LEGO Creator Mystic Witch 3 in 1 40562

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Giordano, R. J., Cardó-Vila, M., Lahdenranta, J., Pasqualini, R. & Arap, W. Biopanning and rapid analysis of selective interactive ligands. Nat. Med. 7, 1249–53 (2001). de Oliveira, H. C. et al. Peptides Derived from a Phage Display Library Inhibit Adhesion and Protect the Host against Infection by Paracoccidioides brasiliensis and Paracoccidioides lutzii. Front. Pharmacol. 7, 509 (2016). de Wit, M. et al. Cell surface proteomics identifies glucose transporter type 1 and prion protein as candidate biomarkers for colorectal adenoma-to-carcinoma progression. Gut 61, 855–864 (2012). Wu, C.-H., Liu, I.-J., Lu, R.-M. & Wu, H.-C. Advancement and applications of peptide phage display technology in biomedical science. J. Biomed. Sci. 23, 8 (2016). Liu, X. et al. Selection and identification of novel peptides specifically targeting human cervical cancer. Amino Acids 50, 577–592 (2018).

Tissue samples from 67 patients treated in Hospital de Braga, North of Portugal, between 1 st January of 2005 and 1 st January of 2010 with CRC diagnosis were collected prospectively. Tumor localization was recorded and classified as colon and rectum (between anal verge and 15 cm at rigid rectoscopy). The histological type of CRC was classified by an experienced pathologist. CRC samples were included into tissue microarrays (TMAs). Prior to TMA construction, haematoxylin and eosin sections were reviewed to select representative areas of the tumor. Normal-adjacent tissue was also included in the TMAs for primary tumors. Each case was represented in the TMA by at least two cores of 0.6 mm. Tissue cores of kidney were used as controls for TMA orientation. The study protocol was approved by the Ethics Committee of Hospital de Braga. The data of CRC series was collected prospectively, patients were informed and signed a written consensus for collecting data and samples collection. All experiments were performed in accordance with relevant guidelines and regulations. Cell Lines and Cell Culture Silencing of MCT1 and MCT4 expression was performed using 5 nM of Silencer Select Pre-Designed & Validated siRNAs from Thermo Fisher (MCT1 siRNA: s580 and MCT4 siRNA: s17417), using an adequate control (Silencer Select Negative Control siRNA: 4390843). Cells were transfected using Lipofectamine RNAiMAX Reagent (Thermo Fisher) according manufacturer’s instructions. Immunocytochemistry for co-localization of FAM-RKOpep and MCT1 Monkie Kid 80054 Megapolis City: 10 Reasons That This Middle-Aged AFOL Is Excited! November 7, 2023 Ferreira, D. & Martins, I. M. Artificial virus particles. In Bioinspired Materials for Medical Applications 427–450, https://doi.org/10.1016/B978-0-08-100741-9.00015-2 (Elsevier, 2017). The human epithelial CRC cell line RKO (ATCC CRL 2577), normal colon cell line CCD-841-CoN (ATCC CRL 1790) and CRC cell line Caco-2 (ATCC HTB-37) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Biochrom). The human CRC cell lines HCT 116 (ATCC CCL-247) and HCT-15 (ATCC CCL-225) were grown in Roswell Park Memorial Institute Medium 1640 (RPMI, Biochrom) supplemented with 10% (v/v) fetal bovine serum (FBS, Biochrom) and 1% (v/v) penicillin-streptomycin (Biochrom). Cells were maintained at 37 °C with 5% CO 2. BRASIL

Verdict

Krag, D. N. et al. Selection of Tumor-binding Ligands in Cancer Patients with Phage Display Libraries. Cancer Res. 66, 7724–7733 (2006). A structural bioinformatics approach was used to identify the potential targets of RKOpep. A complete overview of all cell surface proteins of five CRC cell lines, including RKO, Caco-2 and HCT 116, was provided by de Wit and collaborators 20. Using a high-resolution shotgun proteomics analysis, a total of 2609 proteins were identified in the cell surface fractions. By combining additional selection criteria including the presence in three of the cell lines in study, overexpression in carcinomas when compared to adenomas and subcellular location at the plasma membrane, about 59 candidate proteins for the RKOpep were identified. Miranda-Gonçalves, V. et al. Monocarboxylate transporters (MCTs) in gliomas: expression and exploitation as therapeutic targets. Neuro. Oncol. 15, 172–188 (2013). Interestingly, this upcoming LEGO October 2022 GWP is a Creator 3 in 1 set. Usually, they are other types of LEGO GWP like 18+ LEGO IDEAS or just LEGO Creator.

Before sequencing, the DNA was purified using the Illustra ExoProStar 1-Step kit (GE Healthcare). Sequencing was carried out by GATC Biotech using the M13-pIII primer 5′- TTAACTCCCTGCAAGCCTCA-3′. The SnapGene Version 1.1.3 (GSL Biotech) was used for peptide analysis. The corresponding peptide similarities were identified by Clustal Omega analysis 49. Binding assays using ELISA As RKO-R5-1 phage clone, the free FAM-labelled peptide RKOpep also showed specific selectivity towards RKO cells in comparison to the CCD-841-CoN normal cells. Using microscopy, it was shown for the RKO cells that the fluorescence intensity increased with an increase of FAM-RKOpep concentration, which was not verified for the CCD-841-CoN cells (Fig. 2A). The cytometry analysis also corroborated these results, clearly showing that there is no affinity to the normal cells and that the RKOpep selectively bound to RKO cells (Fig. 2B). Moreover, the FAM-RKOpep showed noteworthy specificity for other three human CRC cells, HCT 116, Caco-2 and HCT-15 (Fig. 3). In addition, the peptide specific targeting of human colorectal cancer tissues, shown by immunofluorescence staining, revealed meaningful results (Fig. 4). Visible and noticeable green fluorescent signal of FAM-RKOpep was detected in human CRC tissues and a weak signal was observed for normal-adjacent ones. This experimental data supports the potential translation of this peptide to clinical oncology. The ability of peptides to recognize human cancer tissues was also demonstrated by Liu and collaborators 33. They identified novel cancer-specific peptides selected through in vivo screening of phage display peptide libraries, that were able to target human cervical cancer tissues. Christiansen, A. et al. High-throughput sequencing enhanced phage display enables the identification of patient-specific epitope motifs in serum. Sci. Rep. 5, 12913 (2015).Sprowl-Tanio, S. et al. Lactate/pyruvate transporter MCT-1 is a direct Wnt target that confers sensitivity to 3-bromopyruvate in colon cancer. Cancer Metab. 4, 20 (2016). Pinheiro, C. et al. Increased expression of monocarboxylate transporters 1, 2, and 4 in colorectal carcinomas. Virchows Arch. 452, 139–146 (2008). Halestrap, A. P. The monocarboxylate transporter family-Structure and functional characterization. IUBMB Life 64, 1–9 (2012). Nobrega, F. L. et al. Screening and characterization of novel specific peptides targeting MDA-MB-231 claudin-low breast carcinoma by computer-aided phage display methodologies. BMC Cancer 16, 881 (2016).