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Wednesday, August 27, 2014

UCB Derived hMSC-MSCGro™ Media-The Wow Factor!

Neuromics-Vitro Biopharma Cells and Media Used to Treat Cerebral Ischemia

I have frequently posted successful outcomes with our Umbilical Cord Blood Derived Human Mesenchymal Stem Cells and MSCGro Expansion Media. These solutions have been tested head to head with other cell and media options and proven superior in cell behavior, doubling time and total number of passages. Competitive testing, until now, was done in culture.

I am pleased to present a study where our cells and media were selected for the the in vivo treatment of Cerebral Ischemia in Rats. This is a key part of building the foundation for human clinical trials: Chelluboina B, Klopfenstein JD, Pinson DM, Wang DZ, Veeravalli KK. Stem cell treatment after cerebral ischemia regulates the gene expression of apoptotic molecules. Neurochemical research. 39(8): 1511-21 DOI: 10.1007/s11064-014-1341-z
Protocol: Cryo-preserved hUCBSCs obtained from Neuromics/Vitro Biopharma (Golden, CO) were used to establish cultures in MSC-GRO low serum complete MSC medium according to the provided instructions. Cultures were maintained at 37 C in a humidified atmosphere containing 5 % CO2 with a change of culture medium twice a week. When the cell cultures were about 80 % to 90 % confluent, cells were split and subcultured. Cells were detached, washed twice with sterile phosphate buffered saline (PBS), counted and suspended in sterile saline prior to intravenous administration. The cells were intravenously injected (0.25 × 10(6) cells or 1 × 10(6) cells) via the tail vein.
Results:

Fig: Stem cell treatment after MCAO procedure reduces caspase-dependent apoptosis and brain damage. a Green fluorescence indicates cleaved caspase 3 protein expression. Representative cleaved caspase3 images were merged with respective DAPI images. Scale bar 100 lm. b Quantification of cleaved caspase-3 protein expression in the ipsilateral hemisphere of untreated [15] and hUCBSCs-treated animals. n C 3. Values are expressed as mean ± SEM; *p\0.05 compared to untreated MCAO subjected animals. c Representative hematoxylin and eosin stained paraffinembedded tissue sections from rat brains. Higher magnification images from the ischemic cortex and striatal regions of MCAOsubjected and untreated animals show interstitial edema and damaged neurons that have a condensed, irregular shaped and darkly stained nuclei which are absent or less frequent in control/hUCBSCs-treated brain sections. Each group consisted of a minimum of three animals. Scale bar value for the magnified images = 100 lm

This provides an in-depth understanding of the molecular mechanisms underlying the neuroprotective effects of mesenchymal stem cells derived from human umbilical cord blood in a rat model of transient focal cerebral ischemia. The study clearly demonstrates the potential of hUCBSCs to regulate various molecules responsible for cell death after transient focal cerebral ischemia followed by reperfusion.

There are some other important factors to consider:

  • Potency and Number of Stem Cells Matter-To move this into clinical applications, Doctors must be allowed to expand Mesenchymal Stem Cells.
  • Media used Matters-it must be best in class and not initiate immune inflammatory response.

We will continue to post studies utilizing Neuromics' Stem Cell Solutions.

Friday, August 22, 2014

We Seek Customer Feedback

Customer Service Matters-A Lot!

We use birdeye.com for customer feedback and input. Yes, it is re-assuring to get the rating and feedback featured in this Video.

We are also interested in hearing when all is not well. If there are issues, we will fix them. We offer all our customers free replacements or full refunds. Testimonials like these matter to us:

"Thank you for working with us in the past year to work out some of our antibody issues (we had run into a bad vial of 2AR ab.) Our work will appear next week online in PNAS- we cite the Neuromics antibodies. Have a great new year!" Dr. Laura Bohn, Associate Professor, OSU

You can contact me directly at any time (direct phone: 612-801-1007) or pshuster@neuromics.com.
I stand ready to serve you. Pete Shuster, CEO and Owner, Neuromics

Monday, August 18, 2014

Stem-Kine and Stem Cell Activation

For Health and Fitness

I have referenced our studies of commercially available Stem Cell Activators or Boosters. Many commercially available supplements make claims of boosting the body's ability increase the production of stem cells.

The catalyst for our researching the active ingredients in these boosters was the desire to find a cost effective way to treat clients with certain autoimmune diseases versus the expensive stem cell transplant options.  We also believed that these supplements could also improve athletic performance.

We have an active internal program to test these in culture using our human stem cell based assays and in actual clients. Stem-Kine is one of the supplements we tested proving it to have therapeutic value. Based on our research and testing, we also believe it is capable of increasing athletic performance by boosting the level of Red Blood Stem Cells.

One way we prove therapeutic value is by testing inflammatory/immune response cytokine, growth factors and oxidative stressors in our client’s blood serum using Quantibody Arrays. We test before prescribing Stem-Kine and other activators. We then re-test every 6 months. As an example,  the graph shows results for TNF-alpha and IL-6 (inflammatory response cytokines)  at initial testing levels (red) and re-tests (green). Here we see clear evidence of these 2 cytokines moderating to healthy levels (lower levels for these are better).


To learn more or order by phone, please call me directly at 612-801-1007 or you can also inquire via my direct e-mail: pshuster@neuromics.com. Pete Shuster, CEO and Owner, CA3 Biosciences, Inc. DBA Neuromics

Thursday, August 07, 2014

Pain Research Pubs

2014 Is Already a Record Year

I am pleased with the acceleration of publications by Pain Researchers using our Pain Research Markers and Gene Expression Analysis Tools.

Here's a sampling of the most recent: Transfection Regent Publications: Lili Hou, Yanfeng Zhang, Yong Yang, Kai Xiang, Qindong Tan, Qulian Guo. Intrathecal siRNA Against GPNMB Attenuates Nociception in a Rat Model of Neuropathic Pain. Journal of Molecular Neuroscience. July 2014...Ten micrograms of siRNA1- GPNMB dissolved in 30 μl i-Fect transfection reagent (Neuromics, Edina, MN, USA) was administered intrathecally once daily for 7 days, starting from 1 day before CCI surgery...

TRPV1 PublicationsCapsaicin-responsive corneal afferents do not contain TRPV1 at their central terminals in trigeminal nucleus caudalis in ratsOriginal Research ArticleJournal of Chemical Neuroanatomy, Volumes 61–62, November 2014, Pages 1-12 Deborah M. Hegarty, Sam M. Hermes, Tally M. Largent-Milnes, Sue A. Aicher

New insights into mechanisms of opioid inhibitory effects on capsaicin-induced TRPV1 activity during painful diabetic neuropathy. Neuropharmacology, Volume 85, October 2014, Pages 142-150 Mohammed Shaqura, Baled.I. Khalefa, Mehdi Shakibaei, Christian Zöllner, Mahmoud Al-Khrasani, Susanna Fürst, Michael Schäfer, Shaaban A. Mousa
TRPV1 IF in Mouse DRGs


Mohammed Shaqura, Baled.I. Khalefa, Mehdi Shakibaei, Christian Zöllner, Mahmoud Al-Khrasani, Desipramine and citalopram attenuate pretest swim-induced increases in prodynorphin immunoreactivity in the dorsal bed nucleus of the stria terminalis and the lateral division of the central nucleus of the amygdala in the forced swimming test. DOI: http://dx.doi.org/10.1016/j.npep.2014.07.001...... After rinsing with PBS, sections were incubated in PBS containing 0.3% Triton X-100 and 5% normal goat serum at room temperature for 30 min and then incubated with polyclonal guineapig anti-prodynorphin antibody (1:1000, Neuromics, Edina, MN, USA) or polyclonal rabbit ...

All Neuromics' Publications

I will continue to post Pain Research Updates!

Monday, July 28, 2014

Peptoid Arrays and Libraries

Designed for Detecting Bio-markers and Small-molecule Ligands
We have been successfully partnering with RayBiotech in providing Medical Testing and Research Lab ELISA Kits and Antibody Arrays. These have proven a cost effective way to detect from one to hundreds of proteins in a single sample. For example we tested 10 ASD Children using a Quantibody Arrays and here's a link to the results: Autism Spectrum Disorder (ASD) Children and Immune/Inflammatory Response Markers.

I am pleased to announce the addition of Peptoid Arrays.

Image: Peptoid Array-How They Work for Novel Ligands and Autoantibodies

The RayBio® Peptoid Array consists of thousands of unique peptoid sequences spotted on a glass slide support. The peptoid array combines the diversity of the bead-based peptoid library with the simplicity and rapid processing of the glass chip array. In this format, 2000+ peptoids can be quickly screened for binding activity to biological targets of interest.

Inage: Peptoid Anchored to Glass Slide

Overview

The Promise of Peptides as Drug Candidates

For nearly 20 years, peptides have been considered a promising class of drugs that possess distinct advantages over small molecule and biologic drugs. Compared to small molecule drugs, peptides can exhibit better efficacy, higher specificity, and lower toxicity. Compared to protein-based biologic pharmaceuticals, peptides are relatively inexpensive to produce and can easily be synthesized to exacting standards of purity and reproducibility between batches. Moreover, their chemistry and small size allows peptides to assume conformations that can mimic portions of full-length proteins or small molecules.

Advantages of Peptoids vs. Peptides

In practice, most peptide drug candidates have not lived up to the promise. Their susceptiblity to proteolytic cleavage, relatively short half-life in the body, and low oral and tissue bioavailability, make peptides less-than-ideal drug candidates. These challenges have hampered the translation of peptide drug candidates into therapeutics.
To overcome these inherent drawbacks, some researchers have investigated the potential for using peptide mimetics instead.  Some of the most promising peptidomimetic compounds are peptoids.
Peptoids are polymers of N-substituted amino acids that mimic many of the properties of peptides, but with distinct advantages. Peptoids have longer half-lives due to their  resistance to proteolysis; they also permeate cell membranes much better than peptides, resulting in greater bioavailability. Peptoids also have a much greater conformational freedom compared to a peptide of similar length, allowing a greater number of possible conformations, which can enhance sensitivity. Questions? Do not hesitate to contact me at pshuster@neuromics.com or 612-801-1007.

Wednesday, July 23, 2014

Pain Research Gene Expression Analysis

Potent and Proven Transfection Kits


Pain Researchers have successfully modulated 25+ genes involved in pain pathways using our Transfection Kits. Highlights include: DOR,The β3 subunit of  Na+,K+-ATPase, NTS1, NAV1.8, Kv 1.1, Kv 9.1, TROY, NOV, β-arrestin, TRPV1, CAV1.2, TLR4 and ASIC and more!  To learn more, check out our Transfection Kit Publications and Blog.

Figures: Intrathecal Kv9.1 siRNA treatment induces pain behaviors in naive rats. A, qRT-PCR quantification of Kv9.1 mRNA in rat PASMC cultures transfected with one of three Kv9.1 siRNA sequences or control siRNA. B, qRT-PCR showing Kv9.1 in vivo knock-down in L5 DRG, 4 d after intrathecal delivery of siRNA #1 compared with vehicle or matched scrambled control.  C, IHC for Kv9.1 in scrambled- and siRNA-treated DRG to determine protein knockdown. Graphs illustrate quantification of number of positive myelinated neurons and mean Kv9.1 signal intensity. D, Kv9.1 siRNA infusion inflicts a reduction in mechanical pain withdrawal thresholds. E, There was no change in heat pain thresholds after siRNA treatment. Vertical arrows on x-axis denote siRNA injections. doi: 10.1523/​JNEUROSCI.3561-12.2012.


We can now add the GPNMB gene to the list of those anaylyzed: Lili Hou, Yanfeng Zhang, Yong Yang, Kai Xiang, Qindong Tan, Qulian Guo. Intrathecal siRNA Against GPNMB Attenuates Nociception in a Rat Model of Neuropathic Pain. Journal of Molecular Neuroscience. July 2014...Ten micrograms of siRNA1- GPNMB dissolved in 30 μl i-Fect transfection reagent (Neuromics, Edina, MN, USA) was administered intrathecally once daily for 7 days, starting from 1 day before CCI surgery...
Abstract: Neuropathic pain is characterized by hyperalgesia, allodynia, and spontaneous pain. Recent studies have shown that glycoprotein nonmetastatic melanoma B (GPNMB) plays a pivotal role in neuronal survival and neuroprotection. However, the role of GPNMB in neuropathic pain remains unknown. The aim of the present study was to assess the role of GPNMB in neuropathic pain. In cultured spinal cord neurons, we used two small interfering RNAs (siRNAs) targeting the complementary DNA (cDNA) sequence of rat GPNMB that had potent inhibitory effects on GPNMB, and siRNA1-GPNMB was selected for further in vivo study as it had the higher inhibitory effect. After sciatic nerve injury in rats, the endogenous level of GPNMB was increased in a time-dependent manner in the spinal cord. Furthermore, the intrathecal injection of siRNA1-GPNMB inhibited the expression of GPNMB and pro-inflammatory factors (TNF-α, IL-1β, and IL-6) and alleviated mechanical allodynia and thermal hyperalgesia in the chronic constriction injury (CCI) model of rats. Taken together, our findings suggest that siRNA against GPNMB can alleviate the chronic neuropathic pain caused by CCI, and this effect may be mediated by attenuated expression of TNF-α, IL-1β, and IL-6 in the spinal cord of CCI rats. Therefore, inhibition of GPNMB may provide a novel strategy for the treatment of neuropathic pain.

If you would like to learn how you can optimize your gene expression analysis studies, do not hesitate to e-mail: pshuster@neuromics.com or direct line: 612-801-1007.

Wednesday, July 16, 2014

Potents Tools for Neuroscience Based Toxicology Assays

Neuromics' Offers Best in Class Cell and Markers

I am always on the hunt for proof that are tools work in the many different applications required by Researchers Studying Neurotoxicology. Success is confirmed to us through Customer Data/Pubs and Testimonials.

I would like to feature here some examples:


Figures: Neurons stained with Neuromics' MAP2 antibody to determine Neurite Damage.

We guarantee results. If you would like to learn more, please contact me directly at pshuster@neuromics.com or direct phone line: 612-801-1007. Thank you.

Friday, July 11, 2014

Immune/Inflammatory Response and Autism

Autism Spectrum Disorder (ASD) Children and Immune/Inflammatory Response Markers

Persistent chronic inflammation/immune response and oxidative stress are hallmarks of ASD. Like autoimmune diseases, an unbalanced immune system leads to unwanted assaults on healthy tissue. In ASD Children, inflammatory/immune response proteins could cross the Blood-Brain Barrier resulting in ongoing neuro-inflammation. This throws out of balance the natural pruning and repair cycle in the brain and could drive the related behaviors and symptoms of ASD Children.

We have been working with ASD children from central Europe residing in areas of heavy industry. All have unhealthy levels of metals in there blood stream and most have persistent viral and/or bacterial infection. This testing is part of our treatment program. Our treatment strategy involves first removing heavy metals and related toxins and the adding stem cell activators (stem cells are key players in modulating the immune system and repairing damage).

Previously I reported results from our testing for neuro growth/repair and oxidative stress markers using our custom ASD array. Here, I report on results from our measuring well known immune/inflammatory markers in 6 of these ASD children. The markers studied were: (IL-2, IL-6 and TNF-alpha). All of the markers were elevated when compared to healthy children. Here're the results:
Graphs: ASD vs Healthy Controls (*Healthy Control Data from Kim et al. Journal of Translational Medicine. 2011: 9:113)
Here's more on these markers:

  •  IL-2 is involved in immune regulation. It is one of the key factors in transplant regulation. It is also elevated in many children suffering from ASD. This elevation could be caused by toxins, chronic infection or genetic abnormalities. It should be also noted that IL-2 can cross the blood brain barrier making it a culprit in the symptoms of ASD. 
  • IL-6 is a potent pro-inflammatory agent that plays a crucial role in the pathogenesis of systemic inflammatory diseases like ASD.
  • TNF-a has been implicated as main effector of the functional consequences of neuroinflammation on neurodegeneration.
We are seeing improvements in symptoms and behaviors in the ASD Children undergoing treatment. We are early in the our Stem Cell Activating treatment phase. Our plans call for a cycles of testing and treating. If the tests yields movement of these markers to healthy levels, this will confirm how well our these treatments are working. Our plans our to openly share data.

If you would like to learn more about our testing and treatment regimes, I can be reached @ pshuster@neuromics.com or 612-801-1007.

Tuesday, June 24, 2014

More Gains on Pain Research

Discovering the Related Pathways

There are multiple descriptions for chronic and acute pain: stabbing, burning, cutting, itching, numbing, crushing and more. Understanding the pathways are important to optimizing the therapies and treatments for the many different forms of pain.

The foundation of Neuromics is providing top notch pain research markers, gene expression analysis tools and cell based assays. Here I share recent publications from customers using these:
Inflammatory Joint Pain: Fiona B Carr, Sandrine M Géranton and Stephen P Hunt. Descending controls modulate inflammatory joint pain and regulate CXC chemokine and iNOS expression in the dorsal horn. Molecular Pain 2014, 10:39 doi:10.1186/1744-8069-10-39...mu opioid receptor, 1:10,000, Neuromics (RA10104)...

Images: Representative single plane confocal images of MOR immunohistochemistry in the 1.5pmole dermorphin-saporin group and saline control. Double labelling with NeuN indicated that although many MOR+ neurons are depleted at this dose, some surviving MOR- neurons remain in the region, indicated by white arrows. Scale bars indicate 25 µm

Temperature Sensation: Marics I, Malapert P, Reynders A, Gaillard S, Moqrich A (2014) Acute Heat-Evoked Temperature Sensation Is Impaired but Not Abolished in Mice Lacking TRPV1 and TRPV3 Channels. PLoS ONE 9(6): e99828. doi:10.1371/journal.pone.0099828. ...TRPV1 antibody (1/1000 dilution, Neuromics)...

Acute PainPaulino Barragán-Iglesias, Hector I. Rocha-González, Jorge Baruch Pineda-Farias, Janet Murbartián, Beatriz Godínez-Chaparro, Peter S. Reinach, Thiago M. Cunha, Fernando Q. Cunha, Vinicio Granados-Soto, Inhibition of peripheral anion exchanger 3 decreases formalin-induced pain, European Journal of Pharmacology, Available online 27 May 2014, ISSN 0014-2999, http://dx.doi.org/10.1016/j.ejphar.2014.05.029. (http://www.sciencedirect.com/science/article/pii/S0014299914003914). ...substance P (guinea pig; 1:200; Cat # GP14110; Neuromics, Edina, MN), and purinergic P2×3 receptor (guinea pig: 1:1000; Cat # GP10108 ...

We are pleased with the many publications and customer provided data proving the quality of our many pain research tools. I am always available to serve you, Pete Shuster (pshuster@neuromics.com) or direct phone: 612-801-1007.

Sunday, June 15, 2014

Neuromics-Vitro Biopharma in Posrche Cup

Racing to Health and Peak Performance.

Neuromics and Vitro Biopharma are proud sponsors of the European Porsche Cup Car driven by our partner, friend and race car driver extraordinaire, Dr. Joe Smarda.
Dr. Smarda, an applied immunologist, is our partner in Europe for testing and treating sufferers of autoimmune diseases, sports injuries and autism with Stem Cell Activators. We are also testing these activating agents for the ability to improve the mental acuity and physical performance of healthy individuals.

He recently won a race at the Nurbring Ring in Germany. Hmmm, I wonder if the Stem Cell Activators he is taking, helped? Stay tuned.

Saturday, June 07, 2014

Stem Cell Therapies without Transplants

Neuromics is partnering with Vitro Biopharma to develop stem cell activating/boosting therapies. 

Stem cell based therapies represent a shining light of hope for sufferers of chronic or life threatening diseases.  Though, for most, realization of this hope is light years into the future.

Dr. Jim Musick, CEO of Vitrobiopharma, recently blogged on the obstacles to approved therapies : "Transplantation of hematopoietic stem cells has been widely used as an approved treatment of leukemia, lymphoma and certain autoimmune conditions for the past fifty years. Other adult stem cells have demonstrated safety and efficacy in pre-clinical research and clinical trials. Mesenchymal stem cell transplants have been most widely studied in animals, especially horses and dogs. Many of these studies have focused on skeletal-muscular effects. There is significant support for safety and efficacy in osteoarthritis, including cartilage regeneration, pain and inflammation reduction as well as recovery of function using intra-articular MSC injections (1). There are fewer studies of neural stem cell and Satellite cell transplants, but these also suggest safety and efficacy in various conditions. There are minimal adverse effects of these stem cell transplants

However, there are significant obstacles to routine clinical use of non-hematopoietic adult stem cell transplantation. First, therapeutic effect is dependent on cell concentration and exhibits characteristic dose-response relationships, necessitating expansion and characterization of MSCs prior to transplantation. While MSCs readily proliferate in vitro, this may result in cellular/genetic modifications and the cell culture conditions necessary for expansion of clinical grade MSCs have not yet been determined. Autologous sources appear superior to allogeneic, but controlled expansion of autologous MSCs could be limited. Also, there are regulatory obstacles including the US FDA that considers expanded stem cells “altered” with associated regulatory burdens prior to approval. Thus, it is likely that MSC transplantation has a long and expensive pathway to attain routine clinical implementation."


We are in the process of developing a new paradigm.
Images: UCB Derived hMSCs Activation Assays

The development of this new model includes assays that enable us to determine optimal dosing for these activators (see above).  Here's more from Dr. Musick: "Stem cell activation awakens innate stem cell systems to optimize healing, cellular regeneration and functionality. This approach has numerous advantages over stem cell transplantation including innately autologous therapy without acute or long-term complications from introduction of allogeneic cellular materials. The approach involves administration of an activating agent or agents and effects are controlled by dosage/pharmacokinetics of the stem cell mobilizing, epigenetic or proliferative agents thus avoiding transplantation issues including possible cellular modification during expansion, contamination, etc. The pathway to regulatory approval is shorter and less costly since certain combinations of generic drugs may be effective and there are natural substances exhibiting apparent efficacy. Also, new drug targets are associated with this paradigm, such as specific combinations of proliferation and epigenetic agents.

Vitro Biopharma has been pursuing this approach for the past six months using patients treated within a clinical network. The primary approach is to resolve toxicity, infectivity and specific deficiencies thus restoring physiological conditions while monitoring clinical status. Optimum cellular functionality is viewed as a necessary condition to elicit therapeutic benefit from activation of endogenous stem cells. We also use proteomic arrays to quantify various cytokines, growth factors and neurotrophins to develop disease profiles. We establish baseline results and are now testing agents known to activate stem cells to determine effects on the biomarker disease profile as well as clinical status. We are also developing additional biometric analysis of stem cell activation including MSC mobilization to peripheral blood, serum content of key stem cell biomarkers and tools for imaging of stem cell activation.

Vitro Biopharma is nearing commercialization of stem cell-based assays utilizing live cell imaging of cell migration, proliferation and reprogramming for drug discovery and support of clinical trials. We are also developing a clinical trial testing of stem cell activation for treatment of TBI and advanced molecular diagnostics while developing new stem cell activators. Vitro Biopharma is committed to further development and testing of the activation of latent, internal stem cells since this approach is widely supported by pre-clinical research and obviates several problematic issues associated with the transplantation of adult stem cells."

I will continue post here developments with our exciting, new approach to stem cell related therapies.

References: 1. Jo, CH. et al., Stem Cells 32: 1254-1266, 2014 2. Sinha, M. et al., Science 344: 649, 2014

Sunday, May 25, 2014

New Human Stress Selected Stem Cells

They Survive to Thrive

I am pleased to announce the addition of Stress Selected Stem Cells to our Cell Based Assays Offerings.

What makes these cells unique? They have:
•High potency, expansion rates and passaging.
•Low telomerase activity=low tumorigenesis properties
•High ability to migrate and differentiate to specific tissue types-High expression of migration/homing chemokines.

Image: Further morphology of these cultures is shown above as phase contrast images of two prominent clusters together with more firmly attached cells of classical mesenchymal morphology. This is day seven following exposure to stress. Scale bar is 50 mm.

We are offering these cells for 600 USD through June 30, 2014 (Save 150 USD)! Ordering options.

Check out our presentation learn more:
We can also conduct contract research to meet your specific requirements. To learn more you can contact me @612-801-1007 or pshuster@neuromics.com.

Tuesday, May 20, 2014

Neurons in 3-D

In Vivo Like Neurite Outgrowth Cultures

Culturing in 3-D requires potent Primary Neurons. Here researchers use our e18 Rat Cortical Neurons in developing their 3-D assays: Chandrasekhar R. Kothapallia, Peyman Honarmandib. Theoretical and experimental quantification of the role of diffusive chemogradients on neuritogenesis within 3D collagen scaffolds. Acta Biomaterialia. Available online 14 May 2014. http://dx.doi.org/10.1016/j.actbio.2014.05.009

Abstract: A critical challenge to regenerating close mimics of native axonal pathways under chronic neurodegenerative disease or injury conditions is the inability to stimulate, sustain and steer neurite outgrowth over a long distance, till they reach their intended targets. Understanding neurite outgrowth necessitates quantitative determination of the role of molecular gradients on growth cone navigation under dynamic physiological conditions. High-fidelity biomimetic platforms are needed to computationally and experimentally acquire and analyze spatio-temporal molecular gradient evolution and the growth cone response across multiple conditions along this gradient pathway. In this study, we utilized a simple microfluidic platform in which diffusive gradients were generated within a 3D porous scaffold in a defined and reproducible manner, and its characteristics (spatio-temporal gradient, steepness, diffusion time, etc.) precisely quantified at every specific location within the scaffold. Using this platform, we show that the cortical neurite response within 3D collagen scaffolds, at both the cellular and molecular level, is extremely sensitive to subtle changes in localized concentration and gradient steepness of IGF-1 within that region. This platform could also be used to study other biological processes such as morphogenesis and cancer metastasis, where chemogradients are expected to significantly regulate the outcomes. Results from this study might be of tremendous use in designing biomaterial scaffolds for neural tissue engineering, axonal pathway regeneration under injury or disease, and in formulating targeted drug delivery strategies.
Image: Neurons in 3-D Assay
Neuromics' provides many Stem and Primary Cell Assay Solutions including tools for 3-D Cultures. We also offer services studying the effects of small molecules and compounds on Stem Cell expansion, differentiation and migration. To learn more contact me at 612-801-1007 or pshuster@neuromics.com.

Sunday, May 11, 2014

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.

Thursday, May 01, 2014

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.

Saturday, April 19, 2014

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.

Thursday, April 17, 2014

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.

Friday, April 04, 2014

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.

Wednesday, March 26, 2014

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.

Sunday, March 23, 2014

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.

Thursday, March 20, 2014

Irritable Bowel Syndrome (IBS) and BDNF

Gene Expression Analysis Determines BDNF's Role in IBS

In this study, Researchers used Neuromics'  i-FectTM 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.

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 pshuster@neuromics.com 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. 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.

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.

Tuesday, January 28, 2014

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.

Tuesday, January 21, 2014

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.

...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.

Monday, January 13, 2014

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.

Monday, December 30, 2013

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.