Genetic manipulation of Anopheles stephensi immunity to increase plasmodium falciparum salivary gland sporozoite infection levels

Human malaria is responsible for over 700,000 deaths a year. To stay abreast of the threat posed by the parasite, a constant stream of new drugs and vector control methods are required. This study focuses on a vaccine that has the potential to protect against parasite infection, but has been hindered by developmental challenges. In malaria prevention, live, attenuated, aseptic, Plasmodium falciparum sporozoites (PfSPZ) can be administered as a highly protective vaccine. PfSPZ are produced using adult female Anopheles stephensi mosquitoes as bioreactors. Production volume and cost of a PfSPZ vaccine for malaria are expected to be directly correlated with Plasmodium falciparum infection intensity in the salivary glands. The sporogonic development of Plasmodium falciparum in A. stephensi to fully infected salivary gland stage sporozoites is dictated by the activities of several known components of the mosquito’s innate immune system. Here I report on the use of genetic technologies that have been rarely, if ever, used in Anopheles stephensi Sda500 to increase the yield of sporozoites per mosquito and enhance vaccine production. By combining the Gal4/UAS bipartite system with in vivo expression of shRNA gene silencing, activity of the IMD signaling pathway downstream effector LRIM1, an antagonist to Plasmodium development, was reduced in the midgut, fat body, and salivary glands of A. stephensi. In infection studies using P. berghei and P. falciparum these transgenic mosquitoes consistently produced significantly more salivary gland stage sporozoites than wildtype controls, with increases in P. falciparum ranging from 2.5 to 10 fold. Using Plasmodium infection assays and qRT-PCR, two novel findings were identified. First, it was shown that 14 days post Plasmodium infection, transcript abundance of the IMD immune effector genes LRIM1, TEP1 and APL1c are elevated, in the salivary glands of A. stephensi, suggesting the salivary glands may play a role in post midgut defense against the parasite. Second, a non-pathogenic IMD signaling pathway response was observed which could suggest an alternative pathway for IMD activation. The information gained from these studies has significantly increased our knowledge of Plasmodium defense in A. stephensi and moreover could significantly improve vaccine production.

Strong shift equivalence, algebraic K-theory, and isolating zero-dimensional dynamics on manifolds

We study the relations of shift equivalence and strong shift equivalence for matrices over a ring $\mathcal{R}$, and establish a connection between these relations and algebraic K-theory. We utilize this connection to obtain results in two areas where the shift and strong shift equivalence relations play an important role: the study of finite group extensions of shifts of finite type, and the Generalized Spectral Conjectures of Boyle and Handelman for nonnegative matrices over subrings of the real numbers. We show the refinement of the shift equivalence class of a matrix $A$ over a ring $\mathcal{R}$ by strong shift equivalence classes over the ring is classified by a quotient $NK_{1}(\mathcal{R}) / E(A,\mathcal{R})$ of the algebraic K-group $NK_{1}(\calR)$. We use the K-theory of non-commutative localizations to show that in certain cases the subgroup $E(A,\mathcal{R})$ must vanish, including the case $A$ is invertible over $\mathcal{R}$. We use the K-theory connection to clarify the structure of algebraic invariants for finite group extensions of shifts of finite type. In particular, we give a strong negative answer to a question of Parry, who asked whether the dynamical zeta function determines up to finitely many topological conjugacy classes the extensions by $G$ of a fixed mixing shift of finite type. We apply the K-theory connection to prove the equivalence of a strong and weak form of the Generalized Spectral Conjecture of Boyle and Handelman for primitive matrices over subrings of $\mathbb{R}$. We construct explicit matrices whose class in the algebraic K-group $NK_{1}(\mathcal{R})$ is non-zero for certain rings $\mathcal{R}$ motivated by applications. We study the possible dynamics of the restriction of a homeomorphism of a compact manifold to an isolated zero-dimensional set. We prove that for $n \ge 3$ every compact zero-dimensional system can arise as an isolated invariant set for a homeomorphism of a compact $n$-manifold. In dimension two, we provide obstructions and examples.

The Real-Quaternionic Indicator of Irreducible Self-Conjugate Representations of Real Reductive Algebraic Groups and A Comment on the Local Langlands Correspondence of GL(2, F )

The real-quaternionic indicator, also called the $\delta$ indicator, indicates if a self-conjugate representation is of real or quaternionic type. It is closely related to the Frobenius-Schur indicator, which we call the $\varepsilon$ indicator. The Frobenius-Schur indicator $\varepsilon(\pi)$ is known to be given by a particular value of the central character. We would like a similar result for the $\delta$ indicator. When $G$ is compact, $\delta(\pi)$ and $\varepsilon(\pi)$ coincide. In general, they are not necessarily the same. In this thesis, we will give a relation between the two indicators when $G$ is a real reductive algebraic group. This relation also leads to a formula for $\delta(\pi)$ in terms of the central character. For the second part, we consider the construction of the local Langlands correspondence of $GL(2,F)$ when $F$ is a non-Archimedean local field with odd residual characteristics. By re-examining the construction, we provide new proofs to some important properties of the correspondence. Namely, the construction is independent of the choice of additive character in the theta correspondence.