Online education is an effective and efficient electronic source for students to gain knowledge. Online education modules allow educators to bridge the gap between theory and practice for students. The modules created during this project tackle several topics in Materials Science & Engineering such as ceramics, hardness of materials, the use of light optical microscopy, corrosion and more. They are designed to encourage students to think critically, form questions and go through the scientific method in order to solve complex problems. They are formatted to combine a variety of learning styles by combining a power point lecture, assigning reading materials and having the hands on experience via demonstration or experimentation. This benefits students immediately in that they will learn how to apply the knowledge in education, research and industry. The educational modules are evaluated several times in order to perfect the lesson plan. In having the modules evaluated by current educators and individuals in industry the modules can be altered and adjusted in order to focus on specific key points. A few of these modules are completed and ready to use in a classroom setting specifically tailored to advanced high school, technical schools, community colleges and four year educational institutions.

A reversible tissue adhesive is a surface that is capable of sticking to tissues repeatedly without losing adhesive ability, similar to how geckos' feet can adhere to smooth and rough dry surfaces. This project seeks to combine the reversible adhesion exhibited by geckos and the bacterium E. coli, a model for receptor-ligand tissue adhesives. This technology has the potential to benefit laparoscopic forceps designs, which are required to manipulate delicate tissues easily punctured or torn during a medical procedure. FimH is an adhesive protein expressed at the end of E. coli fimbriae that binds reversibly to mannose, a sugar expressed in human endothelial tissues. A FimH receptor-ligand adhesive has worked on small length scales (μm²), but adhesion fails at larger scales (cm²), possibly due to the propagation of a single crack at the adhesive interface. My project addresses this failure by 1) fabricating the geometry of the existing adhesive in PDMS allowing an interface size without damaging force instruments and 2) combining E. coli fimbriae with polymer molds of micron-sized pillar arrays that mimic the topology of gecko footpads and arrest crack propagation on dry surfaces. Spherical PDMS slices of increasing size (40μm – 300μm radii) were coated with mannose and brought down to fimbriated cell-culture plates. Results show that PDMS measures higher adhesion (226 nN) than the existing receptor-ligand adhesive (25 nN), indicating a successful increase in adhesive interface size. The PDMS surface can be used to measure and compare the adhesive capabilities of various micropillar array geometries coated with E. coli fimbriae. In conclusion, micropillar geometries that measure higher adhesion than plates coated with fimbraie have the potential to be developed into a tissue adhesive. Future studies may show reversible adhesion is achieved when fimbriated-micropillar substrates are brought in contact with cultured and dissected tissues.

Centromeres in many species are found to reside near origins of replication that are activated early in S-phase, yet the biological significance of this conserved early replication of centromeres is unknown. Previous research has shown that the activation time of an origin is affected by its chromosomal location. A recent study has found that centromeres in the yeast Saccharomyces cerevisiae advance the activation time of nearby origins, and that this effect depends on the centromere’s ability to establish a functional kinetochore. These observations lead to the hypothesis that centromeres need to be replicated early to allow sufficient time for the assembly of functional kinetochores. If kinetochores fail to be established in a timely fashion, chromosomes may not be correctly bi-orient at mitosis. Daughter cells therefore may inherit the incorrect number of chromosomes creating conditions of aneuploidy such as those found in Down syndrome and many cancers. I will test the importance of early centromere replication by deleting origins immediately flanking a yeast centromere to delay that centromere’s replication. I will then conduct an assay to test for the rate of chromosome mis-segregation events in the mutant strain compared to the wild-type strain. I anticipate that the mutant strain may have a higher rate of loss for the modified chromosome, suggesting that early replication of centromeres is important for maintaining genome stability. This project will also provide insights into the regulation of the temporal program of replication origin activation and provide the first functional test for the consequences of disrupting the temporal program of chromosome replication.

We, the researchers in Dr. Wendy Stone’s Lab, examined the developmental trajectory of motor imitation from 12-18 months and its relation to social functioning and language ability at 24 months in younger siblings of children diagnosed with an Autism Spectrum Disorder (ASD; ASD-sibs), compared to younger siblings of typically developing children (TD-sibs). Motor imitation is the reproduction of a modeled presentation of a motor event and ASD refers to neurodevelopmental disorders characterized by impairments in social-communication and the presence of stereotyped, repetitive behaviors. It is important to study ASD-sibs because they are at an increased risk for developing an ASD or sub-clinical symptoms. Young children with ASD display motor imitation deficits, but currently, studies have not examined motor imitation in ASD-sibs within the first 18 months of life. Thus, the age at which impairments emerge remains unanswered. Further investigation of motor imitation is necessary given its importance for learning and social interactions. Using the Screening Tool for Autism in Toddlers (STAT) to measure motor imitation, the Mullen Scales of Early Learning to assess language, and the Autism Diagnostic Observation Schedule to determine ASD severity, we expect to find that: a) motor imitation will show significant growth between 12-18 months of age; b) ASD-sibs will have lower baseline levels and lower rates of growth than TD-sibs; c) baseline levels and rate of growth for motor imitation will predict ASD severity and language ability at 24 months. Hierarchical linear models will be used for the developmental trajectories of motor imitation from 12-18 months of age. Linear regressions will be used to examine whether baseline levels and growth predict later ASD diagnosis and language outcomes. This study may provide information leading to earlier detection of ASD. Earlier diagnosis of ASD allows for earlier intervention, which has been shown to improve children’s social-communicative outcomes.  

 Mycobacterium tuberculosis infection is a leading cause of death worldwide, but the full details of how the human immune system responds to these bacteria are not known. Our lab studies CD1 proteins which allow T-cells to respond to bacterial lipids. We discovered that low expression of CD1A on dendritic cells is associated with an increased susceptibility to tuberculosis; however, the molecular details of CD1A-deficiency are not known. I cloned and analyzed the CD1A promoter from four CD1A-deficient and five control individuals. I identified twelve genetic variations in the CD1A promoter. Of these, eight were single nucleotide polymorphisms (SNPs), three were insertions or deletions, and one was a variable tandem repeat. One SNP (-199 T/C) and the variable tandem repeat showed an association with CD1A-deficiency. A T/T genotype at position -199 from the transcription start site correctly identified five out of five control subjects while a C/C genotype identified three out of four CD1A-deficient subjects. The variable tandem repeat was a string of 20 to 60 thymine bases. The CD1A-deficient subjects showed uniform repeat lengths, while the repeat lengths among control subjects were variable. These data identify two loci within the CD1A promoter that will be the target of future studies aimed at understanding the molecular mechanism of CD1A-deficiency.

Inhibitor of differentiation (Id) is a transcription factor that negatively regulates basic helix-loop-helix (bHLH) transcription factors. Of four Ids identified (Id1-4), Id1 and Id3 have been shown to regulate glial development in the central nervous system. In the retina, there is a special type of glia called the Müller glia. Currently, it is not known whether the Ids are important for Müller glial development. Knockout studies in mice demonstrated that Id1/Id3 double knockout (KO) are embryonic lethal, while either Id1 or Id3 KO mice develop grossly normal. We investigated if Id1/Id3 are necessary for Müller glial development by performing Id1 shRNA knockdown (KD) in Id3 KO retinas. Five Id1 shRNAs were tested for their efficiency in NIH 3T3 cells (mouse fibroblast cell line), and the Western blot results showed over 90% decrease in Id1 expression using three shRNAs. We will electroporate Id3 KO retinal explants with these Id1 shRNAs and determine the effects of Id1/Id3 loss on Muller glial development. We expect to see a reduction in the number of Müller glia if Id1/Id3 play a role in their development.

The nature of Systemic Lupus Erythematosus (SLE) involves a complex network of uncontrolled activations of signals, which leads to a vast range of symptoms. SLE is an autoimmune disease that manifests itself when the immune system attacks the body’s cells and tissues, resulting in inflammations in joints, organs, and skin. The course of the disease is unpredictable and much more is to be understood about the mechanisms of the illness and the background of the patients who are prone to the disease. This study explores both the mechanistic and clinical aspect of SLE in an attempt to gain a broader view of the disease. Using 3 interferon stimulated genes, MX1, PKR, and CXCL10, we evaluated interferon scores of 66 patients. We then utilized cluster analysis to divide patients into two main groups based on interferon scores: Group 1 consisted of controls and SLE patients with low interferon scores (0.4), Group 2 consisted of SLE patients with high interferon scores (10.4). These groups were compared with interferon stimulated genes, RNase1 and Toll-Like Receptors 7 (TLR7), which positively correlated with interferon scores. A system called Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) is often utilized to quantitatively assess patients’ disease activity. The criteria in the evaluation of patients included symptoms often associated with SLE. The score in SLEDAI reflected the disease activity of the patient. In this study, we investigated the relationship between SLEDAI and interferon score, in which we found no correlation. However, when we examined Group 2, there was a trend towards better correlation with SLEDAI. In the clinical aspect, we examined age, race, and gender of SLE patients. In addition, we collected information on autoantibody titers such as double stranded DNA (dsDNA), antiphospholipid antibodies, and antinuclear antibodies (ANA), all of which may be important indicators of disease activity.

E. coli are a species of bacteria that naturally inhabit the mammalian intestinal tract. They can thrive in environments with high fluid shear like the colon or the urinary tract due to their sticky hair-like projections called fimbriae which allow them to grip epithelial lining. These fimbriae are primarily composed of a coiled chain of a repeating protein unit called FimA. This helical coil unwinds in a gradual fashion when pulled and our goal is to see how mutating certain amino acids (all proteins are made of sequences of amino acids) inside FimA can affect the energy required to uncoil the helix. A microscopic cantilever is applied to different mutants for E. coli fimbria and an atomic force microscope records how much force the fimbria exerts back during a pull, indicating the helix uncoiling and how much force it requires. We hope to find mutants that uncoil more easily or not at all which would illuminate which amino acids in FimA are active in maintaining the helix and its uncoiling flexibility. The findings would have many applications practically and for expanding research. Rigid helices are a valuable test group for examining fimbria behavior independent of the effects uncoiling while the amino acids isolated as important can help design target drugs against E. coli infections in urinary tracts by weakening their binding. The protein structure knowledge earned is also of biomedical value as it can help design novel adhesives using bacterial fimbrial protein.

Model membranes are important tools for investigating physical properties of lipid bilayers. Unfortunately, current techniques for producing giant unilamellar vesicles (GUVs) with biologically and physically relevant properties (namely: inclusion of charged lipids, encapsulation of high salt buffer, and compositional asymmetry between the inner and outer leaflets) are difficult, unreliable, and/or low throughput. We examine a method called cDICE, developed by Abkarian et al., that has the potential to address all of these issues. Our results here for charged lipids in high salt buffers are promising, whereas asymmetry has proven difficult to measure.