Hope for Stroke Victims-Transplanting STEMEZ hNP1 Cells
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Hope for Stroke Victims-Transplanting STEMEZ hNP1 Cells
December 11, 2009 – 6:51 am
Featuring Dr. Pat Carr
October 10, 2009 – 10:06 am
Amyotrophic Lateral Sclerosis (ALS)-New Twists on Root Causes
| Teacher, Mentor and Friend Dr. Pat Carr has been a key figure in helping shape the direction of my company. He has a gift for communicating the nuances of his research and coaching me on how to best serve labs like his. Based on these interactions, it came as no surprise to learn of his being Recognized for Excellence in Teaching, Research and Service at University of North Dakota.
“Dr. Carr has a magic way of teaching,” said second-year medical student, Tyson Bolinske. “He is able to take the most difficult topics and, through detailed notes, logically break down the material.”
From a recent dialog, I learned of his growing work on the Ventral Horn and search for root causes of Amyotrophic Lateral Sclerosis (ALS). I wanted to learn more! I would like to thank Pat for agreeing to share his story and giving me the opportunity to feature highlights in “News Behind the Neuroscience News”. Information on ALS
ALS is an insidious disease. It is a progressive neurodenerative disease that is always fatal. Approximately 5600 new cases are diagnosed each year. Average survival is typically 3-5 years from onset. The most common form of ALS in the United States is “sporadic” ALS. It can happen to anyone at anytime. The other is the inherited form named “Familial” ALS (FALS). Only about 5 to 10% of all ALS patients appear to have FALS. As the disease progresses the symptons become more acute. Paralysis spreads through the body affecting speech, swallowing, chewing and breathing. Ventilator support is need in late stages Pat’s Journey Pat took the “road less traveled”. He was a passionate hockey player in Canada. He concluded in his late teens that he was not at a level to take this road to wealth and fame. |
 Pat Carr 06/04–present Associate Professor, Department of Anatomy & Cell Biology, School of Medicine and Health Sciences, University of North Dakota 1996–98 Research Associate/Adjunct Assistant Professor/Auxilliary Assistant Professor, Department of Anatomy;Wright State University 07/98–06/04 Assistant Professor, Department of Anatomy & Cell Biology, School of Medicine and Health Sciences, University of North Dakota Postdoc, National Institutes of Health, Neuroscience, 1994-96 Postdoc, University of Manitoba, Neuroscience, 1992-1994 Ph.D., University of Manitoba, Physiology, 1992 |
Next was a stint as an automechanic in Brandon, Canada. The discipline and logic involved in fixing cars catalyzed an interest in Science which led to him going to Brandon University to study Geology. When the oil market collapsed in 1983, he decided to change his studies to Zoology and earned a BS in 1984.
A passion was sparked when he did field research in the Canadien Rockies studying parasites in Columbian Ground Squirrels. He loved it, but recognized the limited value of continuing thsese studies. This lead to the wide open field of Neuroscience and the opportunity to study and solve problems that could benefit mankind. His graduate work at University of Manitoba and focusing on Neuropathic Pain and the Dorsal Horn. He then moved on to studying Ventral Horn and Motor Control Function for his Post Doc at Wright State.
From Pain to ALS
It was Pat’s work in Pain at the University of North Dakota that brought me into initial contact with him. He generously put some of our key Pain/Inflammation and Neurotransmission Research Antibodies through their paces. These included some of our Neuropeptide and Neuropeptide Receptors , P2X Receptors and TRPV1s (Vanilloids).
His previous work in studying the Ventral Horn combined with a colleagues mouse model of ALS combined to create a prefect opportunity to advance the understanding of ALS. Pat cautioned me with this insight: ”sometimes it is not what you want to study; it is what you can study. The model is SOD1 (superoxide dismutase 1) which is core to FALS.(occurs in only about 10% of the ALS cases).
Pat is broadening the play field by looking at what else is happening in sporadic ALS vs FALS. Specifically, he is looking at modulation of alpha Motor Neurons and how the activity of adjacent Renshaw Cells impact signaling and modulation. Renshaw Cells act as a “governor” on the activity of these alpha Motor Neurons.
He is drilling down by studying the signaling of ChAT (Choline Acetyltransferase), VAChT (Vesicular acetylcholine transporter) and related molecules. By gaining a deeper understanding of how Renshaw Cells signaling changes the activity of alpha Motor Neurons in ALS, Pat and his team are taking steps towards discovering roots causes.
As these root causes are further illuminated, I will be reporting specifics in my blog.
Spinal Cord Injury Repair
August 4, 2009 – 8:21 pm
I wanted to feature yet more research on spinal cord injury repair.
Dr. Mark Tuszynski and his team of researchers at USCD recently published work that tested their hypothesis that chemotropic mechanisms would guide regenerating spinal cord axons to appropriate brainstem targets.
Laura Taylor Alto, Leif A Havton, James M Conner, Edmund R Hollis II, Armin Blesch & Mark H Tuszynski. Chemotropic guidance facilitates axonal regeneration and synapse formation after spinal cord injury. Nature Neuroscience. Published online: 2 August 2009 | doi:10.1038/nn.2365.
Their study included use of Neuromics’ NT-3 Antibody.
STEMEZ hN2 Human Neurons Data
July 26, 2009 – 12:33 pm
I have been working with Dr. Steve Stice and Aruna Biomedical to deliver human stem and neural cells to identified niche research areas related to drug discovery. Neuromics rolled out STEMEZTM hN2 Human Neurons Discovery Kits several months ago. Applications for these include: cellular model studies, high content screening, developmental studies, RNAi studies and genetic manipulation.
Drilling down further, I am pleased to present Electro-physiology and related data generated by Aruna and collaborators: hN2 Cells-Electro Phys Data Supplement
hN2-Whole Cell Voltage Clamp
Figure. hN2 cells can produce inward currents that generate action potentials. (A) Isolated hN2 with significant neurite growth 1 week after plating . This cell was subjected to whole cell voltage clamp utilizing a potassium gluconate based intracellular solution. (B) Voltage gated inward and outward currents were elicited from this cell with depolarizing voltage steps. (C) Inward currents from another cell (potassium gluconate intracellular) were abolished by local application of 1 µM tetrodotoxin (red trace) while outward currents remained. Inward current recovered as TTX washed out of the region (green trace). (D) A different cell which exhibited voltage activated inward currents that inactivated in response to a 50 ms prepulse at different membrane potentials. The experiment was done 27 days after the removal of bFGF. A cesium gluconate based intracellular solution was used for this experiment to block outward potassium currents. The membrane potential for half maximal inactivation by standard Boltzman fitting (red line) was -40.1 mV with a slope of 4.7. (E) Recovery from fast inactivation utilizing a paired pulse protocol in the same cell as C. The single exponential time constant for recovery of inactivation was 1.7 ms (red line). (F) A different cell which elicited an overshooting action potential upon current injection under whole cell current clamp utilizing a potassium gluconate based intracellular solution. Inset: Response of the same cell under voltage clamp to a change in membrane potential from -80 mV to -10 mV elicited a peak current of 457 pA. Scale bars for inset: 5 ms, 0.2 nA.
Gary Johnson-Apoptosis Ace
June 27, 2009 – 9:45 am
About Gray
 Gary Johnson 1994-present-President, ICT 1993-1996-Conjugation Chemist, R&D Systems 19989-1993-Supervisor Protein Conjugation & ELISA Development Group, Solvay Animal Health 1986-1989-Immunologists, Biosciences Lab, 3M 1976-1986-Various Lab, U of MN Gary’s Conatct Info:
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Inventing Better Ways to Measure Apoptosis
This profile features another Scientist Entrepreneur. Dr Gary Johnson is the Founder and President of Immunochemistry Technologies LLC (ICT). His company manufactures kits that have the capabilities to quantitatively measure apoptosis effects. This is important to Neuromics, because these are core to many diseases of research interest to our customers. These range from Cancer where apoptosis detection can be used to to visualize the efficacy of tumor killing therapies to Neuroscience where apoptosis could be a root cause of many cognitive and neuro-muscular diseases.
I am excited about featuring Gary. I have been working with him and his team over the past 5 years. They have actively supported my company in providing Apoptosis Research Kits. The strength in our relationship is built on his company supplying best of breed reagents. The feedback I receive from users is overwhelmingly positive. In addition to these kits, ICT is also recoginized for their rock solid ELISA Buffers and Diluents.
It takes a unique blend of business and scientific acumen to build a company like ICT. So let’s start with Gary’s background and experience and then on to the specifics on his company and products and what sets ICT apart from competitors.
Gary’s Background
Gary’s began his career at the University of Minnesota in 1978 where he worked in a variety of labs. There he gained a wealth of experience and expertise in research techniqes. These included chromatography, immunoelectrophoresis, radiolabeling, mass spectrometry, proton NMR spectroscopy and western blotting.
He leveraged his abilities and became more deeply involved in immunobiology. He joined Dr. Harry Orr’s lab in 1981. There he used recombinant DNA techniques to study the class I genes of the major histocompatibility complex and he also supervised the tissue culture work. This provided the stepping stone to Dr. David Klein’s lab in 1984. There he studied the difference between diabetic and non-diabetic glomerular basement membrane proteoglycans in kidney disease. In order to do this research Gary developed in vivo or in vivo labeling techniques.
Gary then moved from University to commercial labs. We will see how his growing expertise morphed into the founding of ICT and why his broad knowledge and experise enabled a successful launch of the company.
From 1986 until founding ICT Gary worked at 3M, Solvay Animal Health and R&D Systems. Over his tenure, he worked as an Immunologist, Supervised an ELISA and Protein Purification and was a Conjugation Chemist. Having mastered a unique range of basic and commercial bio-research techniques, the evolution to Scientist-Entreprenuer was a natural next step.
In 1994, Dr. Brain Lee and Gary launched ICT. The company’s early success was in contract assay development. The revenue generated from these programs, has enabled ICT to manufacture and release a growing catalog of Apoptosis Detection Kits.
ICT’s Products and Capabilties
ICT’s provides proprietary probes for measuring apoptosis in vitro and in vivo. These probes are used by researchers to detect caspases, cathepsins, serine proteases, cholinesterase enzymes, and assess mitochondrial health.Applications include: assessing the efficacy of chemotherapy, to quantifying neurodegeneration, and early detectionof eye disease, to name a few.
Specific Products Include:
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- Fast!-Use Caspase kits to quantitate apoptosis via active caspases in whole, living cells. These kits do not use ELISA or any antibodies for detection
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Measures chymotrypsin-like protease
activation in whole living cells.
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Measures apoptosis in whole living, intact cells – no lysis required
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Quantitate mitochondrial functionality and apoptosis
Images: Normal (left) and keratoconus (right) corneal fibroblasts were labeled with Caspase 3 & 7 Assay Kit, green.
Pacing the Field
ICT is setting the pace in Apoptosis Detection by recognizing and resolving issues inherent in competitive offerings. These include:
- Difficulty permeating cells.
- High background problems.
- Does not bind to early stage apoptotic cells.
- Not as sensitive as a cell permeant inhibitor probe.
- Does not bind to all apoptotic tumor cells (Dicker, Cancer Biol. Ther., 2005. 9:1014-1017).
- Binds positively to normal and healthy bone marrow derived cells (Dillon, J. of Immunol., 2001. 166:58-71).
- Many in vitro protocols involve lysing the red blood cells before running flow cytometry, this method results in the binding of Annexin V to all of the cells in the sample (Tait, Blood, Cells, Molecules, and Diseases., 1999. 25:271-278). The inversion of PS and cells containing large amounts of PS may not be related to apoptosis and this adds to the background issues.
- Does not measure a process of apoptosis, but rather an effect of apoptosis.
Capabilities that will enable them strengthen their leadership position include:
- Uses a cell permeant probe that can easily penetrate tissues and cells.
- Very sensitive.
- Specific, no reported false positives.
- It is a direct measurement of an intracellular process of apoptosis, detects only active caspases and caspase active cells are always apoptotic.
- Passage through the blood-brain barrier has been demonstrated.
- Passage through the blood-retinal barrier has been demonstrated.
- No background problems when injected intravenously.
- Detects very early through late stage apoptosis.
ICT is continuing to invest heavily in developing new capabilties. Gary highlighlighted some of the breakthroughs that are on the horizon. I plan on announcing these as they become public.Stay tuned.
Advancing the Study of Apoptosis
May 25, 2009 – 10:32 am
Featuring Gary Johnson and his team at Immunochemistry Technologies LLC.
The ability to accurately measure apoptosis processes is a core research component for many of our customers and colleagues. Neuromics has leveraged our growing partnership with Gary and ICT to meet the exacting requirements of Researchers studying apoptosis.
I am excited to be featuring Gary and his company in our June News Behind the News. Gary and his team have
Polycaspase Apoptsis
proven to me time and again the ability to deliver methods and kits that meet our customers’ needs. I can count on the feedback to be positive and use to expand in user labs.
In the feature, I will provide details of Gary’s unique background. The path that lead him to founding ICT and the development of current capabilities. Most importantly, I will provide a glimpse of coming new methods and products. These could significantly improve the development of therapies for diseases that involve aptotosis.
Image: Jurkat cells dually stained with Hoechst and Polycaspase Assay Kit, green-FAM-VAD-FMK. Caspase activity is revealed by green fluorescence in cell #2, indicating that only this cell is apoptotic. Cell #1 is also dying (scattered blue), but is not apoptotic because it is not green. Cell #3 is healthy (concentrated blue nucleus).
Knockdown of rSNSR1 in vivo
April 20, 2009 – 10:54 am
I have featured successes with delivering siRNA in vivo in this blog. These included stories on Dr. Philipe Serrat and his team at the University of Sherbrooke and Dr. Mark Behlke’s work at Integrated DNA and Dicerna.
I am pleased to report the parade of success with use our i-FectTM in vivo grows.
Here’s the most recent study:
Christian Ndong, Amynah Pradhan, Carole Puma, Jean-Pierre Morello, Cyrla Hoffert, Thierry Groblewski , Dajan O’Donnell, Jennifer M.A. Laird. Role of rat sensory neuron-specific receptor (rSNSR1) in inflammatory pain: Contribution of TRPV1 to SNSR signaling in the pain pathway. PAIN 143 (2009) 130–137.
…For experiments in which siRNA was delivered by bolus injections, 10 ul of siRNA or vehicle was injected directly into the intrathecal catheter once daily for 4 days. In this case, siRNAs were prepared immediately prior to administration by mixing the RNA solution (200 uM in annealing buffer) with the transfection reagent i-FectTM (Neuromics) at a ratio of 1:4 (w:v) for a final siRNA/ lipid complex concentration of 2 ug/10 ul…
Related Data:
Images: in vivo characterization of knockdown produced by rSNSR1 siRNA. (A) A dose-dependent decrease in rSNSR1 mRNA levels measured in lumbar L3/L4/L5 DRGs was
observed when rSNSR1 siRNA (n = 7–14/group) or MM siRNA (n = 6/group) was delivered by four daily bolus injections. *p < 0.05; **p < 0.01; ***p < 0.001 as determined by oneway analysis of variance followed by sequential testing. (B) rSNSR1 immunoreactivity in dorsal horn of the spinal cord was visibly reduced in rSNSR1 siRNA-treated animals (5 lg/day, left panel). Immunoreactivity with neuron-specific isolectin B4 (IB4; right panel) did not change between treatment groups, showing the integrity of each dorsal horn analyzed (n = 6/group). (C) A semi-quantitative score of rSNSR1 immunoreactivity showed that siRNA treatment greatly decreased rSNSR1 protein levels compared to MM and control groups. A blinded observer scored 9–12 individual sections taken from a 1 cm segment of the spinal cord.
Primary Neurons Culturing Expertise
April 6, 2009 – 2:03 pm
I recently featured Dr. Evanna Gleason. As part of this, we highlighted her lab’s epertise in culturing our E18 Primary Rat Hippocampal Neurons.
I recently received impressive data and protocols from Emily Mcmains, a lab member.
Please note the excellent image of the cells 1 week after culturing and images taken after 4 days.
Courtesy of Emily McMains (Gleason Lab), LSU.
Consistent Human Neurons
March 25, 2009 – 10:16 am
We have featured Dr. Steve Stice here. He and his team at UGA and Aruna Biomedical are developing products that are highly desired by Neuroscience Researchers.
We are in the process of finalizing details for distributing their human neuron cultures. Here is the related press release:
| ArunA Biomedical, Inc. announces alliance with Neuromics for distribution of normal human neural cells. |
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| Athens, Georgia – - March 23, 2009 – - ArunA Biomedical, Inc., announced today an agreement with Neuromics, Inc. of Edina, MN, giving Neuromics the right to non-exclusively market and sell the ArunA hN2™ Human Neural Cells and Neural Culture Medium to support applications in neurological research.ArunA has an exclusive worldwide license to develop and commercialize neural cells derived from human embryonic stem cells (hESC), and hN2 is a second generation product from this technology. These cells offer a consistent population of normal human neural cells that the neural research and pharmaceutical market highly desires.
“ArunA has further developed its adherent monolayer technology by creating hN2™, a normal human neural cell ideal for drug screening, toxicology studies and basic neural research, and we are pleased to have Neuromics as a distribution partner,” said David Ray, Chief Executive Officer of ArunA Biomedical “Neuromics growth is catalyzed by offering the unique products and expertise our customers require for research success through strategic alliances with companies like ArunA Biomedical. This relationship represents a growth opportunity for us. Their hN2™ cells fill a stated research need of the Neuroscience Community and we look forward to our customers having these cells and the related new discoveries they will help generate,” said Pete Shuster, CEO and Owner of Neuromics. Founded in 2003, ArunA Biomedical, Inc. is a privately held biotechnology corporation dedicated to the discovery, manufacturing and commercialization of emerging new technologies in human embryonic stem cell research for use in drug discovery and neuroscience research. Founded in 2003, Neuromics is a privately held Bio-regents Company focusing on providing research ready and proven products and methods expertise to Neuroscience, Diabetes/Obesity, Immunology and Researchers. |
