Showing posts with label TNF-alpha. Show all posts
Showing posts with label TNF-alpha. Show all posts

Wednesday, February 11, 2015

Proven and Published Sandwich ELISA Kits

Growing Catalog of ELISA Kits

We are partnering with RayBiotech to manufacture our ELISA Kits. They have proven valuable additions to our Cell Based Assay solutions.

Neuromics has been built on a foundation of quality. RayBiotech's commitment to quality includes compliance to GLP/GMP FDA and ISO9000 regulations.
ELISA Kits Sample Data

Figure: Induction of proinflammatory cytokines is attenuated in CX3CL1−/− mice expressing sFKN. TNFα and IL-1β concentrations were measured using standard ELISA techniques for VM lysates. a, TNFα concentrations were upregulated following MPTP administration (three-way ANOVA; F(1, 23) = 18.36, ★★★p < 0.001). Comparatively, CX3CL1−/− mice expressing sFKN in the SNpc had significantly lower concentrations of TNFα relative to mFKN (Tukey's HSD; ***p < 0.001) and GFP (Tukey's HSD; ###p < 0.001) expressing mice. There were no significant differences between sFKN and WT-MPTP (Tukey's HSD; p = 0.384) or mFKN and GFP (Tukey's HSD; p = 0.773). b, The IL-1β concentrations in the VM were significantly upregulated for mice exposed to MPTP (three-way ANOVA; F(1, 23) = 11.97, ★★★p = 0.002). Similar to the pattern of TNFα, IL-1β concentrations in CX3CL1−/− mice expressing sFKN were significantly blunted compared to both mFKN (Tukey's HSD; ***p = 0.001) and GFP (Tukey's HSD; ###p < 0.001). As was observed with TNFα, there were no significant differences in IL-1β concentrations between WT-MPTP and sFKN (Tukey's HSD; p = 0.785) or mFKN and GFP (Tukey's HSD; p = 0.845) expressing mice. Data are presented as the mean ± SEM.

Questions? Please contact me directly, Pete Shuster, CEO and Owner, 612-801-1007 or pshuster@neuromics.com. Thank you.

Friday, July 11, 2014

Immune/Inflammatory Response and Autism

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

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

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

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

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

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

Sunday, June 06, 2010

STEMEZ(TM) hNP1 Cells and Neuroprotection Studies

Neuroinflammation has been shown to be a potent negative regulator of stem cell and progenitor cell proliferation in the neurogenic regions of the brain.

Here researchers used our STEMEZ(TM) hNP1 Human Neural Progenitors Expansion Kit to study the neuroprotection capabilities of a propriety nutraceutical formulation.

Adam D. Bachstetter, Jennifer Jernberg, Andrea Schlunk, Jennifer L. Vila, Charles Hudson, Michael J. Cole, R. Douglas Shytle, Jun Tan, Paul R. Sanberg, Cyndy D. Sanberg, Cesario Borlongan, Yuji Kaneko, Naoki Tajiri, Carmelina Gemma, Paula C. Bickford. Spirulina Promotes Stem Cell Genesis and Protects against LPS Induced Declines in Neural Stem Cell Proliferation. PLoS ONE 5(5): e10496. doi:10.1371/journal.pone.0010496.

Abstract:Adult stem cells are present in many tissues including, skin, muscle, adipose, bone marrow, and in the brain. Neuroinflammation has been shown to be a potent negative regulator of stem cell and progenitor cell proliferation in the neurogenic regions of the brain. Recently we demonstrated that decreasing a key neuroinflammatory cytokine IL-1β in the hippocampus of aged rats reversed the age-related cognitive decline and increased neurogenesis in the age rats. We also have found that nutraceuticals have the potential to reduce neuroinflammation, and decrease oxidative stress. The objectives of this study were to determine if spirulina could protect the proliferative potential of hippocampal neural progenitor cells from an acute systemic inflammatory insult of lipopolysaccharide (LPS). To this end, young rats were fed for 30 days a control diet or a diet supplemented with 0.1% spirulina. On day 28 the rats were given a single i.p. injection of LPS (1 mg/kg). The following day the rats were injected with BrdU (50 mg/kg b.i.d. i.p.) and were sacrificed 24 hours after the first injection of BrdU. Quantification of the BrdU positive cells in the subgranular zone of the dentate gyrus demonstrated a decrease in proliferation of the stem/progenitor cells in the hippocampus as a result of the LPS insult. Furthermore, the diet supplemented with spirulina was able to negate the LPS induced decrease in stem/progenitor cell proliferation. In a second set of studies we examined the effects of spirulina either alone or in combination with a proprietary formulation (NT-020) of blueberry, green tea, vitamin D3 and carnosine on the function of bone marrow and CD34+ cells in vitro. Spirulina had small effects on its own and more than additive effects in combination with NT-020 to promote mitochondrial respiration and/or proliferation of these cells in culture. When examined on neural stem cells in culture spirulina increased proliferation at baseline and protected against the negative influence of TNFα to reduce neural stem cell proliferation. These results support the hypothesis that a diet enriched with spirulina and other nutraceuticals may help protect the stem/progenitor cells from insults. Figure 7. Spirulina increases proliferation of human neural stem cells in vitro and protects against a TNFα insult.

Figure: Human neural progenitors grown under proliferation conditions were assessed by MTT assay (A) or BrdU (B) for the effects of spirulina (125 ng/ml) or NT-020 (500 ng/ml) or the two treatments combined in the presence or absence of TNFα (20 ng/ml) for 72 hours. (A) The MTT assay shows that spirulina alone or NT-020 alone increase proliferation; surprising, the in combination proliferation is decrease compare to control ** p less tan 0.005.


Related Reagents:

STEMEZ(TM) hNP1 Human Neural
Progenitors Expansion Kit

Primary Neurons and Astrocytes-Primary
human, rat and mouse neurons and astrocytes

Neuron/Glial Marker Antibodies

Neurotrophins and Growth Factor
Antibodies


Stem Cell Research Reagents
-includes
cells, antibodies, proteins, media and FACS kits.

Monday, August 18, 2008

MMP-9, E. Coli and Breast Cancer

The application of live bacteria for cancer therapies holds promise. In this study, Escherichia coli K-12 colonization on the tumour microenvironment by immunohistochemistry and fluorescence microscopy in the murine 4T1 breast carcinoma model. MMP-9 and TNF-alpha expression is altered in the sites of tumors.

Interestingly, the authors observed a postive change in the expression of these proteins after colonization.

Stephanie Weibel, Jochen Stritzker, Matthias Eck, Werner Goebel, and Aladar A. Szalay. Colonization of experimental murine breast tumours by Escherichia coli K-12 significantly alters the tumour microenvironment. 10.1111/j.1462-5822.2008.01122.x. Cellular Microbiology. © 2008 Blackwell Publishing Ltd
...goat anti-mouse MMP-9 antibody (Neuromics, Edina, MN)...