immunoLink Therapies and Treating ASD Children

6-Months of Therapy and Testing Results

I have previously posted our testing and treatment regime for ASD Children. For some of the children, we are seeing marked improvement after 6 months of our treatment regime using solutions aimed at balance immunity and stem cell vitality.

We find that for the children we treat all have varying dysregulated levels of immune/inflammatory response, oxidative stress and growth factor/neurotrophin biomarkers.  We customize treatment plans based on pre-treatment testing results. We then re-test after 6 months to see the level of moderation in these biomarkers/factors. The re-tests are used to show the parents how well these children are responding to treatment and to further adjust plans.

Here I would like to share the results from our more traditional medical testing. These are representative results from one of the Children we are treating.

Graph: ASD Child Pre-treament and 6 month treatment analyte levels vs healthy sweet spots

If you are interested in learning more, please contact me directly (612-801-1007) or pshuster@neuromics.com. Pete Shuster, CEO and Owner of Neuromics and ImmunoLink^TMTherapies.

immunoLink Therapies-Initial Products

Feeling and performing your best depends on balanced immunity and stem cell vitality

Your current investment in exercise, healthy diet and supplements are aiming you towards what we call the immunoLink sweet spot. If you have hit the bull’s eye, you would be seeking ways to stay there, because this is where you feel awesomely great and are most productive. If you have never been there, now is the time.

Illness, injury and medical procedures push you from the spot, but if you are near or on it, your recovery will be faster and more complete. The far reaches of the immunoLink Spectrum represent a cold wasteland of misery and despair.

How do we Know These Solutions Work?-Our Clients have autoimmune and degenerative diseases or are seeking peak performance. Prior to customizing their treatment strategies, we measure the levels of immune/inflammatory response, oxidative stress, allergens and stem cell health factors in our Clients' blood serum. The levels of these factors are compared with healthy controls. We then re-test after 6 months of treatments. For many the improvement in health and performance are startling. This is confirmed in the re-tests.

Stem-Kine and StemTrophin are central to our treatment strategies. Stem-Kine boosts the levels of circulating blood stem cells. These cells differentiate into immune factors and red blood cells. This boosts oxygen delivery and immunity resulting in greater endurance, mental acuity and cardiovascular health. StemTrophin activates mesenchymal stem cell migration. These cells are essential for immune suppression and then initiating the process of repair and regeneration at the sites of injury or degeneration. To learn more or order by phone, please call me directly at 612-801-1007 or you can also inquire via my e-mail: pshuster@neuromics.com. Pete Shuster, CEO and Owner, Neuromics and immunoLink Therapies.

Stem Cell Based Therapy For Tendinosis

Using Collagen I Producing Cells from Hair Follicles

Learn how these cells can be these cells can be harvested, purified and expanded to treat a chronic, degeneration of tendons.

Treating Autism-Immune System Balancing and Stem Cell Activating Therapies

Linking the Immune System with Stem Cell Health to Treat ASD Children

We have been testing immune/inflammatory response, oxidative stress and growth factor/neurotrophin biomarkers in the blood sera of Children with Autism Spectrum Disorder (ASD). We did this testing using our Quantibody Arrays.

The purpose of these tests were to establish a baseline for the state of these children's immune system and stem cell health. In this baseline testing, we found that all had high immune/inflammatory response, oxidative stress and low levels of the growth factors/neurotrophins needed for repairing and regenerated damaged cells/tissue (stem cell health).

We then treated these children with customized combinations of immune balancing and stem cell activating agents with the goal of showing that any improvement in ASD related symproms would be confirmed by positive modulations of biomarkers that are dysregulated in this disorder. We re-test at 6 months after initiation of the treatment plans.

The goal of our treatment strategies is to balance the immune system and activate the natural cell/tissue repair and regeneration (stem cell) processes. We call this the immunoLink^TM.

We are seeing improvement in the symptoms of the first 2 children recently retested and a marked improvement in the levels of their biomarkers. Here's data from one of the children. Note that key immune/inflammatory response markers have now moderated to the levels of healthy controls.

Graphs: Comparison of initial testing (red) and 6 month re-test (green) vs healthy controls (blue) for one of our ASD Children

We are developing a website that will have a wealth of information to help parents determine treatment strategies for ASD Children. We are also making our treatment plans and strategies available to interested parents. The website ecosystem will be named immunoLink^TM Therapies.

In the meantime, should you want to investigate if we could help your child, do not hesitate to contact me e-mail: pshuster@neuromics.com or cell: 612-801-1007. Pete Shuster, CEO and Owner.

Good Axon; Bad Axon: Regeneration in the CNS and Age

"The incapacity of the central nervous pathways to regenerate is a dogma accepted by science..." - Ramon y Cajal


Harvard University has opened access to Michio Wendell Painter's Dissertation: Regeneration in the aging peripheral nervous system. Will Spinal Cord Injury (SCI) and Neurodegenerative Diseases of the PNS become treatable with regenerative therapies? This work provides important insights.

Age plays a central role in regenerative capacities.

Neuropathy Research Solutions

New Publications and Past Postings

Here're are several key postings on Neuropathy and Neuropathic Pain:

• Cytokines and Neuropathic Pain
• Epidermal Nerve Fibers in Neuropathic Pain Model
• Pain Research and Gene Expression Analysis

Here're some "hot of the press" publications referencing use of Neuron-Glial Markers: 

Bethany L. Johnson-Kerner, Faizzan S. Ahmad, Alejandro Garcia Diaz, J. Palmer Greene, Steven J. Gray, R. Jude Samulski, Wendy K. Chung, Rudy Van Coster, Paul Maertens, Scott A. Noggle, Christopher E. Henderson and Hynek Wichterle. Intermediate filament protein accumulation in motor neurons derived from giant axonal neuropathy iPSCs rescued by restoration of gigaxonin. Hum. Mol. Genet. (2014) doi: 10.1093/hmg/ddu556. First published online: November 4, 2014.
...MAP2 (1:2,000, Neuromics, CH22103), NF-H (1:2000, Neuromics, CH22104), NF-L (1:200, Neuromics, MO22104)...

Images: Cells grown from adult rat brain. Large cell in middle is stained with mouse monoclonal to NF-L clone DA2 (green). Another type of neuronal lineage cell was stained with rabbit polyclonal to alpha-internexin (red). These cells were mitotic but had several characteristics of neurons. Rat spinal cord homogenate showing the major intermediate filament proteins of the nervous system (lane 1). The remaining lanes show blots of this material stainted with various antibodies including: NF-H, NF-M,, NF-L,, NF66 and GFAP 

Jianfei Guo, Xudong Fu, Xia Cui, Minhua Fan. Contributions of purinergic P2X3 receptors within the midbrain periaqueductal gray to diabetes-induced neuropathic pain. The Journal of Physiological Sciences November 2014.
...An equal volume of total and membrane samples was applied to SDS-PAGE. Membranes were incubated with the rabbit anti-P2X 3 primary antibody (1:1000, Neuromics, Edina, MN, USA) and goat anti-rabbit secondary antibody (1:200, Neuromics, Edina, MN, USA).

Neuropathy Research Solutions Include:

Antibodies Antibodies by Category

Apoptosis Research Reagents Detection kits, antibodies and proteins by category

ELISA Solutions ELISA kits, buffers, diluents and stabilizers by Category

Labeling Kits and Reagents Labeling kits, Cloning-Purification kits, Fluoro-tracers, i-Brite+, GFP and Related Markers

Cells, Cell Culturing Systems and Cell Based Assay Reagents Cells, Petaka Cell Culturing Systems, Biolumonics Kits, Media, 3-D Scaffolds and Related Reagents Designed to Optimize Cell Based Assays

Primary Neurons and Astrocytes Primary human, rat and mouse neurons and astro-glia by category

Proteins, Growth/Differentiation Factors and Small Molecules Recombinant/natural proteins plus small molecule/peptide agonists, antogonists, inihibitors and ligands

Stem Cell Research Reagents Cell, Media, Markers and Growth/Differentiation Factors

Transfection Reagents Transfection kits and reagents by Category

Immune System/Stem Cell Health and Aging

Looking, feeling and  performing your best depends on balanced immunity and stem cell vitality

As we age we encounter:

• Slower healing
• Longer recovery time from vigorous exercise
• More general aches and pains
• Longer recovery from illness
• Wrinkled, dry and thinning skin

Why is that? What are the causes? Answers can be found by a better understanding of immune/inflammatory response and stem cell systems. As we age our immune system weakens or becomes dysregulated (autoimmunity) and our stem cell "bank balances" deplete. My friend and world class stem cell therapies expert, Dr. Neil Riordan, gives an excellent description of the process: Your Body’s Stem Cell Bank Account.

To oversimplify, the immune system is responsible for cleansing the body of pathogens, toxins, allergens and damaged tissues/cells. This creates a healthy environment for stem cells to repair and regenerate new, healthy cells and tissue. We call this process the immunoLink^TM.

immunoLink Therapies is a new company of mine slated to launch in the spring of 2015. Our goal is to slow the aging process by offering solutions that balance your immunity and increase your stem cell vitality. Our vanguard product (currently available) is Stem-Kine. It is a blood stem cell booster which increases the level of immune factors and red blood cells in your cardiovascular system. The result is faster healing/recovery and increased endurance via better oxygen delivery.

If you desire to learn more, do not hesitate to call (612-801-1007) or e-mail: pshuster@neuromics.com. Thank you. Pete Shuster

Synchronized Bursting-Mouse Motor Neurons

GFP-Labeled Mouse Motor Neurons in Action

Video shows stunning bursting activity. Detected using Axion Biosystems MEAs. Check it out!

GFP Labeled Mouse Motor Neurons-Buy Now Save 100 USD

Designed for Motor Neuron Disease Research

I am pleased to add (finally) to our catalog potent, pure and ready to culture motor neurons. I know from the many request I have gotten for these that demand could be out stripping supply.

These are designed foruse in High-throughput fluorescent screening applications. Derived from transgenic mice expressing eGFP using Hb9 motorneuron promoter enables easy tracking and vi sualization of spinal motor neurons without the need for additional fluorescent markers and extended cultures.

Image: GFP+ mMN Mouse Motor Neurons at 2 days post thaw 20X

If you want to learn more about our any of our Neuron-Glial-Astrocyte Based Assay Solutions, do not hesitate to contact me (612-801-1007) or pshuster@neuromics.com. Pete Shuster, Owner and CEO, Neuromics.

100% Serum-Xeno Free Media for Expanding hMSCs

Works as well as our Serum Containing Media

 Selection of Growth and Differentaition media for Mesenechymal Stem Cell Assays is important for the ultimate performance of your cell based assays. The better the media the better the cutures & the lower your costs.

Images: Human mesenchymal stem cells (Catalog no. SC00A1) were plated at 5,000/cm² in a Falcon BD TC-coated T-25 flasks and maintained in serum containing media (Cat. No. SC00B1) and serum-free/xeno-free media (Cat. No. SC00B3) in reduced O2 environment (1% O2, 5% CO2, 90% N2) at 37°C in a humidified chamber. When cells became 80-90% confluent, they were subcultured, counted on a Beckermen Z2 particle counter (range 10-30uM), and passaged. Doubling time of 20-25hrs were calculated in each pass using ln(2*dT)/ln(Cf/Ci), where dT is the time, in hours, from inoculation to detachment; Ci is the initial number of cells plated and Cf is the final number of cells recovered from subculture. There were no apparent differences in doubling time with TC-coated flasks and Laminin/Fibronectin treated flasks.

Testimonial: “We tested the effects of MSCGroTM defined medium using several different lots of human adult primary stem cells and found that MSCGro supports a more robust proliferation rate than normal undefined media. This provided shorter doubling times and increased cellular yield, and maintained the cells in an undifferentiated state. We also found that MSCGro medium is stable under normal laboratory conditions for an extended time period compared to other defined media.” Ben Buehrer, VP and CSO, Zen-Bio

Neurite Outgrowth Assays

Cells, Media and Markers

I have previously posted use of our Neurons in Live Content Assays for the study of Neurite Outgrowth: Neurons-Live Content Assays. These assays are critical for the study of repair and regeneration.

A recent publication featured several of our Neuron Markers: Serena Quarta, Bastian E. Baeumer, Nadja Scherbakov1, Manfred Andratsch, Stefan Rose-John, Georg Dechant3, Christine E. Bandtlow, and Michaela Kress: Peripheral Nerve Regeneration and NGF-Dependent Neurite Outgrowth of Adult Sensory Neurons Converge on STAT3 Phosphorylation Downstream of Neuropoietic Cytokine Receptor gp130. The Journal of Neuroscience, 24 September 2014, 34(39): 13222-13233; doi: 10.1523/JNEUROSCI.1209-13.2014.
Live labeling of neuron cultures: After 20 or 48 h, neurons were live-labeled with α-gp130 antibody diluted in cold TNB medium for 30 min on ice. After washing, neurons were incubated with the secondary antibody diluted in cold TNB medium for 30 min and washed with PBS. Cells were fixed either with 4% PFA for 20 min at room temperature (RT) or with methanol at −20°C for 2 min. After permeabilization with 0.01% TX-100 (Pierce) unspecific binding was blocked for 30 min with 10% normal goat serum (Sigma-Aldrich) in PBS. Cells were incubated with the first antibody for 1 h, washed three times for 10 min with PBS and incubated with the appropriate secondary antibody for 30 min, counterstained with 4′, 6-diamidino-2-phenylindole (1:10,000; Sigma-Aldrich) and embedded in Mowiol (Calbiochem). As primary antibodies, α-gp130 (1:50; Neuromics), α-β-III-tubulin clone TuJ-1 (1:1000; R&D Systems), and α-neurofilament-H (α-NF-H; 1:200; Neuromics) were used. Secondary antibodies used were α-goat AlexaFluor 594 (1:1000; Invitrogen), chicken α-mouse AlexaFluor 594 or donkey α-mouse AlexaFluor 488 (1:1000; Invitrogen), and goat α-chicken AlexaFluor 568 (1:10,000; Invitrogen) for fluorescence microcopy.

Images: Reduced density of TuJ-1+ nerve endings in the epidermis in SNS-gp130−/− mice after lesion. A, Representative cross sections of hindpaw glabrous skin of naive and 12 dpl gp130fl/fl and SNS-gp130−/− mice stained with the pan neuronal marker TuJ-1. The dotted line indicates the border between dermis and epidermis. Scale bar, 40 μm. B, Quantification of the total number of TuJ-1+ fibers (NE) per 1000 μm2 of epidermal area shows a significant decrease in density in SNS-gp130−/− mice after lesion compared with control animals (*p < 0.05; n = 4 for each group). Data are presented as mean ± SEM and analyzed by Mann–Whitney U test. C, 3D reconstruction of the deeper layer of the dermis shows fewer nerve bundles in SNS-gp130−/− dermis compared with controls. D, No NF-H+ proprioceptive fibers were detectable in the epidermis of gp130fl/fl animals at 12 dpl. Scale bar, 40 μm.

If you want to learn more about our Neuron-Glial-Astrocyte based assay solutions do not hesitate to contact me (612-801-1007) or pshuster@neuromics.com. Pete Shuster, Owner and CEO, Neuromics.

The Growing Value of Cancer Associated Fibroblasts or CAFS

Balancing Supply and Demand

There is reciprocal communication between Cancer Associated Fibroblasts (CAFs) and tumor cells. They are now recognized as playing a key role in promoting the malignant process and blocking immune surveillance  They are becoming increasing important in Cancer Drug Discovery Programs.

Their growing value is making it harder for us to source resected tumors. We have been receiving advanced orders from clients who need to insure they have a stock. We have ordered tumors for the below CAFs and will be shipping to those who have place advance orders by the end of November.

Name	Catalog #	Type	Species	Applications	Size	Price
Human Colorectal Tumor CAFs Kit	CAF05	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Endometrial Adenocarcinoma CAFs	CAF01	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Ovarian Serous Tumor CAFs	CAF02	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Lung Adenocarcinoma CAFs	CAF07A	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Lung Squamous Cell Tumor CAFs	CAF07SC	Human Primary CAFs	H	Cell Assays	1,000,000 cells	$799
Human Immortalized Pancreatic CAF-Stellate Cells	CAF08	Human Primary CAFs	H	Cell Assays	500,000 Immortized Cells	$4,335
Human Pancreatic Tumor Cells Kit	PXPC3A1	Cells+Media Kit	H	Cell Assays	1,000,000 cells	$499
Conditioned Media for Ovarian Serous Tumor CAFS	CAF04	Cell Growth Media	H	Cell Assays	5 ml	$250
Conditioned Media from Endometrial CAFS	CAF03	Cell Growth Media	H	Cell Assays	5 ml	$250
VitroPlus III, low serum, complete	PC00B1-100	Cell Growth Media	H	Cell Assays	100 ml 500 ml	$65 $165

Image: Endometrial CAFs courtesy of Tiana Tonrey, Vitro Biopharma

Should have questions about availability and ship dates for any of these please call (612-801-1007) or e-mail: pshuster@neuromics.com. Thank you.
Pete Shuster, CEO, Neuromics

The Dance Between The Immune System and Stem Cells

We named it the  immunoLink^TM 

We have been testing a growing number of Clients with our Quantibody Arrays. Many of of these clients have Autoimmune Disorder Diseases. These range from Rheumatoid Arthritis to Multiple Sclerosis.

These arrays are designed to precisely measure factors or markers (proteins) that are dysregulated by these diseases. We measure the levels of these biomarkers in our Clients' Blood serum. The arrays have also been used to measure the levels of markers in plasma and cell culture supernatants.

Based on results, we are finding links between immune system and stem cell health. We call this the immunoLink. The link shows that when immune/inflammatory response markers are elevated, markers related to stem cell health are depleted.

Here we see the immune/inflammatory response markers IL-6, MCP-1 and TNF-alpha are elevated in our Clients with autoimmunity (A) vs Healthy Controls (HC). We also see lower levels of G-CSF and GM-CSF in these Clients.

G-CSF and GM-CSF are know to play a role in increasing circulating stem cells. GM-CSF is also know to be secreted by Mesenchymal Stem Cells (hMSCs) AND GM-CSF has anti-apoptotic functions on neurons, and is neuroprotective in animal stroke models while G-CSF has a prominent effect on the differentiation of adult neural stem cells (see: BMC Neuroscience 2007, doi:10.1186/1471-2202-8-88).

To us, the immunoLink means achieving a balance between immune system and stem cell health.

We provide immune system balancing and stem cell activating therapies for our Autoimmune Disease Clients and Children with Autism. We first do baseline and follow on testing (each 6 months) to determine how well the therapies are working. Our goal is to bring the many markers we test to healthier levels. As stem cell transplants become more common, moderating levels of immune/inflammatory response in patients could improve outcomes. If you would like to learn more, you can contact me at pshuster@neuromics.com or 612-801-1007.

Potent and Pure HUVECs

Only $199 for 500,000 Cells Through Sept. 30

HUVECs (Human Umbilical Vein Endothelial Cells) assays are important for studying the pathology of many diseases and cancers. We provide fresh cells at P1 in T25 flasks. They come to you ready to culture. Initial customer feedback has been favorable regarding ease of culturing and doubling cell populations (up to 16 passages). Now is the time to invest and build your stock of HUVECs!

Image: HUVECs cultured using our Collagel Hydrogel.

Characterization of the cells:

• Cytoplasmic VWF / Factor VIII: 95% positive by immunofluorescence 
• Cytoplasmic uptake of Di-I-Ac-LDL: 95% positive by immunofluorescence 
• Cytoplasmic PECAM1: 95% positive by immunofluorescence 

HUVECs are negative for HIV-1, HBV, HCV, and mycoplasma.

Questions? Contact information: Pete Shuster, CEO and Owner, Neuromics-phsuster@neuromics.com or direct phone: 612-801-1007.

Cytokines and Neuropathic Pain

I have been frequently posting results of testing blood serum of our clients suffering from Autoimmune (including Neuroimmuno) Diseases and Autistic Children. Our testing includes both standard medical testing (serum levels of toxins, metals, pathogen loads and markers like Cortisol, CRP, CDs, ALPS, NSE, S100b). In addition, we also doing testing using our Quantibody® Antibody.

Many of the clients report chronic mild to severe neuropathic pain.

By definition, this pain encompasses a series of heterogeneous conditions with some similar clinical manifestations. Peripheral examples include traumatic nerve injury, diabetic peripheral neuropathy and chemotherapy-induced peripheral neuropathy or Multiple Sclerosis. (see: Front Pharmacol. 2013; 4: 142.Published online Nov 22, 2013. doi: 10.3389/fphar.2013.00142). It is hard to unravel the cause and effects of this pain,

However, in analyzing the results from our clients' test we notice a relationship between dysregulation of some key cytokines and levels of pain (more data is required to confirm this). Here're our results (n=5)

Figure: Clients reporting pain vs healthy controls.

We will continue posting results here. We are releasing a Custom Quantibody Neuroimmuno-Pain Array which will include more bio-markers suspected to play a role in pain signaling.

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.

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

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

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 Publications: Capsaicin-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!

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.

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 Pain: Paulino 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.

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.

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

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.