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.

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.

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:

• Neuron-Glial-Astrocyte-Progentitor Markers-Bin Xing, Adam D. Bachstetter, Linda J. Van Eldik. Inhibition of Neuronal p38α, but not p38β MAPK, Provides Neuroprotection Against Three Different Neurotoxic Insults. Journal of Molecular Neuroscience. July 2014...Cells were fixed with 3.7 % formaldehyde containing 0.1 % Triton X-100 in PBS for 10 min at room temperature. After washing three times with PBS, the coverslips were incubated with blocking buffer (PBS containing 6 % goat serum, 3 % bovine serum albumin (fraction V), 0.1 % Triton X-100) for 30 min at room temperature. Primary chicken anti-MAP2 antibody (1:1,000, Neuromics, Cat. no. CH22103) was diluted in blocking buffer and incubated with the cells at room tem- perature for 2 h. For detection of MAP2 staining, the cells were incubated with secondary biotin SP-conjugated goat antichicken antibody (1:1,000, Jackson ImmunoResearch) for 1 h, followed by streptavidin Alexa Fluor® fluorescent 488 (1:1,000, Invitrogen) incubation in blocking buffer at room temperature for 1 h. Wide field fluorescent photomicro- graphs were obtained using a Nikon Eclipse Ti microscope with an Axiocam MRc5 digital camera (Carl Zeiss)...

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

• Primary Neurons and Astrocytes-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 ...Primary hN2™ Human Neurons (Neuromics) were grown in hN2™ human neuron culture media (Neuromics)..
• Xiugong Gao, Hsiuling Lin, Radharaman Ray, Prabhati Ray. Toxicogenomic Studies of Human Neural Cells Following Exposure to Organophosphorus Chemical Warfare Nerve Agent VX. Neurochemical Research. February 2013. ...Human hN2 neurons were obtained from Neuromics...

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.

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.