Life Sciences Commons^™

Characterization Of The Reconstituted And Native Pseudomonas Aeruginosa Type Iii Secretion System Translocon, Kathryn R. Monopoli

Masters Theses

The Type III Secretion (T3S) system is a system utilized by many pathogenic bacteria to inject proteins into host cells during an infection. Effector proteins enter the host cell by passing through the proteinaceous T3S translocon, which forms a pore on the host cell membrane. Pseudomonas aeruginosa is an opportunistic pathogen that utilizes the T3S system, and very little is known about how the P. aeruginosa translocon forms.

The proteins PopB and PopD are believed to assemble into the P. aeruginosa translocon. A pore-forming heterocomplex of PopB and PopD has been reconstituted in model membranes, however this heterocomplex has not …

Promoting Extracellular Matrix Crosslinking In Synthetic Hydrogels, Marcos M. Manganare

Masters Theses

The extracellular matrix (ECM) provides mechanical and biochemical support to tissues and cells. It is crucial for cell attachment, differentiation, and migration, as well as for ailment-associated processes such as angiogenesis, metastases and cancer development. An approach to study these phenomena is through emulation of the ECM by synthetic gels constructed of natural polymers, such as collagen and fibronectin, or simple but tunable materials such as poly(ethylene glycol) (PEG) crosslinked with short peptide sequences susceptible to digestion by metalloproteases and cell-binding domains. Our lab uses PEG gels to study cell behavior in three dimensions (3D). Although this system fosters cell …

Energetic Limitations Of Thermophilic Methanogens And Thiosulfate Reducers In The Subsurface Biosphere At Deep-Sea Hydrothermal Vents, Lucy C. Stewart

Doctoral Dissertations

This dissertation examined the substrate and energetic limitations of hydrogenotrophic thermophiles from deep-sea hydrothermal vents. Thermophilic and hyperthermophilic organisms in diffuse hydrothermal venting are thought to represent a hot subsurface biosphere associated with deep-sea hydrothermal vents, where primary production is dominated by hydrogenotrophy rather than sulfide oxidation as at the vent/seawater interface of hydrothermal sulfide chimneys. Methanogens and sulfur-reducers are known to compete for hydrogen in mesophilic, freshwater systems, and likely do so in deep-sea hydrothermal vent environments as well. However, the exact size and biomass of the subsurface biosphere is difficult to determine through direct sampling. Firstly, the distribution …

Engineering Novel Detection And Treatment Strategies For Bacterial Therapy Of Cancer, Jan T. Panteli

Doctoral Dissertations

Finding and treating cancer is difficult due to limited sensitivity and specificity of current detection and treatment strategies. Many chemotherapeutic drugs are small molecules that are limited by diffusion, making it difficult to reach cancer sites requiring high doses that lead to systemic toxicity and off-target effects. Tomographic detection techniques, like PET, MRI and CT, are good at identifying macroscopic lesions in the body but are limited in their ability to detect microscopic lesions. Biomarker detection strategies are extremely sensitive and able to identify ng/ml concentrations of protein, but are poor at discriminating between healthy and disease state levels due …

Dissolution, Ocean Acidification And Biotic Extinctions Prior To The Cretaceous/Paleogene (K/Pg) Boundary In The Tropical Pacific, Serena Dameron

Masters Theses

The several million years preceding the Cretaceous/Paleogene (K/Pg) boundary has been the focus of many studies. Changes in ocean circulation and sea level, extinctions, and major volcanic events have all been documented for this interval. Important research questions these changes raise include the climate dynamics during the warm, but not hot, time after the decay of the Late Cretaceous greenhouse interval and the stability of ecosystems prior to the mass extinctions at the end-Cretaceous.

I document several biotic perturbations as well as changes in ocean circulation during the Maastrichtian stage of the latest Cretaceous that question whether the biosphere was …

Eneterotoxigenic Bacillus Cereus And Bacillus Thuringiensis Spores In U.S. Retail Spices, Upasana Hariram

Masters Theses

Bacillus cereus is a ubiquitous organism and a potential foodborne pathogen that can cause two types of gastrointestinal diseases: emesis and diarrhea. The emetic syndrome is caused by a heat and acid stable peptide toxin that is pre-formed in food, while the diarrheal syndrome is associated to two 3-protein, heat labile enterotoxin complexes that are formed in the intestine after ingestion of the organism. There are many reports on the isolation and characterization of Bacillus cereus from various foods, however there are no studies on the levels, toxigenicity and physical characteristics of B. cereus isolated from U.S. retail spices. …

Two Regulatory Aspects Of Ino1 Transcription In Yeast, Tschen-Wei Chang

Masters Theses

This study focuses on understanding the mechanisms of expression control of a phospholipid biosynthetic gene, INO1. This study also includes investigation into transcriptional regulation of SNA3, a gene in tandem upstream of INO1.

INO1 expression is a prevailing model for transcription studies. INO1 is repressed under growth conditions with inositol and derepressed by two transcription activators, Ino2 and Ino4, when inositol is absent. Coordination of the centromeric binding factor, Cbf1, with Ino2 and Ino4 is required for efficient derepression of INO1. Transcription of the INO1 adjacent SNA3 gene is also influenced by inositol. INO1 and SNA3 are …

Mechanisms For Extracellular Electron Exchange By Geobacter Species, Jessica A. Smith

Doctoral Dissertations

Understanding the mechanisms for microbial extracellular electron exchange are of interest because these processes play an important role in the biogeochemical cycles of both modern and ancient environments, development of bioenergy strategies, as well as for bioremediation applications. Only a handful of microorganisms are capable of extracellular electron exchange, one of the most thoroughly studied being the Geobacter species. Geobacter species are often the predominant Fe(III) reducing microorganisms in many soils and sediments, can exchange electrons directly via interspecies electron transfer, and can both donate or accept electrons with a wide variety of extracellular substrates including the electrode of a …

Two Decades Of Warming Increases Diversity Of A Potentially Lignolytic Bacterial Community, Grace Pold, Jerry M. Melillo, Kristen Deangelis

Microbiology Department Faculty Publication Series

As Earth's climate warms, the massive stores of carbon found in soil are predicted to become depleted, and leave behind a smaller carbon pool that is less accessible to microbes. At a long-term forest soil-warming experiment in central Massachusetts, soil respiration and bacterial diversity have increased, while fungal biomass and microbially-accessible soil carbon have decreased. Here, we evaluate how warming has affected the microbial community's capability to degrade chemically-complex soil carbon using lignin-amended BioSep beads. We profiled the bacterial and fungal communities using PCR-based methods and completed extracellular enzyme assays as a proxy for potential community function. We found that …

Genome Sequence And Description Of The Anaerobic Lignin-Degrading Bacterium Tolumonas Lignolytica Sp. Nov., Andrew F. Billings, Julian L. Fortney, Terry C. Hazen, Blake Simmons, Karen W. Davenport, Lynne Goodwin, Natalia Ivanova, Nikos C. Kyrpides, Konstantinos Mavromatis, Tanya Woyke, Kristen Deangelis

Microbiology Department Faculty Publication Series

Tolumonas lignolytica BRL6-1T sp. nov. is the type strain of T. lignolytica sp. nov., a proposed novel species of the Tolumonas genus. This strain was isolated from tropical rainforest soils based on its ability to utilize lignin as a sole carbon source. Cells of Tolumonas lignolytica BRL6-1T are mesophilic, non-spore forming, Gram-negative rods that are oxidase and catalase negative. The genome for this isolate was sequenced and returned in seven unique contigs totaling 3.6Mbp, enabling the characterization of several putative pathways for lignin breakdown. Particularly, we found an extracellular peroxidase involved in lignin depolymerization, as well as several enzymes involved …

Long-Term Forest Soil Warming Alters Microbial Communities In Temperate Forest Soils, Kristen Deangelis, Grace Pold, Begüm D. Topçuoğlu, Linda T.A. Van Diepen, Rebecca M. Varney, Jeffrey L. Blanchard, Jerry Melillo, Serita D. Frey

Microbiology Department Faculty Publication Series

Soil microbes are major drivers of soil carbon cycling, yet we lack an understanding of how climate warming will affect microbial communities. Three ongoing field studies at the Harvard Forest Long-term Ecological Research (LTER) site (Petersham, MA) have warmed soils 5°C above ambient temperatures for 5, 8, and 20 years. We used this chronosequence to test the hypothesis that soil microbial communities have changed in response to chronic warming. Bacterial community composition was studied using Illumina sequencing of the 16S ribosomal RNA gene, and bacterial and fungal abundance were assessed using quantitative PCR. Only the 20-year warmed site exhibited significant …