Thursday, February 27, 2014

Autism, Heavy Metals, BDNF and HSP-70

Autism Spectrum Disorder (ASD) Testing using Custom Quantibody® Array

We recently completed testing blood serum from ASD children. These children were all from Central Europe and lived in cities with heavy industries. All had elevated levels of heavy metals like Aluminum and Copper. We believe that these levels may be contributing to the pathogenesis of the ASD symptoms.

Though the sample was small, the results were striking. The elevated levels of Heat Shock Protein 70 (HSP 70) is consistent with high heavy metal toxicity. Studies have reported both elevated and decreased levels of Brian Derived Neurotrophic Factor (BDNF) vs healthy controls, but is always reduced in subjects with high heavy metal blood serum levels. Could these elevated heavy metals be a root factor ASD Children?

Based on these findings, we have proposed a 2 pronged treatment strategy:
  1. Bring heavy metal serum levels into acceptable balance.
  2. Treat ASD children with natural stem cell enhancing substances
Stem Cells are immune/inflammation supressive and capable of repairing tissue damage. Our theory is that this could result in improvement in ASD related symptoms. Testing serum levels of BDNF, HSP-70 and other select bio-markers would confirm therapeutic efficacy.

I will be posting results updates here so stay tuned.

Tuesday, February 25, 2014

Kinetic Assays and Stem Cell Toxicology Studies

UCB Derived hMSCs and Cobalt

Here're results from a recent Kinetic Assay study we conducted using our Umbilical Cord Blood Derived Stem Cells:

Images and Figure: Images: Dose-Response curve for Co++ toxicity to Hoechst-stained hMSCs (UCB Derived catalog number SC00A1-HC). The bar graph on the lower right shows cell counts verses [Co++] at 24, 48 & 72 hours exposure to Co++. Results at 144 hours showed massive cell death. The initial increase in cell counts at low concentration may reflect the well-known activation of HIFs by Co++. Counts were determined by Hoechst-stained MSCs and simultaneous propidium iodide staining shows increasing numbers of permeable (presumably dead) cells at 24, 48 & 72 hours post-Co++ during exposure to 10% DMSO. Images acquired on Biotek Cytation 3 imaging system.

We are currently designing  assays for testing small molecules and compounds. These are customized for Musculoskeletel Diseases Drug Discovery. They will be released in Q2 2014.

We also offer CRO Services. We have the ability to test different analytes and their impact on: Cell Migration, Differentiation and Expansion. This could include the study of toxic analytes on these behaviors. We are also assaying various supplements that claim to endogenously boost stem cells or other natural substances like Li-VPA. We can do these studies using your stem cells or ours.

Questions or interest? I can be reached at or cell: 612-801-1007.

Thursday, February 20, 2014

More MOR Publications

Latest Mu Opioid Publications

Our Mu Opioid Receptor Antibodies have set the standard for the study of Pain Mechanisms. We have posted >40 publications referencing use of these antibodies.

Here's several published in 2014:

Charlie H.T. Kwok, Ian M. Devonshire, Andrew J. Bennett, Gareth J. Hathway. Postnatal maturation of endogenous opioid systems within the periaqueductal grey and spinal dorsal horn of the rat. PAIN - January 2014 (Vol. 155, Issue 1, Pages 168-178, DOI: 10.1016/j.pain.2013.09.0220. ...rabbit anti-MOR (Neuromics, Edina, MN, USA; 1:1000 with tyramide signal amplification protocol)...

Images: Immunohistochemical expression of opioid peptides and receptors in the DH (spinal cord dorsal horn) during postnatal development. (A) POMC (pro-opiomelanocortin) immunoreactivity in the dorsal horn in postnatal day (P)10, P21, and adult rats. White arrows depict where cell staining was found. Interestingly, fibre staining was also observed in the superficial dorsal horn (lamina I) of adult rats, but not in the younger ages. (B) Since both cell and fibre staining were observed, staining intensity was used to quantify the immunoreactivity of POMC in the DH. Quantified staining intensity for POMC in the DH significantly decreased as the animals aged, with highest immunoreactivity found at P10. (C) Enkephalin immunoreactivity in the DH was restricted to the superficial laminae and only fibre staining was observed. (D) Quantified staining intensity for enkephalin illustrate an age-dependent increase in immunoreactivity, which was highest at adult P10.(C) Enkephalin immunoreactivity in the DH was restricted to the superficial laminae and only fibre staining was observed. (D) Quantified staining intensity for enkephalin illustrate an age-dependent increase in immunoreactivity, which was highest at adult. (E) MOR (μ-opioid receptor) immunoreactivity in the DH, cell staining was found throughout the superficial and deeper laminae in all ages. (F) Cell count of MOR staining in the DH, which showed a significant increase as the animals aged (∗∗P<0 .01="" adult="" i="" p21="" vs="">

J. Desroches, J.-F. Bouchard, L. Gendron, P. Beaulieu. Involvement of cannabinoid receptors in peripheral and spinal morphine analgesia ☆ Neuroscience, Volume 261, 7 March 2014, Pages 23–42. ...The floating sections were then incubated in 1% sodium borohydride in PBS for 30 min, rinsed twice with PBS, and incubated for 30 min at room temperature in a blocking solution containing 3% normal goat serum (NGS) and 0.3% Triton X-100 in PBS. The sections were then incubated overnight at 4 °C with the guinea pig anti-MOP primary antibody (cat# GP10106; Neuromics, Minneapolis, MN, USA) diluted 1:1000 in the blocking solution. The floating sections were then washed in PBS and incubated with a goat anti-guinea pig secondary antibody conjugated with Alexa Fluor 488 (Molecular Probes, Invitrogen, Carlsbad, CA, USA) at a concentration of 1:1000 in PBS for 2 h at room temperature...

We will continue posting customer data and publications that give new insights into the mechanisms of pain.

Monday, February 10, 2014

Quantibody Arrays for Tracking Patient Health

Neuromics is working with Dr. Joe Smarda, a renowned Immunologist, to track levels of cytokines in the blood serum of his clients. We have selected RayBiotech's Quantibody® Arrays for these assays. The Clinics in Joe's network treat his clients for autoimmune related disorders.

Our regime is:
  1. Test clients pre-treatment
  2. Treat
  3. Test
  4. Refine treatment
  5. Test 
The specified treatment regime is continued until clients have blood serum cytokine levels that are in the range of our healthy controls. Here's data from our Quantibody® T-helper cell Cytokine Arrays (pre-treatment).

Figures IL-6, IL-1 beta, MCP-1 and PAI1 Array results in 4 clients with Autoimmune related Diseases.

We plan on posting these serial  testing results. They are designed to monitor status and indicate therapeutic effectiveness.

We will also be sharing some of the specific therapies being used. These will include treatments aimed at mobilizing endogenous stem cells. These cells have natural immune suppression/anti-inflammatory properties. Stay tuned.