Life Sciences Commons^™

Development And Application Of A Gis-Based Long Island Sound Eelgrass Habitat Suitability Index Model, Jamie M.P. Vaudrey, Justin Eddings, Christopher Pickerell, Lorne Brousseau, Charles Yarish

Department of Marine Sciences

The primary objectives of the Eelgrass Habitat Suitability Index Model (EHSI Model) are to assist in the evaluation of sites being considered for eelgrass restoration efforts in the Long Island Sound (LIS) area and to identify areas where water quality issues reduce or eliminate the potential for natural eelgrass colonization. To achieve this goal, geospatial processing of data available from the Long Island Sound area was conducted using ArcGIS v10.0 including the 3D Analyst and Spatial Analyst extensions. The result is a series of maps presented in this report and a GIS-based model available for users to interact with the …

Expression Of Phosphodiesterase 6 (Pde6) In Human Breast Cancer Cells, Hongli Dong, Kevin P. Claffey, Stefan Brocke, Paul M. Epstein

UCHC Articles - Research

Considerable epidemiological evidence demonstrates a positive association between artificial light at night (LAN) levels and incidence rates of breast cancer, suggesting that exposure to LAN is a risk factor for breast cancer. There is a 30-50% higher risk of breast cancer in the highest LAN exposed countries compared to the lowest LAN countries, and studies showing higher incidence of breast cancer among shift workers exposed to more LAN have led the International Agency for Research on Cancer to classify shift work as a probable human carcinogen. Nevertheless, the means by which light can affect breast cancer is still unknown. In …

One-Day Treatment Of Small Molecule 8-Bromo-Cyclic Amp Analogue Induces Cell-Based Vegf Production For In Vitro Angiogenesis And Osteoblastic Differentiation, Kevin W.-H. Lo, Ho Man Kan, Keith A. Gagnon, Cato T. Laurencin

UCHC Articles - Research

Small molecule based regenerative engineering is emerging as a promising strategy for regenerating bone tissue. Small molecule cAMP analogues have been proposed as novel biofactors for bone repair and regeneration, and while promising, the effect that these small molecules have on angiogenesis, a critical requirement for successful bone regeneration, is still unclear. Our previous research demonstrated that the small molecule cAMP analogue 8-bromoadenosine-3’,5’-cyclic monophosphate (8-Br-cAMP) was able to promote initial osteoblast adhesion on a polymeric scaffold via cAMP signaling cascades. Here, we report that 8-Br-cAMP is capable of inducing in vitro cell-based VEGF production for angiogenesis promotion. We first demonstrated …

The General Qapp For Long Island Sound Volunteer Coastal Monitoring (With Adoption Form), Jamie M.P. Vaudrey, Kimberly Gallagher

Department of Marine Sciences

The General QAPP for Long Island Sound Volunteer Coastal Monitoring is designed to streamline the process of writing a QAPP for Monitoring Programs in the Long Island Sound area. This document does not replace guidance on developing a program and is not sufficient as a stand-alone document to guide the initial development and sample design process for a monitoring program.

Evaluation Of Current Community-Based Monitoring Efforts And Recommendations For Developing A Cohesive Network Of Support For Monitoring Long Island Sound Embayments., Jamie M.P. Vaudrey, Jamie Alonzo, Adrienne Esposito, Curt Johnson, Maureen Dolan Murphy, Charles Yarish

Department of Marine Sciences

This project focuses on Long Island Sound embayments and represents an exploration of the potential for coordination among community-based water quality monitoring groups, and coordination between community-based groups and data end users, including scientists and managers. Community-based monitoring groups represent a potential valuable source of water quality information. The development of standardized protocols, training methods and reporting procedures will serve to provide end users with the confidence necessary to utilize the data collected as part of community-based monitoring programs. Both groups of stakeholders stand to benefit from this type of coordination with the ultimate benefit being a healthier, better understood …

Preosteocytes/Osteocytes Have The Potential To Dedifferentiate Becoming A Source Of Osteoblasts, Elena Torreggiani, Brya G. Matthews, Slavica Pejda, Igor Matic, Danka Grcevic, Mark C. Horowitz, Ivo Kalajzic

UCHC Articles - Research

Presently there is no clear evidence for the ability of mature osteogenic lineage cells to dedifferentiate. In order to identify and trace mature osteogenic lineage cells, we have utilized transgenic mouse models in which the dentin matrix protein 1 (Dmp1) promoter drives expression of GFP (active marker) or Cre recombinase (historic label) in preosteocytes/osteocytes. In long bone chip outgrowth cultures, in which cells on the bone surface were enzymatically removed, cells with previous activity of the Dmp1 promoter migrated onto plastic and down-regulated Dmp1-GFP expression. Dmp1Cre-labeled cells from these cultures had the potential to re-differentiate into the osteogenic lineage, while …

Osterix-Cre Labeled Progenitor Cells Contribute To The Formation And Maintenance Of The Bone Marrow Stroma, Yaling Liu, Sara Strecker, Liping Wang, Mark S. Kronenberg, Wen Wang, David W. Rowe, Peter F. Maye

UCHC Articles - Research

We have carried out fate mapping studies using Osterix-EGFPCre and Osterix-CreERt animal models and found Cre reporter expression in many different cell types that make up the bone marrow stroma. Constitutive fate mapping resulted in the labeling of different cellular components located throughout the bone marrow, whereas temporal fate mapping at E14.5 resulted in the labeling of cells within a region of the bone marrow. The identity of cell types marked by constitutive and temporal fate mapping included osteoblasts, adipocytes, vascular smooth muscle, perineural, and stromal cells. Prolonged tracing of embryonic precursors labeled at E14.5dpc revealed the continued …

Utilization Of Transgenic Models In Evaluation Of Osteogenic Differentiation Of Embryonic Stem Cells, Dario Repic, Elena Torreggiani, Tiziana Franceschetti, Brya G. Matthews, Alexander C. Lichtler, Ivo Kalajzic

UCHC Articles - Research

Previous studies reported that embryonic stem cells (ESCs) can be induced to differentiate into cells showing a mature osteoblastic phenotype by culturing them under osteo-inductive conditions. It is probable that osteogenic differentiation requires that ESCs undergo differentiation through an intermediary step involving a mesenchymal lineage precursor. Based on our previous studies indicating that adult mesenchymal progenitor cells express αSMA, we have generated ESCs from transgenic mice in which an αSMA promoter directs the expression of red fluorescent protein (RFP) to mesenchymal progenitor cells. To track the transition of ESC-derived MSCs into mature osteoblasts, we have utilized a bone-specific fragment of …

Crystal Structure Of The Ubiquitin-Like Small Archaeal Modifier Protein 2 From Haloferax Volcanii, Yunfeng Li, Mark W. Maciejewski, Kai Jin, Yuhang Zhang, Bing Hao

UCHC Articles - Research

The discovery of ubiquitin-like small archaeal modifier protein 2 (SAMP2) that forms covalent polymeric chains in Haloferax volcanii has generated tremendous interest in the function and regulation of this protein. At present, it remains unclear whether the Hfx. volcanii modifier protein SAMP1 has such polyubiquitinating-like activity. Although SAMP1 and SAMP2 use the same conjugation machinery to modify their target proteins, each can impart distinct functional consequences. To better understand the mechanism of SAMP2 conjugation, we have sought to characterize the biophysical and structural properties of the protein from Hfx. volcanii. SAMP2 is only partially structured under mesohalic solution conditions …

Post-Injury Calcium Chelation Rescues Skeletal Muscle Regeneration In Mice, Matthew D. Magda

Honors Scholar Theses

During aging the ability of skeletal muscle to regenerate after injury wanes until muscle integrity cannot be maintained. In this study it is shown that calcium chelation can restore young-like regenerative ability in an aged mouse model of skeletal muscle regeneration.

Ptpn11 Deletion In A Novel Cartilage Cell Causes Metachondromatosis By Activating Hedgehog Signaling, Qian Wu

UCHC Articles - Research

SHP2, encoded by PTPN11, is required for survival, proliferation and differentiation of various cell types^1,2. Germ line activating mutations in PTPN11 cause Noonan Syndrome, while somatic PTPN11 mutations cause childhood myeloproliferative disease and contribute to some solid tumors. Recently, heterozygous inactivating mutations in PTPN11 were found in metachondromatosis, a rare inherited disorder featuring multiple exostoses, endochondromas, joint destruction and bony deformities^3,4. The detailed pathogenesis of this disorder has remained unclear. Here, we used a conditional knockout allele (Ptpn11^fl) and Cre recombinase (Cre) transgenic mice to delete Ptpn11 specifically in …

The Plastid Genome Of Najas Flexilis: Adaptation To Submersed Environments Is Accompanied By The Complete Loss Of The Ndh Complex In An Aquatic Angiosperm, Elena L. Peredo, Ursula M. King, Donald H. Les

Open Access Author Fund Awardees' Articles

The re-colonization of aquatic habitats by angiosperms has presented a difficult challenge to plants whose long evolutionary history primarily reflects adaptations to terrestrial conditions. Many aquatics must complete vital stages of their life cycle on the water surface by means of floating or emergent leaves and flowers. Only a few species, mainly within the order Alismatales, are able to complete all aspects of their life cycle including pollination, entirely underwater. Water- pollinated Alismatales include seagrasses and water nymphs ( Najas ), the latter being the only freshwater genus in the family Hydrocharitaceae with subsurface water-pollination. We have determined the complete …

The Transcriptomics To Proteomics Of Hair Cell Regeneration: Looking For A Hair Cell In A Haystack, Gopinath Rajadinakaran

UCHC Articles - Research

Mature mammals exhibit very limited capacity for regeneration of auditory hair cells, while all non-mammalian vertebrates examined can regenerate them. In an effort to find therapeutic targets for deafness and balance disorders, scientists have examined gene expression patterns in auditory tissues under different developmental and experimental conditions. Microarray technology has allowed the large-scale study of gene expression profiles (transcriptomics) at whole-genome levels, but since mRNA expression does not necessarily correlate with protein expression, other methods, such as microRNA analysis and proteomics, are needed to better understand the process of hair cell regeneration. These technologies and some of the results of …

Subtelomeric Hotspots Of Aberrant 5-Hydroxymethylcytosine-Mediated Epigenetic Modifications During Reprogramming To Pluripotency, Stormy J. Chamberlain, I-Ping Chen

UCHC Articles - Research

Mammalian somatic cells can be directly reprogrammed into induced pluripotent stem cells (iPSCs) by introducing defined sets of transcription factors. Somatic cell reprogramming involves epigenomic reconfiguration, conferring iPSCs with characteristics similar to embryonic stem cells (ESCs). Human ES cells contain 5-hydroxymethylcytosine (5hmC), which is generated through the oxidation of 5-methylcytosine by the TET enzyme family. Here we show that 5hmC levels increase significantly during reprogramming to human iPSCs mainly due to TET1 activation, and this hydroxymethylation change is critical for optimal epigenetic reprogramming, but does not compromise primed pluripotency. Compared with hES cells, we find iPS cells tend to form …

Dikar-Induced Synthetic Lethality In A Drosophila Model Of Cag Repeat Diseases Does Not Result From An Expression Feedback Loop, Daniel Camacho

Honors Scholar Theses

Human CAG repeat diseases manifest themselves through the common pathology of neurodeneration. This pathological link is attributed to the property shared by all nine of these diseases: an expanded polyglutamine (polyQ) tract. The most evident result of polyQ expansion is protein aggregation, and it is believed that this phenomenon is partly responsible for conferring cytotoxic properties on the mutated protein. Apart from sequestering the mutated protein, cellular aggregates are able to incorporate native proteins via polyQ-mediated aggregation, thus disrupting important cellular pathways. Using Drosophila melanogaster as a disease model, researchers have been able to compile collections of these so-called disease …

Pharmacological Inhibition Of Histone Methyltransferase G9a Affects Expression Of Citron Kinase Target Genes In Neural Stem Cells, Nicholas Heliotis

Honors Scholar Theses

Regulated repression of gene expression by post-translational modification of histones is required for normal development. The histone methyltransferase G9a is essential for embryonic development, and we have shown that phosphorylation of G9a by the CitK, a gene required for normal CNS development, gates gene repression and dimethylation of histone H3 at lysine 9 (H3K9me2) in neural progenitors. CitK and G9a co-localize to promoter regions of genes up-regulated in CitK null cells. CitK mutant progenitors lack H3K9me2 at promoter regions of up-regulated genes, and re-expression of CitK restores both repression of gene expression and H3K9me2 occupancy. In this thesis we examine …

Investigations In Enhancing The Reproducibility Of Implantable Amperometric Glucose Biosensors For Mass Production, Dipesh Manharbhai Patel

Honors Scholar Theses

Implantable glucose sensors for Diabetes management should possess several attributes such as linearity, sensitivity, selectivity, long life time and should elicit no negative tissue response. Based on this, the UConn implantable glucose sensor team has developed a 5-layer sensor architecture that simultaneously affords the aforementioned requirements. However, the large scale production of these 5-layer sensors is inhibited by the lack of high sensor-to-sensor reproducibility. Herein, we investigate the origin of sensor-to-sensor irreproducibility and develop methodologies for large-scale fabrication of these devices.

A Detergent-Free Strategy For The Reconstitution Of Active Enzyme Complexes From Native Biological Membranes Into Nanoscale Discs, Ashley R. Long, Catherine C. O’Brien, Ketan Malhotra, Christine T. Schwall, Arlene D. Albert, Nathan N. Alder

Open Access Author Fund Awardees' Articles

Background

The reconstitution of membrane proteins and complexes into nanoscale lipid bilayer structures has contributed significantly to biochemical and biophysical analyses. Current methods for performing such reconstitutions entail an initial detergent-mediated step to solubilize and isolate membrane proteins. Exposure to detergents, however, can destabilize many membrane proteins and result in a loss of function. Amphipathic copolymers have recently been used to stabilize membrane proteins and complexes following suitable detergent extraction. However, the ability of these copolymers to extract proteins directly from native lipid bilayers for subsequent reconstitution and characterization has not been explored.

Results

The styrene-maleic acid (SMA) copolymer effectively …

The Effects Of Maternal Diet During Gestation On Postnatal Growth Of Lambs And On Metabolic Hormones In Lambs And Ewes, Amanda R. Fox

Honors Scholar Theses

Fetal programming is defined as when a stimulus at a vital stage of development results in permanent effects that persist throughout the lifetime of the individual (Lucas, 1991). There is evidence that poor maternal diet, consisting of both undernutrition and overnutrition, during gestation has permanent negative effects on the growth and development of offspring through fetal programming. Fetal programming enhances the ability of the fetus to absorb and use nutrients while in-utero, ensuring postpartum survival (Hales and Barker, 1992). However, in a postnatal environment where nutrition is plentiful, fetal programming results in negative health consequences (Wu et al., 2006). We …

Biophysical Studies Of Styrene-Maleic Acid Copolymer Stabilized Membrane Mimetics, Diane Yu

University Scholar Projects

Traditional methods of isolating membrane proteins involve the use of detergents that destabilize the membrane environment. Biophysical studies of membrane proteins are challenged by the requirement of a stable membrane platform. Lipodisq® nanoparticles are novel membrane mimetics that consist of 3:1 styrene-maleic acid copolymer encircling phospholipids to form discoidal species on the order of 10 nm in diameter. Bacteriorhodopsin was previously incorporated into Lipodisq® nano-particles from its native purple membrane without the addition of detergent. In this study, dynamic light scattering was used to investigate the stability of empty Lipodisq® nano-particles over time. These data show that Lipodisq® nano-particles are …

The Effects Of Aging And Injury On The Murine Ependymal Barrier And Subventricular Zone, John Jeffrey Peters

University Scholar Projects

Previous research has shown that numerous age-related changes occur in the subventricular zone (SVZ) and diminish the neurogenic output of the region. Here, various agerelated changes in the structure of the lateral ventricle, the ependymal barrier, and the SVZ were examined. It was found that the walls of the lateral ventricles move together, adhere, and eventually fuse. This results in a degradation of the SVZ. Other changes associated with aging were found, including ependymal cell stretching, heightened, ependymal repair, changes in CSF flow velocity and dynamics, and altered ependymal cell polarity. These changes may be related to the process of …

Mmp-3 Mediates Psychosine-Induced Globoid Cell Formation: Implications For Leukodystrophy Pathology, Kumiko Ijichi, Graham D. Brown, Craig S. Moore, Paige N. Winokur, Roberto Pagarigan, Stephen J. Crocker

UCHC Articles - Research

Globoid cell leukodystrophy (GLD) or Krabbe disease, is a fatal demyelinating disease attributed to mutations in the galactocerebrosidase (GALC) gene. Loss of function mutations in GALC result in accumulation of the glycolipid intermediate, galactosylsphingosine (psychosine). Due to the cytotoxicity of psychosine, it has been hypothesized that accumulated psychosine underlie the pathophysiology of GLD. However, the cellular mechanisms of GLD pathophysiology remain unclear. Globoid cells, multinucleated microglia/macrophages in the central nervous system (CNS), are a defining characteristic of GLD. Here we report that exposure of primary glial cultures to psychosine induces the expression and the production of matrix metalloproteinase …

In Vitro Characterization Of Avian Influenza Virus Isolates With A Truncated Ns1 Gene Segment, Aaron Dick

Honors Scholar Theses

Avian Influenza Virus represents a significant threat to the world poultry population, and is a potential threat to humans due to the possibility of cross-species AIV infection. Our approach is to characterize a number of avian virus populations with respect to their content of biologically active particles that include hemagglutinating particles (HAP), plaque forming particles (PFP), interferon inducing particles (IFP), interferon induction-suppressing particles (ISP), defective-interfering particles (DIP), cell-killing particles (CKP) and non-infectious cell killing particles (niCKP) using unique in vitro assays developed for avian influenza virus in the Marcus-Sekellick Laboratory. Specifically, we will use a strain of Avian influenza virus, …

Effects Of Poor Maternal Nutrition On Gh, Igf-I, Igfbp-2 And -3, Insulin, And Leptin Concentrations In Pregnant Ewes, Michelle E. Forella

Honors Scholar Theses

Maternal hormones impact fetal growth and development through their effect on nutrient partitioning and placental development. Maternal diet is one factor regulating maternal hormone concentrations. We hypothesized that during gestation, diet alters concentrations of maternal metabolic hormones. Pregnant ewes (n=36; 12/treatment) were individually fed 1 of 3 diets [100% (CON), 140% (OVER), or 60% (RES) NRC requirements] from wk 5 of gestation until parturition. Weekly blood samples (20 mL) were obtained via jugular venipuncture and BW was measured. Concentrations of GH, IGF-I and leptin were determined by RIA. Insulin was determined by ELISA. Data were analyzed using ANOVA. By the …

Direct Reprogramming Of Mouse Embryonic Fibroblasts Into Gabaergic Neurons, Nicholas J. Wasko

Honors Scholar Theses

Recent studies in which mouse and human fibroblasts were directly reprogrammed into functional neurons have demonstrated that lineage-specific transcription factors can override differentiation processes previously thought to be irreversible. Notably, three factors have been shown to be required for the induction of neural cell fate. Subsequent experiments have identified additional factors capable of generating specific neuronal subtypes, including motor neurons and dopaminergic neurons, but the factors necessary for inducing GABAergic neuronal fate have yet to be elucidated. Here, factors linked to GABAergic fate were transfected into mouse fibroblasts in order to identify candidate factors for inhibitory interneuron direct reprogramming. Results …

The Effects Of Aging And Injury On The Murine Ependymal Barrier And Subventricular Zone, John J. Peters

Honors Scholar Theses

Previous research has shown that numerous age-related changes occur in the subventricular zone (SVZ) and diminish the neurogenic output of the region. Here, various age-related changes in the structure of the lateral ventricle, the ependymal barrier, and the SVZ were examined. It was found that the walls of the lateral ventricles move together, adhere, and eventually fuse. This results in a degradation of the SVZ. Other changes associated with aging were found, including ependymal cell stretching, heightened, ependymal repair, changes in CSF flow velocity and dynamics, and altered ependymal cell polarity. These changes may be related to the process of …

Chemical And Biological Triggers For The Improved Intercalation And Release Of Glucose Oxidase (Go) In The Galleries Of Α-Zirconium Phosphate Nanolayers For A Noninvasive Oral Alternative To Insulin Drug Therapies, Momina Kamal Afrede

Honors Scholar Theses

Noninvasive treatment alternatives for diseases from diabetes to hypercholerestemia are at the cutting edge of research. Although many noninvasive drug delivery routes, including oral nasal, buccal, dermal, and pulmonary have been investigated no one route has been fully approved for patient use by the Food and Drug Administration (FDA) standards as an alternative to current needle dependent subcutaneous insulin delivery. 10 12 This thesis focuses on two novel oral protein drug delivery systems α-Zr(IV) phosphate (α-ZrP)-protein-Divalent Metal Ion and α-Zr(IV) phosphate (α-ZrP)-protein- cationized Bovine Serum Albumin BSA. Glucose oxidase (GO) serves as a model protein in this thesis for noninvasive …

Characterizing The Role Of Cortactin In Actin Pedestal Assembly By Enterohemorrhagic Escherichia Coli (Ehec), Sarah E. Grout

Honors Scholar Theses

Enterohemorrhagic Escherichia coli (EHEC) is a major foodborne cause of bloody diarrhea and renal failure. During colonization of the intestine, EHEC injects the transmembrane receptor protein Tir and the cytoplasmic effector protein EspFU into host cells to reorganize the actin cytoskeleton into adhesion “pedestals.” EspFU has been shown to bind and activate the actin nucleation factor N-WASP to drive actin polymerization into pedestals. However, EspFU can still assemble pedestals in cells lacking N-WASP, suggesting that this effector protein is able to also trigger N-WASP-independent pathways of actin polymerization during infection. Cortactin is an atypical nucleation factor that localizes to pedestals, …

The Effect Of Landlocking On Intestinal Aquaporin 1 Expression In Alosa Pseudoharengus (Alewife), Emily Funk

Honors Scholar Theses

To determine the role of intestinal aquaporin 1 (AQP1), a passive water absorption channel, in the evolution of osmoregulatory physiology, I examined the differential expression of intestinal AQP1 between anadromous and landlocked ecotype of alewife in response to seawater challenges. I cloned and sequenced AQP1 from intestinal tissue of the alewife, following which I quantified the relative expression of AQP1 in each ecotype using Real-Time qPCR. In response to an acute seawater (30ppt) challenge, the anadromous alewives showed an upregulation of intestinal AQP1, while the landlocked alewives did not show a significant increased in AQP1 expression. After …

Modulation Of Synaptic Transmission By Adenosine In Layer 2/3 Of The Rat Visual Cortex In Vitro, Nicholas M. Bannon

Master's Theses

No abstract provided.