Life Sciences Commons^™

Identification Of Compounds Causing Cellular Autofluorescence In Touch Samples, Elora C. Wall

Master of Science in Forensic Science Directed Research Projects

As DNA analysis has advanced and produced tests with higher sensitivities, attention has turned toward obtaining DNA profiles from cells left with fingermarks. Recent studies have reported that cells deposited within fingermarks can exhibit differences in autofluorescence emission in the ‘red’ region of the visible spectrum (e.g., between 650-670 nm), which can be used to differentiate contributor cell population and separate them before DNA profiles. Interestingly, this emission was not consistent to the individual day-to-day and likely not a genetically-controlled attribute of the contributor. Instead, this emission signature results from extended exposure of the skin to certain materials such as …

Gold/Qds-Embedded-Ceria Nanoparticles: Optical Fluorescence Enhancement As A Quenching Sensor, Nader Shehata, Effat Samir, Ishac Kandas

Biology Student Research

This work focuses on improving the fluorescence intensity of cerium oxide (ceria) nanoparticles (NPs) through added plasmonic nanostructures. Ceria nanoparticles are fluorescent nanostructures which can emit visible fluorescence emissions under violet excitation. Here, we investigated different added plasmonic nanostructures, such as gold nanoparticles (Au NPs) and Cadmium sulfide/selenide quantum dots (CdS/CdSe QDs), to check the enhancement of fluorescence intensity emissions caused by ceria NPs. Different plasmonic resonances of both aforementioned nanostructures have been selected to develop optical coupling with both fluorescence excitation and emission wavelengths of ceria. In addition, different additions whether in-situ or post-synthesis have been investigated. We found …

Quantifying Anthropogenic Indicators And Changes In Dissolved Organic Matter In Coastal Urban Aquatic Ecosystems Exposed To High Tidal Flooding, Gonzalo E. Eyzaguirre

Department of Biological Sciences

Sea-level rise is causing an increase in tidal flooding in coastal urban areas. Extreme high tides, also known as king tides, are peak tide moments in which tidal amplitude is increased and shallow groundwater flows from the underlying water table are introduced. During tidal flooding in urban areas, accumulated anthropogenic indicators of different water sources are released from groundwater to surface waters, but how these tidal events affect the contributions of different water sources to urban flood waters is uncertain. We quantified tracers of anthropogenic origin including fluoride, fecal coliform bacteria, as well as dissolved organic carbon (DOC) concentrations and …

Cpef Is The Bilin Lyase That Ligates The Doubly Linked Phycoerythrobilin On Phycoerythrin In The Cyanobacterium Fremyella Diplosiphon, Wendy M. Schluchter, R. B. Cole, D. M. Kehoe, M. N. Boutaghou, J. A. Karty, A. Gutu, L. S. Hernandez, J. P. Frick, C. V. Hernandez, C. M. Kronfel

Biological Sciences Faculty Publications

Phycoerythrin (PE) is a green light-absorbing protein present in the light-harvesting complex of cyanobacteria and red algae. The spectral characteristics of PE are due to its prosthetic groups, or phycoerythrobilins (PEBs), that are covalently attached to the protein chain by specific bilin lyases. Only two PE lyases have been identified and characterized so far, and the other bilin lyases are unknown. Here, using in silico analyses, markerless deletion, biochemical assays with purified and recombinant proteins, and site-directed mutagenesis, we examined the role of a putative lyase-encoding gene, cpeF, in the cyanobacterium Fremyella diplosiphon. Analyzing the phenotype of the cpeF deletion, …

Plasmonic-Ceria Nanoparticles As Fluorescence Intensity And Lifetime Quenching Optical Sensor, Nader Shehata, Effat Samir, Ishac Kandas

Biology Faculty Publications

Ceria nanoparticles have been recently used as an optical fluorescent material with visible emission under ultraviolet excitation, due to the formation of trivalent cerium ions with corresponding oxygen vacancies. This paper introduces the enhancement of both fluorescence emission and lifetime through adding gold nanoparticles. The reason is due to possible coupling between the plasmonic resonance of gold nanoparticles and the fluorescence emission of ceria that has been achieved, along with enhanced formation of trivalent cerium ions. Both factors lead to higher fluorescence intensity peaks and shorter fluorescence lifetimes. As an application, gold-ceria nanoparticles have been used as an optical sensing …

Electrospun Pva Polymer Embedded With Ceria Nanoparticles As Silicon Solar Cells Rear Surface Coaters For Efficiency Improvement, Effat Samir, Mohamed Salah, Ali Hajjiah, Nader Shehata, Marwa Fathy, Aya Hamed

Biology Faculty Publications

This paper introduces electrospun nanofibers embedded with ceria nanoparticles as silicon solar cells coaters, showing their influence on the solar cells efficiency. Ceria nanoparticles can be synthesized to have formed oxygen vacancies (O-vacancies), which are associated with converting cerium ions from the Ce4+ state ions to the Ce3+ ones. These O-vacancies follow the rule of improving silicon solar cellconductivity through the hopping mechanism. Besides, under violet excitation, the reduced trivalent cerium Ce3+ ions are directly responsible for down-converting the un-absorbed violet or ultra-violet (UV) wavelengths to a resulted green fluorescence emission at ~520 nm. These are absorbed through the silicon …

Nuclear Features Of The Heterotrich Ciliate Blepharisma Americanum: Genomic Amplification, Life Cycle, And Nuclear Inclusion, Megan M. Wancura, Ying Yan, Laura A. Katz, Xyrus X. Maurer-Alcalá

Biological Sciences: Faculty Publications

Blepharisma americanum, a member of the understudied ciliate class Heterotrichea, has a moniliform somatic macronucleus that resembles beads on a string. Blepharisma americanum is distinguishable by its pink coloration derived from the autofluorescent pigment blepharismin and tends to have a single somatic macronucleus with 3–6 nodes and multiple germline micronuclei. We used fluorescence confocal microscopy to explore the DNA content and amplification between the somatic and germline nuclei of B. americanum through its life cycle. We estimate that the DNA content of the macronucleus and micronucleus are 43 ± 8 Gbp and 83 ± 16 Mbp respectively. This correlates with …

Effects Of Cold Acclimation On Rectal Macromorphology, Ultrastructure, And Cytoskeletal Stability In Gryllus Pennsylvanicus Crickets., Lauren E Des Marteaux, Joseph R Stinziano, Brent J Sinclair

Biology Publications

Cold-acclimated insects maintain ion and water balance in the cold, potentially by reducing permeability or increasing diffusion distance across ionoregulatory epithelia such as the rectum. We explored whether cold acclimation induces structural modifications that minimize water and ion diffusion across the rectum and maintain rectal cell integrity. We investigated rectal structure and cytoskeletal stability in chill-susceptible adult Gryllus pennsylvanicus crickets acclimated for one week to either warm (25 °C) or cold (12 °C) conditions. After acclimation, we used light and transmission electron microscopy to examine rectal macromorphology and rectal pad paracellular ultrastructure. We also used fluorescence microscopy and a filamentous-actin …

Septin Assemblies Form By Diffusion-Driven Annealing On Membranes, Andrew A. Bridges, Huaiying Zhang, Shalin B. Mehta, Patricia Occhipinti, Tomomi Tani, Amy S. Gladfelter

Dartmouth Scholarship

Septins assemble into filaments and higher-order structures that act as scaffolds for diverse cell functions including cytokinesis, cell polarity, and membrane remodeling. Despite their conserved role in cell organization, little is known about how septin filaments elongate and are knitted together into higher-order assemblies. Using fluorescence correlation spectroscopy, we determined that cytosolic septins are in small complexes, suggesting that septin filaments are not formed in the cytosol. When the plasma membrane of live cells is monitored by total internal reflection fluorescence microscopy, we see that septin complexes of variable size diffuse in two dimensions. Diffusing septin complexes collide and make …

Septin Phosphorylation And Coiled-Coil Domains Function In Cell And Septin Ring Morphology In The Filamentous Fungus Ashbya Gossypii, Rebecca A. Meseroll, Patricia Occhipinti, Amy S. Gladfelter

Dartmouth Scholarship

Septins are a class of GTP-binding proteins conserved throughout many eukaryotes. Individual septin subunits associate with one another and assemble into heteromeric complexes that form filaments and higher-order structures in vivo. The mechanisms underlying the assembly and maintenance of higher-order structures in cells remain poorly understood. Septins in several organisms have been shown to be phosphorylated, although precisely how septin phosphorylation may be contributing to the formation of high-order septin structures is unknown. Four of the five septins expressed in the filamentous fungus, Ashbya gossypii, are phosphorylated, and we demonstrate here the diverse roles of these phosphorylation sites …

Plant Calcium Content: Ready To Remodel, Jian Yang, Tracy Punshon, Mary Lou Guerinot, Kendal D. Hirschi

Dartmouth Scholarship

By identifying the relationship between calcium location in the plant cell and nutrient bioavailability, the plant characteristics leading to maximal calcium absorption by humans can be identified. Knowledge of plant cellular and molecular targets controlling calcium location in plants is emerging. These insights should allow for better strategies for increasing the nutritional content of foods. In particular, the use of preparation-free elemental imaging technologies such as synchrotron X-ray fluorescence (SXRF) microscopy in plant biology may allow researchers to understand the relationship between subcellular location and nutrient bioavailability. These approaches may lead to better strategies for altering the location of calcium …

Phycoerythrin-Specific Bilin Lyase-Isomerase Controls Blue-Green Chromatic Acclimation In Marine Synechococcus, Wendy M. Schluchter, Avijit Biswas, Frédéric Partensky, J. A. Karty, Laurence Garczarek, A. Gutu, David M. Kehoe, A. Shukla, N. Blot, L. A. Hammad

Biological Sciences Faculty Publications

The marine cyanobacterium Synechococcus is the second most abundant phytoplanktonic organism in the world's oceans. The ubiquity of this genus is in large part due to its use of a diverse set of photosynthetic light-harvesting pigments called phycobiliproteins, which allow it to efficiently exploit a wide range of light colors. Here we uncover a pivotal molecular mechanism underpinning a widespread response among marine Synechococcus cells known as type IV chromatic acclimation (CA4). During this process, the pigmentation of the two main phycobiliproteins of this organism, phycoerythrins I and II, is reversibly modified to match changes in the ambient light color …

Septin Filaments Exhibit A Dynamic, Paired Organization That Is Conserved From Yeast To Mammals, Bradley S. Demay, Xiaobo Bai, Louisa Howard, Patricia Occhipinti, Rebecca A. Meseroll, Elias T. Spiliotis, Rudolf Oldenbourg, Amy S. Gladfelter

Dartmouth Scholarship

The septins are conserved, GTP-binding proteins important for cytokinesis, membrane compartmentalization, and exocytosis. However, it is unknown how septins are arranged within higher-order structures in cells. To determine the organization of septins in live cells, we developed a polarized fluorescence microscopy system to monitor the orientation of GFP dipole moments with high spatial and temporal resolution. When GFP was fused to septins, the arrangement of GFP dipoles reflected the underlying septin organization. We demonstrated in a filamentous fungus, a budding yeast, and a mammalian epithelial cell line that septin proteins were organized in an identical highly ordered fashion. Fluorescence anisotropy …

Determining The Composition Of The Dwelling Tubes Of Antarctic Pterobranchs, Lukasz J. Sewera

Honors Projects

Pterobranchs are a group of marine invertebrates within the Hemichordata, which share characteristics with both chordates and echinoderms. Pterobranchs live in colonies of secreted tubes, coenicia, which are composed of a gelatinous material of unknown composition. Visually, the tubes appear similar to the tunic of tunicates, a group of invertebrates within the Chordata. The nonproteinaceous tunic of tunicates is composed of cellulose, which is unusual. The goal of this study was to determine the composition of the pterobranch coenicium. Some aspects of pterobranch phylogeny are still unclear even after multiple molecular and morphological studies. Identification of any new shared characteristics …

Development Of A Fluorescent Anti-Factor Xa Assay To Monitor Unfractionated And Low Molecular Weight Heparins, Leanne Harris, Vanessa Castro-Lopez, Nissrin Hammadi, James S. O'Donnell, Tony Killard

Articles

Fluorogenic assays have many potential advantages over traditional clot-based and chromogenic assays such as the absence of interference from a range of factor deficiencies as well as offering the possibility of assays in platelet rich plasma or whole blood. A fluorogenic anti-factor Xa (anti-FXa) assay has been developed for the determination of heparin-like anticoagulants including unfractionated heparin (UFH), low-molecular weight heparins (LMWHs), namely enoxaparin and tinzaparin, and the synthetic heparinoid danaparoid, in commercial human pooled plasma. The assay was based on the complexation of heparin-spiked plasmas with exogenous FXa at a concentration of 4 nM in the presence of 0.9 …

Mutational Studies Uncover Non-Native Structure In The Dimeric Kinetic Intermediate Of The H2a–H2b Heterodimer, Matthew R. Stump, Lisa M. Gloss

Faculty Publications - Department of Biological & Molecular Science

The folding pathway of the histone H2A–H2B heterodimer minimally includes an on-pathway, dimeric, burst-phase intermediate, I2. The partially folded H2A and H2B monomers populated at equilibrium were characterized as potential monomeric kinetic intermediates. Folding kinetics were compared for initiation from isolated, folded monomers and the heterodimer unfolded in 4 M urea. The observed rates were virtually identical above 0.4Murea, exhibiting a log-linear relationship on the final denaturant concentration. Below ∼0.4 M urea (concentrations inaccessible from the 4-M urea unfolded state), a rollover in the rates was observed; this suggests that a component of the I2 ensemble contains non-native structure that …

Chloroplast Fe(Iii) Chelate Reductase Activity Is Essential For Seedling Viability Under Iron Limiting Conditions, Jeeyon Jeong, Christopher Cohu, Loubna Kerkeb, Marinus Pilon, Erin L. Connolly, Mary Lou Guerinot

Dartmouth Scholarship

Photosynthesis, heme biosynthesis, and Fe-S cluster assembly all take place in the chloroplast, and all require iron. Reduction of iron via a membrane-bound Fe(III) chelate reductase is required before iron transport across membranes in a variety of systems, but to date there has been no definitive genetic proof that chloroplasts have such a reduction system. Here we report that one of the eight members of the Arabidopsis ferric reductase oxidase (FRO) family, FRO7, localizes to the chloroplast. Chloroplasts prepared from fro7 loss-of-function mutants have 75% less Fe(III) chelate reductase activity and contain 33% less iron per microgram of chlorophyll than …

Mutational Analysis Of The Stability Of The H2a And H2b Histone Monomers, Matthew R. Stump, Lisa M. Gloss

Faculty Publications - Department of Biological & Molecular Science

The eukaryotic histone heterodimer H2A–H2B folds through an obligatory dimeric intermediate that forms in a nearly diffusion-limited association reaction in the stopped-flow dead time. It is unclear whether there is partial folding of the isolated monomers before association. To address the possible contributions of structure in the monomers to the rapid association, we characterized H2A and H2B monomers in the absence of their heterodimeric partner. By far-UV circular dichroism, the H2A and H2B monomers are 15% and 31% helical, respectively—significantly less than observed in X-ray crystal structures. Acrylamide quenching of the intrinsic Tyr fluorescence was indicative of tertiary structure. The …

The Cohesion Protein Ord Is Required For Homologue Bias During Meiotic Recombination, Hayley A. Webber, Louisa Howard, Sharon E. Bickel

Dartmouth Scholarship

During meiosis, sister chromatid cohesion is required for normal levels of homologous recombination, although how cohesion regulates exchange is not understood. Null mutations in orientation disruptor (ord) ablate arm and centromeric cohesion during Drosophila meiosis and severely reduce homologous crossovers in mutant oocytes. We show that ORD protein localizes along oocyte chromosomes during the stages in which recombination occurs. Although synaptonemal complex (SC) components initially associate with synapsed homologues in ord mutants, their localization is severely disrupted during pachytene progression, and normal tripartite SC is not visible by electron microscopy. In ord germaria, meiotic double strand breaks appear …

Small Heat Shock Protein Responses Of A Closely Related Pair Of Desert And Coastal Encelia, Charles A. Knight, David D. Ackerly

Biological Sciences

Evolutionary variation for accumulation of small heat shock protein (sHsp) may contribute to thermal niche differentiation between species. Here we examine temperature and time-course-dependent variation for sHsp accumulation in a recently diverged pair of Encelia raised in a common environment: Encelia farinosa, common in the Mojave desert, and Encelia californica, which is found along the cool coastal bluffs of southern North America. Both species exhibit peak sHsp accumulation at 42^oC. Encelia californica accumulated greater levels of sHsp at temperatures below 42^oC, while E. farinosa had greater levels above 42^oC. Encelia farinosa accumulates …

Gag Proteins Of The Two Drosophila Telomeric Retrotransposons Are Targeted To Chromosome Ends, Svetlana Rashkova, Sarah E. Karam, Rebecca Kellum, Mary-Lou Pardue

Biology Faculty Publications

Drosophila telomeres are formed by two non-LTR retrotransposons, HeT-A and TART, which transpose only to chromosome ends. Successive transpositions of these telomeric elements yield arrays that are functionally equivalent to the arrays generated by telomerase in other organisms. In contrast, other Drosophila non-LTR retrotransposons transpose widely through gene-rich regions, but not to ends. The two telomeric elements encode very similar Gag proteins, suggesting that Gag may be involved in their unique targeting to chromosome ends. To test the intrinsic potential of these Gag proteins for targeting, we tagged the coding sequences with sequence of GFP and expressed the constructs …

An Ecological And Evolutionary Analysis Of Photosynthetic Thermotolerance Using The Temperature-Dependent Increase In Fluorescence, Charles A. Knight, David D. Ackerly

Biological Sciences

The hypothesis that species inhabiting warmer regions have greater photosynthetic tolerance of high temperatures was tested using the temperature-dependent increase in fluorescence (T-F_o). Congeneric species pairs of Atriplex, Salvia, Encelia, and Eriogonum with desert versus coastal distributions were studied in a common environment and in the field. In addition, 21 species with contrasting microclimate distributions were studied at a field site in a northern California chaparral community. The average July maximum temperature within the current distributions of species was quantified using a geographic information system. Four parameters (T_crit, T_S20, …

The Kinetics Of Solvent Reorientation In Hydroxylated Solvents From The Exciting-Wavelength Dependence Of Chromophore Emission Spectra, John Milton, Robert M. Purkey, William C. Galley

WM Keck Science Faculty Papers

The disappearance of the exciting-wavelength dependence of the phosphorescence spectra of polar, aromatic chromophores in supercooled glycol–water mixtures is utilized to monitor the kinetics of solvent reorientation. Reorientation times in the nanosecond to second range are obtained for (3:2 v/v) glycerol–water and (1:1 v/v) ethylene glycol–water at 140–240 °K. The results suggest that the process is one involving a cluster of solvent molecules and in which the chromophore plays a relatively passive role. Steady-state data and direct measurements of phosphorescence shifts as a function of time indicate that the solvent reorientation process is nonexponential in nature. The decay function derived …

Petroleum Hydrocarbons From Effluents: Detection In Marine Environment, John T. Tanacredi Ph.D.

Faculty Works: CERCOM

The marine environment has become the primary disposa ground for an increasing quantity of petroleum wastes. Mushrooming demands for petroleum products and the lack of economic incentive to recycle waste oil will increase the concentrations of detrimental petroleum hydrocarbons in the marine environment

Although a continuous, low-level discharge of waste petroleum hydrocarbons into the marine environment may not be as dramatic as a major oil spill, the consequences could be more devastating over an extended period. As noted by Blumer, earlier interpretations of the environmental effects of oil must not be reevaluated in the light of recent evidence of its …