Autism Spectrum Disorder (ASD) Custom Array Results

Blood Serum Levels and Key Markers

We have been testing immune/inflammatory response, oxidative stress and growth factor markers in ASD children from central Europe. Most showed high levels of  related cytokines and chemokines.

From our initial testing and published results, we have developed a custom Quantibody Array to test ASD children. The markers in this array are: BDNF, HSP-70, Leptin, RAGE and TGF-beta1.

Figure: Serum Levels of Key Markers in Tested ASD Children

Here's more on each marker:

• BDNF is a protein involved in making healthy new neurons. This protein is dis-regulated in autism. It is shown to be decreased in some studies and increased in others. These variations could be a function of age. During brain development, BDNF regulates the birth and differentiation of brain cells, or neurons. Some of BDNF’s target cells, such as cortical interneurons, which transmit information between different layers of the brain cortex, have been implicated in autism. BDNF is also a regulator of brain growth, and children with the disorder tend to have abnormally large brains during early development. Vigorous exercise, for example, increases BDNF levels in blood and studies have linked this increase to growth of new healthy neurons in the hippocampus region of the brain. We found all ASD children tested showed low levels of BDNF. All had moderate to high levels of heavy metals and viral/bacterial pathogens. Most also had evidence of leaky blood brain barriers. Could this low level be due to consumption of BDNF demanded by the chronic need for neuro-repair or is genetic in origin or perhaps both?
• HSP70 is strongly upregulated by heat stress and toxic chemicals, particularly heavy metals such as: aluminum, arsenic, cadmium, copper, mercury, etc. This upregulation in ASD could be linked to difficulty is in clearing toxins. Children with highest tested HSP70 levels also had the highest concentration of heavy metals. The 3 children with healthy levels have been undergoing ongoing treatment to clear metals.
• Leptin modulates appetite and energy tough elevated levels of leptin present in cases of autism might be an important sign of immune processes, particularly those related to inflammation. It is also suggested leptin may be a link between autism and epilepsy that provides an avenue for novel or better management of autistic children with epilepsy. All were in the range of published healthy controls (mean=2065 pg/ml).
• RAGE is hypothesised to have a causative effect in a range of inflammatory diseases such as diabetic complications, Alzheimer's Disease and even some tumors. In ASD, it is thought to be a master switch for chronic inflammation. Including in the brain. RAGE is a receptor for S100B so it is also elevated in ASD and is reflective of neurological damage. All showed elevated serum levels of rage.
• TGF-beta1 is a protein that controls proliferation, cellular differentiation, and other functions in most cells. This control includes tissue repair in the nervous system. Decreased transforming growth factor beta1 in autism is a potential link between immune dysregulation and impairment in clinical behavioral outcomes. 6 of 8 of the children had low TGF-beta1 levels. Though the correlation between symptoms and behaviors is not conclusive.

We plan to significantly increase the samples of ASD children tested using this custom Quantibody Array. . 

Our ultimate goal is to use this array as a tool to determine the efficacy of therapies we are developing. Our goals is to develop natural products based therapies that are proven to activate stem cells. These cells could catalyze immune response modulation and tissue repair. The process is to first clear metals and pathogens, then treat with stem cell activators and test and fine tune treatments and test again. 

I will be posting results on an ongoing basis. I also welcome any and all comments. I am available for direct contact at pshuster@neuromics.com or 612-801-1007.

Pancreatic Cancer Tumor Cell Assays

Providing Pancreatic Tumor Cells or Related Custom Assays

We are pleased to announce the addition of Pancreatic Tumor Cells to our Cell Based Assay offerings. We are also generating a wealth of internal data with our Tumor Cells and Cancer Associated Fibroblasts. We have now opened up our capabilities to Cancer Researchers via our new CRO services. These included customized assays for studying toxicity using compounds/small molecules. Here's an example:

In addition, we can do kinetic, live cell screens using these cells plus our hMSC and CAFs. To learn more call 612 -801-1007 e-mail me: pshuster@neuromics.com. We stand ready to serve you in a way that fits your specific requirements.

Extracellular Matrix Environment and Chemotherapeutics

Substrate Matters!

Neuromics' has been promoting a variety of 3-D Cell Based Assay Solutions for the past several years. These include: Nanofibers, Hydrogels and Extracellular Matrix (ECM) Proteins. We have found that the adoption rate for these as standard tools for drug discovery is slower that we anticipated.

We believe substrate matters so I am pleased to share a recent publication that references use of our Collagen IV and other ECM proteins. This confirms the importance of using a more in vivo like environment in testing chemotherapeutics: Thuy V. Nguyen,Marianne Sleiman,Timothy Moriarty,William G. Herrick,Shelly R. Peyton. Sorafenib resistance and JNK signaling in carcinoma during extracellular matrix stiffening. Publication: Biomaterials. Elsevier. 13 April 2014. http://dx.doi.org/10.1016/j.biomaterials.2014.03.058.

Abstract: Tumor progression is coincident with mechanochemical changes in the extracellular matrix (ECM). We hypothesized that tumor stroma stiffening, alongside a shift in the ECM composition from a basement membrane-like microenvironment toward a dense network of collagen-rich fibers during tumorigenesis, confers resistance to otherwise powerful chemotherapeutics. To test this hypothesis, we created a high-throughput drug screening platform based on our poly(ethylene glycol)-phosphorylcholine (PEG-PC) hydrogel system, and customized it to capture the stiffness and integrin-binding profile of in vivo tumors. We report that the efficacy of a Raf kinase inhibitor, sorafenib, is reduced on stiff, collagen-rich microenvironments, independent of ROCK activity. Instead, sustained activation of JNK mediated this resistance, and combining a JNK inhibitor with sorafenib eliminated stiffness-mediated resistance in triple negative breast cancer cells. Surprisingly, neither ERK nor p38 appears to mediate sorafenib resistance, and instead, either ERK or p38 inhibition rescued sorafenib resistance during JNK inhibition, suggesting negative crosstalk between these signaling pathways on stiff, collagen-rich environments. Overall, we discovered that β1 integrin and its downstream effector JNK mediate sorafenib resistance during tumor stiffening. These results also highlight the need for more advanced cell culture platforms, such as our high-throughput PEG-PC system, with which to screen chemotherapeutics.

Figure: High-throughput biomaterial platform for drug screening. (A) The high-throughput platform consists of a black-walled, glass bottom plate, with PEG-PC gels cast in each of the inner 6x10 wells. (B) Gels can be functionalized with any protein or peptide of interest, and they support the adhesion and growth of carcinoma cells. We used this platform to test carcinoma cell response to a kinase inhibitor (sorafenib) as a function of underlying gel stiffness and ECM adhesive protein cocktail. (C) A representative graph of SkBr3 proliferation (y-axis) in response to sorafenib (x-axis) across a range of gel stiffness (colors) demonstrates the IC-50 calculation.

This confirms the importance of considering your substrate environment when developing your in vitro assays for High Content and High Throughput Drug Discovery. We will continue to provide updates.

HIV-1R Viral Protein R and Memory Impairment

Using Synpatophysin as Marker for Synaptic Loss

Our Neuron-Glial Markers continue to shine in challenging applications. Here researchers examined whether infusion of the Vpr-expressing astrocytes affected synaptophysin expression in the hippocampus. The authors of the study, using Neuromics' Mouse Monoclonal Synaptophysin Antibody,  found a significant reduction in synaptophysin staining in CA3: Lilith Torres and Richard J Noel. Astrocytic expression of HIV-1 viral protein R in the hippocampus causes chromatolysis, synaptic loss and memory impairment. Journal of Neuroinflammation 2014, 11:53 doi:10.1186/1742-2094-11-53.

Images: Astrocytic HIV-1 viral protein R (Vpr) expression decreased synaptophysin immunoreactivity (A) Representative light photomicrograph showing the distribution of synaptophysin immunoreactivity in the rat hippocampal CA3 formation. Green fluorescent protein (GFP) right side. Vpr shows both left and right. Magnification 100×. (B) Densitometric analysis revealed significantly decreased mean value for the Vpr group compare to control.


Protocol: To examine changes in synaptophysin between control and HIV-1 Vpr exposed rats, tissue sections from each group were processed for immunocytochemistry. The samples were cut at 4 μm thickness with a microtome (Microm HM340, Microm International) and fixed to positively charged microscope slides. Fixed tissues were deparaffinized in xylene substitute for 30 minutes, rehydrated through graded alcohols and neutralized with 3% hydrogen peroxide (Sigma-Aldrich), followed by a rinse under running tap water and immersion in antigenretrieval solution (0.01 M citrate, pH 6.0) for 1 minute at 98°C. Then sections were washed in TBS for 5 minutes and treated with blocking solution containing normal goat serum (BioGenex, cat# HK112-9KE). Sections were incubated for 24 hrs at 4°C in mouse monoclonal antisynaptophysin antibody (Neuromics, cat # MO20000, 1:500 dilutions). Negative controls with TBS instead of primary antibody were run in each slide. Primary antibody was washed in TBS buffer for 2 × 5 minutes and incubated with Multi Link secondary antibody (Super Sensitive Link-Label IHC Detection System, cat# LP000- ULE, BioGenex, San Ramon, CA, USA). Secondary antibody was washed in TBS and incubated in ABC-HRP, washed in TBS buffer and incubated in 3,3′-diaminobenzidine (cat# HK153-5KE, Biogenex, San Ramon, CA, USA). Slides were rinsed in water and counterstained with hematoxylin for 30 sec. The sections were rinsed, dehydrated and and mounted with Cytoseal XYL (cat# 8312-4, Richard Allan Scientific, Kalamazoo, MI, USA). For quantitative densitometry, images of regions of interest (ROI) in the CA3 were captured from 5 rats in each group using NIH Image J 1.50 software.

We will continue to work hard to fill all your Neuroscience Research Needs.

SOX that Rocks

Excellent Adult, iPSC and eSC Markers 

Our SOX Markers consistently deliver in helping determine differentiation states of Stem Cells.
Here are some recent pubs referencing use of these important antibodies:

Xiaoling Wang, Yingjia Wang, He Huang, Buyuan Chen, Xinji Chen, Jianda Hu, Tammy Chang, Ren-Jang Lin, Jiing-Kuan Yee Precise Gene Modification Mediated by TALEN and Single-Stranded Oligodeoxynucleotides in Human Cells. Published: April 01, 2014DOI: 10.1371/journal.pone.0093575... Sox2 (1:100, Neuromics, Minneapolis, MN)...

Megumi Kowno, Kanako Watanabe-Susaki, Hisako Ishimine, Shinji Komazaki, Kei Enomoto, Yasuhiro Seki, Ying Ying Wang, Yohei Ishigaki, Naoto Ninomiya, Taka-aki K. Noguchi, Yuko Kokubu, Keigoh Ohnishi, Yoshiro Nakajima, Kaoru Kato, Atsushi Intoh, Hitomi Takada, Norio Yamakawa, Pi-Chao Wang, Makoto Asashima, Akira Kurisaki. Prohibitin 2 Regulates the Proliferation and Lineage-Specific Differentiation of Mouse Embryonic Stem Cells in Mitochondria. Research Article | published 07 Apr 2014 | PLOS ON...Sox17 (Neuromics, GT15094)...

We will continue to update you on new developments with our Stem Cell Markers.

Osteoblast Activators and Musculoskeletal Disease Assays

We now have the ability to precisely test human Osteoblast activating agents. Tools available include:

1.  Live cell screening
2. Quantibody Bone Metabolism Arrays
3. PCR
4. Flow Cytometry
5. Immunostaining
6. Western Blotting

This means we can generate data that provides you a clear picture of the effects of small molecules/compounds on the activation, expansion and migration of Osteoblasts. Here's an example.

Image: Activin A expression from Osteoblasts treated with Activators vs Controls

We will soon be releasing a turn-key assay for Osteoporosis Drug Discovery. We can also run custom assays that fit your specific requirements. To learn more, you can call (612-801-1007) or e-mail: pshuster@neuromics.compshuster@neuromics.com). Thank you. Pete Shuster, CEO and Owner, Neuromics.

hN2 Primary Human Neurons and Toxicity Assays

Neuromics' hN2 Cells Continue to Shine

I previously posted our Human hN2 Neurons being used for studying the mechanisms of  Nerve Agent VX: hN2 Human Neurons for Toxicity Screening. The versatility of these neurons enabled the researchers to perform a microarray study in which cultured human neural cells were exposed to 0.1 or 10 μM of VX for 1 h. Global gene expression changes were analyzed 6, 24, and 72 h post exposure. Solid primary cell based assay results start with healthy and well behaving cells.

I am pleased to announce success in exposing the cells to a specific cytotoxicity inducing compound (small molecule): Mark RichardsChee Wee PhoonGwendoline Tze Wei GohEng Khuan SengXu Ming GuoCherine Mei Fong TanWoon-Khiong ChanJoel Mun Kin Lee. A New Class of Pluripotent Stem Cell Cytotoxic Small Molecules. Research Article | published 19 Mar 2014 | PLOS ONE 10.1371/journal.pone.0085039.

Image: Cytotoxic agent used in dose-response curve

Figure: Dose response curves for 3 specialized somatic cell lines (MRC-5, human primary neurons and human neonatal cardiomyocytes) treated with JC011.

This curve illustrates the sensitivity of human neurons to toxic agents. 

We plan on continuing to use these and our hNP1 Human Neural Progenitors in kinetic, "in vivo like" assays. These assays will give quantitative data on both growth and differentiation inducing agents as well as specifics on how the cells behave when exposed to toxic agents. I will be posting results here.

This data should be of interested to neuro-disease/disorders basic and drug discovery researchers. We plan on making the assays available to researchers. We currently also do small molecule testing and gene expression analysis studies a CRO offering. To learn more, I can be reached at pshuster@neuromics.com or 612-801-1007.

Autism, Inflammation and Stem Cell Enhancers

Proving the Therapeutic Value

We have been running Quantibody® Antibody Arrays on blood serum of children diagnosed with Autism Spectrum Disorder (ASD). All reside in areas of heavy industry in Central Europe. All have elevated levels of one or several heavy metals.

These assays are being run as part of our strategy of treating these children with natural stem cell enhancing supplements. Here are the average serum levels of 2 cytokines (IL-6 and TNF-alpha) and 1 related chemokine (CCL3). All of the children had elevated levels vs healthy controls:

Figure: Serum ASD levels vs Healthy Controls (pg/ml)

TNF-alpha and IL-6 promotes the immune/inflammatory response. These two cytokines are guided to sites (including the CNS) of infection or tissue damage by the chemokine CCL-3 and others. In the normal process, the site(s) of immune response are cleaned (the response) of infection and/or damaged tissue and then repaired. The key with autoimmune diseases and disorders, is that that this process becomes a continuous loop; hence, these cytokines and chemokines are elevated. If the loop is broken or down modulated, the the levels of these should decrease.

Mesenchymal stem cells (MSCs) are immunomodulating and anti-inflammatory. We plan on testing candidate substances (all our currently available as natural supplements) on kinetic assays using our umbilical cord blood human mesenchymal stem cells. These will enable us to quantify  cell growth and expansion. We also plan on testing the best candidates on our human neurons to see the effects on cell behavior.

We will then determine safe dosing working with experts in the U.S. and Europe. During treatment, we will again be testing serum to see if the levels of these and other key Cytokines, Chemokines and Growth Factors. This will give proof as to to whether or not the treatments are working. If so, we should see the serum levels of these move toward to those of healthy controls.

I plan on making more data as it becomes available.

Irritable Bowel Syndrome (IBS) and BDNF

Gene Expression Analysis Determines BDNF's Role in IBS

In this study, Researchers used Neuromics'  i-Fect^TM Transfection Kit to deliver BDNF to determine effect on visceral hypersensitivity (VHS): J. H. Winston1 Q. Li1, S. K. Sarna1. Chronic prenatal stress epigenetically modifies spinal cord BDNF expression to induce sex-specific visceral hypersensitivity in offspring. Article first published online: 4 MAR 2014. Neurogastroenterology & Motility. DOI: 10.1111/nmo.12326. The Journal of Pain, Volume 14, Issue 11, November 2013, Pages 1485–1491 http://dx.doi.org/10.1016/j.jpain.2013.07.007

Intrathecal treatment with brain-derived neurotrophic factor (BDNF) antagonists reduced VMR to colorectal distension (CRD) in female chronic prenatal stress (CPS)+HeICS rats. (A) Graph shows that intrathecal administration of BDNF antagonist trkBFc in female CPS rats significantly decreased VMR to CRD, 24 h following adult 29 HeICS (two-way repeated measures ANOVA found a significant main effect of treatment, F1,53 = 10.4, p = 0.015; post hoc tests found significant differences at 30, 40, 50, and 60 mmHg, n = 4). (B) Graph shows that intrathecal administration of BDNF siRNA in female CPS rats significantly decreased VMR to CRD, 24 h following adult 29 HeICS (two-way repeated measures ANOVA: treatment 9 pressure interaction, F1,77 = 3.49, p = 0.008, tukey post hoc tests found significance at 30 mmHg, p = 0.013 and at 40, 50 50, 80 mmHg, p < 0.001, n = 7 Ctr., n = 6 BDNF siRNA). (C) Western blot shows a significant decrease in spinal cord BDNF protein expression in rats treated with BDNF siRNA (*p < 0.05).

Conclusion: Chronic prenatal stress followed by a second exposure to HeICS in adult offspring exacerbated visceral hypersensitivity (VHS) greater in female offspring that persisted longer than in male offspring. Chronic prenatal stress up-regulated BDNF expression in the lumbar-sacral dorsal horn that correlated with the exacerbation of VHS in female, but not in male offspring by increasing RNA Pol II binding and histone H3 acetylation, and decreasing histone deacetylase 1 association with the core promoter of BDNF in female offspring.

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.

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 pshuster@neuromics.com or cell: 612-801-1007.

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. http://dx.doi.org/10.1016/j.neuroscience.2013.12.030. ...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.

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.

Pot and Pain

The pressure for states to legalize Marijuana for medical and recreational use is building. The tax benefits are self evident.

The debate for many centers of "true medical benefits". That's why research on understanding analgesic pathways is so important. Ironically, this study was conducted by my friends at Université de Montréal and Université de Sherbrooke in Quebec Canada: 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. http://dx.doi.org/10.1016/j.neuroscience.2013.12.030.

These teams have proven expert is using our Opioid Receptor Antibodies in their pain research. Here's a synopsis:
•Analgesia is the most common feature shared by the cannabinoid and opioid systems.
•The role of the cannabinoid system in the morphine-induced analgesia is uncertain.
•Peripheral and intrathecal morphine analgesia is altered in cnr1KO and cnr2KO mice.
•This attenuation is neither caused by a MOP malfunction nor by its downregulation.

Images: Deletion of the CB1 or CB2 receptors has no effect on the expression of MOP in the spinal cord. Immunofluorescence of spinal MOP revealed that the expression of MOP in laminae I and II of the dorsal horn of the spinal cord did not differ between cnr1WT (A) and cnr1KO (B) mice or between cnr2WT (C) and cnr2KO mice (D).


Observations here further support the existence of interactions between the cannabinoid and opioid systems. The loss of peripheral and spinal morphine analgesia is apparently caused neither by a decrease in MOP spinal expression nor by altered binding properties or G protein coupling of this receptor in cnr1KO and cnr2KO mice. The mechanisms underlying the loss of morphine analgesia are not clear but could include the release of endogenous cannabinoids in structures along the pain pathway or a disrupted endocannabinoid tone.

It is important funding that enables researchers to understand the analgesic pathways of marijuana continues to grow. This research could yield better control of pain with reduced side effects.

FGF and Stem Cells-Options Matter

Proven, Potent and Cost Effective Fibroblast Growth Factors (FGF).

Neuromics have a wealth of expertise in Stem Cells, Media and Growth Factors. FGF is an important component of Stem Cell Based Assays. Our goal is to provide an FGF that fits your requirements like "hand in glove".

Here's a small sampling:
ISO-kine bFGF-100% animal free and serum free-is produced in the endosperm tissue of barley grain (Hordeum vulgare), that exhibits up to 50 times less protease activity than E.coli or mammalian cells. Barley seed is void of any human or animal viral contaminants that could jeopardize your cell culture.

Images A: Expression of OCT4 (green) in the ORF group. B: Expression of TRA-1-60 (green) in the ORF group.

FGF basic (146 aa)-Anika Chowdhury, Louis W Bezuidenhout, Aillette Mulet-Sierra, Nadr M Jomha and Adetola B Adesida. Effect of interleukin-1β treatment on co-cultures of human meniscus cells and bone marrow mesenchymal stromal cells. BMC Musculoskeletal Disorders 2013, 14:216 doi:10.1186/1471-2474-14-216

...15 million MNCs were seeded per 150 cm2 tissue culture flask. Culture media was alpha MEM supplemented with 10% heat inactivated fetal bovine serum, 1 mM sodium pyruvate, 100 mM HEPES buffer, 1 mM sodium pyruvate, 100 U/ml penicillin, 100 μg/ml streptomycin, 0.29 mg/ml L-glutamine (all from Invitrogen, Mississauga, Ontario, Canada) and 5 ng/ml of basic FGF or FGF-2 (Neuromics, Edina, MN, USA). Plastic adherent MNCs were allowed to attach and proliferate for 7 days before the first media change under normal oxygen tension (21% O2; 95% air) at 37°C in a humidified incubator with 5% CO2...

Images: Histological characteristics of pellets formulated from mono-cultured MCs, mono-cultured BMSCs and co-cultures of MC and BMSCs after a total of 17 days culture in defined serum-free chondrogenic media. (A-B) Safranin O and collagen II immuno-histochemical staining of representative pellets from cells derived from the same donor. Magnification lens × 20; scale bar is 100 μm. Chowdhury et al. BMC Musculoskeletal Disorders 2013 14:216 doi:10.1186/1471-2474-14-216

We are working hard to provide unique and cost effective solutions for your Stem Cell Based Assay Requirements.

TRPV1 and Osteoarthritis Related Pain

Our TRPV Antibodies are widely used and frequently published. Many of these feature TRPVs' role in nociceptive pain. Specifically they play important roles in the detection of noxious stimuli and inflammatory hyperalgesia.

TRPV1 has been implicated in OA pain, both in animal models and by the finding that TRPV1 genetic variants are associated with the risk of symptomatic knee OA in humans: S Kelly, R J Chapman, S Woodhams, D R Sagar, J Turner, J J Burston, C Bullock, K Paton, J Huang, A Wong, D F McWilliams, B N Okine, D A Barrett, G J Hathway, D A Walsh, V Chapman. Increased function of pronociceptive TRPV1 at the level of the joint in a rat model of osteoarthritis pain. Ann Rheum Dis doi:10.1136/annrheumdis-2013-203413.
Methods: Rat spinal cord sections from MIA- and saline-treated rats (n=5/group) (see online supplemental methods) were incubated with a polyclonal guinea pig anti-TRPV1 antibody (1 : 500, Neuromics, Edina, Minnesota, USA catalogue number GP14100) and then with Alexa 568-conjugated goat anti-guinea pig secondary antibody (1:300, Molecular Probes). TRPV1 immunostaining was visualised with a Leica DMRB/DM4000 B fluorescence microscope and images were acquired using Openlab software (PerkinElmer)...

Images: Transient receptor potential vanilloid 1 (TRPV1) immunoreactivity in the spinal cord. TRPV1 immunofluorescence detected in superficial dorsal horn (10× magnification) in rat lumbar (L3–L5) spinal cord at day 28 post-intra-articular injection of saline (A) or mono-iodoacetate (MIA) (B). Minimum and maximum brightness values were altered (32.01 min and 90.14 max) using Image J so as to highlight the area of TRPV1 positive staining. (C) Quantification of TRPV1 immunofluorescence in superficial dorsal horn of spinal cord taken from rats at 14 or 28 days following intra-articular injection of MIA and at day 28 following intra-articular injection of saline. Data are expressed as mean and SEM (n=5 per group).

Clinical trials of oral TRPV1 antagonists have been limited by on-target-induced hyperthermia. Here experimental evidence for increased functional role of TRPV1 at the level of the joint in a model of OA pain and the demonstration that blockade of joint TRPV1 ablates sensory afferent sensitization and pain behaviour support future targeted site-specific investigations of the therapeutic potential of TRPV1 for OA pain associated with synovitis. This could be good news for OA sufferers.

Autism Spectrum Disorder and Biomarkers

We used our IFN-gamma and BDNF Sandwich Elisa Kits to test the levels of these biomarkers in the blood serum of 2 individuals with Austism Spectrum Disorder. We anticipated  that BDNF would be significantly up or down regulated and IFN-gamma up regulated as confirmed by the literature. see 10.1371/journal.pone.0020470 , DOI: 10.1371/journal.pone.0020470 and DOI: 10.1111/acps.12071.

Our testing results showed:

Figure 1: ASD and Biomarker Levels

Based on these initial findings we are developing a custom Quantibody® Antibody Array designed for the fine tuned testing of individuals with ASD. The biomarkers included in the array will be: HSP70, TGF-beta2, 

Caspase-7, IFN-gamma and BDNF.

Figure 2: ASD Markers vs Controls

These panels will enable us to determine the severity of ASD. From the results, we will be working with our collaborators to determine potential cell based therapies for improving the symptom and behaviors of ASD sufferers. As part of the therapeutic regimes the panels can then be used check on progress towards "wellness". 

This a key initiative for us in 2014 so stay tuned.

hMSCs and Mesen-X Media in Actions

Customer Feedback on Neuromics' Human Mesenchymal Stem Cells

The demand for these UCB derived hMSCs continues to grow. We do everything we can to insure users' success. This includes replacing cells if there are any issues.

It is always great to get documented feedback. Dr. Rodney Nash, CEO of Javeen Biosciences used their new Mesen-X media to grow the cells. This GMP manufactured media is TOTALLY serum and animal free. It is shipped at room temperature and requires no attachment agents.

Here's Dr. Nash's feedback: We recently used the hMSCs derived from Umbilical Cord Blood. Their performance was nothing less than excellent. We were highly impressed with their morphology and their doubling rate. In addition the cells respond very well to accutase and maintain their performance after passaging. We highly recommend this cell line."

Neuromics' hMSCs grown using Mesen-X Media

The poperties of this Media makes it an excellent solution for drug discovery. Neuromics will be distributing in early 2014 so there will be much more data to follow.

Biochemistry of Hair Loss

Hair Follicle Stem Cells and the WNT Pathway

This publication hit my Radar because it referenced use of our Goat Polyclonal GFRA1 Antibody.


In this study, Researchers use Tbx18Cre knockin mouse line to ablate the Wnt-responsive transcription factor β-catenin specifically in  at E14.5 during the first wave of guard hair follicle formation. In the absence of β-catenin, canonical Wnt signaling is effectively abolished in cell clusters of precursors for the hair follicle dermal papilla (DP). Sox2+ dermal condensates initiate normally; however by E16.5 guard hair follicle numbers are strongly reduced and by E18.5 most whiskers and guard hair follicles are absent, suggesting that active Wnt signaling in dermal condensates is important for hair follicle formation to proceed after induction. To explore the molecular mechanisms by which Wnt signaling in dermal condensates regulates hair follicle formation, we analyze genome-wide the gene expression changes in embryonic β-catenin null DP precursor cells. We find altered expression of several signaling pathway genes, including Fgfs and Activin: Su-Yi Tsai, Rachel Sennett, Amélie Rezza, Carlos Clavel, Laura Grisanti, Roland Zemla, Sara Najam, Michael Rendl Wnt/β-catenin signaling in dermal condensates is required for hair follicle formation. Developmental Biology, Available online 3 December 2013. http://dx.doi.org/10.1016/j.ydbio.2013.11.023

Could manipulating PDs be the answer for halting hair loss or better yet reverse the process? This research moves us closer.

In vivo Like Cell Based Assays!

Save 100 USD on our Potent, Pure and Easy to Culture Stem and Progenitor Cells

We continue to design in vivo like behaviors into our  Cell Based Assays.
Our goal is to enable you to make better research decisions earlier
in your discovery process.

These assays are engineered using our proven, potent and easy to culture eSC and Adult Human Stem Cells and Progenitors.

UCB Derived hMSC differentiated to Osteoblast. Note the calcium depositions.

***December Promotions-New Products-Save 100 USD+ 
on Primary and Stem Cells, Media, Markers and Growth Factors***

• Human Mesenchymal Stem Cells-Passage up to 12X-395 USD 
• Native Human Chondrocytes-HCS-449 USD
• Native Human Osteoblasts-449 USD
• Cancer-Associated Fibroblasts (CAFs)-499USD
• Save 70 USD-only 225 USD for new Neuron, Astroglial and Progenitor Markers

I'll be posting videos of our in vivo like assays with specific quantitative data here soon.