Heidi Sleister - Profile on Academia.edu (original) (raw)

Papers by Heidi Sleister

Why Research On the Pharmacogenetics of Atypical Antipsychotic-Induced Weight Gain in Individuals With Intellectual Disabilities Is Warranted

Journal of Mental Health Research in Intellectual Disabilities, 2011

... DOI: 10.1080/19315864.2011.555600 Heidi M. Sleister a & Maria Gabriela Valdovinos b * pag... more ... DOI: 10.1080/19315864.2011.555600 Heidi M. Sleister a & Maria Gabriela Valdovinos b * pages 65-78. ... Journal of Clinical Psychiatry , 64: 6062. [CrossRef], [PubMed], [Web of Science ®], [CSA] View all references; McKee, Bodfish, Mahorney, Heeth, & Ball, 200526. ...

Analysis of mRNA changes as a function of learning and methamphetamine exposure

Methamphetamine is a powerful, addictive drug that is of great concern in Iowa and the Midwest. P... more Methamphetamine is a powerful, addictive drug that is of great concern in Iowa and the Midwest. Powerful methamphetamine abuse is correlated with depression and learning disabilities (e.g. attention deficit disorder). Although the mechanism of action of methamphetamine in the brain has been well characterized at the cellular and synaptic level, the effects of this drug at the genetic level are not well understood. To analyze the possible effects of methamphetamine on learning, we are using real-time PCR to investigate changes in gene expression (i.e., messenger RNA levels) in rat brains as a function of learning and exposure to methamphetamine. Specifically, we are analyzing relative mRNA levels from genes whose products have been implicated in learning and/or addiction: GABA receptors, dopamine receptors, and glutamate (NMDA) receptors. Comparison of these data sets is expected to reveal potential cellular and molecular mechanisms involved in producing drug-induced learning disabil...

Inhibition of drug efflux by pluronics in S. cerevisiae

Overexpression of P-gp efflux pumps is believed to be responsible for multidrug resistance of mam... more Overexpression of P-gp efflux pumps is believed to be responsible for multidrug resistance of mammalian cancer cells. Inhibition of these pumps is expected to increase the efficacy and decrease the toxicity of chemotherapeutic agents. The non-ionic triblock copolymer Pluronic P85 is reported to inhibit mammalian P-gp efflux pumps, leading to higher intracellular drug concentrations. An analogous, well-characterized efflux transporter, Pdr5p, has been identified in the yeast Saccharomyces cerevisiae, but the effect of Pluronic copolymers on this transporter has not been studied. We have examined the inhibitory effects of P85 on three strains of S. cerevisiae: a pdr5 deletion strain, a PDR5 over-expressing strain, and the PDR5 wild-type strain using the hydrophilic Pdr5p substrate cycloheximide (CHX) as a model antifungal “drug”. Yeast cells were grown in the presence of a range of CHX (0-0.3 mcg/ml) and P85 (0-10 mg/ml). After incubation for 24hrs at 30°C, the ability of P85 to inhib...

Virology journal, Jan 30, 2014

BackgroundInfluenza A viruses (IAVs) are important pathogens that affect the health of humans and... more BackgroundInfluenza A viruses (IAVs) are important pathogens that affect the health of humans and many additional animal species. IAVs are enveloped, negative single-stranded RNA viruses whose genome encodes at least ten proteins. The IAV nucleoprotein (NP) is a structural protein that associates with the viral RNA and is essential for virus replication. Understanding how IAVs interact with host proteins is essential for elucidating all of the required processes for viral replication, restrictions in species host range, and potential targets for antiviral therapies.MethodsIn this study, the NP from a swine IAV was cloned into a yeast two-hybrid ¿bait¿ vector for expression of a yeast Gal4 binding domain (BD)-NP fusion protein. This ¿bait¿ was used to screen a Y2H human HeLa cell ¿prey¿ library which consisted of human proteins fused to the Gal4 protein¿s activation domain (AD). The interaction of ¿bait¿ and ¿prey¿ proteins resulted in activation of reporter genes.ResultsSeventeen po...

Journal of Immunological Methods, 2002

Antibodies specific for a protein of interest are invaluable tools for monitoring the protein's s... more Antibodies specific for a protein of interest are invaluable tools for monitoring the protein's structure, location and activity. Due to the tendency of an immune system to mount a response toward the abundant, immunodominant epitopes in a protein mixture, difficulties are inherent in the isolation of antibodies specific for proteins that are rare or poorly immunogenic. Likewise, isolation of antibodies specific for a protein with significant sequence similarity to other proteins, such as those derived from protein engineering, may be challenging. Subtractive immunization is a technique proven to facilitate efforts to produce monoclonal antibodies specific for antigens that are present in low abundance in a protein mixture, poorly immunogenic and/or similar in sequence or structure to other proteins. This protocol provides a detailed, stepwise procedure for the isolation of antibodies specific for a protein with sequence similarity to other proteins. As an example, we describe methods established to isolate antibodies specific to a methionine-enriched variant of soybean vegetative storage protein b (VSPb-Met) that shares 91.8% amino acid sequence identity to the wild-type protein (VSPb-WT). These methods include cyclophosphamide-induced immunosuppression of mice for the wild-type protein followed by immunization with VSPb-Met. As a result of this procedure, mouse polyclonal antibodies that exhibited 10-fold greater reactivity with VSPb-Met than VSPb-WT in an ELISA were generated. It is anticipated that this strategy will have utility for generating antibodies specific to protein variants derived from protein engineering.

Journal of Immunological Methods, 2001

Protein engineering is a common strategy for the generation of protein variants with new properti... more Protein engineering is a common strategy for the generation of protein variants with new properties. The engineered Ž . variants often have a high degree of similarity with the wild-type progenitor protein, necessitating a tool e.g., antibody to distinguish the wild-type and variant protein forms. As part of an overall effort to understand the process of incorporation of amino acids into storage proteins during seed fill in soybean, we have engineered a variant of soybean vegetative storage Ž . protein b VSPb that is 91.8% identical in amino acid sequence to the wild-type protein, but contains 10% methionine Ž . VSPb-Met, unpublished results . Thus, it would be desirable to have antibodies that specifically recognize VSPb-Met over the endogenously expressed wild-type protein in transgenic plants. To this end, we compared three strategies for the isolation Ž . Ž . of VSPb-Met-specific antibodies: 1 hybridoma production using VSPb-Met protein as the antigen, 2 polyclonal antibody Ž . production in rabbits using a peptide antigen corresponding to a methionine-rich region of VSPb-Met, and 3 subtractive immunization in mice using VSPb-WT as the tolerogen, cyclophosphamide for immunosuppression and VSPb-Met as the immunogen. While the first strategy generated antibodies cross-reactive to both antigens, the second strategy generated polyclonal antibodies that preferentially recognized the variant protein in immunoblots. However, using subtractive immunization, we were able to generate mouse polyclonal antibodies that exhibited 10-fold greater reactivity with VSPb-Met than VSPb-WT in an ELISA.

Nucleic Acids Research, 1992

In order to construct a human chromosome 4-specific YAC library, we have utilized pYAC4 and a mou... more In order to construct a human chromosome 4-specific YAC library, we have utilized pYAC4 and a mouse/human hybrid cell line HA(4)A in which the only human chromosome present is chromosome 4. From this cell line, approximately 8Mb of chromosome 4 have been cloned. The library includes 65 human-specific clones that range in size from 30kb to 290kb, the average size being 108kb. In order to optimize the manipulation of YAC libraries, we have begun to investigate the stability of YACs containing human DNA in yeast cells; these studies will also determine if there are intrinsic differences in the properties of chromosomes containing higher eukaryotic DNAs. We are examining two kinds of stability: 1 ] mitotic stability, the ability of the YAC to replicate and segregate properly during mitosis, and 2] structural stability, the tendency of the YAC to rearrange. We have found that the majority of YACs examined are one to two orders of magnitude less stable than authentic yeast chromosomes. Interestingly, the largest YAC analyzed displayed a loss rate typical for natural yeast chromosomes. Our results also suggest that increasing the length of an artificial chromosome improves its mitotic stability. One YAC that showed a very high frequency of rearrangement by mitotic recombination proved to be a mouse/human chimera. In contrast to studies using total human DNA, the frequency of chimeras (i.e., mouse/human) in the YAC pool appeared to be low.