Organisms Commons^™

Aav Serotype Influences Gene Transfer In Corneal Stroma In Vivo, Ajay Sharma, Jonathan C. K. Tovey, Arkasubhra Ghosh, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

This study evaluated the cellular tropism and relative transduction efficiency of three AAV serotypes, AAV6, AAV8 and AAV9, for corneal gene delivery using mouse cornea in vivo and donor human cornea ex vivo. The AAV6, AAV8 and AAV9 serotypes having AAV2 plasmid encoding for alkaline phosphatase (AP) gene were generated by transfecting HEK293 cell line with pHelper, pARAP4 and pRep/Cap plasmids. Viral vectors (109 vg/μl) were topically applied onto mouse cornea in vivo and human cornea ex vivo after removing the epithelium. Human corneas were processed for transgene delivery at day 5 after viral vector application. Mouse corneas were harvested …

Different Training Volumes Yield Equivalent Increases In Bmd, R. A. Pierce, L. C. Lee, C. P. Ahles, S. M. Shdo, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

The purpose of this study was to determine if an exercise threshold existed in stimulating an elevation in bone mineral density (BMD), via resistance training, during the growth period in male rats. 27 male rats were randomly divided into Control (Con, n = 9), 3 ladder climb resistance trained group (3LC, n = 9), and 6 ladder climb resistance trained group (6LC, n = 9). The 3LC and 6LC groups were conditioned to climb a vertical ladder with weights appended to their tail 3 days/wk for a total of 6 wks, but the 6LC group performed significantly more work than …

Equal Bmd After Daily Or Triweekly Exercise In Growing Rats, B. D. Kayser, J. K. Godfrey, R. M. Cunningham, R. A. Pierce, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

The purpose of this study was to examine the efficacy of continuous resistance training (3 days/wk) compared to interrupted resistance training where 20-24 h separated an exercise bout (i.e. 6 days/wk) for enhancing bone mineral density (BMD) in growing male rats. The total volume of work performed per week between the two resistance training programs was equivalent by design. Young male rats were randomly divided into Control (Con, n = 9), 3 days/wk resistance trained group (RT3, n = 9), and 6 days/wk resistance trained group (RT6, n = 9). The RT3 and RT6 groups were conditioned to climb a …

The Isolation Of Viruses Infecting Archaea, Kenneth M. Stedman, Kate Porter, Mike L. Dyall-Smith

Biology Faculty Publications and Presentations

A mere 50 viruses of Archaea have been reported to date; these have been investigated mostly by adapting methods used to isolate bacteriophages to the unique growth conditions of their archaeal hosts. The most numerous are viruses of thermophilic Archaea. These viruses have been discovered by screening enrichment cultures and novel isolates from environmental samples for their ability to form halos of growth inhibition, or by using electron microscopy to screen enrichment cultures for virus-like particles. Direct isolation without enrichment has not yet been successful for viruses of extreme thermophiles. On the other hand, most viruses of extreme halophiles, the …

The Transition Zone: Impact Of Riverbanks On Emergent Dragonfly Nymphs. Implications For Riverbank Restoration And Management, Kirsten Hope Martin

Antioch University Dissertations & Theses

The use of riprap in the restoration and stabilization of riverine landscapes is an issue of concern for many ecologists. While current methods of bank stabilization, especially those involving the placement of rocks (riprap) along the waterline, are effective in controlling erosion their presence changes habitat components (slope, substrate composition, near-shore river velocity) at the river-land interface. The additional impacts of river current, water temperature, soil composition, slope, and water level fluctuation, may further imperil emerging nymphs. The purpose of this research is to document the effects of riprap, location (upriver or downriver of hydroelectric intake/outtake facilities), water level fluctuation, …

Female Humpback Whale (Megaptera Novaeangliae) Reproductive Class And Male-Female Interactions During The Breeding Season, Meagan E. Jones

Antioch University Dissertations & Theses

This study examined whether female reproductive class (i.e., presence or absence of a calf) in humpback whales (Megaptera novaeangliae) affects female behavior and male-female interactions on the Hawaiian breeding grounds. From 2003-2008, 295 female-calf groups (with or without a male escort) and 256 female no-calf groups (with a male) were observed. Forty-one female no-calf groups were confirmed male-female pairs by genetic sexing or photo-identification. Focal follows of 36 male-female pairs and 50 female-calf groups (29 escorted and 21 unescorted female-calf groups) were analyzed for time budget and movement estimates. Relative fluke size (fluke photogrammetry, n=30) and sighting histories …

Rearrangement Of The Rna Polymerase Subunit H And The Lower Jaw In Archaeal Elongation Complexes, Sebastian Grünberg, Christoph Reich, Mirijam E. Zeller, Michael S. Bartlett, Michael Thomm

Biology Faculty Publications and Presentations

The lower jaws of archaeal RNA polymerase and eukaryotic RNA polymerase II include orthologous subunits H and Rpb5, respectively. The tertiary structure of H is very similar to the structure of the C-terminal domain of Rpb5, and both subunits are proximal to downstream DNA in pre-initiation complexes. Analyses of reconstituted euryarchaeal polymerase lacking subunit H revealed that H is important for open complex formation and initial transcription. Eukaryotic Rpb5 rescues activity of the ΔH enzyme indicating a strong conservation of function for this subunit from archaea to eukaryotes. Photochemical cross-linking in elongation complexes revealed a striking structural rearrangement of RNA …

Killing Of Treponema Denticola By Mouse Peritoneal Macrophages, P. Gaibani, C. Vocale, S. Ambretti, F. Cavrini, Jacques Izard, L. Miragliotta, M. T. Pellegrino, V. Sambri

Department of Food Science and Technology: Faculty Publications

Treponema denticola has been identified as an important cause of periodontal disease and hypothesized to be involved in extra-oral infections. The objective of this study was to investigate the role of T. denticola cell length and motility during mouse peritoneal macrophages in vitro uptake. Macrophages, incubated under aerobic and anaerobic conditions, produced a similar amount of TNF-α when stimulated with Escherichia coli LPS. The uptake of FlgE- and CfpA-deficient mutants of T. denticola was significantly increased compared with the wild-type strain, due to cell size or lack of motility. Opsonization with specific antibodies considerably improved the treponemes’ uptake. …

Native Cellular Architecture Of Treponema Denticola Revealed By Cryo-Electron Tomography, Jacques Izard, Chyong-Ere Hsieh, Ronald J. Limberger, Carmen A. Mannella, Michael Marko

Department of Food Science and Technology: Faculty Publications

Using cryo-electron tomography, we are developing a refined description of native cellular structures in the pathogenic spirochete Treponema denticola. Tightly organized bundles of periplasmic flagella were readily observed in intact plunge-frozen cells. The periplasmic space was measured in both wild-type and aflagellate strains, and found to widen by less than the diameter of flagella when the latter are present. This suggests that a structural change occurs in the peptidoglycan layer to accommodate the presence of the flagella. In dividing cells, the flagellar filaments were found to bridge the cytoplasmic cylinder constriction site. Cytoplasmic filaments, adjacent to the inner membrane, …

Trichostatin A Inhibits Corneal Haze In Vitro And In Vivo, Ajay Sharma, Maneesh M. Mehan, Sunilima Sinha, John W. Cowden, Rajiv R. Mohan

Pharmacy Faculty Articles and Research

PURPOSE. Trichostatin A (TSA), a histone deacetylase inhibitor, has been shown to suppress TGF- –induced fibrogenesis in many nonocular tissues. The authors evaluated TSA cytotoxicity and its antifibrogenic activity on TGF- –driven fibrosis in the cornea with the use of in vitro and in vivo models.

METHODS. Human corneal fibroblasts (HSFs) were used for in vitro studies, and New Zealand White rabbits were used for in vivo studies. Haze in the rabbit cornea was produced with photorefractive keratectomy (PRK) using excimer laser. Trypan blue exclusion and MTT assays evaluated TSA cytotoxicity to the cornea. Density of haze in the rabbit …

Interrupted Resistance Training And Bmd In Growing Rats, J. K. Godfrey, B. D. Kayser, G. V. Gomez, Jason Bennett, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

A resistance training program, where the exercise was uninterrupted (UT, i.e. continuous repetitions) was compared against another resistance training program where the exercise was interrupted (IT, i.e. 2 exercise sessions during a training day) for enhancing bone modeling and bone mineral density (BMD) in maturating animals. The total volume of work performed between the two resistance training programs was equivalent by design. Young male rats (similar to 8 weeks old) were randomly divided into Control (Con, n = 8), UT (n = 8) and IT (n = 7) resistance trained groups. The UT and IT groups were conditioned to climb …

Cryo-Electron Tomography Elucidates The Molecular Architecture Of Treponema Pallidum, The Syphilis Spirochete, Jacques Izard, Christian Renken, Chyong-Ere Hsieh, Daniel C. Desrosiers, Star Dunham-Ems, Carson La Vake, Linda L. Gebhardt, Ronald J. Limberger, David L. Cox, Michael Marko, Justin D. Radolf

Department of Food Science and Technology: Faculty Publications

Cryo-electron tomography (CET) was used to examine the native cellular organization of Treponema pallidum, the syphilis spirochete. T. pallidum cells appeared to form flat waves, did not contain an outer coat and, except for bulges over the basal bodies and widening in the vicinity of flagellar filaments, displayed a uniform periplasmic space. Although the outer membrane (OM) generally was smooth in contour, OM extrusions and blebs frequently were observed, highlighting the structure’s fluidity and lack of attachment to underlying periplasmic constituents. Cytoplasmic filaments converged from their attachment points opposite the basal bodies to form arrays that ran roughly …

The Guinea Pig Ileum Lacks The Direct, High-Potency, M2-Muscarinic, Contractile Mechanism Characteristic Of The Mouse Ileum, Michael T. Griffin, Minoru Matsui, Rennolds S. Ostrom, Frederick J. Ehlert

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

We explored whether the M2 muscarinic receptor in the guinea pig ileum elicits a highly potent, direct-contractile response, like that from the M3 muscarinic receptor knockout mouse. First, we characterized the irreversible receptor-blocking activity of 4-DAMP mustard in ileum from muscarinic receptor knockout mice to verify its M3 selectivity. Then, we used 4-DAMP mustard to inactivate M3 responses in the guinea pig ileum to attempt to reveal direct, M2 receptor-mediated contractions. The muscarinic agonist, oxotremorine-M, elicited potent contractions in ileum from wild-type, M2 receptor knockout, and M3 receptor knockout mice characterized by negative log EC50 (pEC 50 ) values ± …

Pseudomonas Aeruginosa-Candida Albicans Interactions: Localization And Fungal Toxicity Of A Phenazine Derivative, Jane Gibson, Arpanah Sood, Deborah A. Hogan

Dartmouth Scholarship

Phenazines are redox-active small molecules that play significant roles in the interactions between pseudomonads and diverse eukaryotes, including fungi. When Pseudomonas aeruginosa and Candida albicans were cocultured on solid medium, a red pigmentation developed that was dependent on P. aeruginosa phenazine biosynthetic genes. Through a genetic screen in combination with biochemical experiments, it was found that a P. aeruginosa-produced precursor to pyocyanin, proposed to be 5-methyl-phenazinium-1-carboxylate (5MPCA), was necessary for the formation of the red pigmentation. The 5MPCA-derived pigment was found to accumulate exclusively within fungal cells, where it retained the ability to be reversibly oxidized and reduced, and its …

The Effects Of Host Physiological Conditions On The Expression Of Icsp In Shigella Flexneri, Karen Levy, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Shigella flexneri is a gram-negative bacterium capable of causing diarrhea and dysentery known as shigellosis. It is estimated there are 167.4 million shigellosis episodes throughout the world each year causing 1.1 million deaths. Shigella invades cells in the lower intestine through an induced phagocytosis. Once in the cytoplasm, bacteria move from one cell to another using actin-based motility. The Shigella outer membrane protease IcsP regulates actin-based motility and cell-to-cell spread by cleaving the actin assembly protein IcsA from the bacterial cell surface. We hypothesize that IcsP may serve additional functions during infection. By examining which environmental signals trigger icsP expression, …

Characterization Of The Omptin Protease, Ompt, In Escherichia Coli, Amanda Yates, Eun-Hae Kim, Helen Wing

Undergraduate Research Opportunities Program (UROP)

Omptins are outer membrane proteases found in gram negative bacteria that cause diseases in humans, such as pathogenic Escherichia coli, Shigella flexneri, Salmonella typhimurium, and Yersinia pestis. Bacterial species that express omptins cause diseases such as highly fatal plague and severe diarrhea and dysentery. The genes that encode these proteases are ompT, icsP, pgtE, and pla, respectively. These proteases are highly related in structure and share approximately 50% sequence identity. In S. flexneri, IcsP has been shown to cleave a key virulence determinant, IcsA (Egile et al., 1997). IcsA recruits host actin and allows for intracellular movement within host cells …

Transmembrane Domain Length Of Viral K+ Channels Is A Signal For Mitochondria Targeting, Jorg Balss, Panagiotis Papatheodorou, Mario Mehmel, Dirk Baumeister, Brigitte Hertel, Nicolas Delaroque, Frank C. Chatelain, Daniel L. Minor Jr., James L. Van Etten, Joachim Rassow, Anna Moroni, Gerhard Thiel

James Van Etten Publications

K⁺ channels operate in the plasma membrane and in membranes of organelles including mitochondria. The mechanisms and topogenic information for their differential synthesis and targeting is unknown. This article describes 2 similar viral K⁺ channels that are differentially sorted; one protein (Kesv) is imported by the Tom complex into the mitochondria, the other (Kcv) to the plasma membrane. By creating chimeras we discovered that mitochondrial sorting of Kesv depends on a hierarchical combination of N- and C-terminal signals. Crucial is the length of the second transmembrane domain; extending its C terminus by >2 hydrophobic amino acids redirects Kesv …

Effect Of Short Exposure Of Naoci/Edta On E. Faecalis In Extracted Human Teeth, Rylan N. Gustafson

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this project was to study the antibacterial effect of the short exposure of 5.25% NaOCl and 17% EDTA on eight strains of Enterococcus faecalis in extracted human teeth. The crowns of 240 single rooted teeth were removed and the canals were instrumented to ISO 0.36. The canals were irrigated with 5.25% NaOCl and 17% EDTA, autoclaved and separated into eight groups of 30 teeth consisting of 20 experimental teeth, 5 positive and 5 negative controls. Each group was inoculated with one of eight different strains of E. faecalis in BHl broth and incubated for four weeks. The …

Differential Role Of Nadp+ And Nadph In The Activity And Structure Of Gdp-D-Mannose 4,6-Dehydratase From Two Chlorella Viruses, Floriana Fruscione, Laura Sturla, Garry Duncan, James L. Van Etten, Paola Valbuzzi, Antonio De Flora, Eleonopra Di Zanni, Michela Tonetti

James Van Etten Publications

GDP-D-mannose 4,6-dehydratase (GMD) is a key enzyme involved in the synthesis of 6-deoxyhexoses in prokaryotes and eukaryotes. Paramecium bursaria chlorella virus-1 (PBCV-1) encodes a functional GMD, which is unique among characterized GMDs because it also has a strong stereospecific NADPH-dependent reductase activity leading to GDP-D-rhamnose formation (Tonetti, M., Zanardi, D., Gurnon, J., Fruscione, F., Armirotti, A., Damonte, G., Sturla, L., De Flora, A., and Van Etten, J.L. (2003) J. Biol. Chem. 278, 21559–21565). In the present study we characterized a recombinant GMD encoded by another chlorella virus, Acanthocystis turfacea chlorella virus 1 (ATCV-1), demonstrating that it has the expected dehydratase …

Algae In Agricultural Fields From St. Francis County, Arkansas, T. Smith

Journal of the Arkansas Academy of Science

On August 9th, 2007, two agriculture fields (rice and sorghum fields) were sampled for freshwater algae in St. Francis County. The purpose of this study was to document the algal species in the rice and sorghum fields and compare the similarities of species composition. There were a total of 53 species identified. Overall, diatoms and cyanobacteria were equally dominant with both represented by 21 species (39.6% of the total) and 11 green algal species (20.8%) were present. The sorghum field was dominated by Chlorogloeopsis fritschii and Chroococcus limneticus, while Anabeana cylindrica was abundant in the rice field.

Dr. Maximillian Schmidt's 1884 Review Of The Growth Rates Of Asian Elephants, Robert H.I. Dale, Fred Yaniga

Scholarship and Professional Work - LAS

Rarely is there a scientific article of such creativity, insight and importance that it is fascinating even 125 years after it was written. The following article by Max Schmidt, translated in English for the first time, as far as we know, is one of these. As in the original article, an illustration precedes the text.

Dr. Schmidt addressed a topic of considerable interest in recent times (for example, Sukumar, 2003, Appendix 2): The growth curves of elephants. He combined data on multiple elephants from several sources to generate a table of shoulder heights from birth to the age of 34 …

Calf Development: Most Births At Night, Robert H.I. Dale

Scholarship and Professional Work - LAS

For many years, field researchers studying both African (Loxodonta africana) and Asian (Elephas maximas) elephants have indicated that they have observed relatively few births in situ, suggesting that most elephant dams give birth at night. For example, according to Cynthia Moss, "Possibly the majority of births occur at night and perhaps those that do take place in the daytime happen in secluded places" (1988, p. 151). Others, for example, Clive Spinage, have referred to "the old beliefs that the cows retreated to 'calving grounds' or that birth took place at night." (Spinage, 1994, p. 90). Although …

Resistance Training And Bone Mineral Density During Growth, M. Z. Smith, B. M. Goettsch, R. D. Van Ramshorst, J. A. O'Brien, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

This study examined the efficacy of two different resistance training programs in enhancing bone modeling and bone mineral density (BMD) in maturating rats. One exercise mode involved lifting a lighter weight with more repetitions (LI), while the other regimen involved lifting a heavier weight with fewer repetitions (HI) where the total volume of work between exercise programs was equivalent by design. Twenty-three male rats were randomly divided into control (Con, n = 8), LI (n = 7), and HI (n = 8) groups. The LI and HI groups were conditioned to climb a vertical ladder with weights appended to their …

Interrupted Vs. Uninterrupted Training On Bmd During Growth, B. M. Goettsch, M. Z. Smith, J. A. O'Brien, G. V. Gomez, S. V. Jaque, Ken D. Sumida

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

This study compared a resistance training program where the exercise was uninterrupted (UT, i.e., continuous repetitions) against a resistance training program where the exercise was interrupted (IT, i.e., 3 exercise sessions during a training day) for enhancing bone modeling and bone mineral density (BMD) in maturating animals. The total volume of work performed between the two resistance training programs was equivalent by design. 24 young male rats were randomly divided into Control (Con, n = 8), UT (n = 8) and IT (n=8) resistance trained groups. The UT and IT groups were conditioned to climb a vertical ladder with weights …

N-Glycan Modification In Aspergillus Species, Elke Kainz, Andreas Gallmetzer, Christian Hatzl, Juergen H. Nett, Huijuan Li, Thorsten Schinko, Robert Pachlinger, Harald Berger, Yazmid Reyes-Dominguez, Andreas Bernreiter, Tillmann Gerngross, Stefan Wildt, Joseph Strauss

Dartmouth Scholarship

The production by filamentous fungi of therapeutic glycoproteins intended for use in mammals is held back by the inherent difference in protein N-glycosylation and by the inability of the fungal cell to modify proteins with mammalian glycosylation structures. Here, we report protein N-glycan engineering in two Aspergillus species. We functionally expressed in the fungal hosts heterologous chimeric fusion proteins containing different localization peptides and catalytic domains. . This strategy allowed the isolation of a strain with a functional -1,2-mannosidase producing increased amounts of N-glycans of the Man 5 GlcNAc 2 type. This strain was further engineered by the introduction of …

Antimicrobial Activity Screening Of Recombinant And Synthetic Varasin A Defensin From The Hard Tick Dermacentor Variabilis Against Various Bacteria, Julia A. Sharp

Biological Sciences Theses & Dissertations

Hematophagous arthropods, such as ticks and mosquitoes, rely on their innate immune system for defense against pathogens ingested in a blood meal as well as those acquired through injury. In response to pathogen recognition, the production of antimicrobial peptides, such as defensin, is typically upregulated. Varisin, a defensin, is thought to be a key component in the immunocompetence of the hard tick D. variabilis against Borrelia burgdorferi, the causative agent of Lyme disease. To study the antimicrobial effects of varisin, recombinant varisin was expressed by both insect cells and E coli. Purification of the protein followed by enterokinase treatment yielded …

The Effect Of Mtad On Eight Strains Of E. Faecalis In Contaminated Root Canals, Bradley M. Newberry

Loma Linda University Electronic Theses, Dissertations & Projects

The purpose of this investigation was to determine the antimicrobial effect of Biopure MTAD (a mixture of a tetracycline isomer, an acid, and a detergent) as a final irrigant on eight strains of Enterococcus faecalis (E. faecalis) in contaminated root canals of extracted human teeth. In addition, the Minimum Inhibitory Concentration (MIC) and Minimum Lethal Concentration (MLC) of MTAD were also measured on the same strains of bacteria. Two-hundred and forty extracted human teeth were instrumented using 1.3% NaOCl followed by 17% EDTA to remove the smear layer. The teeth were randomly contaminated with one of eight strains of E. …

Structure And Function Of A Chlorella Virus Encoded Glycosyltransferase, Ying Zhang, Ye Xiang, James L. Van Etten, Michael G. Rossmann

James Van Etten Publications

Paramecium bursaria chlorella virus-1 encodes at least 5 putative glycosyltransferases that are probably involved in the synthesis of the glycan components of the viral major capsid protein. The 1.6 Å crystal structure of one of these glycosyltransferases (A64R) has a mixed α/β fold containing a central, six-stranded β-sheet flanked by α-helices. Crystal structures of A64R, complexed with UDP, CMP, or GDP, established that only UDP bound to A64R in the presence of Mn2+, consistent with its high structural similarity to glycosyltransferases which utilize UDP as the sugar carrier. The structure of the complex of A64R, UDP-glucose, and Mn2+ showed that …

Sequence And Annotation Of The 288-Kb Atcv-1 Virus That Infects An Endosymbiotic Chlorella Strain Of The Heliozoon Acanthocystis Turfacea, Lisa A. Fitzgerald, Michael V. Graves, Xiao Li, James Hartigan, Artur J.P. Pfitzner, Ella Hoffart, James L. Van Etten

James Van Etten Publications

Acanthocystis turfacea chlorella virus (ATCV-1), a prospective member of the family Phycodnaviridae, genus Chlorovirus, infects a unicellular, eukaryotic, chlorella-like green alga, Chlorella SAG 3.83, that is a symbiont in the heliozoon A. turfacea. The 288,047-bp ATCV-1 genome is the first virus to be sequenced that infects Chlorella SAG 3.83. ATCV-1 contains 329 putative protein-encoding and 11 tRNA-encoding genes. The protein-encoding genes are almost evenly distributed on both strands and intergenic space is minimal. Thirty-four percent of the viral gene products resemble entries in the public databases, including some that are unexpected for a virus. For example, these …

Chlorella Viruses Contain Genes Encoding A Complete Polyamine Biosynthetic Pathway, Sascha Baumann, Adrienne Sander, James Gurnon, Giane M. Yanai-Balser, James L. Van Etten, Markus Piotrowski

James Van Etten Publications

Two genes encoding the putative polyamine biosynthetic enzymes agmatine iminohydrolase (AIH) and N-carbamoylputrescine amidohydrolase (CPA) were cloned from the chloroviruses PBCV-1, NY-2A and MT325. They were expressed in Escherichia coli to form C-terminal (His)6-tagged proteins and the recombinant proteins were purified by Ni2+- binding affinity chromatography. The biochemical properties of the two enzymes are similar to AIH and CPA enzymes from Arabidopsis thaliana and Pseudomonas aeruginosa. Together with the previously known virus genes encoding ornithine/arginine decarboxlyase (ODC/ADC) and homospermidine synthase, the chloroviruses have genes that encode a complete set of functional enzymes that synthesize the rare polyamine homospermidine from arginine …