Microbiology Commons^™

Final Report For The 2015 Meg Entitled: Development Of Real-Time Polymerase Chain Reaction (Q-Pcr) Assays For The Detection And Identification Of Human Tick-Borne Pathogens, Richard Robison

Journal of Undergraduate Research

1. Develop singleplex q-PCR assays to identify the tick-borne pathogens Borrelia burgdorferi, Borrelia hermsii, Bartonella henselae, and Babesia microti. Primer generation software will be used to design primer and probe sequences that will theoretically perform optimally in PCR reactions. All sequences will be subjected to BLAST searches to confirm their specificity for a particular organism. Once primer sequences have been obtained, they will be synthesized by Integrated DNA Technologies, Inc. (IDT). These primers will be evaluated in q-PCR reactions using target DNA from multiple isolates and SyberGreen to detect nucleic acid amplification. Parameters for each reaction will be optimized using these …

Autoimmunity-Promoting B Cell Effects Of The Risk Factor Irf5, Brian Poole

Journal of Undergraduate Research

The academic objectives of this proposal were met. Using the funding provided by the MEG, we generated data that were used to publish two peer-reviewed journal articles:

The Regulation Of Snail: On The Ubiquitin Edge, Qian Yu, Binhua P. Zhou, Yadi Wu

Pharmacology and Nutritional Sciences Faculty Publications

Metastasis accounts for a majority of cancer death. One key feature during metastasis is epithelial-mesenchymal transition (EMT), which is regulated by transcription factors such as Snail and Twist. In non-malignant cells, Snail has a short half-life and is degraded via ubiquitination, but its stability is increased in cancer cell. However, the mechanism by which Snail escapes ubiquitination and degradation remains unknown. Recently, we found that Dub3 is a deubiquinase of Snail. Most importantly, we determined that Dub3 responded to extracellular signals such as IL-6, and that the resultant signaling prevented Snail degradation, and promoted cancer growth, invasion, and migration. In …

Parasite Microbiome Project: Systematic Investigation Of Microbiome Dynamics Within And Across Parasite-Host Interactions., Nolwenn M Dheilly, Daniel Bolnick, Seth Bordenstein, Paul J Brindley, Cédric Figuères, Edward C Holmes, Joaquín Martínez Martínez, Anna J Phillips, Robert Poulin, Karyna Rosario

Microbiology, Immunology, and Tropical Medicine Faculty Publications

Understanding how microbiomes affect host resistance, parasite virulence, and parasite-associated diseases requires a collaborative effort between parasitologists, microbial ecologists, virologists, and immunologists. We hereby propose the Parasite Microbiome Project to bring together researchers with complementary expertise and to study the role of microbes in host-parasite interactions. Data from the Parasite Microbiome Project will help identify the mechanisms driving microbiome variation in parasites and infected hosts and how that variation is associated with the ecology and evolution of parasites and their disease outcomes. This is a call to arms to prevent fragmented research endeavors, encourage best practices in experimental approaches, and …

Mutations In The Transmembrane Domain And Cytoplasmic Tail Of Hendra Virus Fusion Protein Disrupt Virus-Like-Particle Assembly, Nicolás P. Cifuentes-Muñoz, Weina Sun, Greeshma Ray, Phuong Tieu Schmitt, Stacy Webb, Kathleen Gibson, Rebecca Ellis Dutch, Anthony P. Schmitt

Molecular and Cellular Biochemistry Faculty Publications

Hendra virus (HeV) is a zoonotic paramyxovirus that causes deadly illness in horses and humans. An intriguing feature of HeV is the utilization of endosomal protease for activation of the viral fusion protein (F). Here we investigated how endosomal F trafficking affects HeV assembly. We found that the HeV matrix (M) and F proteins each induced particle release when they were expressed alone but that their coexpression led to coordinated assembly of virus-like particles (VLPs) that were morphologically and physically distinct from M-only or F-only VLPs. Mutations to the F protein transmembrane domain or cytoplasmic tail that disrupted endocytic trafficking …

Functional Analysis Of Microrna Pathway Genes In The Somatic Gonad And Germ Cells During Ovulation In C. Elegans, Carmela Rios

Dissertations (1934 -)

MicroRNAs (miRNAs) are post-­transcriptional regulators of gene expression that play critical roles in animal development and physiology, though functions for most miRNAs remain unknown. Worms with reduced miRNA biogenesis due to loss of Drosha or Pasha/DGCR8 activity are sterile and fail to ovulate, indicating that miRNAs are required for the process of oocyte maturation and ovulation. Starting with this penetrant sterile phenotype and using new strains created to perform tissue-­specific RNAi, we characterize the roles of the C. elegans Pasha, pash-­1, and two miRNA-­specific Argonautes, alg-­1 and alg-­2, in somatic gonad cells and in germ cells in the regulation of …

Evaluation Of 'Ckd-Epi Pakistan' Equation For Estimated Glomerular Filtration Rate (Egfr): A Comparison Of Egfr Prediction Equations In Pakistani Population, Sibtain Ahmed, Lena Jafri, Ayesha Habib Khan

Department of Pathology and Laboratory Medicine

Objective: To evaluate the results of 24-hour urinary creatinine clearance (CrCl) with estimated glomerular filtration rate (eGFR) using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI), CKD-EPI Pakistan (CKD-EPI Pak), Cockcroft Gault (CG) and 4-variable Modification of Diet in Renal Disease (MDRD) equations.
Study Design: Descriptive, cross-sectional study.
Place and Duration of Study: Section of Clinical Chemistry, Department of Pathology and Laboratory Medicine, The Aga Khan University, Karachi, from June to October 2013. Methodology: Laboratory data of subjects ≥18 years ordering 24-hour urinary CrCl from June to October 2013 was retrieved. Statistical comparison of eGFR using CKD-EPI, CKD-EPI Pak, CG and …

Geographic Strain Differentiation Of Schistosoma Japonicum In The Philippines Using Microsatellite Markers, Kharleezelle J. Moendeg

Biology Faculty Publications

Background

Microsatellites have been found to be useful in determining genetic diversities of various medically-important parasites which can be used as basis for an effective disease management and control program. In Asia and Africa, the identification of different geographical strains of Schistosoma japonicum, S. haematobium and S. mansoni as determined through microsatellites could pave the way for a better understanding of the transmission epidemiology of the parasite. Thus, the present study aims to apply microsatellite markers in analyzing the populations of S. japonicum from different endemic areas in the Philippines for possible strain differentiation.

Methodology/ Principal …

Comparing The Ecological Stoichiometry In Green And Brown Food Webs: A Review And Meta-Analysis Of Freshwater Food Webs, Michelle A. Evans-White, Halvor M. Halvorson

Faculty Publications

The framework of ecological stoichiometry was developed primarily within the context of “green” autotroph-based food webs. While stoichiometric principles also apply in “brown” detritus-based systems, these systems have been historically understudied and differ from green ones in several important aspects including carbon (C) quality and the nutrient [nitrogen (N) and phosphorus (P)] contents of food resources for consumers. In this paper, we review work over the last decade that has advanced the application of ecological stoichiometry from green to brown food webs, focusing on freshwater ecosystems. We first review three focal areas where green and brown food webs differ: (1) …

Sulfolobus Spindle-Shaped Virus 1 Growth Kinetics, Setarah Mohammad Nader

PSU McNair Scholars Online Journal

Geothermal and hypersaline environments are rich in viral particles, among which spindle-shaped morphologies predominate. Currently, viruses with spindle- or lemon-shaped virions are unique to Archaea and belong to two distinct viral families. The larger of the two families, the Fuselloviridae, encompasses spindle-shaped viruses with very short tails, which infect Sulfolobus solfataricus and close relatives. Sulfolobus spindle-shaped virus 1 (SSV1) is the best-known member of the family and was one of the first hyperthermophilic archaeal viruses to be isolated. However, our knowledge of fuselloviral life cycles and the relationships between these viruses and their hosts is still limited. As a …

Genetic Diversity Of Mycobacteriophages And The Unique Abilities Of Cluster K, Shea A. Morris

The Corinthian

Mycobacteriophage are a diverse set of viruses that infect a broad host range of mycobacterium. Genome analysis of this phage group reveals its diversity and provides evidence that they have experienced substantial horizontal gene transfer Collectively they have a large range of host bacteria they can infect, although each viral type may have a limited host range. However, this range is easily expanded and constantly changing in nature by mutation within and between virus types. These phages can lytically destroy their host mycobacteria, and it is this viral ability that have many mycobacteriophage researchers excited about the potential that phage …

Colonization Density Of The Upper Respiratory Tract As A Predictor Of Pneumonia-Haemophilus Influenzae, Moraxella Catarrhalis, Staphylococcus Aureus, And Pneumocystis Jirovecii., Daniel E Park, Henry C Baggett, Stephen R C Howie, Qiyuan Shi, Nora L Watson, W Abdullah Brooks, Perch Study Group

Epidemiology Faculty Publications

Background.

There is limited information on the association between colonization density of upper respiratory tract colonizers and pathogen-specific pneumonia. We assessed this association for Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus, and Pneumocystis jirovecii. Methods.

In 7 low- and middle-income countries, nasopharyngeal/oropharyngeal swabs from children with severe pneumonia and age-frequency matched community controls were tested using quantitative polymerase chain reaction (PCR). Differences in median colonization density were evaluated using the Wilcoxon rank-sum test. Density cutoffs were determined using receiver operating characteristic curves. Cases with a pathogen identified from lung aspirate culture or PCR, pleural fluid culture or …

The Chemotherapeutic Potential Of Resveratrol-Based Treatments, Connor Peck, Kim O'Neill

Journal of Undergraduate Research

Cancer is one of the most prevalent and deadly diseases in the world. In 2012 alone, there were approximately 14 million new cases of cancer and over 8 million cancer-related deaths worldwide. Despite extensive research in past decades, the burden of cancer continues to increase.

One of the most common treatments for cancer is chemotherapy, the use of various chemical substances that exhibit anti-cancer activity. Unfortunately, chemotherapy can be quite toxic and often produces inadequate results. There remains a great need for improved chemotherapeutic treatment strategies in order to improve outcomes for cancer patients.

The purpose of my project was …

Effect Of High Expression Of Irf5 On B Cell Activation After Antigen Stimulation, Kalarelyn Eberting, Brian Poole

Journal of Undergraduate Research

While autoimmune diseases are very prevalent in the United States, little is understood about the cause of autoimmunity, making it difficult to take preventative measures and develop treatments. Previous studies have determined that a common genetic risk factor across many autoimmune diseases leads to B cell overexpression of the transcription factor IRF5 (1,2,4). Researchers have also discovered that high IRF5 levels are strongly associated with autoantibody production, suggesting that IRF5 may play a role in actually causing autoimmunity. At this point, however, it is unclear how IRF5 promotes the production of autoantibodies.

Wolbachia-Host Interactions And The Implications To Insect Conservation And Management, Amy Michelle Truitt

Dissertations and Theses

Parasitic reproductive endosymbionts are emerging as formidable threats to insect biodiversity. Wolbachia are prevalent maternally inherited intra-cellular bacteria found in >50% of arthropod species. These symbiotic bacteria interact with their hosts in diverse ways, most often they alter host reproduction causing four conditions that all selectively favor infected females: feminization, male killing, parthenogenesis, and cytoplasmic incompatibility (CI). Furthermore, depending on strain-type and host genetic background, Wolbachia are known to affect insect behavior, expand or shift host thermal tolerance ranges, and confer anti-viral protection to their hosts. Because Wolbachia both reside in and are transmitted with host cell cytoplasm, mitochondria and …

Cellular Mechanism Of Thymidine Kinase 1 Secretion, Wei Meng, Kim O'Neill

Journal of Undergraduate Research

Cancer remains the second most common cause of death in the US, accounting for nearly 1 of every 4 deaths. Studies have shown that the early detection of cancer leads to better patient prognosis and a greater five-year survival rate. [1] Diagnostic and prognostic markers play a key role in classifying tumors and determining the best treatment plan for a patient. Among these biomarkers, Thymidine kinase 1 (TK1) has been studied extensively, primarily as a diagnostic biomarker for a variety of cancer types. TK1 plays a role in regulating the intracellular thymidine pools throughout the cell cycle, and thus regulates …

Phenome-Wide Association Study Of Y Chromosome Genetic Markers, Matthew Durrant, Mary Davis

Journal of Undergraduate Research

The purpose of this project was to analyze genetic variants on the Y chromosome for significant association with various diseases. In Dr. Mary Davis’ lab we gained access to a unique data set that allows us to analyze many single-nucleotide polymorphisms (SNPs) on the Y chromosome. This was a hypothesis-generating study that has the potential to revolutionize the currently accepted hypothesis that genetic variants on the Y chromosome do not play a significant role in common disease.

Expression Changes Of Dopamine Type 2 Receptor On The Surface Of Human Monocytes And Lymphocytes After Exposure To Dopamine In Vitro., Brandon Thomas Garcia, Sandra Hope, Scott Steffensen

Journal of Undergraduate Research

Dopamine (DA) is a neurotransmitter in the brain that plays a role in several functions such as reward, motivation, and dependence. The imbalance of DA in the central nervous system (CNS) can lead to several debilitating diseases like Parkinson’s Disease, Alzheimer’s, and Schizophrenia as well as drug dependence. Levels of DA in the brain are often determined by measuring the expression of Dopamine type 2 (D2) receptors by positron emission topography (PET) scans in the Nucleus Accumbens (NAc). This test which is often expensive and unavailable to many people without access to hospitals or regular healthcare.

Development Of Chimeric Antigen Receptor T Cells To Target Cancer, Justin Crandall, Dr. Scott Weber

Journal of Undergraduate Research

Cancer negatively affects the lives of millions of individuals, and remains a difficult ailment to treat. In 2014, approximately 585,720 deaths and 1,665,540 new cases of cancer were reported in the USA alone.1 Treatment of cancer is challenging due to cancer mutating to combat the body’s immune system. An example is shown in major histocompatibility complexes (MHCs) located on the surface of nearly every cell. These are patrolled by T cells to ensure that cells are properly functioning and healthy. With cancer however, these become downregulated allowing the cancer to grow and proliferate since T cells are unable to detect …

Development Of A Model For Adult T Cell Leukemia In Htlv-1 Infected Humanized Mice, Matthew Smith, Dr. Brad Berges

Journal of Undergraduate Research

Millions of people in the world are infected with human T-lymphotropic virus type-1 (HTLV-1). As the name states, this virus attacks the T cells of the infected organism. Though many of these individuals will remain asymptomatic throughout life, a small percentage will develop adult T cell leukemia/lymphoma (ATLL), a rare and understudied form of cancer. The latency period, or amount of time between infection and detection of cancer, is generally 20-40 years before infected patients begin developing cancer.

Creation Of Twelve Member Plasmid Library For Promoter Swapping To Control Chromosomal Gene Expression In E. Coli, Chris Nielson, William R. Mccleary

Journal of Undergraduate Research

Metabolic engineering is becoming a very important area of research, allowing researchers to harness metabolic pathways to either eliminate or synthesize desired compounds. Understanding metabolic pathways by altering expression of proteins involved in the pathway helps to uncover thermodynamic bottlenecks which render certain pathways inefficient or infeasible. Altering expression of genes through the use of promoter swapping is a useful research technique used to understand important metabolic and physiologic pathways. Engineered promoters contained in plasmids can be inserted into chromosomal DNA through the utilization of promoter swapping to alter the expression of a given gene in order to better understand …

The Role Of Dietary Highly Unsaturated N-3 Fatty Acids In Northern Quahogs, Mercenaria Mercenaria, Through Decreasing Temperature, Sixto E. Portilla

Dissertations, Theses, and Capstone Projects

The series of investigations described in this thesis were designed to assess the effect of diet on mortality through decreasing temperature in two genetic varieties of M. mercenaria important to the natural environment and the aquaculture industry: “wild” from natural stocks and “notata” selected for fast growth. Data from these novel investigations demonstrate a temperature specific sensitivity to each dietary component designating 20:5n-3 as homeoviscously favorable in the upper thermal range above 12°C, and 22:6n-3 as favorable in the lower. Statistical analyses of variance demonstrate distinct cumulative mortalities among the 5 dietary treatments (P < 0.05) for each genetic variety at each thermal range, upper (>12°C) and lower (2 = 0.9326 …

Plasmonic Gold Nanoparticle For Detection Of Fungi And Human Cutaneous Fungal Infections, Tobiloba Sojinrin, Kangze Liu, Joao Conde, Hugh Byrne, James Curtin, Daxiang Cui, Furong Tian

Articles

Fungi, which are common in the environment, can cause a multitude of diseases. Warm, humid conditions allow fungi to grow and infect humans via the respiratory, digestive and reproductive tracts, genital area and other bodily interfaces. Fungi can be detected directly by microscopy, using the potassium hydroxide (KOH) test, which is the gold standard and most popular method for fungal screening. However, this test requires trained personnel operating specialist equipment, including a fluorescent microscope and culture facilities. As most acutely infected patients seek medical attention within the first few days of symptoms, the optimal diagnostic test would be rapid and …

Thymidine Kinase 1: The Key To Predicting Cancer Recurrence, Rachel Brog, Dr. Kim L. O'Neill

Journal of Undergraduate Research

Currently it is estimated that over 230,000 women will be diagnosed with breast cancer this year in the United States alone. Although treatments are improving, an alarming number of these cases will show cancer recurrence after treatment. A method to consistently predict cancer recurrence is crucial to better treat cancer and improve patient care. Thymidine Kinase 1 (TK1) has been found in abnormally high levels in serum and tissue of cancer patients compared to healthy patients. There is question to why TK1 is present in such high quantities when normally it aids in the DNA salvage pathway during S phase …

Molecular Assisted Identification Reveals Hidden Red Algae Diversity From The Burica Peninsula, Pacific Panama, David Wilson Freshwater, Jennifer N. Idol, Seth L. Parham, Cindy Fernández-García, Noemi León, Paul W. Gabrielson, Brian Wysor

Arts & Sciences Faculty Publications

The marine flora of Panama harbors a rich diversity of green, red and brown algae, and despite chronic understudy, it is reported as the second most diverse marine flora along the Pacific Central American coast, with 174 macroalgal species. Extensive new collections and molecular assisted identification (MAI) by an international team of researchers has revealed an even greater diversity for this country. Here, the intertidal and shallow subtidal marine flora of the remote Burica Peninsula is introduced. This area is characterized by an uplifted extensive intertidal flat composed of firm, sedimentary benthos known as mudrock, on which abundant algal communities …

The 'Pseudomonas Aeruginosa' Psl Polysaccharide Is A Social But Noncheatable Trait In Biofilms, Yasuhiko Irie, Aled E. Roberts, Kasper N. Kragh, Vernita D. Gordon, Jaime B. Hutchison, Rosalind J. Allen, Gavin Melaugh, Thomas Bjarnsholt, Stuart A. West, Stephen P. Diggle

Biology Faculty Publications

Extracellular polysaccharides are compounds secreted by microorganisms into the surrounding environment, and they are important for surface attachment and maintaining structural integrity within biofilms. The social nature of many extracellular polysaccharides remains unclear, and it has been suggested that they could function as either cooperative public goods or as traits that provide a competitive advantage. Here, we empirically tested the cooperative nature of the PSL polysaccharide, which is crucial for the formation of biofilms in Pseudomonas aeruginosa. We show that (i) PSL is not metabolically costly to produce; (ii) PSL provides populationlevel benefits in biofilms, for both growth and antibiotic …

Serious Fungal Infections In Pakistan, Kauser Jabeen, Joveria Farooqi, Sajjad Mirza, D Denning, Afia Zafar

Department of Pathology and Laboratory Medicine

The true burden of fungal infection in Pakistan is unknown. High-risk populations for fungal infections [tuberculosis (TB), diabetes, chronic respiratory diseases, asthma, cancer, transplant and human immunodeficiency virus (HIV) infection] are numerous. Here, we estimate the burden of fungal infections to highlight their public health significance. Whole and at-risk population estimates were obtained from the WHO (TB), BREATHE study (COPD), UNAIDS (HIV), GLOBOCAN (cancer) and Heartfile (diabetes). Published data from Pakistan reporting fungal infections rates in general and specific populations were reviewed and used when applicable. Estimates were made for the whole population or specific populations at risk, as previously …

Pas Signaling Mechanisms In Aer And Aer2, Darysbel Garcia

Loma Linda University Electronic Theses, Dissertations & Projects

PAS domains are widespread signal sensors that share a conserved three-dimensional αβ fold that consists of a central β-sheet flanked by several α- helices. The aerotaxis receptor Aer from Escherichia coli and the Aer2 chemoreceptor from Pseudomonas aeruginosa both contain PAS domains. Aer senses oxygen (O2) indirectly via an FAD cofactor bound to its PAS domain, while Aer2 directly binds O2 to its PAS b-type heme cofactor. The Aer and Aer2 PAS domains both interact with a signal transduction domain known as a HAMP domain. The PAS-HAMP arrangement differs between Aer and Aer2, with Aer- PAS residing adjacent to its …

Genome Sequence Variation In The Constricta Strain Dramatically Alters The Protein Interaction And Localization Map Of Potato Yellow Dwarf Virus, Chanyong Jang, Renyuan Wang, Joseph Wells, Fabian Leon, Mark Farman, John Hammond, Michael M. Goodin

Plant Pathology Faculty Publications

The genome sequence of the constricta strain of Potato yellow dwarf virus (CYDV) was determined to be 12 792 nt long and organized into seven ORFs with the gene order 3′-N-X-P-Y-M-G-L-5′, which encodes the nucleocapsid, phospho, movement, matrix, glyco, and RNA-dependent RNA polymerase proteins, respectively, except for X, which is of unknown function. Cloned ORFs for each gene, except L, were used to construct a protein interaction and localization map (PILM) for this virus, which shares greater than 80 % amino acid similarity in all ORFs except X and P with the sanguinolenta strain of this species (SYDV). Protein localization …

The Effects Of Nucleosome Positioning And Chromatin Architecture On Transgene Expression, Colton E. Kempton

Theses and Dissertations

Eukaryotes use proteins to carefully package and compact their genomes to fit into the nuclei of their individual cells. Nucleosomes are the primary level of compaction. Nucleosomes are formed when DNA wraps around an octamer of histone proteins and a nucleosome's position can limit access to genetic regulatory elements. Therefore, nucleosomes represent a basic level of gene regulation. DNA and its associated proteins, called chromatin, is usually classified as euchromatin or heterochromatin. Euchromatin is transcriptionally active with loosely packed nucleosomes while heterochromatin is condensed with tightly packed nucleosomes and is transcriptionally silent. In order to become active, heterochromatin must first …