The Logical Problem of Language Change

This paper considers the problem of language change. Linguists must explain not only how languages are learned but also how and why they have evolved along certain trajectories and not others. While the language learning problem has focused on the behavior of individuals and how they acquire a particular grammar from a class of grammars ${cal G}$, here we consider a population of such learners and investigate the emergent, global population characteristics of linguistic communities over several generations. We argue that language change follows logically from specific assumptions about grammatical theories and learning paradigms. In particular, we are able to transform parameterized theories and memoryless acquisition algorithms into grammatical dynamical systems, whose evolution depicts a population's evolving linguistic composition. We investigate the linguistic and computational consequences of this model, showing that the formalization allows one to ask questions about diachronic that one otherwise could not ask, such as the effect of varying initial conditions on the resulting diachronic trajectories. From a more programmatic perspective, we give an example of how the dynamical system model for language change can serve as a way to distinguish among alternative grammatical theories, introducing a formal diachronic adequacy criterion for linguistic theories.

Electrical Design: A Problem for Artificial Intelligence Research

This report outlines the problem of intelligent failure recovery in a problem-solver for electrical design. We want our problem solver to learn as much as it can from its mistakes. Thus we cast the engineering design process on terms of Problem Solving by Debugging Almost-Right Plans, a paradigm for automatic problem solving based on the belief that creation and removal of "bugs" is an unavoidable part of the process of solving a complex problem. The process of localization and removal of bugs called for by the PSBDARP theory requires an approach to engineering analysis in which every result has a justification which describes the exact set of assumptions it depends upon. We have developed a program based on Analysis by Propagation of Constraints which can explain the basis of its deductions. In addition to being useful to a PSBDARP designer, these justifications are used in Dependency-Directed Backtracking to limit the combinatorial search in the analysis routines. Although the research we will describe is explicitly about electrical circuits, we believe that similar principles and methods are employed by other kinds of engineers, including computer programmers.

Cell-line Engineering of Chinese Hamster Ovary Cells for Low-temperature Culture

Developments in mammalian cell culture and recombinant technology has allowed for the production of recombinant proteins for use as human therapeutics. Mammalian cell culture is typically operated at the physiological temperature of 37°. However, recent research has shown that the use of low-temperature conditions (30-33°) as a platform for cell-culture results in changes in cell characteristics, such as increased specific productivity and extended periods of cell viability, that can potentially improve the production of recombinant proteins. Furthermore, many recent reports have focused on investigating low-temperature mammalian cell culture of Chinese hamster ovary (CHO) cells, one of the principal cell-lines used in industrial production of recombinant proteins. Exposure to low ambient temperatures exerts an external stress on all living cells, and elicits a cellular response. This cold-stress response has been observed in bacteria, plants and mammals, and is regulated at the gene level. The exact genes and molecular mechanisms involved in the cold-stress response in prokaryotes and plants have been well studied. There are also various reports that detail the modification of cold-stress genes to improve the characteristics of bacteria or plant cells at low temperatures. However, there is very limited information on mammalian cold-stress genes or the related pathways governing the mammalian cold-stress response. This project seeks to investigate and characterise cold-stress genes that are differentially expressed during low-temperature culture of CHO cells, and to relate them to the various changes in cell characteristics observed in low-temperature culture of CHO cells. The gene information can then be used to modify CHO cell-lines for improved performance in the production of recombinant proteins.

Over Expression of the CMP-sialic Acid Transporter in Chinese Hamster Ovary Cells Leads to Increased Sialylation

Most glyco-engineering approaches used to improve quality of recombinant glycoproteins involve the manipulation of glycosyltransferase and/or glycosidase expression. We investigated whether the over expression of nucleotide sugar transporters, particularly the CMP-sialic acid transporter (CMP-SAT), would be a means to improve the sialylation process in CHO cells. We hypothesized that increasing the expression of the CMP-SAT in the cells would increase the transport of the CMP-sialic acid in the Golgi lumen, hence increasing the intra-lumenal CMP-sialic acid pool, and resulting in a possible increase in sialylation extent of proteins being produced. We report the construction of a CMP-SAT expression vector which was used for transfection into CHO-IFNγ, a CHO cell line producing human IFNγ. This resulted in approximately 2 to 5 times increase in total CMP-SAT expression in some of the positive clones as compared to untransfected CHO-IFNγ, as determined using real-time PCR analysis. This in turn concurred with a 9.6% to 16.3% percent increase in site sialylation. This engineering approach has thus been identified as a novel means of improving sialylation in recombinant glycoprotein therapeutics. This strategy can be utilized feasibly on its own, or in combination with existing sialylation improvement strategies. It is believed that such multi-prong approaches are required to effectively manipulate the complex sialylation process, so as to bring us closer to the goal of producing recombinant glycoproteins of high and consistent sialylation from mammalian cells.

Variability in the Stability and Productivity of Transfected Genes in Chinese Hamster Ovary (CHO) cells

In the field of biologics production, productivity and stability of the transfected gene of interest are two very important attributes that dictate if a production process is viable. To further understand and improve these two traits, we would need to further our understanding of the factors affecting them. These would include integration site of the gene, gene copy number, cell phenotypic variation and cell environment. As these factors play different parts in the development process, they lead to variable productivity and stability of the transfected gene between clones, the well-known phenomenon of “clonal variation”. A study of this phenomenon and how the various factors contribute to it will thus shed light on strategies to improve productivity and stability in the production cell line. Of the four factors, the site of gene integration appears to be one of the most important. Hence, it is proposed that work is done on studying how different integration sites affect the productivity and stability of transfected genes in the development process. For the study to be more industrially relevant, it is proposed that the Chinese Hamster Ovary dhfr-deficient cell line, CHO-DG44, is used as the model system.