Cell Cycle Analysis (PI / RNASE) solution.

An article published this year in “NANO RESEARCH” using our Cell Cycle Analysis (PI / RNASE) solution, by our customers from Department of Organic Chemistry, Faculty of ScienceUniversity of Málaga and Biosanitary Institute of Granada (IBS. GRANADA) SAS-Universidad de Granada, Spain, in the study of how Paclitaxel-loaded hollow-poly (4-vinylpyridine) nanoparticles enhance drug chemotherapeutic efficacy in lung and breast cancer cell lines. Congrats and Thanks.

Summary:

Paclitaxel (PTX), one of the most effective cytotoxins for the treatment of breast and lung cancer, is limited by its severe side effects and low tumor selectivity. In this work, hollow-poly(4-vinylpyridine) (hollow-p4VP) nanoparticles (NPs) have been used for the first time to generate PTX@p4VP NPs, employing a novel technique in which a gold core in the center of the NP is further oxidized to produce the hollow structure into which PTX molecules can be incorporated. The hollow-p4VP NPs exhibit good physicochemical properties and displayed excellent biocompatibility when tested on blood (no hemolysis) and cell cultures (no cytotoxicity). Interestingly, PTX@p4VP NPs significantly increased PTX cytotoxicity in human lung (A-549) and breast (MCF-7) cancer cells with a significant reduction of PTX IC50 (from 5.9 to 3.6 nM in A-549 and from 13.75 to 4.71 nM in MCF-7). In addition, PTX@p4VP caused a decrease in volume of A-549 and MCF-7 multicellular tumor spheroids (MTS), an in vitro system that mimics in vivo tumors, in comparison to free PTX. This increased antitumoral activity is accompanied by efficient cell internalization and increased apoptosis, especially in lung cancer MTS. Our results offer the first evidence that hollow-p4VP NPs can improve the antitumoral activity of PTX. This system can be used as a new nanoplatform to overcome the limitations of current breast and lung cancer treatments.

Reference:

http://link.springer.com/article/10.1007/s12274-016-1340-2

Product link:

http://immunostep.com/solutions-buffers-chemicals/1289-pi-rnase-solution.html

Annexin V FITC.

An article published this year in “CHEMISTRY AND PHYSICS OF LIPIDS” using our Annexin V FITC, by our customers from Unidad de Biofísica (CSIC, UPV/EHU), and Departamento de Bioquímica, Universidad del País Vasco, Bilbao, Spain, in the analysis of Lipidomic profile of GM95 cell death induced by Clostridium perfringens alpha-toxin. Congrats and Thanks.

Summary:

Clostridium perfringens alpha-toxin (ATX) is considered as a prototype of cytotoxic bacterial phospholipases C, and is the major virulence factor in C. perfringens-induced gas gangrene. It is known that, depending on the dose, ATX causes membrane disruption and cytolysis or only limited hydrolysis of its substrates. In the latter case, toxin activity leads to the unregulated generation of bioactive lipids that can ultimately induce cell death. We have characterized apoptosis and necrosis in highly ATX-sensitive, ganglioside-deficient cells exposed to different concentrations of ATX and we have studied the lipidomic profile of cells treated with ATX as compared to native cells to detect the main changes in the lipidomic profile and the possible involvement of lipid signals in cell death. ATX causes both apoptosis and necrosis, depending on dose and time. ATX activates cell death, stimulating the release of cytochrome C from mitochondria and the consequent activation of caspases-3. Moreover GM95 cells treated with ATX showed important lipidomic alterations, among them we detected a general decrease in several phospholipid species and important changes in lipids involved in programmed cell death e.g. ceramide. The data suggest two different mechanisms of cell death caused by ATX, one leading to (mainly saturated) glycerophospholipid hydrolysis related to an increase in diacylglycerols and associated to membrane damage and necrosis, and a second mechanism involving chiefly sphingomyelin hydrolysis and generation of proapoptotic lipidic mediators such as ceramide, N-acylethanolamine and saturated non-esterified fatty acids.

Reference: 

http://www.sciencedirect.com/science/article/pii/S0009308416301177

Product link:

http://immunostep.com/apoptosis-tools/3731-anxvf-200t.html

AnnexinV-DY634.

An article published this year in “PLOS ONE” using our AnnexinV-DY634, by our customers from Fundación Inbiomed, Foundation for Stem Cell Research, Mesenchymal Stem Cell Laboratory, San Sebastián, Spain, in the study of how IFNγ Regulates Activated Vδ2+ T Cells through a Feedback Mechanism Mediated by Mesenchymal Stem Cells. Congrats and Thanks.

Summary:

γδ T cells play a role in a wide range of diseases such as autoimmunity and cancer. The majority of circulating human γδ T lymphocytes express a Vγ9Vδ2+ (Vδ2+) T cell receptor (TCR) and following activation release pro-inflammatory cytokines. In this study, we show that IFNγ, produced by Vδ2+ cells, activates mesenchymal stem cell (MSC)-mediated immunosupression, which in turn exerts a negative feedback mechanism on γδ T cell function ranging from cytokine production to proliferation. Importantly, this modulatory effect is limited to a short period of time (<24 hours) post-T cell activation, after which MSCs can no longer exert their immunoregulatory capacity. Using genetically modified MSCs with the IFNγ receptor 1 constitutively silenced, we demonstrate that IFNγ is essential to this process. Activated γδ T cells induce expression of several factors by MSCs that participate in the depletion of amino acids. In particular, we show that indolamine 2,3-dioxygenase (IDO), an enzyme involved in L-tryptophan degradation, is responsible for MSC-mediated immunosuppression of Vδ2+ T cells. Thus, our data demonstrate that γδ T cell responses can be immuno-modulated by different signals derived from MSC.

Reference:

http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0169362

Product Link:

http://immunostep.com/apoptosis-tools/3730-anxvdy-200t.html?search_query=DY634&results=6

Annexin V FITC.

An article published 1st January 2017 in “ONCOMEDICINE” using our Annexin V kit, by our customers from Medical Clinic, Hospital Coburg, University of Wurzburg, Coburg, Germany and Pulmonary Department-Oncology Unit, “G. Papanikolaou” General Hospital, Aristotle University of Thessaloniki, Greece, in the study of COX-2 Inhibitors, a Potential Synergistic Effect with Antineoplastic Drugs in Lung Cancer. Congrats and Thanks

Summary:

Lung cancer represents the leading cause of cancer-related deaths worldwide and novel therapeutic approaches targeting crucial pathways are urgently needed to improve its treatment. Inflammation plays a critical role in multistage tumor development and increased evidence has supported the involvement of cyclooxygenase-2 expression in carcinogenesis. We investigated the potential use of COX-2 inhibitors in cancer proliferation and apoptosis.

Reference:

http://www.oncm.org/v02p0028.htm

Product link:

http://immunostep.com/apoptosis-tools/3731-anxvf-200t.html

CELL FIXATION & PERMEABILIZATION (INTRA)

An article published last 27 December 2016 in “AGING CELL” using our Fixation/Permeabilization kit, by our customers from Department of Immunology, Hospital Universitario Central de Asturias, Oviedo, Spain, in the study of Phenotypic characteristics of aged CD4⁺ CD28^null T lymphocytes are determined by changes in the whole-genome DNA methylation pattern. Congrats and Thanks.

Summary. 

Aging is associated with a progressive loss of the CD28 costimulatory molecule in CD4+ lymphocytes (CD28null T cells), which is accompanied by the acquisition of new biological and functional properties that give rise to an impaired immune response. The regulatory mechanisms that govern the appearance and function of this cell subset during aging and in several associated inflammatory disorders remain controversial. Here, we present the whole-genome DNA methylation and gene expression profiles of CD28null T cells and its CD28+ counterpart. A comparative analysis revealed that 296 genes are differentially methylated between the two cell subsets. A total of 160 genes associated with cytotoxicity (e.g. GRZB, TYROBP, and RUNX3) and cytokine/chemokine signaling (e.g. CX3CR1, CD27, and IL-1R) are demethylated in CD28null T cells, while 136 de novo-methylated genes matched defects in the TCR signaling pathway (e.g. ITK, TXK, CD3G, and LCK). TCR-landscape analysis confirmed that CD28null T cells have an oligo/monoclonal expansion over the polyclonal background of CD28+ T cells, but feature a Vβ family repertoire specific to each individual. We reported that CD28null T cells show a preactivation state characterized by a higher level of expression of inflammasome-related genes that leads to the release of IL-1β when activated. Overall, our results demonstrate that CD28null T cells have a unique DNA methylation landscape, which is associated with differences in gene expression, contributing to the functionality of these cells. Understanding these epigenetic regulatory mechanisms could suggest novel therapeutic strategies to prevent the accumulation and activation of these cells during aging.

Reference:

http://onlinelibrary.wiley.com/doi/10.1111/acel.12552/full

Product link:

http://immunostep.com/accesory-reagents/1278-intracell.html