Thursday 15 February 2018


An article published this year in European Journal of Medicinal Chemistry using some of our products, FITC Apoptosis Detection Kit” & “PI/RNASE Solution” by our customers present at the University of Coimbra, Portugal, in the analysis of how Advances on photodynamic therapy of melanoma through novel ring-fused 5,15-diphenylchlorins. Congrats and Thanks.


Summay:
The synthesis, photophysical behaviour and photosensitization ability of novel 4,5,6,7-tetrahydropyrazolo[1,5-a]pyridine-fused 5,15-diphenylchlorins against melanoma cells are described. All studied chlorins were found to be extremely active against melanoma cell lines A375 showing IC50 values below 20 nM. Furthermore, a dihydroxymethyl diphenylchlorin was identified as an excellent candidate to allow modulating of different types of cell death, apoptosis vs. necrosis, by varying its concentration. This can be explored as a tool to improve the effectiveness of PDT since inflammatory response resulting from necrotic cell death after PDT can activate the antitumor immune response with implications also regarding the vascular damage. This feature combined with very low cytotoxicity against human melanoma cells in the absence of light activation and against human fibroblast HFF-1 cells makes this chlorin a candidate of choice as a photosensitizer for PDT. A comprehensive photophysical investigation including the determination of quantum yields for fluorescence, singlet oxygen sensitization and internal conversion, lifetimes and rate constants of all the excited state deactivation processes has been undertaken.


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An article published this year in “Plos One” using some of our products, FITC Apoptosis Detection Kit”, by our customers present at the Institute of Health Research & Hospital Lozano Blesa from Zaragoza as well as University and Hospital San Jorge from Huesca, Spain, in the analysis of Pharmacological activation of TRPV4 produces immediate cell damage and induction of apoptosis in human melanoma cells and HaCaT keratinocytes. Congrats and Thanks.


Summay:
TRPV4 channels are calcium-permeable cation channels that are activated by several physicochemical stimuli. Accordingly, TRPV4 channels have been implicated in the regulation of osmosensing, mechanotransduction, thermosensation, and epithelial/endothelial barrier functions. Whether TRPV4 is also mechanistically implicated in melanoma cell proliferation is not clear. Here, we hypothesized that TRPV4 is expressed in human melanoma and that pharmacological activation interferes with cell proliferation.

A) Representative flow cytometry dot plots with double Annexin V-FITC/PI staining for control cells (DMSO 0,2%), cells exposed to GSK1016790A (10 nM), and cells exposed to GSK1016790A and HC067047 (1 μM) at 1 h, 24 h, and 72 h. B) Summary data. C) Induction of apoptosis in HaCaT cells and protective effects of HC067047. D) Summary data. *P<0.05 vs. Control, #P<0.05 vs. GSK1016790A, ANOVA, n = 3). Data are means ± SEM (number of independent experiments, n = 3.


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An article published this year inRevista Española de Patología torácicausing some of our products, CF-Blue AnnexinV”, by our customers present at the IBIS Research Center, Sevilla, Spain in the analysis of CELULAR ORIGIN OF CIRCULATING MICROPARTICLES IN PATIENTS WITH VENOUS THROMBOEMBOLISM AND CANCER. Congrats and Thanks.


Summay:
Micro particles (MPs) are extracellular vesicles considered to be powerful cellular effectors. They are present in healthy individuals and are elevated in pathological conditions such as inflammatory and neoplastic diseases, and thrombosis. The relationship between venous thromboembolism (VTE) and cancer has been well established. MPs are thought to be a pathogenic connection between the two entities. If confirmed, they could be used as biomarkers. Our aim was to characterize the MPs in both diseases according to their cellular origin (cellular, endothelial, platelet, leukocyte and those that exhibited mucin 1 on their surface). Functional parameters such as D-dimer (DD) and soluble P-selectin (sPsel) were also studied. 96 patients with idiopathic VTE and 85 with advanced lung, stomach or pancreatic neoplasia were considered. All of them were followed clinically for two years and those who were diagnosed with cancer in the VTE group or those who developed thrombosis in the group of neoplastic patients were excluded from the study. Finally, 82 VTE patients and 68 cancer patients were analyzed.
In our results, we found that total MPs and platelet-derived MPs differentiated both patient groups. Additionally, significantly greater numbers of DD and sPsel   (p<0.001) were determined in the VTE group. The differences found between both groups, taking into account the origin of the MPs, could be caused by the prothrombotic characteristics of the neoplastic group and their sequestration within active clots in the VTE group



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An article published this year inProteomesusing some of our products, FITC Mouse Anti-Human CD14, APC Mouse Anti-Human CD19 and OC-515 Mouse Anti-Human CD45”, by some of our customers present some Proteomics Units from Universities of Navarra, Valencia, Madrid, Basque Country and Salamanca, in the analysis of In-Depth Proteomic Characterization of Classical and Non-Classical Monocyte Subsets Antitumor Efficacy of Human Monocyte-Derived Dendritic Cells: Comparing Effects of two Monocyte Isolation Methods. Congrats and Thanks.

Summay:
Monocytes are bone marrow-derived leukocytes that are part of the innate immune system. Monocytes are divided into three subsets: classical, intermediate and non-classical, which can be differentiated by their expression of some surface antigens, mainly CD14 and CD16.
These cells are key players in the inflammation process underlying the mechanism of many diseases. Thus, the molecular characterization of these cells may provide very useful information for understanding their biology in health and disease. We performed a multicentric proteomic study with pure classical and non-classical populations derived from 12 healthy donors. The robust workflow used provided reproducible results among the five participating laboratories. Over 5000 proteins were identified, and about half of them were quantified using a spectral counting approach.
The results represent the protein abundance catalogue of pure classical and enriched non-classical blood peripheral monocytes, and could serve as a reference dataset of the healthy population. The functional analysis of the differences between cell subsets supports the consensus roles assigned to human monocytes.




Protein abundance correlation between purified monocyte populations and public datasets obtained from PaxDB. Protein abundances determined in the present work (y-axis) are plotted against public datasets (x-axis). CD14 (classical) and CD16 (non-classical), are shown in left and right panels, respectively. Complete monocyte population and liver datasets are shown in the top panels and bottom panels, respectively.



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An article published this year in “Biological Procedures Online” using one of our products, “APC Mouse anti-Human HLA-DR”, by our customers from  NOVA Medical School, Faculdade de Ciências Médicas, Universidade NOVA de Lisboa, Portugal, in the analysis of Antitumor Efficacy of Human Monocyte-Derived Dendritic Cells: Comparing Effects of two Monocyte Isolation Methods. Congrats and Thanks.


Summay:
Dendritic cells (DCs), which can be used as anti-cancer vaccines, are generally obtained in vitro from isolated CD14+ monocytes (MoDCs). This generates high cell numbers and allows instructing DCs to guarantee effective antitumor responses. However, the impact of the monocyte isolation step in the antitumor effectiveness of the generated MoDCs is still unknown. Here, we compared the most used immunomagnetic technologies for monocyte isolation: magnetic activated cell sorting (MACS) from Miltenyi Biotec and EasySep from STEM CELL.



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An article published this year in “Biomaterials” using one of our products, “FITC ANNEXIN V”, by our customers from the Research Center Principe Felipe, Valencia, Spain, in the analysis of how Metabolomics facilitates the discrimination of the specific anti-cancer effects of free- and polymer-conjugated doxorubicin in breast cancer models. Congrats and Thanks.


Summay:
Metabolomics is becoming a relevant tool for understanding the molecular mechanisms involved in the response to new drug delivery systems. The applicability of this experimental approach to cell cultures and animal models makes metabolomics a useful tool for establishing direct connections between in vitro and in vivo data, thus providing a reliable platform for the characterization of chemotherapeutic agents. Herein, we used metabolomic profiles based on nuclear magnetic resonance (NMR) spectroscopy to evaluate the biochemical pathways involved in the response to a chemotherapeutic anthracycline drug (Doxorubicin, Dox) and an N-(2-hydroxypropyl) methacrylamide (HPMA) copolymer-conjugated form (HPMA-Dox) in an in vitro cell culture model and an in vivo orthotopic breast cancer model. We also used protein expression and flow cytometry studies to obtain a better coverage of the biochemical alterations associated with the administration of these compounds. The overall analysis revealed that polymer conjugation leads to increased apoptosis, reduced glycolysis, and reduced levels of phospholipids when compared to the free chemotherapeutic drug. Our results represent a first step in the application of integrated in vitro and in vivo metabolomic studies to the evaluation of drug delivery systems.

(G), levels of apoptosis after treating MCF7 cells with Dox and HPMA-Dox at 7.5 μg/ml Dox equiv. for 72 h. Cells were stained with AnnexinV and the results are shown as the percentage of positive cells for staining.

 
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FITC AnnexinV