A James-Bondesque View at the Real Situation

Stephanie Hoeppner, BSc, MBA, PhDCandidate

Deib A. Birkholz, MSc, PhD, P.Chem.

Milan Ralitsch, PhD, P.Chem.

What We’ll Talk About Today

Who Am I And Why Am I Even Here?

Environmental Determinants of Chronic Disease

The WHY

What We Found And What we do now - The AHA!

Water Sampling James Bond Style - The HOW

Bear with me…I’m going a different

way today than our usual talks…a

formal thank-you to the Friends of

Paracel for making this work

About Us…And Who’s This Paracelsus Guy?

He is credited as the founder of toxicology - Paracelsus pioneered the use of chemicals and minerals in medicine.

Open for over 25 years, we have more locations in Ontario than any other lab, and opened a Western Canadian lab in Calgary in 2014 – 3 years! We are in a partnership with Life Science Forensics. It is routine chemistry (Paracel) with specialty chemistry (LSF) under one roof. We have 6 PhDs, 1MD, 1DC and 3 MScs in our group.

We love crazy projects!

We start ‘em young

We are philanthropists

Cowboy chemistry isthe best chemistry!

Bailey the “Lab” Lab.

LABORATORY MANAGEMENT WITH CANCER AND OFFSPRING BORN 2004-

2007Rob Deverall, ALS– 2014, 59 – Lung Cancer

Don LaBerge, President of ETL – 2005, 57 – Brain Cancer

Type of Cancer Status

Brain Cancer - 2 Both Deceased <60

Paranasal Sinus Cancer - 2 Remission

Prostate Cancer - 1 Remission

Melanoma - 1 Remission

Lung Cancer - 2 Both Deceased <60

Breast Cancer – 3 1 Deceased <60, 1 metastatic – liver cancer

Cervical Cancer - 1 Remission

WE WILL ADAPT, RESISTANCE IS FUTILE! YOUR LIFE, AS IT HAS BEEN - IS OVER. FROM THIS TIME FORWARD,

YOU WILL SERVICE US.

=

EPIDEMIOLOGY

The branch of medicine that deals

with the incidence, distribution, and

possible control of diseases and

other factors relating to health.

THE GENOME OF A POPULATION

DOES NOT CHANGE IN 100 YEARS

We deal with environmental

determinants of chronic disease in

addition to dealing with infectious

disease.

OUR GOAL IS TO MARRY TOGETHER

THE FIELDS OF ENVIRONMENTAL

CONSULTING AND ENVIRONMENTAL

MEDICINE/TOXICOLOGY – AND THIS

IS TODAY’S POINT!

OK, SO WHAT?

• So what…

• Let’s go back to our example and look at it as an epidemiologist…

• 4 large multinational labs in Canada (plus Paracel, of COURSE!): not a huge amount of managers in the last 20 years.

• Cancer incidence rate: 24%

• Children born between 2004 and 2007: 16

• Children with chronic disease incidence rate: 87.5%

• Obviously this is not a big enough sample size to evaluate this tiny population…but it’s still worth thinking about, don’t you think?

CHRONIC DISEASE IN NORTH AMERICA

• Asthma in children under age five has increased by 160%

• Autism has increased by 1000% since the mid-1980s – NOT because of changes to the DSM IV to V.

• Congenital misplacement of the urinary opening in the penis, has increased by 100% and now affects 1 of 125 male babies born

• Cancer in children has increased by 26%

• Acute lymphocytic leukemia increased by 62%

• Brain and nervous system cancers increased by 50%

• Testicular cancer in young men has increased by 85% and is now the most common cancer in men ages 15–35

• 1 in 6 children are born with a neurodevelopment disability and the rate of ASD has increased from 1 in 10,000 in the 80’s to 1 in 66 (CDC, 2014).

As per the American Cancer Society, only 5–10% of all cancers can be attributed to inherited factors; the rest occur from environmental exposures and other damage throughout our own lifetimes.

The genome loads the gun but the environment pulls the trigger…

EPIDEMIOLOGY AND TOXICOLOGY

• Let’s add toxicology to the mix…

• Most people are aware that metals are an issue, but is that it?

• NO! Not even close.

• Metals bad, BPA bad…hexamethyl-chicken-wire?

• Acute toxicity is well understood

• Regulations – how are they made?

GET TO IT ALREADY, WHAT’S YOUR POINT…

JAMES BOND-ESQUE SAMPLING• We are not disclosing where we got our samples from BECAUSE

we are not here to get people in trouble, we are in it for the science only.

NOT

JAMES BOND-ESQUE SAMPLING

8 samplestaken over S,Central and N-Central AB

2 N-Central2 Central4 Southern

2 MunicipalDrinking Water6 Natural Source

•Chemicals that are usually found when doing this kind of analysis include:

• Antibiotics

• Steroids

• Antacids

• Antidepressants

• Anti-inflammatories

• Antipyretics

• Beta blockers

• Lipid lowering drugs

• Tranquillizers

• Stimulants

• Hormones

WHAT DID WE FIND?

(Magenta means we found them in both natural sources and in treated drinking water sources)-17B-estradiol (E2) - an estrogen made by humans and some other organisms.-Acetaminophen - (Tylenol). OTC Analgesic-Amitriptyline - (Elavil). Tricyclic antidepressant-Amoxicillin - Antibiotic-Amphetamine - could be an illicit drug, could be a by-product of ADHD drugs.-Bezafibrate - (Bezalip). Lipid-lowering agent to treat hyperlipidaemia-Caffeine - a common marker of human activity.-Carbamazepine - (Tegretol). Used to treat seizures and nerve pain such as trigeminal neuralgia and diabetic neuropathy, also Bipolar Disorder.-Codeine* - Weak opiate pain reliever.-Diazepam - benzodiazepine (Valium). Used to treat anxiety disorders, alcohol withdrawal symptoms, or muscle spasms.

WHAT DID WE FIND?

-Ephedrine - Decongestant and bronchodilator.-Erythromycin - Antibiotic.-Fenoprofen - Used for treating rheumatoid arthritis, osteoarthritis, mild/ moderate pain.-Gabapentin - (Neurontin). Anti-epileptic, for restless leg syndrome, epilepsy, neuropathic pain, shingles pain.-Ibuprofen - (Advil). OTC Analgesic-Gemfibrozil - helps reduce cholesterol and triglycerides (fatty acids) in the blood.-Metoprolol - (Lopressor, Betaloc). Beta-blocker. Used to treat angina and hypertension. It is also used to treat or prevent heart attacks.-*Nicotine - a common marker of human activity. In cigarette smoke.-Norfentanyl* - metabolite of opiate Fentanyl.-Norfloxacin - antibiotic. Usually used for urinary tract infections.

WHAT DID WE FIND?-Oxymorphone* - Moderate to severe pain opiate.-Pentoxifylline - (Trental). Used to treat claudication (pain from reduced blood flow) cramping, numbness, or weakness in the arms or legs.-Ranitidine - (Zantac). H2 receptor agonist - histamine-2 blockers. Reduces the amount of acid created in stomach.-Sildenafil (Viagra) - relaxes muscles found in the walls of blood vessels and increases blood flow to particular areas of the body. Used to treat erectile dysfunction in men.-Sulfamethazine - a veterinary antibiotic.Sulfamethoxazole - used for bacterial infections - urinary tract infections, bronchitis, and prostatitis - effective against both gram negative and positive bacteria (Listeria monocytogenes/E. coli).-Tamoxifen - A drug used to treat certain cancers.-Tetracycline - antibiotic for skin infections and Lyme disease.-Trimethoprim - antibiotic. Used to treat bladder/kidney/ear infections/C.Diff infections. *Note that opiates and some compounds were hard to distinguish and these are the databases’ “best guess” more so than the others. They are opiates; however, specific characterization needs work.+Also note that for the most part, if there was a metabolite that was picked up, we translated it into the parent analyte for ease of use.

WHAT DID WE FIND?

Solely Environmental Parameters- Di-n-butylphthalate - used to make plastics more flexible and is also in carpet backings, paints, glue, insect repellents, hair spray, nail polish, and rocket fuel.-Indole - Indole-containing compounds are best known for their medicinal properties in the pharmaceutical industry. Natural jasmine oil, used in the perfume industry, contains around 2.5% of indole.-3,4-Methylphenol - widely used, cosmetics and oral care applications.-Phenol - Phenols are widely used in household products and as intermediates for industrial synthesis. For example, phenol itself is used (in low concentrations) as a disinfectant in household cleaners and in mouthwash.

WHAT DIDN’T WE FIND?

• Benzophone - a sunscreen agent, but duh. Common in other similar studies and previous studies of ours.

• Glyphosate - again, it was winter though, so duh. Found in previous studies.

• Any of the major pesticides or herbicides.

• These samples were mostly frozen and taken from the shoreline; therefore, circulation was extremely limited. No sediment samples were taken. No tissue samples were obtained. Future studies will include this.

WHAT ABOUT TOXICITY THOUGH?

Petrie, B., Barden, R., & Kasprzyk-Hordern, B. (2014

ANTI-DEPRESSION INTERESTING FACTOIDS

Consumption of antidepressants in Canada is the third-highest among 23 developed countries surveyed by the organization, with 86 doses consumed daily per 1,000 people. Only Australia (89 doses per 1,000 people) and Iceland (106 doses per 1,000 people) ranked higher.

In 2013 the Canadian Pediatric Society officially approved the use of one class of antidepressants -- which includes Prozac -- for children.

- There are NO long-term studies on how chemicals that affect neurochemistry affect humans still undergoing neurogenesis.

Good Use of Money?

Was that correct?

When Relying on Point Source Data Alone:

Extremely hard, if not impossible, to identify if there is a human health concern.

There are few, if any, reference ranges for these compounds, even within the large bio-monitoring projects, which are in the wrong matrix anyway.

There is no way to know what parameters are having an effect if it is a multi-analyse mixture.

Possibly incorrect assignment of liability, if that is the goal.

USEPA’s Toxcast Program

• ToxCast has evaluated over 2,000 chemicals from a broad range of sources including: industrial and consumer products, food additives, and potentially "green" chemicals that could be safer alternatives to existing chemicals.

• Chemicals were evaluated in over 700 high-throughput assays that cover a range of high-level cell responses and approximately 300 signalling pathways.

High-Throughput ScreeningUSEPA Approved Methods Through Toxcast

Several Different Types Possible

Endocrine Disruption

Neurotoxicity

DNA Damage and Oxidative Stress

Mutagenicity

Toxicological Specificity of Additive Mixtures

Results of Preliminary ScreeningEndocrine Disruption

Mutagenicity Neurotoxic Effects

Drinking Waters Yes No No

Southern Alberta Yes No No

Central Alberta Yes No Yes

Northern Central Alberta

Yes No Yes

Soil, Water, Blood…it’s all just small molecule chemistry...

IBM’s Watson shows us what’s actually important…

IBM’s Watson shows us what’s actually important…

IBM’s Watson shows us what’s actually important…

Cost Savings:If you did the sampling like was done here, it would cost you ~$60,000.

Looking at Toxcast data, we could have narrowed that way down to ~<$15,000.

Forensic Analysis:

If we had done this for a forensic investigation, we would have significantly more clout looking at both point-source and ToxCast data together than just one alone.

Final Conclusions!

Next Steps:

Determine LC50 and reference values for implicated analytes

Clinical samples: Blood/urine/stool for persistent organic pollutants, vitamins, genomics and many more parameters “Personalized Medicine”

Fecal Sterols

SPMDs

Adverse Health Effects in Air Screen

Graphene nanoparticles and PFCs

Deformulation – I want to test for “everything”

Pharmaceuticals and personal care products in environmental samples

Human Biomonitoring

Emerging contaminants of concern

Novel food sensitivities screen

Metabolomics and other “omics” testing

Diagnostic Screens for MS and Autism

The list goes on…

Lastly, Other Cool Stuff: We don’t DO no.

Bailey the Lab Lab Says Thank-You! Questions?

Life Science Forensics www.lifescienceforensics.com

HASAT™: Allergy & Sensitivity Testing

Antitissue

Trans-glutaminase

lgAAntibodies

Antigliadin

Completely novel to Canada: High Accuracy Stool Allergy Testing (HASAT™) can be completed regardless of previous non-gluten consumption. Significantly more robust and sensitive over other available options. Stool analysis is the best way of assessing for gluten sensitivity before celiac disease develops. We combine a single malabsorption test with HLA gene testing, without reintroducing gluten into the diet for a simple, non-invasive and accurate test.

Life Science Forensics www.lifescienceforensics.com

Xeno75: Environmental Toxicant Testing

Tests for the 75 most ubiquitous foreign chemicals and their metabolites, including:

• Polyaromatic Hydrocarbons (PAHs)• Polybrominated diphenylethers (PBDEs)• Pesticides• Parabens• Polychlorinated biphenyls• Phthalates

Heavy Metals

Pesticides

HerbicidesPlasticizers

Preservatives

Life Science Forensics www.lifescienceforensics.com

MetaComplete: Optimal Nutrition Testing

Vitamins

Minerals

Amino Acids

Fatty Acids

Metabolites

Tests for important vitamins, nutrients, and other important metabolites required for optimal health, including:

• Amino acids• Vitamins• Minerals• Fatty acids• Metabolic intermediates

Life Science Forensics www.lifescienceforensics.com

Heavy Metal Testing

Life Science Forensics can determine metals concentration in:

• Blood• Urine• Hair

Our metals test looks at a number of metals including:

• Gadolinium• Mercury• Thallium• Lead• Cadmium

Biomarkers included: auto immune dysfunction, immune response and related inflammation, body burdens of xenobiotics (persistent organic pollutants and metals) and related ability to detoxify naturally, oxidative stress, methionine dysfunction, gastrointestinal dysfunction, abnormal gut flora, abnormal protein functions, abnormal levels of amino acids and other organic acids and associated metabolites, specific genetic polymorphisms, mitochondrial dysfunction, and abnormal cytokine, neurotransmitter and hormone levels.

Our screen is unique and the most complete of anypotential testing to date. To test for all theparameters above would cost thousands of dollars.Our screen focuses on the most commonly foundbiomarkers and encompasses the likely dysfunctionsthat occur in autism AND many other chronicdiseases.

Contact one of our scientists today for moreinformation and pricing details.

We are ASD moms who are also scientists. It is ourmission to help, heal, and teach.

BioHF