20 ug/dL Borderline
[HPL] ≥ 20 ug/dL Elevated

Reference Ranges – Adults
[uHPL] ≤ 40 ug/dL Normal
40 ug/dL < [HPL] < 150 u/dL Mild Elevation
150 ug/dL < [HPL] < 400 ug/dL Moderate Elevation
[HPL] ≥ 400 ug/dL Severe Elevation

The majority of results release is done by up-load through secure portal to practice software using an encryption service. As soon results are available, they’re uploaded to your system. If a result has not been uploaded to your practice within 6-10 workings days of collection, it means we have not received the sample – so please contact us urgently for follow up. All discrepancies are documented in Non-conformance registers.

In summary
The major difference medical practitioners should note is that it is now clear the “Pyrrole” test measures a BIOMARKER of oxidative stress and is not a confirmation of a mental health condition called “Pyrrole disorder”. This actually makes the test more important and more useful to practitioners.

By understanding that there is a direct correlation between severity of symptoms and the larger measurement range and that the product is a definite measure of oxidative stress there can be a better understanding of treatment response. The extra information provided to practitioners when the results are very low or very high give a very useful tool to investigate other conditions.

Currently it appears the treatment process should not markedly change except to be able to provide future illuminations as per the comments above. Please re-read the references cited above so you understand the research behind this clarification. Please be assured we are continuing to research to improve the understanding of this important mental health biomarker.

Feel free to contact us if you have any queries.
Yours Faithfully
Brett Lambert (MAppSc, BAppSc(Chem)), Principle Scientist and Director, AAL; October 2018
Shop 6, 11 Logandowns Drive,
Meadowbrook Qld 4131
Ph 07 3133 1615
E – info_apan@bigpond.com

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