Microbiology Commons^™

Sara Of Staphylococcus Aureus Binds To The Sara Promoter To Regulate Gene Expression, Ambrose L. Cheung, Koren Nishina, Adhar C. Manna

Dartmouth Scholarship

The 375-bp sarA open reading frame is driven by three promoters, P1, P3, and P2. Using gel shift and DNase I footprinting assays, we found that SarA binds to two 26-bp sequences and one 31-bp sequence within the P1 and P3 promoters, respectively. Together with the results of transcription analyses, our data indicate that SarA binds to its own promoter to down-regulate sarA expression.

Oral Immunization Of Rhesus Macaques With Adenoviral Hiv Vaccines Using Enteric-Coated Capsules, George T. Mercier, Pramod N. Nehete, Marco F. Passeri, Bharti N. Nehete, Eric A. Weaver, Nancy Smyth Templeton, Kimberly Schluns, Stephanie S. Buchl, K. Buchl, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Targeted delivery of vaccine candidates to the gastrointestinal (GI) tract holds potential for mucosal immunization, particularly against mucosal pathogens like the human immunodeficiency virus (HIV). Among the different strategies for achieving targeted release in the GI tract, namely the small intestine, pH sensitive enteric coating polymers have been shown to protect solid oral dosage forms from the harsh digestive environment of the stomach and dissolve relatively rapidly in the small intestine by taking advantage of the luminal pH gradient. We developed an enteric polymethacrylate formulation for coating hydroxy-propyl-methyl-cellulose (HPMC) capsules containing lyophilized Adenoviral type 5 (Ad5) vectors expressing HIV-1 gag …

The Isolation And Characterization Of The Microbial Flora In The Alimentary Canal Of Gromphadorhina Portentosa Based On Rdna Sequences., Amy Renee Robertson

Electronic Theses and Dissertations

Multicellular organisms are not single individuals but carry a complex natural microflora with them. This complex's diversity and function can be considered a distinct ecosystem. Traditional methods of isolation and identification miss >90% of the actual diversity. This study uses the gut microflora of the Madagascar hissing roach, Gromphadorhina portentosa, as a model to examine this ecosystem. Isolated cultured bacteria were used to establish methods for identifying members of the microflora based on ribosomal RNA sequences. Universal primers for Eubacterial, Archaeal, and Eukaryotic 16s/18s rRNA were then used for PCR amplification of total DNA isolated from gut contents. Sequences …

Direct Inhibition Of Cdk9 Blocks Hiv-1 Replication Without Preventing T Cell Activation In Primary Human Peripheral Blood Lymphocytes, Dominic Salerno, Muneer G Hasham, Renée Marshall Demarest, Judit Garriga, Alexander Y Tsygankov, Xavier Graña

Rowan-Virtua School of Osteopathic Medicine Faculty Scholarship

HIV-1 transcription is essential for the virus replication cycle. HIV-1 Tat is a viral transactivator that strongly stimulates the processivity of RNA polymerase II (RNAPII) via recruitment of the cyclin T1/CDK9 positive transcription elongation factor, which phosphorylates the C-terminal domain (CTD) of RNAPII. Consistently, HIV-1 replication in transformed cells is very sensitive to direct CDK9 inhibition. Thus, CDK9 could be a potential target for anti-HIV-1 therapy. A clearer understanding of the requirements for CDK9 activity in primary human T cells is needed to assess whether the CDK9-dependent step in HIV-1 transcription can be targeted clinically. We have investigated the effects …

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 …

Fungal Conversion And Supercritical Carbon Dioxide Processing For Value-Added Canola Oil, Meidui Dong

All Dissertations

There is a growing awareness of health benefits of polyunsaturated fatty acids (PUFAs) e.g. arachidonic acid (ARA, C20:4n6) and eicosapentaenoic acid (EPA, C20:5n3). In general, plant seed oil and animal fat do not contains long-chain PUFAs with more than eighteen carbons. This study investigated the feasibility for addition of ARA and EPA to canola oil through the fermentation of fungi, Mortierella alpina, using canola flake, cake or meal as substrates. Supercritical CO2 extraction was also investigated for lipid extraction. In addition, a supercritical CO2 explosion process was characterized and applied to enhance oil extraction using supercritical CO2 and to potentially …

The Effect Of Febrile Temperature On Plasmodium Falciparum, Heidi Sue Porter

Theses and Dissertations

Previously it has been shown that cultures of Plasmodium falciparum died following exposure to a febrile temperature of 40°C, as demonstrated by a decrease in parasitemia of the following generation. In the current study, the effect of 40°C treatment on culture media, erythrocytes, and parasite glucose consumption, were ruled out as possible influences on parasite death, demonstrating that 40°C impacted the parasites directly. Metabolic profiling of DNA synthesis, protein synthesis, and glucose utilization during exposure to 40°C clearly indicated that febrile temperatures had direct effect on major metabolic pathways and parasite development, beginning 20-24 hr after erythrocyte invasion. The ring …

U.S.-Korea Cooperative Research: Carbon Monoxide As A Substrate For Microbial Maintenance, Gary M. King

University of Maine Office of Research Administration: Grant Reports

Bacteria play an important role in the global budget of carbon monoxide (CO). Largely unknown bacterial populations in soils and the water column of aquatic systems oxidize hundreds of teragrams per year, or about 10%-20% of the estimated annual flux to the atmosphere. In spite of their biogeochemical significance, relatively little is known about the identity of CO-oxidizing populations active in situ, their phylogenic and physiological diversity or the importance of CO as substrate for their basic metabolic needs. of CO oxidizers. It is clear that CO at high concentrations (> 1000 ppm) can serve as a sole source of …

Prevention Of Staphylococcus Epidermidis Biofilm Formation Using A Low-Temperature Processed Silver-Doped Phenyltriethoxysilane Sol–Gel Coating, Niall Stobie, Brendan Duffy, Declan Mccormack, John Colreavy, Martha Hidalgo, Patrick Mchale, Steven Hinder

Articles

Sol–gel coatings which elute bioactive silver ions are presented as a potential solution to the problem of biofilm formation on indwelling surfaces. There is evidence that high-temperature processing of such materials can lead to diffusion of silver away from the coating surface, reducing the amount of available silver. In this study, we report the biofilm inhibition of a Staphylococcus epidermidis biofilm using a low-temperature processed silver-doped phenyltriethoxysilane sol–gel coating. The incorporation of a silver salt into a sol–gel matrix resulted in an initial high release of silver in de-ionised water and physiological buffered saline (PBS), followed by a lower sustained …

Systemic And Mucosal Infection Program Protective Memory Cd8 T Cells In The Vaginal Mucosa., Thandi M. Onami, P. K. Suvas, H. M. Dech, J. Zeng

Microbiology Publications and Other Works

Whether mucosal immunization is required for optimal protective CD8 T cell memory at mucosal surfaces is controversial. In this study, using an adoptive transfer system, we compare the efficacy of two routes of acute lymphocytic choriomeningitis viral infection on the generation, maintenance, and localization of Ag-specific CD8 T cells in tissues, including the vaginal mucosa. Surprisingly, at day 8, i.p. infection results in higher numbers of Ag-specific CD8 T cells in the vaginal mucosa and iliac lymph node, as well as 2-3x more Ag-specific CD8 T cells that coexpress both IFN-gamma and TNF-alpha in comparison to the intranasal route of …

Systemic And Mucosal Infection Program Protective Memory Cd8 T Cells In The Vaginal Mucosa., Thandi M. Onami, P. K. Suvas, H. M. Dech, J. Zeng

Thandi M. Onami

Whether mucosal immunization is required for optimal protective CD8 T cell memory at mucosal surfaces is controversial. In this study, using an adoptive transfer system, we compare the efficacy of two routes of acute lymphocytic choriomeningitis viral infection on the generation, maintenance, and localization of Ag-specific CD8 T cells in tissues, including the vaginal mucosa. Surprisingly, at day 8, i.p. infection results in higher numbers of Ag-specific CD8 T cells in the vaginal mucosa and iliac lymph node, as well as 2-3x more Ag-specific CD8 T cells that coexpress both IFN-gamma and TNF-alpha in comparison to the intranasal route of …

Genome-Wide Transcriptional Profiling Of The Cyclic Amp-Dependent Signaling Pathway During Morphogenic Transitions Of Candida Albicans, Yong-Sun Bahn, Matthew Molenda, Janet F. Staab, Courtney A. Lyman, Laura J. Gordon, Paula Sundstrom

Dartmouth Scholarship

Candida albicans is an opportunistic human fungal pathogen that causes systemic candidiasis as well as superficial mucosal candidiasis. In response to the host environment, C. albicans transitions between yeast and hyphal forms. In particular, hyphal growth is important in facilitating adhesion and invasion of host tissues, concomitant with the expression of various hypha-specific virulence factors. In previous work, we showed that the cyclic AMP (cAMP) signaling pathway plays a crucial role in morphogenic transitions and virulence of C. albicans by studying genes encoding adenylate cyclase-associated protein (CAP1) and high-affinity phosphodiesterase (PDE2) (Y. S. Bahn, J. Staab, and P. Sundstrom, Mol. …

Jhe In Gryllus Assimilis: Cloning, Sequence-Activity Associations And Phylogeny, E. J. Crone, Anthony J. Zera, A. Anand, J. G. Oakeshott, T. D. Sutherland, R. J. Russell, Lawrence G. Harshman, Federico G. Hoffmann, C. Claudianos

Anthony Zera Publications

The 458 amino acid sequence of a mature JHE protein from the cricket Gryllus assimilis was identified after isolating the partial cDNA sequence encoding this protein from a fat body and midgut cDNA library. This hemimetabolan JHE sequence shows over 40% amino acid similarity to the known JHE sequences of several holometabolous insects. It also includes previously determined peptide sequences for G. assimilis JHE as well as two other motifs associated with JHE enzymes in holometabolous insects. The predicted molecular weight of the protein agrees with that of the JHE previously purified from G. assimilis. Partial genomic sequence encoding …

Glucose-6-Phosphate Dehydrogenase (G6pd) Screening In Pakistani Neonates: To Be Or Not To Be, Bushra Moiz

Department of Pathology and Laboratory Medicine

No abstract provided.

Longitudinal Tracking Of Cytokines After Acute Exposure To Tuberculosis: Association Of Distinct Cytokine Patterns With Protection And Disease Development, Rabia Hussain, Najeeha Talat, Firdaus Shahid, Ghaffar Dawood

Department of Pathology and Laboratory Medicine

Household contacts (HCs) of patients with tuberculosis (TB) are at higher risk of infection as well as the development of active disease. Longitudinal tracking of antigen-specific cytokines after acute exposure may significantly advance our understanding of the dynamic changes in cytokine patterns associated with disease establishment. To achieve this objective, we carried out a prospective cohort study with healthy HCs after exposure to TB. The patterns of cytokines (gamma interferon [IFN-gamma] and interleukin 10 [IL-10]) in response to mycobacterial antigens (culture filtrate [CF] proteins) and nonspecific mitogens (phytohemagglutinin [PHA] and lipopolysaccharide [LPS]) were assessed at 0, 6, 12, and 24 …

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Serg/Gorduj Gordienko

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Growth Of Thermophilic And Hyperthermophilic Fe(Iii)-Reducing Microorganisms On A Ferruginous Smectite As The Sole Electron Acceptor, Derek Lovley, Kazem Kashefi, Evgenya S. Shelobolina, W. Crawford Elliott

Derek Lovley

Recent studies have suggested that the structural Fe(III) within phyllosilicate minerals, including smectite and illite, is an important electron acceptor for Fe(III)-reducing microorganisms in sedimentary environments at moderate temperatures. The reduction of structural Fe(III) by thermophiles, however, has not previously been described. A wide range of thermophilic and hyperthermophilic Archaea and Bacteria from marine and freshwater environments that are known to reduce poorly crystalline Fe(III) oxides were tested for their ability to reduce structural (octahedrally coordinated) Fe(III) in smectite (SWa-1) as the sole electron acceptor. Two out of the 10 organisms tested, Geoglobus ahangari and Geothermobacterium ferrireducens, were not able …

Stenoxybacter Acetivorans Gen. Nov., Sp. Nov., An Acetate-Oxidizing Obligate Microaerophile Among Diverse O2-Consuming Bacteria From Termite Guts, John T. Wertz, John A. Breznak

University Faculty Publications and Creative Works

In termite hindguts, fermentative production of acetate - a major carbon and energy source for the insect - depends on efficient removal of inwardly diffusing oxygen by microbes residing on and near the hindgut wall. However, little is known about the identity of these organisms or about the substrate(s) used to support their respiratory activity. A cultivation-based approach was used to isolate O2-consuming organisms from hindguts of Reticulitermes flavipes. A consistently greater (albeit not statistically significant) number of colonies developed under hypoxia (2% [vol/vol] O 2) than under air, and the increase coincided with the appearance of morphologically distinct colonies …

Genomic Resources For Myzus Persicae: Est Sequencing, Snp Identification, And Microarray Design, John S. Ramsey, Alex C. C. Wilson, Marin De Vos, Qi Sun, Cecilia Tamborindeguy, Agnese Winfield, Gaynor Malloch, Dawn M. Smith, Brian Fenton, Stewart M. Gray, Georg Jander

Biology Articles and Papers

Backgorund

The green peach aphid, Myzus persicae (Sulzer), is a world-wide insect pest capable of infesting more than 40 plant families, including many crop species. However, despite the significant damage inflicted by M. persicae in agricultural systems through direct feeding damage and by its ability to transmit plant viruses, limited genomic information is available for this species.

Results

Sequencing of 16 M. persicae cDNA libraries generated 26,669 expressed sequence tags (ESTs). Aphids for library construction were raised on Arabidopsis thaliana, Nicotiana benthamiana, Brassica oleracea, B. napus, and Physalis floridana (with and without Potato leafroll virus infection). The …

Physiological Ecology Of Stenoxybacter Acetivorans, An Obligate Microaerophile In Termite Guts, John T. Wertz, John A. Breznak

University Faculty Publications and Creative Works

Stenoxybacter acetivorans is a newly described, obligately microaerophilic β-proteobacterium that is abundant in the acetate-rich hindgut of Reticulitermes. Here we tested the hypotheses that cells are located in the hypoxic, peripheral region of Reticulitermes flavipes hindguts and use acetate to fuel their O2-consuming respiratory activity in situ. Physical fractionation of R. flavipes guts, followed by limited-cycle PCR with S. acetivorans-specific 16S rRNA gene primers, indicated that cells of this organism were indeed located primarily among the microbiota colonizing the hindgut wall. Likewise, reverse transcriptase PCR of hindgut RNA revealed S. acetivorans-specific transcripts for acetate-activating enzymes that were also found in …

Detecting Shifts In Soil Microbial Community Structure And Function Post Landspread Of Manure Or Biosolids Containing Antimicrobial Chemicals, Kelly Lehnert, Sharon A. Clay, Susan Gibson, Volker Brozel

Agronomy, Horticulture and Plant Science Faculty Publications

Soil microbial diversity and community interaction play an indispensable role in 2,4-dichorophenoxyacetic acid (2,4-D) herbicide degradation. The addition of manure or municipal waste biosolids through landspreading may alter soil community structure and function if these materials contain antimicrobial chemicals like chlortetracycline (CTC), administered in livestock feed to promote animal growth and health or tetracycline (TET) utilized in human health. In this study, soil applied with manure collected from pigs fed standard CTC levels was compared to soil containing manure from control pigs fed no CTC, and a comparison of soil applied with biosolids containing TET or without TET to distinguish …

Design, Synthesis, And Evaluation Of 10-N-Substituted Acridones As Novel Chemosensitizers In Plasmodium Falciparum, Jane X. Kelly, Martin J. Smilkstein, Roland A. Cooper, Kristin D. Lane, Robert A. Johnson, Aaron Janowsky, Rozalia A. Dodean, Rolf Winter, Michael Riscoe

Collected Faculty and Staff Scholarship

A series of novel 10-N-substituted acridones, bearing alkyl side chains with tertiary amine groups at the terminal position, were designed, synthesized, and evaluated for the ability to enhance the potency of quinoline drugs against multidrug-resistant (MDR) Plasmodium falciparum malaria parasites. A number of acridone derivatives, with side chains bridged three or more carbon atoms apart between the ring nitrogen and terminal nitrogen, demonstrated chloroquine (CQ)-chemosensitizing activity against the MDR strain of P. falciparum (Dd2). Isobologram analysis revealed that selected candidates demonstrated significant synergy with CQ in the CQ-resistant (CQR) parasite Dd2 but only additive (or indifferent) interaction in the CQ-sensitive …

Influence Of The Hydrodynamic Environment On Quorum Sensing In Pseudomonas Aeruginosa Biofilms, Jeffrey J. Margolis, Mary Jo Kirisits, Boloroo L. Purevdorj-Gage, Benjamin Vaughan, David L. Chopp, Paul Stoodley, Matthew R. Parsek

Jeffrey Margolis

The Potential Role Of Probiotics In Reducing Poverty-Associated Infections In Developing Countries, Kingsley C. Anukam

Kingsley C Anukam

Probiotics are defined by the Food and Agricultural Organization/ World Health Organization as “live microorganisms which when administered in adequate amounts confer a health benefit on the host” [1]. The potential benefits of their use have not been adequately investigated, especially in the developing world. Japan introduced Yakult, a probiotic fermented food drink in 1935, and in the Northern hemisphere, research and use of probiotics has gained an unprecedented momentum in the last decade [2]. Use of probiotics is not uncommon in Europe [3], but in many developing countries use of probiotics in its present definition is a foreign concept. …

Characterization Of Mazfsa, An Endoribonuclease From Staphylococcus Aureus, Zhibiao Fu, Niles P. Donegan, Guido Memmi, Ambrose L. Cheung

Dartmouth Scholarship

The mazEF homologs of Staphylococcus aureus, designated mazEF(sa), have been shown to cotranscribe with the sigB operon under stress conditions. In this study, we showed that MazEF(Sa), as with their Escherichia coli counterparts, compose a toxin-antitoxin module wherein MazF(Sa) leads to rapid cell growth arrest and loss in viable CFU upon overexpression. MazF(Sa) is a novel sequence-specific endoribonuclease which cleaves mRNA to inhibit protein synthesis. Using ctpA mRNA as the model substrate both in vitro and in vivo, we demonstrated that MazF(Sa) cleaves single-strand RNA preferentially at the 5' side of the first U or 3' side of the second …

Sheeppox Virus Kelch-Like Gene Sppv-019 Affects Virus Virulence, C. A. Balinsky, Gustavo A. Delhon, C. L. Afonso, G. R. Risatti, M. V. Borca, R. A. French, E. R. Tulman, S. J. Geary, D. L. Rock

Nebraska Center for Virology: Faculty Publications

Sheeppox virus (SPPV), a member of the Capripoxvirus genus of the Poxviridae, is the etiologic agent of a significant disease of sheep in the developing world. Genomic analysis of pathogenic and vaccine capripoxviruses identified genes with potential roles in virulence and host range, including three genes with similarity to kelch-like genes of other poxviruses and eukaryotes. Here, a mutant SPPV with a deletion in the SPPV-019 kelch-like gene, ΔKLP, was derived from the pathogenic strain SPPV-SA. ΔKLP exhibited in vitro growth characteristics similar to those of SPPV-SA and revertant virus (RvKLP). ΔKLP-infected cells exhibited a reduction in Ca^{2+ …}

Impact Of Carbon Sources On Growth And Oxalate Synthesis By The Phytopathogenic Fungus Sclerotinia Sclerotiorum, Bryan J. Culbertson, Jaymie Krone, Erastus Gatebe, Norbert C. Furumo, Steven L. Daniel

Steven L. Daniel

The impact of various supplemental carbon sources (oxalate, glyoxylate, glycolate, pyruvate, formate, malate, acetate, and succinate) on growth and oxalate formation (i.e., oxalogenesis) by Sclerotinia sclerotiorum was studied. With isolates D-E7, 105, W-B10, and Arg-L of S. sclerotiorum, growth in an undefined broth medium (0.1% soytone; pH 5) with 25 mM glucose and 25 mM supplemental carbon source was increased by the addition of malate and succinate. Oxalate accumulation occurred in the presence of glucose and a supplemental carbon source, with malate, acetate, and succinate supporting the most oxalate synthesis. With S. sclerotiorum Arg-L, oxalate-to-biomass ratios, an indicator of oxalogenic …

Impact Of Carbon Sources On Growth And Oxalate Synthesis By The Phytopathogenic Fungus Sclerotinia Sclerotiorum, Bryan J. Culbertson, Jaymie Krone, Erastus Gatebe, Norbert C. Furumo, Steven L. Daniel

Faculty Research & Creative Activity

The impact of various supplemental carbon sources (oxalate, glyoxylate, glycolate, pyruvate, formate, malate, acetate, and succinate) on growth and oxalate formation (i.e., oxalogenesis) by Sclerotinia sclerotiorum was studied. With isolates D-E7, 105, W-B10, and Arg-L of S. sclerotiorum, growth in an undefined broth medium (0.1% soytone; pH 5) with 25 mM glucose and 25 mM supplemental carbon source was increased by the addition of malate and succinate. Oxalate accumulation occurred in the presence of glucose and a supplemental carbon source, with malate, acetate, and succinate supporting the most oxalate synthesis. With S. sclerotiorum Arg-L, oxalate-to-biomass ratios, an indicator of oxalogenic …

Impact Of Carbon Sources On Growth And Oxalate Synthesis By The Phytopathogenic Fungus Sclerotinia Sclerotiorum, Steven L. Daniel, Bryan J. Culbertson, Jaymie Krone, Norbert Furumo

Faculty Research & Creative Activity

The impact of various supplemental carbon sources (oxalate, glyoxylate, glycolate, pyruvate, formate, malate, acetate, and succinate) on growth and oxalate formation (i.e., oxalogenesis) by Sclerotinia sclerotiorum was studied. With isolates D-E7, 105, W-B10, and Arg-L of S. sclerotiorum, growth in an undefined broth medium (0.1% soytone; pH 5) with 25 mM glucose and 25 mM supplemental carbon source was increased by the addition of malate and succinate. Oxalate accumulation occurred in the presence of glucose and a supplemental carbon source, with malate, acetate, and succinate supporting the most oxalate synthesis. With S. sclerotiorum Arg-L, oxalate-to-biomass ratios, an indicator of oxalogenic …

Impact Of Carbon Sources On Growth And Oxalate Synthesis By The Phytopathogenic Fungus Sclerotinia Sclerotiorum, Bryan Culbertson, Jaymie Krone, Erastus Gatebe, Norbert Furumo, Steven Daniel

Faculty Research & Creative Activity

The impact of various supplemental carbon sources (oxalate, glyoxylate, glycolate, pyruvate, formate, malate, acetate, and succinate) on growth and oxalate formation (i.e., oxalogenesis) by Sclerotinia sclerotiorum was studied. With isolates D-E7, 105, W-B10, and Arg-L of S. sclerotiorum, growth in an undefined broth medium (0.1% soytone; pH 5) with 25 mM glucose and 25 mM supplemental carbon source was increased by the addition of malate and succinate. Oxalate accumulation occurred in the presence of glucose and a supplemental carbon source, with malate, acetate, and succinate supporting the most oxalate synthesis. With S. sclerotiorum Arg-L, oxalate-to-biomass ratios, an indicator of oxalogenic …