Journal: Journal of Neurosurgery

Article Title: Face, content, and construct validity of human placenta as a haptic training tool in neurointerventional surgery

doi: 10.3171/2015.1.jns141583

Figure Lengend Snippet: Fig. 5. Coil embolization of the placental aneurysm. A: Angiogram showing the small-neck aneurysm that was created in the placenta. B: Catheter navigation toward the aneurysm using road map views. C: Coil embolization of the aneurysm.

Article Snippet: While this paper was under review, a report on the use of the HP in interventional neuroradiology was issued.9 The focus in that report was the use of HP in endovascular research, especially with the use of the Pipeline Embolization Device (ev3/Covidien).

Techniques:

Journal: Scientific Reports

Article Title: EV-3, an endogenous human erythropoietin isoform with distinct functional relevance

doi: 10.1038/s41598-017-03167-0

Figure Lengend Snippet: Depiction of the EV-3 specific peptide used for nano-LC-ESI-MS/MS analysis. According to the obtained sequence of the two EPO -PCR amplification products from human adult kidney and foetal brain, aminoacid sequences of EPO and hEPOΔ3 derived protein EV-3 differ only by 29 aminoacids. The deleted region contains one N-glycosylation site and the AB loop including the minihelix B’. The underlined amino acids in the EV-3 sequence represent the EV-3 specific peptide used in nano-LC-ESI-MS/MS. The representation of EPO’s primary structure is taken from Boissel et al . .

Article Snippet: Recombinant EV-3 and EPO proteins were produced by IBA GmbH (Germany) as Twin-Strep-Tag fusion proteins via transient transfection in CHO cells.

Techniques: Tandem Mass Spectroscopy, Sequencing, Amplification, Derivative Assay

Journal: Scientific Reports

Article Title: EV-3, an endogenous human erythropoietin isoform with distinct functional relevance

doi: 10.1038/s41598-017-03167-0

Figure Lengend Snippet: Isolation of hEPOΔ3 from HepG2 cells and detection of hEPOΔ3 derived protein EV-3 and EPO in HepG2 cell culture supernatant. ( a ) The nested PCR approach amplifies EPO full-length and hEPOΔ3 transcripts from HepG2 cells. ( b ) Captured proteins of HepG2 supernatants (α-EPO IP, using biotinylated anti-EPO antibody BAF959) are visualized by western blotting using a polyclonal rhEPO-antibody (H-162). The Western Blot shows two EPO-antibody reactive proteins (prot. 1 and prot. 2) running at the same size of recombinantly produced EV-3 (rEV-3) and EPO (two different batches of rEPO). ( c ) Densitometry analysis was done on the Western Blot image (jpeg) shown in Fig. 4b. The plot profile of the HepG2 immunoprecipitate in lane 5 (ROI3) reveals a faint but distinct second peak, corresponding to the plot profile of EV-3 loaded in lane 1 (ROI1). ( d ) EV-3, indicated by the specific peptide ‘DITVGQQAVE’, was detected in HepG2 supernatants by nano-LC-ESI-MS/MS. Quantification of EV-3 specific peptide (fmol/µg total protein of analysed sample) in sample H1, the immunoprecipitate (α-EPO-IP) of HepG2 supernatant, or sample H2, HepG2 supernatant enriched for glycoproteins by WGA chromatography.

Article Snippet: Recombinant EV-3 and EPO proteins were produced by IBA GmbH (Germany) as Twin-Strep-Tag fusion proteins via transient transfection in CHO cells.

Techniques: Isolation, Derivative Assay, Cell Culture, Nested PCR, Western Blot, Produced, Tandem Mass Spectroscopy, Chromatography

Journal: Scientific Reports

Article Title: EV-3, an endogenous human erythropoietin isoform with distinct functional relevance

doi: 10.1038/s41598-017-03167-0

Figure Lengend Snippet: EV-3 protein identification in human serum from healthy donors. ( a ) Depiction of the optimized multi-step purification strategy of serum for the detection and quantification of the EV-3 specific peptide by nano-LC-ESI-MS/MS. ( b ) In serum samples of the healthy human donors (S1, S2), the EV-3 specific peptide ‘DITVGQQAVE’ was detected by using nano-LC-ESI-MS/MS. Applying two size exclusion chromatography steps to serum S2 enriched EV-3 in the sample. Compared to S1, higher levels of the EV-3 specific peptide (fmol/µg total protein) were quantified in two fractions of S2.

Article Snippet: Recombinant EV-3 and EPO proteins were produced by IBA GmbH (Germany) as Twin-Strep-Tag fusion proteins via transient transfection in CHO cells.

Techniques: Purification, Tandem Mass Spectroscopy, Size-exclusion Chromatography

Journal: Scientific Reports

Article Title: EV-3, an endogenous human erythropoietin isoform with distinct functional relevance

doi: 10.1038/s41598-017-03167-0

Figure Lengend Snippet: EPO but not EV-3 promotes formation of erythroid progenitors from human CD34+ haematopoietic stem cells. CD34+ cells (500 cells/well) from two donors were cultured in EPO-free MethoCult medium supplemented either with EPO or EV-3 at different concentrations (0–30 U/ml) for 14 days. Analysis of the different colony-forming units (CFU) revealed no erythroid progenitors (CFU-GEMM or BFU-E) in control or EV-3 treated wells but in wells treated with EPO or positive control H84434 (medium includes EPO). n = 2 per treatment; CFU-G/M/GM, colony-forming unit-granulocyte or -macrophage or mix of granulocyte and macrophage; CFU-GEMM, colony-forming unit granulocyte, erythrocyte, monocyte, megakaryocyte; BFU-E, burst-forming unit erythroid.

Article Snippet: Recombinant EV-3 and EPO proteins were produced by IBA GmbH (Germany) as Twin-Strep-Tag fusion proteins via transient transfection in CHO cells.

Techniques: Cell Culture, Positive Control

Journal: Scientific Reports

Article Title: EV-3, an endogenous human erythropoietin isoform with distinct functional relevance

doi: 10.1038/s41598-017-03167-0

Figure Lengend Snippet: EV-3 and EPO protect primary hippocampal neurons from cell death. Pretreatment with EPO or EV-3 reduced cell death, induced by starvation (48 h) followed by hypoxia (20 h), of rat primary hippocampal neurons (n = 5/treatment; hypoxia effect set to 100%; one-tailed paired t-test, EPO and EV-3 [U weight equivalents] treatment compared to placebo). Mean ± SEM presented.

Article Snippet: Recombinant EV-3 and EPO proteins were produced by IBA GmbH (Germany) as Twin-Strep-Tag fusion proteins via transient transfection in CHO cells.

Techniques: One-tailed Test

Journal: Scientific Reports

Article Title: EV-3, an endogenous human erythropoietin isoform with distinct functional relevance

doi: 10.1038/s41598-017-03167-0

Figure Lengend Snippet: Cognitive performance in a touchscreen-test and haematocrit levels in placebo, EV-3 and EPO treated mice. Kaplan-Meier presentation of group performance during ( a ) pretraining 1–3, ( b ) visual discrimination and ( c ) overall task (pretraining and visual discrimination). Curves represent group performance (n = 10–11 per group) and indicate the cumulative percentage of animals not reaching performance criteria of the touch-screen paradigm. The EPO- and the EV-3 treated mice finished the overall task ( c ) 10 days earlier than the placebo group (EV-3 vs. placebo, p = 0.060, EPO vs placebo, p = 0.041*). ( d ) Following 3-weeks treatment with EPO (5000 IU/kg, NeoRecormon) versus EV-3 (weight-equivalent dose) and placebo, haematocrit is significantly enhanced in the EPO-treated but not in the EV-3 treated group.

Article Snippet: Recombinant EV-3 and EPO proteins were produced by IBA GmbH (Germany) as Twin-Strep-Tag fusion proteins via transient transfection in CHO cells.

Techniques: