FITC Annexin V Apoptosis Detection Kit with PI.

An article published this year in “STEM CELLS INTERNATIONAL” using our Annexin V–fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) double staining, by our customers from Department of Endodontics, Faculty of Medicine and Dentistry, Catholic University of Valencia “San Vicente Mártir”, Valencia, Spain, in the analysis of Cellular Responses in Human Dental Pulp Stem Cells Treated with Three Endodontic Materials. Congrats and Thanks.

Summary:

Human dental pulp stem cells (HDPSCs) are of special relevance in future regenerative dental therapies. Characterizing cytotoxicity and genotoxicity produced by endodontic materials is required to evaluate the potential for regeneration of injured tissues in future strategies combining regenerative and root canal therapies. This study explores the cytotoxicity and genotoxicity mediated by oxidative stress of three endodontic materials that are widely used on HDPSCs: a mineral trioxide aggregate (MTA-Angelus white), an epoxy resin sealant (AH-Plus cement), and an MTA-based cement sealer (MTA-Fillapex). Cell viability and cell death rate were assessed by flow cytometry. Oxidative stress was measured by OxyBlot. Levels of antioxidant enzymes were evaluated by Western blot. Genotoxicity was studied by quantifying the expression levels of DNA damage sensors such as ATM and RAD53 genes and DNA damage repair sensors such as RAD51 and PARP-1. Results indicate that AH-Plus increased apoptosis, oxidative stress, and genotoxicity markers in HDPSCs. MTA-Fillapex was the most cytotoxic oxidative stress inductor and genotoxic material for HDPSCs at longer times in preincubated cell culture medium, and MTA-Angelus was less cytotoxic and genotoxic than AH-Plus and MTA-Fillapex at all times assayed.

Reference:

https://www.hindawi.com/journals/sci/2017/8920356/

Product link:

FITC Annexin V Apoptosis Detection Kit with PI: http://www.immunostep.com/apoptosis-tools/3727-anxvkf-100t.html

FITC Annexin V Apoptosis Detection Kit with PI.

An article published this year in “NANOMATERIALS” using our Annexin V–fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) double staining by our customers from Dipartimento di Scienze Biomediche e Cliniche L. Sacco, Università di Milano, Italy, in the analysis of Silver Nanoparticles in Orthopedic Applications: New Insights on Their Effects on Osteogenic Cells. Congrats and Thanks.

Summary:

Infections of orthopedic implants are associated with high morbidity. The emergence of antibiotic resistant strains and the tendency of microbes to form biofilms on orthopedic devices prompt the individuation of novel antimicrobial agents. Silver nanoparticles represent an interesting alternative, but their effects on bone cells need to be clarified. We focused on osteoblast-like cells and on bone marrow-mesenchymal stem cells and found that these cells are rather resistant to the cytotoxic effects of silver nanoparticles, with a half maximal inhibitory concentration around 25 µg/mL as detected by MTT assay. Within a month of treatment, osteoblast-like cells adapt to the presence of the nanoparticles by upregulating hsp70 as shown by western blot. Hsp70 overexpression correlates with the restoration of normal cell proliferation. No alterations in the extent and time requirements were detected in mesenchymal stem cell induced to differentiate in osteoblasts in the presence of silver nanoparticles. Because the concentrations of silver nanoparticles which show antimicrobial activity are lower than those exerting toxic effects on bone-forming cells in vitro, we suggest that silver nanoparticles might represent a challenging tool to fight infections in orthopedic implants.

Reference:

http://www.mdpi.com/2079-4991/7/6/124/htm

Product link:

FITC Annexin V Apoptosis Detection Kit with PI: http://www.immunostep.com/apoptosis-tools/3727-anxvkf-100t.html

FITC Annexin V Apoptosis Detection Kit with PI and PI/RNASE Solution.

An article published this year in “FRONTIERS IN ONCOLOGY” using our Annexin V–fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI) double staining and our PI/RNase staining solution, by our customers from Cancer Research Center-IBMCC (USAL-CSIC), Institute of Biomedical Research of Salamanca (IBSAL), Hematology Department, University Hospital of Salamanca, Salamanca, Spain, in the analysis of Targeting Ongoing DNA Damage in Multiple Myeloma: effects of DNA Damage response inhibitors on Plasma cell survival. Congrats and Thanks.

Summary:

Human myeloma cell lines (HMCLs) and a subset of myeloma patients with poor prognosis exhibit high levels of replication stress (RS), leading to DNA damage. In this study, we confirmed the presence of DNA double-strand breaks (DSBs) in several HMCLs by measuring γH2AX and RAD51 foci and analyzed the effect of various inhibitors of the DNA damage response on MM cell survival. Inhibition of ataxia telangiectasia and Rad3-related protein (ATR), the main kinase mediating the response to RS, using the specific inhibitor VE-821 induced more cell death in HMCLs than in control lymphoblastoid cells and U266, an HMCL with a low level of DNA damage. The absence of ATR was partially compensated by ataxia telangiectasia-mutated protein (ATM), since chemical inhibition of both kinases using VE-821 and KU-55933 significantly increased the death of MM cells with DNA damage. We found that ATM and ATR are involved in DSB repair by homologous recombination (HR) in MM. Inhibition of both kinases resulted in a stronger inhibition that may underlie cell death induction, since abolition of HR using two different inhibitors severely reduced survival of HMCLs that exhibit DNA damage. On the other hand, inhibition of the other route involved in DSB repair, nonhomologous end joining (NHEJ), using the DNA-PK inhibitor NU7441, did not affect MM cell viability. Interestingly, we found that NHEJ inhibition did not increase cell death when HR was simultaneously inhibited with the RAD51 inhibitor B02, but it clearly increased the level of cell death when HR was inhibited with the MRE11 inhibitor mirin, which interferes with recombination before DNA resection takes place. Taken together, our results demonstrate for the first time that MM cells with ongoing DNA damage rely on an intact HR pathway, which thereby suggests therapeutic opportunities. We also show that inhibition of HR after the initial step of end resection might be more appropriate for inducing MM cell death, since it prevents the occurrence of a compensatory NHEJ repair mechanism. These preclinical observations provide the rationale for its clinical evaluation.

Reference:

http://journal.frontiersin.org/article/10.3389/fonc.2017.00098/pdf

Product link:

FITC Annexin V Apoptosis Detection Kit with PI: http://www.immunostep.com/apoptosis-tools/3727-anxvkf-100t.html

PI/RNASE Solution: http://www.immunostep.com/solutions-buffers-chemicals/1289-pi-rnase-solution.html

FITC Annexin V and PI Solution.

An article published this year in “ALCOHOLISM CLINICAL AND EXPERIMENTAL RESEARCH” using our FITC-conjugated annexin-V and cell impermeant DNA fluorophorepropidium iodide, by our customers from Department of Molecular and Cellular Pathology of Alcohol, Príncipe Felipe Research Center, Valencia, Spain, in the analysis of Nalmefene prevents alcohol-induced neuroinflammation and alcohol drinking preference in adolescent female mice: role of TLR4. Congrats and Thanks.

Summary:

Background: We previously showed that, by activating innate immune receptors toll-like 4 (TLR4), adolescent intermittent ethanol exposure causes neuroinflammation, myelin damage and behavioral dysfunctions. Recent findings reveal that clinically-used opioid antagonists naltrexone (NT) and naloxone (NX) inhibit opioid-induced TLR4 signaling, and that NT, NX and nalmefene (NF), the 6-methylene derivative of NX, are able to reduce alcohol drinking escalation.

Methods: NF (0.1 mg/kg, i.p.) was injected 1 h prior to ethanol (3 g/kg, i.p.) following intermittent treatment in female (PND35) adolescent mice. Inflammatory molecules, myelin proteins and apoptotic markers were assessed in the prefrontal cortex (PFC) and striatum/nucleus accumbens (STR/NAcc). The effect of NF on alcohol drinking preference was evaluated in both the WT and TLR4-knock out adolescent mice. Using astroglial cells, the inhibitory potential of NT, NX and NF on LPS, or the ethanol-triggered TLR4 response, was compared.

Results: Our findings indicate that NF prevents the up-regulation of cytokines (IL-1β, IL-17A, TNF-α), chemokines (MCP-1, MIP-1, KC) and pro-inflammatory mediators (iNOS, COX-2), along with myelin damage and apoptotic events, in both PFC and STR/NAcc. NF also abolishes ethanol-induced escalation of alcohol preference/consumption, but has no effect when administered to TLR4-KO mice. In vitro experiments indicate that NX and NF inhibit TLR4 activation upon LPS or ethanol stimulation. Immunofluorescence studies and lipid rafts isolation show that NF is able to prevent TLR4 translocation to lipid rafts/caveolae in astrocytes.

Conclusions: These results suggest that NF prevents neuroinflammation and brain damage by blocking the TLR4 response, and also support the role of central pro-inflammatory immune signaling in the modulation of alcohol consumption/addiction

Reference:

http://onlinelibrary.wiley.com/doi/10.1111/acer.13416/full

Product link:

FITC Annexin V: http://www.immunostep.com/apoptosis-tools/3731-anxvf-200t.html

PI Solution: http://www.immunostep.com/solutions-buffers-chemicals/3872-pi-solution.html

CD29 FITC

An article published this year in “STEM CELLS INTERNATIONAL” using our CD29 FITC, by our customers from Virotherapy and Gene therapy Group, ProCure Program, Translational Research Laboratory, Instituto Catalan de Oncología IDIBELL, Barcelona, Spain, in the analysis os Human menstrual blood-derived mesenchymal stem cells as potential cell carriers for oncolytic adenovirus. Congrats and Thanks.

Summary:

T cells (in gray) of the immune system surround the tumor cells (in green)
thanks to the action of BiTE antibodies secreted by the oncolytic adenovirus.

Antitumor efficacy of systemically-administered oncolytic adenoviruses (OAdv) is limited due to diverse factors such as liver sequestration, neutralizing interactions in blood, elimination by the immune system, and physical barriers in tumors. It is therefore of clinical relevance to improve OAdv bioavailability and tumor delivery. Among the variety of tumor targeting strategies, the use of stem cells and specifically bone marrow-derived mesenchymal stem cells (BM-MSCs) is of particular interest due to their tumor tropism and immunomodulatory properties.

Nonetheless, the invasive methods to obtain these cells, the low number of MSCs present in the bone marrow, and their restricted in vitro expansion represent major obstacles for their use in cancer treatments, pointing out the necessity to identify an alternative source of MSCs. Here we have evaluated the use of menstrual blood-derived mesenchymal stem cells (MenSCs) as cell carriers for regional delivery of an OAdv in the tumor. Our results indicate that MenSCs can be isolated without invasive methods, they have an increased proliferation rate compared to BM-MSCs, and they can be efficiently infected with different serotype 5-based capsid-modified adenoviruses, leading to viral replication and release. In addition, our in vivo studies confirmed the tumor-homing properties of MenSCs after regional administration.

Reference:

https://www.hindawi.com/journals/sci/aip/3615729/

Product link:

CD29 FITC: http://www.immunostep.com/antibodies/2619-cd29.html

Kit for Paroxysomal Nocturnal Hemoglobinuria (PNH).

An article published this year in “JOURNAL OF THROMBOSIS AND THROMBOLYSIS” using our Kit for Paroxysomal Nocturnal Hemoglobinuria (PNH), by our customers from Hematology Laboratory (Division of Thrombosis and Hemostasis), ICO-Badalona, Hospital Germans Trias i Pujol, Josep Carreras Leukemia Research InstituteUniversitat Autonoma de Barcelona, Badalona, Spain, in the study of Retinal vein occlusion and paroxysmal nocturnal hemoglobinuria. Congrats and Thanks.

Summary:

Paroxysmal nocturnal hemoglobinuria (PNH) is a rare disorder associated with increased risk for thrombosis and reduced life expectancy. Retinal vein occlusion (RVO) is a frequent cause of vision loss but its relationship with PNH has not been studied systematically. Patients followed up for RVO in our ophthalmology department were screened for the presence of a PNH clone in peripheral blood by means of flow cytometry. The presence of other well-documented risk factors for RVO was also analyzed. In a series of 110 patients (54 males, median age of 67) we found no evidence of PNH. Most patients (97/110) had cardiovascular risk factors and/or hyperhomocysteinemia (67/110). Inherited thrombophilias were rare (three confirmed cases). Therefore, PNH does not appear to play a role in the development of RVO. However, this finding does not necessarily apply to young patients and/or those with no conventional risk factors for RVO, due to the low number of patients in these subgroups in our population.

Reference:

https://link.springer.com/article/10.1007/s11239-017-1502-4

Product link:

Kit for Paroxysomal Nocturnal Hemoglobinuria (PNH): http://www.immunostep.com/diagnostic-kits/2335-pnh-kit.html