Dmitri Shalin Interview with Peter Archibald about Erving Goffman entitled “If You Are Such a Great Sociologist, Why Are You Still in Canada?” My Encounter with Erving Goffman

Dr. Peter Archibald, Professor Emeritus of Sociology at the McMaster University, wrote this memoir at the suggestion of Tony Puddephatt and the request of Dmitri Shalin and gave his permission to post the present version in the Erving Goffman Archives

The nature of freedom

https://stars.library.ucf.edu/prism/1771/thumbnail.jp

Comparability & reimbursement for the translation of scalable, automated stem cell cultures

The research in this thesis focuses primarily on two critical challenges that inhibit the late stage translation of cell-based therapies and Regenerative Medicines (RMs). These include product comparability after a change in manufacturing process or site; and the reimbursement of RMs, in particular those which target multiple simultaneous indications, or Multimorbidity . The automation and standardisation of stem cell cultures also represent key themes of this thesis, which may facilitate the development of scalable, reproducible manufacturing processes for cell-based therapies. Furthermore, given the current uncertainty regarding the characterisation and potency of Human Mesenchymal Stromal (or Stem) Cells (hMSCs) that has inhibited the successful clinical translation of hMSC-based products, understanding the characterisation and putative modes of action of these cells was also a priority throughout this research. Also, due to the increasing number of Human Embryonic Stem Cell (hESC) derived therapies progressing towards market, and the industry-wide shift towards Human Induced Pluripotent Stem Cells (hiPSC) as an alternative to hESCs, the measurement of the growth and characterisation of these cells types represents an important method of demonstrating product comparability after alternative manufacturing process steps in the present thesis. Finally, due to the potential of hiPSCs as a source of large numbers of hMSCs, the culture conditions required to direct the differentiation of hiPSCs to hMSCs are explored

Development of a novel polyamide-based agent to inhibit EVI1 function

The EVI1 gene at chromosome 3q26 is associated with acute myeloid leukemogenesis, due to both chromosomal rearrangement and to overexpression in the absence of rearrangement. Some rearrangements such as t(3;3) and inv(3) result in overexpression of EVI1 protein, while translocation t(3;21) yields an AML1-MDS1-EVI1 (AME) fusion protein. EVI1 possesses two zinc finger domains, an N-terminal domain with fingers 1–7, which binds to GACAAGATA, and a C-terminal domain (fingers 8–10) which binds GAAGATGAG. Inhibition of EVI1 function with a small molecule compound may provide a targeted therapy for EVI1-expressing leukemias. As a first step towards inhibiting the leukemogenic function of EVI1, we performed structure-function studies on both EVI1 and AME protein to determine what domains are critical for malignant transformation activity. Assays were Rat1 fibroblasts in a soft agar colony forming assay for EVI1; primary bone marrow cells in a serial replating assay for AME. Both assays revealed that mutation of arginine 205 in zinc finger 6 of EVI1, which completely abrogates sequencespecific DNA binding via the N-terminal zinc finger domain, resulted in complete loss of transforming activity; mutations in other domains, such as the C-terminal zinc finger domain, CtBP binding domain, and the domains of AML1 had less of an effect or no effect on transforming activity. In an effort to inhibit EVI1 leukemogenic function, we developed a polyamide, DH-IV-298, designed to block zinc fingers 1–7 binding to the GACAAGATA motif. DNAseI footprinting revealed a specific interaction between DH-IV-298 and the GACAAGATA motif; no significant interaction was observed elsewhere; a mismatch polyamide failed to footprint at equivalent concentrations; and DH-IV-298 failed to bind to a control DNA lacking the GACAAGATA motif. Electromobility shift assay showed that, at a 1:1 polyamide:DNA ratio, DH-IV-298 lowered EVI1:DNA affinity by over 98%, while mismatch was significantly less effective (74% reduction). To assess the effect of DH-IV-298 on EVI1 binding to DNA in vivo, we performed CAT reporter assays in a NIH-3T3-derived cell line with a chromosome-embedded tet-inducible EVI1-VP16 as well as a EVI1-responsive CAT reporter. Removal of tetracycline resulted in a four-fold increase in CAT activity that was completely blocked by DH-IV-298. The mismatch polyamide was significantly less effective than DH-IV-298. Further studies are being performed to assess the effect on endogenous gene expression, and on growth of leukemic cells that express EVI1. These studies provide evidence that a cell permeable small molecule compound may effectively block the activity of a leukemogenic transcription factor

A Quality-Preserving Increase in Four-Year College Attendance

We use the National Longitudinal Study of the High School Class of 1972 and the Education Longitudinal Study of 2002 data sets to evaluate changes in the college matching process. Rising attendance rates at 4-year institutions have not decreased average preparedness of college goers or of college graduates, and further attendance gains are possible before diminishing returns set in. We use multinomial logic models to demonstrate that measures of likely success (grade point average) became more predictive of college attendance over time, while other student characteristics such as race and parents’ education became less predictive. Our evidence suggests that schools have become better at sorting while students have efficiently responded to changes in the return to higher education

Developing an effective scale-down model for a suspension adapted Hek293t-derived lentiviral vector stable producer cell line

Lentiviral vectors (LVV) represent an important tool for cell and gene therapy applications. However, inefficiencies in LVV manufacturing processes, such as the inability to achieve high cell densities with HEK293T cell lines in a fed batch process, have resulted in poor upstream yields. Optimisation of cell culture conditions is needed to improve upstream yields, which can be expedited by high-throughput screening (HTP). In this work, we describe the use of the 24 deep square well (24-DSW) microwell platform to develop a scale-down mimic of GSK’s established stable suspension LVV production process model at 2 L bioreactor scale. We found that matched mixing time was an effective basis for scale-translation between the stirred tank reactor (STR) and microwells. The growth kinetics and LVV productivity profile in the microwell were reproducible and comparable to the 2 L bioreactor process model. In both vessels, a 6-fold increase in cell density was achieved at the harvest time point and high cell viability (i.e. \u3e 90 %) was also maintained throughout the entirety of the cultures. The 24-DSW model, therefore, is an effective scale-down model for larger-scale stirred-tank bioreactor culture and provides an important tool for rapid, high-throughput optimization of the LVV production process