Blogs for May 2017

The role of STING/TMEM173 in gamma and encephalitis Herpes Simplex Virus (HSV)

Wednesday, May 31, 2017 - 10:57

Stimulator of interferon genes (STING), also known as TMEM173, promotes the production of the interferon’s IFN-alpha and IFN-beta.  STING possesses three functional domains: a cytoplasmic C-terminal tail, a central globular domain, and four N-terminal transmembrane motifs that attach it to the ER.  The role of STING in the immune response is specific to its ability to sense nucleic acids, particularly dsDNA.  When STING is over induced, the protein IRF3 undergoes a nuclear translocation resulting in IFN induction, which in turn activates the innate immune response. The herpes virus is a DNA based virus targeted to DNA-sensing pathways that has the capacity to elicit latent recurrent infections on a host. Because of the overlap between the herpes virus affecting DNA-sensing and the ability of STING to sense nucleic acids, using...

Apoptosis and Necroptosis Part I: Important factors to identify both types of programmed cell death

Friday, May 26, 2017 - 11:22

Different types of cell death have classically been identified by discrete morphological changes. The hallmarks of apoptosis include cell shrinkage, nuclear fragmentation and membrane blebbing whereas necroptosis is characterized by cell swelling and plasma membrane breakdown. While these two forms of cell death are clearly distinct, substantial crosstalk occurs between them.  Accordingly, it is becoming increasingly important to understand how these processes differ and to understand ways to differentiate them in cellular populations. 

Apoptosis is a genetically programmed mechanism of cell death that is activated in response to cell stress, infection or developmental cues.   Apoptosis is split into two main pathways, the intrinsic and extrinsic pathways, and can be triggered by granzymes from natural killer (NK) cells and cytotoxic T lymphocytes.  The intrinsic pathway is regulated by the ...

Rules for Selection of Fluorochromes in Multicolor ICC/IF

Monday, May 22, 2017 - 10:24

Immunocytochemistry/immunofluorescence (ICC/IF) involves visualization of antigens in cultured or smeared cells with the use of fluorochrome-labeled antibodies. Various combinations of fluorochrome-labeled antibodies make it possible to simultaneously detect multiple antigens in the same sample. However, every experiment involving multiplex or multicolor ICC/IF requires selection of an optimal set of fluorochromes. Poor selection of fluorochrome combinations can make it difficult to distinguish individual antigens, especially when the targets under consideration are co-localized at the sub-cellular level. Here we have outlined key rules for selecting fluorochromes to minimize potential errors and to simplify the selection process.

1. Check for fluorochrome(s) compatibility with your microscope’s filter sets

Use spectra viewers and fluorochrome reference charts to determine the maximum excitation (Ex) and emission (Em) wavelengths...

Article Review: Effects of the administration of high-dose interleukin-2 on immunoregulatory cell subsets in patients with advanced melanoma and renal cell cancer

Friday, May 19, 2017 - 10:38

The immune system is composed of a portion of T cells that express an invariant T cell receptor (TCR) alpha chain known as V alpha 24 J alpha 18. These highly conserved populations are referred to as iNKT populations and have the ability to rapidly produce cytokines following activation, making them hot targets for therapeutic research initiatives. In this article, we will review van derVilet et al’s study of the effects of administering high dose interleukin-2 to immunoregulatory cell subsets in patients presenting advanced melanoma and renal cell cancer.  This group uses antibodies in a variety of applications to determine whether these cells can rescue skin and kidney malignancies.  In this paper, dendritic cell subsets (DC), CD1d-reactive invariant natural killer T cells (iNKT) and CD4+CD25+ regulatory-type T cells are exposed to high-dose IL-2 therapy. 


Article Review: Dual effects of carbon monoxide on pericytes and neurogenesis in traumatic brain injury

Tuesday, May 9, 2017 - 08:58

Traumatic brain injury (TBI) currently contributes to nearly 30% of all injury deaths in the United States.  Characterized by an abrasive head injury that interrupts normal brain function, TBI can range from mild to severe.  Mild symptoms can present themselves as excessive tiredness, difficulty concentrating and lack of clear thinking.  Severe cases of TBI are hallmarked by unusual behavior, seizures and loss of consciousness.  Research has shown that on a molecular level TBI triggers various mechanisms of cell death alongside attempted tissue recovery, therefore Choi et al sought out to test the treatment of carbon monoxide on rescuing the effects of TBI.  Their article titled “Dual effects of carbon monoxide on pericytes and neurogenesis in traumatic brain injury” tests the effects of CO-releasing molecular 3 (CORM-3) and N-tert-butyl-a-phenylnitrone (PBN) on a mouse model of controlled cortical impact TBI.


Recent advances in CRISPR-Cas9

Thursday, May 4, 2017 - 09:49

The CRISPR-Cas9 genome-engineering tool is a powerful opportunity for researchers to study individual gene function. CRISPR-Cas9, abbreviated for Clustered Regularly Interspaced Short Palindromic Repeats, is a bacterial defense system that can be reprogrammed to target specific areas of DNA followed by precise editing.  Essentially, CRISPR sequences are transcribed into short RNA sequences that will match the desired DNA sequence of interest. From here, the DNA is bound and cut, turning off its function.  While this recent discovery opened up doors for gene-targeted therapies, the mechanisms in which DNA cutting was achieved are somewhat debated. Simply put, when DNA is damaged, it sets out to repair itself, resulting in randomized nucleic acid insertions that are not needed. 


Pathway Highlight: Three key factors that contribute to cellular heterogeneity in apoptosis

Monday, May 1, 2017 - 08:54

Have you ever wondered why cells in the same population respond differently to an apoptotic stimulus? Apoptosis, a form of programmed cell death, is vital for the removal of unwanted or damaged cells. As with most cellular processes, too much or too little activation can be detrimental and lead to various diseases including autoimmune disorders and cancer. While this process is tightly regulated, cells undergo apoptosis in a non-synchronized manner, which complicates the analysis of apoptotic events.  Here are 3 keys factors that determine how an individual cell will respond to an apoptotic stimulus.

caspase-3 antibody

Processed caspase 3 was detected in immersion fixed anti-FAS treated Jurkat human acute T cell leukemia cell line using Human/Mouse Active Caspase 3 Polyclonal Antibody (...

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