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The article discusses the efforts of the Albertans Heading for Peace to make social justice an expectation. It notes that Albertans Heading for Peace of students at High Park School in Alberta is part of the PeaceJam initiative that aims to empower other students to get involved and promote a more peaceful world. Furthermore, Albertans Heading for Peace is a highly anticipated expectation that continues to open up possibilities for people to take action and make a difference.

Introduction: Human spermatozoa have unique, well-organized nuclear architecture different from somatic cells. Improperly packaged sperm chromatin will have a high probability of disrupting the particularly structured sequence of fertilization. The evaluation of nuclear organization in spermatozoa is challenging due to the extreme compactness of chromatin and the absolute necessity for induced nuclear decondensation (ND), which itself alters native sperm architecture. The purpose of our study was to identify sperm ND protocol that results in minimal disturbance of sperm architecture. Materials and Methods: Three widely used ND protocols for fluorescence in situ hybridization (FISH) were evaluated in this study: protocol 1 was combined denaturation and decondensation with 0.5N NaOH; in protocol 2, decondensation was achieved with 10mM dithiothreitol (DTT) and lithium diiodosalicylate (LIS); in protocol 3, 2.5mM DTT and heparin was used for ND. Protocols 2 and 3 were followed by denaturation with formamide at 75°C. The effect of denaturation and hybridization on spontaneous ND was also evaluated. Samples were counterstained with DAPI. Images were acquired using unbiased sampling and digital image morphometry analysis by Visiopharm Integrator System. Results were compared to neat samples using SPSS 16.0 statistical computer software. Results: Comparison of ND protocols is provided in the table. Mild decondensation was defined as 1.5 3 fold increase in nuclear area. Spontaneous ND occurred in an unexpectedly large proportion of spermatozoa during denaturation (74%) and hybridization (39%). Conclusions: Combined denaturation and decondensation with 0.5N NaOH demonstrated superior results providing uniform mild ND with over 80% of nucleus suitable for FISH sperm architecture assessment. The influence of spontaneous ND should be considered in any FISH preparation protocols. [ABSTRACT FROM AUTHOR]

Aim We present our experience using the Holotype™ Kit by Omixon for next generation sequencing (NGS) of HLA genes in a clinical setting for one and a half years, starting December 2013. Methods Samples were prepared using the X4 Holotype™ kit by Omixon for up to 7 loci (full length HLA-A, B, C, DQA1 and DQB1, intron 1 to intron 4 of DRB1, and intron 1–3’ UTR of DPB1) and were sequenced in both an individual and pooled amplicon approach using 2 × 251 bp reads on the Illumina MiSeq. Samples were also characterized in parallel with a second method (Sanger SBT, SSO, or SSP) to confirm the genotyping results by NGS and identify possible causes of discrepancies, if any. Two analysis programs were used to genotype the NGS data, Target by Omixon (v 1.7/1.8) and NGSengine by GenDX (v 1.3/1.6). Results A total of 1046 samples have been sequenced: 497 (47%) related to bone marrow transplant cases, 250 (24%) related to solid organ transplantation, and 299 (29%) samples being either proficiency, outreach or disease association studies. Approximately 975 typings were performed for HLA-A, B, C, DRB1 and DQB1, and 600 typings for DQA1 and DPB1. On average, one sequencing run was performed per week with 12 samples on the MiSeq for a total of 83 runs utilizing both nano and full flow cells. At best, the turnaround time from amplification to analysis completion was 4 days, but averages 10 days due to the laboratory workflow and a single sequencing run per week. Concordance was 99.7% between NGS and secondary typing methods (SBT/SSP/SSOP). Discordance was due to incorrect or incomplete (ambiguous) SBT/SSP typing, allele imbalance, NGS genotyping software errors, incorrect reporting of results and new alleles. Two genotyping programs were used for analysis to safeguard against systematic analysis errors and agree for 94.8% of the best match allele calls. Ambiguous results are most common at the DPB1 locus due to a lack of phasing between exons 2 and 3 or the unsequenced exon 1 (17% of alleles) and the DRB1 locus due to not sequencing exon 1 (3.8% of alleles). No ambiguities were detected among the other loci. Conclusions We have genotyped over 1000 samples using NGS, with a very high concordance rate with other HLA typing methods and low ambiguity rates for DRB1 and DPB1 as described above. This technology is therefore very appropriate for HLA typing in a clinical setting. D. Monos: Grant/Research Support; Company/Organization; Omixon. 7. Other (Identify); Company/Organization; Royalty from Omixon. [ABSTRACT FROM AUTHOR]

Aim Provide complete, unambiguous characterization of 6 HLA loci (HLA-A, B, C, DPB1, DQB1, and DRB1) for 109 publicly available cell lines from Coriell Cell Repository, in collaboration with the Center for Disease Control and Prevention’s Genetic Testing Reference Material program (GeT-RM). These cell lines have previously been characterized for 5 pharmacogenetic (PGx) genes (Pratt, V et al J Mol Diag 2010 12(6):835–846) and HLA genes using multiple assays and are intended for use as reference materials for clinical genetic testing laboratories. Method DNA samples were amplified at 6 HLA loci using Omixon Holotype primers delineating full length genes (5’UTR to 3’UTR) with the exception of DRB1 (intron 1 to intron 4). Sequencing libraries were prepared using Holotype X4 kits for NGS on the Illumina MiSeq. Target (Omixon) and NGSengine (GenDX) were used to analyze the NGS data and results were compared to HLA typing generated by Sanger sequencing (SBT) using SSO and SSP to resolve ambiguities. Results 187 unique alleles were identified. The table below indicates the percent of people in the given population that will carry one of the alleles included in this study (Maiers, M et al Hum Imunol 2010 68(9):779–788). DPB1 frequencies were not available from this source. We observed 98 5% concordance between HLA typing results using SBT and NGS, with discordance caused by sequence differences outside regions characterized by SBT ( n = 21), and errors in SBT/SSP/SSO typing ( n = 2). Ambiguities persist in 32/1308 allele calls, with the majority due to alternative cis/trans combinations of exons 2 and 3 in DPB1 ( n = 30) and 2 caused by polymorphisms in the unsequenced exon 1 of the DRB1 gene. Conclusion NGS enhanced the characterization of alleles in this study by identifying errors and allowing analysis of regions not examined by other typing methods. These samples will be useful for assay development and validation, quality control and proficiency testing, and should help to improve the accuracy of PGx and HLA testing in clinical laboratories. D. Monos: Other (Identify); Company/Organization; Royalty. [ABSTRACT FROM AUTHOR]

Objective: Our objective was to analyze MRI contrast-enhancement patterns in arthritic and nonarthritic knees and the relationship of those patterns with clinical, laboratory, and histologic synovium markers., Materials and Methods: Dynamic contrast-enhanced MRI was performed in nine arthritic and three nonarthritic knees of juvenile rabbits. A two-compartment pharmacokinetic model of signal intensity-time data was implemented to generate parametric maps of signal slope, maximal percentage of signal change, capillary permeability, leakage space volume, and time-to-peak. MRI values were compared with clinical, laboratory, and histologic markers for evaluation of synovial changes during the progression of arthritis., Results: Parametric maps of capillary permeability and signal slope depicted significant differences between arthritic and nonarthritic knees. Arthritic knees showed increased capillary permeability (p = 0.006) and signal slope (p = 0.01) with time after onset of disease as opposed to nonarthritic knees (permeability, p = 0.65; slope, p = 0.56). Significant correlations were found between temporal changes in capillary permeability (p = 0.002), signal slope (p = 0.003), and serum concentrations of amyloid A. No relationship was noted between any MRI parameters and histologic scores. The discriminative power of MRI indexes varied according to the stage of arthritis: time-to-peak was most accurate for differentiation of presence versus absence of arthritis in early arthritis (day 1, p = 0.0002), and signal slope was most accurate in midterm arthritis (day 14, p = 0.001)., Conclusion: In vivo capillary permeability and signal slope have distinctive dynamic MRI properties. The accuracy of MRI parameters for diagnostic evaluation of experimental arthritis differs according to the stage of disease.