Fatty Acids and IBD and Colitis

The Dangers of a High Fatty Acid Diets
Our westernized high-fat diet (HFD) is associated with the development of inflammatory bowel disease (IBD), Colitis and other intestinal related diseases and cancers. Here one the root causes is identified to elevated production of pro-inflammatory cytokine-IL-1β by macrophages. It is suspected that HFD sets off an autoimmune like cascade.

This cascade is triggered by NLRP3 inflammasomes. Our Apoptosis Kit-Magic Red-Real Time Cathepsin B is used to demonstrate these inflammasomes are activated via Cathepsin B Release: Shengnan Zhao, Zizhen Gong1, Jiefei Zhou1, Chunyan Tian, Yanhong Gao, Congfeng Xu, Yingwei Chen, Wei Cai and Jin Wu1Deoxycholic Acid Triggers NLRP3 Inflammasome Activation and Aggravates DSS-Induced Colitis in Mice. Front. Immunol., 28 November 2016 | http://dx.doi.org/10.3389/fimmu.2016.00536 ...Lipopolysaccharide-primed J774A.1 cells were incubated with or without DCA (100 μM, 24 h); then, the cells were stained with Lyso Tracker Green DND-26 (Invitrogen) or magic red cathepsin B fluorogenic substrate z-Arg-Arg cresyl violet (Neuromics) for 1 h, followed by Hoechst staining for half an hour. Fluorogenic signals were captured by inverted fluorescence microscope (Leica)...

Image: Lipopolysaccharide-primed J774A.1 cells were incubated with or without DCA (100 μM, 24 h); then, the cells were stained with Lyso Tracker Green DND-26 (Invitrogen) or Magic RedTM Cathepsin B fluorogenic substrate z-Arg-Arg cresyl violet (Neuromics) for 1 h, followed by Hoechst staining for half an hour. Fluorogenic signals were captured by inverted fluorescence microscope (Leica). Front. Immunol., 28 November 2016 http://dx.doi.org/10.3389/fimmu.2016.00536

We will continue to post how our customers are using our Apoptosis Kits for their discoveries here.

hNP1 Neural Progenitors and HTS Tox Assays

Sensitivity of Neural Progenitor Cells to Chemical Induced Apoptosis

This article demonstrates the sensitivity of Neuromics' hNP1TM Neural Progenitors to chemically induced apoptosis: Ingrid Druwea, Theresa M. Freudenrich, Kathleen Wallace, Timothy J. Shafer, William R. Mundy. Sensitivity of neuroprogenitor cells to chemical-induced apoptosis using a multiplexed assay suitable for high-throughput screening. doi:10.1016/j.tox.2015.03.011.

Summary: The results demonstrate that, (1) all three commercially available models generated a robust source of proliferating neuroprogenitor cells, and that the assay was sensitive and reproducible when used in a multi-well plate format; (2) there were differences in the response of the rodent and human neuroprogenitor cells to a set of chemicals previously shown to induce apoptosis in vitro; and (3) proliferating neuroprogenitor cells were more sensitive to chemical-induced apoptosis than differentiated neurons, suggesting that neuroprogenitor cells are one of the cell models that should be considered for use in a developmental neurotoxicity screening battery.

Here're more publications of referencing use of hNP1 Neural Progenitors:

Yuji Kaneko, Hideki Shojo, Jack Burns, Meaghan Staples, Naoki Tajiri, Cesar V. Borlongan, DJ-1 ameliorates ischemic cell death in vitro possibly via mitochondrial pathway, Neurobiology of Disease, Available online 21 September 2013, ISSN 0969-9961, http://dx.doi.org/10.1016/j.nbd.2013.09.007...Cell culture and oxygen-glucose deprivation (OGD) hNPCs were obtained from Neuromics...

Xiufang Guo, Severo Spradling, Maria Stancescu, Stephen Lambert, James J. Hickman. Derivation of sensory neurons and neural crest stem cells from human neural progenitor hNP1. Biomaterials, In Press, Corrected Proof,Mar 2013.doi:10.1016/j.biomaterials.2013.02.061 ...hNP1, were obtained from Neuromics (Edina, Minnesota)...

Questions? Do not hesitate to contact me, Pete Shuster: pshuster@neuromics.com or direct phone: 612-801-1007.

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.

Chronic Type II Diabetes Mellitus and Cardiovascular Disease

Our Neuropeptide and Neuropeptide Receptors and Leptin and Leptin Receptor Antibdodies are frequently used to study pathologies and biology specific to Obesity and Diabetes.

Here's a new publication studying the relationship between diabetic neuropathy and altered neuropeptide Y and its receptor expression levels in myocardium and plasma.

Robina Matyal, Feroze Mahmood, Michael Robich, Hiliary Glazera, Kamal Khabbaza, Philip Hessa, Cesario Bianchia, Robert Hagberga, Shu-Xu Hua, and Frank W. Sellkea. Chronic type II diabetes mellitus leads to changes in neuropeptide Y receptor expression and distribution in human myocardial tissue. European Journal of Pharmacology. Volume 665, Issues 1-3, 31 August 2011, Pages 19-28.

Abstract: Neuropeptide Y is one of the most abundant neurotransmitters in the myocardium, and is known to influence cardiovascular remodeling. We hypothesized that diabetic neuropathy could possibly be associated with altered neuropeptide Y and its receptor expression levels in myocardium and plasma. Plasma neuropeptide Y levels in diabetic (n = 24, HgbA1c 7.9 ± 1.1%) and non-diabetic (n = 27, HgbA1c 5.8 ± 0.5%) patients undergoing cardiac surgery utilizing cardiopulmonary bypass were analyzed. Right atrial tissue of these patients was used to determine the expression of neuropeptide Y, the receptors 1–5, and leptin by immunoblotting, real-time PCR and immunofluorescence. Apoptosis signaling and endostatin and angiostatin were measured to determine the effects of leptin.


Plasma neuropeptide Y levels were significantly increased in patients with Type II diabetes mellitus as compared to non-diabetic patients (P = 0.026). Atrial tissue neuropeptide Y mRNA levels were lower in diabetic patients (P = 0.036). There was a significant up-regulation of myocardial Y2 and Y5 receptors (P = 0.009, P = 0.01 respectively) in the diabetic patients. Leptin, involved with apoptosis and angiogenesis, was down regulated in diabetic patients (P = 0.05). The levels of caspase-3, endostatin and angiostatin were significantly elevated in diabetic patients (P = 0.003, P = 0.008, P = 0.01 respectively). Y1 receptors were more likely to be localized within the nuclei of cardiomyocytes and vascular smooth muscle cells.


Neuropeptide expression is altered differentially in the serum and myocardium by diabetes. Altered regulation of this system in diabetics may be in part responsible for the decreased angiogenesis, increased apoptosis, and increased vascular smooth muscle proliferation leading to coronary artery disease and heart failure in this patient population.

Neurotoxicity Testing

We combine our expertise in providing fresh and healthy:

E18 and E20 Rat Primary Neuronal Tissue -NEURON CULTURES

E18 Rat Primary Neuronal Tissue - ASTROCYTE CULTURES

E18 Mouse Neuronal Tissue -NEURON CULTURES

E18 Mouse Neuronal Tissue -ASTROCYTE CULTURES

AND Apoptosis Research Reagents

to help researchers more effectively study Neurotoxicity.

Images: Polycaspase Assay Kit, green was used to assess cell death in primay rat hippocampal neurons.Cells were plated on 25-mm poly-l-lysine-coated coverslips at 300,000 cells per coverslip. Cells were used at 4 or 8 days in vitro. Composite imagae (A) 3 out of 4 cells are apoptotic (green). No cells were necrotic as both of the PI-positive cells were FLICA-positive; they had compromised membranes and were probably in the late stages of apoptosis rather than necrosis. (B) 3 Caspase-positive cells fluoresce green.