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Microbiology Commons

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2017

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Articles 331 - 360 of 411

Full-Text Articles in Microbiology

Characterization Of Inhibitory Molecules Produced By Lactobacillus Crispatus, Giuseppe Anthony Pistone Jan 2017

Characterization Of Inhibitory Molecules Produced By Lactobacillus Crispatus, Giuseppe Anthony Pistone

Master's Theses

Lactobacillus species are widely accepted as beneficial bacteria of the human microbiota1-8. Lactobacilli spp. are well documented to inhibit pathogens by production and secretion of hydrogen peroxide (H2O2), organic acids, and/or proteinaceous bacteriocins into their environment. Most research attributes bactericidal activity of cell free supernatant (CFS) to H2O2 and/or lactic acid. Here, I demonstrate that CFS from a clinical isolate of Lactobacillus crispatus contains a molecule(s) that inhibits uropathogenic Escherichia coli (UPEC) colony formation independently of H2O2 and organic acids. Physiologic concentrations of H2O2 and organic acids produced by L. crispatus do not inhibit UPEC colony formation. Incubation of UPEC …


The Role Of The Dosage Compensation Complex As A Pathway For Spiroplasma To Induce Male Lethality In Drosophila Melanogaster, Becky Cheng Jan 2017

The Role Of The Dosage Compensation Complex As A Pathway For Spiroplasma To Induce Male Lethality In Drosophila Melanogaster, Becky Cheng

CMC Senior Theses

Drosophila melanogaster and many other insects harbor intracellular bacterial symbionts that are transmitted vertically from infected host mothers to their offspring. Many of these bacteria alter host reproductive developmental processes in order to increase their transmission success. For example, Spiroplasma, a spirochete that naturally infects D. melanogaster, selectively kills males during mid-embryogenesis while sparing females. Previous studies suggested that Spiroplasma interacts genetically with the male-specific dosage compensation pathway, which causes ~2-fold up-regulation of most genes located on the male’s single X chromosome so that their expression matches the levels found in females who have two Xs. To further …


Tissue Specific Microenvironments: A Key Tool For Tissue Engineering And Regenerative Medicine, Patrick C. Sachs, Peter A. Mollica, Robert D. Bruno Jan 2017

Tissue Specific Microenvironments: A Key Tool For Tissue Engineering And Regenerative Medicine, Patrick C. Sachs, Peter A. Mollica, Robert D. Bruno

School of Medical Diagnostics & Translational Sciences Publications

The accumulated evidence points to the microenvironment as the primary mediator of cellular fate determination. Comprised of parenchymal cells, stromal cells, structural extracellular matrix proteins, and signaling molecules, the microenvironment is a complex and synergistic edifice that varies tissue to tissue. Furthermore, it has become increasingly clear that the microenvironment plays crucial roles in the establishment and progression of diseases such as cardiovascular disease, neurodegeneration, cancer, and ageing. Here we review the historical perspectives on the microenvironment, and how it has directed current explorations in tissue engineering. By thoroughly understanding the role of the microenvironment, we can begin to correctly …


Expression Of Matrix Metalloproteinases In Naegleria Fowleri And Their Role In Degradation Of The Extracellular Matrix, Charlton Lam Jan 2017

Expression Of Matrix Metalloproteinases In Naegleria Fowleri And Their Role In Degradation Of The Extracellular Matrix, Charlton Lam

Theses and Dissertations

Naegleria fowleri is a free-living amoeba found in freshwater lakes and ponds that is the causative agent of Primary Amoebic Meningoencephalitis (PAM). Matrix metalloproteinases (MMPs) have been described in protozoa, such as Plasmodium falciparum, Trypanosoma brucei, and Balamuthia mandrillaris, and have been linked to their increased motility and invasive capability by degrading components of the extracellular matrix (ECM). In addition, MMPs are often upregulated in tumorigenic cells and have been attributed as responsible for the metastasis of certain cancers. In the present study, in vitro experiments indicated that MMPs are linked functionally to the ECM degradation process. …


Profiling Resistance To P450-Activated Food Carcinogens Using Toxicogenomic Approaches In Budding Yeast, Nicholas Stjohn Jan 2017

Profiling Resistance To P450-Activated Food Carcinogens Using Toxicogenomic Approaches In Budding Yeast, Nicholas Stjohn

Legacy Theses & Dissertations (2009 - 2024)

The human response to environmental carcinogens, some of which require metabolic activation, is highly variable. Factors such as environment, lifestyle, and genetics all influence the rates of exposure to and ultimate bioactivation of these compounds. Genetic factors include mutations to cell-cycle regulation, cell proliferation, and DNA repair genes; however, epidemiological studies may lack significance due to inadequate patient numbers. We used budding yeast as a model organism to determine genetic susceptibility to food-associated carcinogens, including aflatoxin (AFB1) and heterocyclic aromatic amines (HAAs). Budding yeast does not contain P450s that activate these compounds, so expression vectors were induced that contain human …


Novel Approaches To Mitigating Bacterial Biofilm Formation And Intercellular Communication, Stephen Kasper Jan 2017

Novel Approaches To Mitigating Bacterial Biofilm Formation And Intercellular Communication, Stephen Kasper

Legacy Theses & Dissertations (2009 - 2024)

Long thought of as solitary single-cell organisms, it is now widely accepted that bacteria can act and cooperate as social organisms. Phenomena such as biofilm formation and quorum sensing (QS) are two intimately intertwined cooperative behaviors that significantly contribute to the pathogenesis of many bacteria. Biofilms are surface associated communities of bacteria encased in a secreted extracellular matrix, which provides several advantages over an individualized lifestyle, such as increased protection from antimicrobial agents as well as enhanced opportunity for the exchange of genetic material. Bacterial QS is a system of population-based communication through the production, sensing, and response to chemical …


Structural And Functional Characterization Of An Unusual Camp Responsive Transcription Factor, Cmr, In Tb Complex Mycobacteria, Sridevi Ranganathan Jan 2017

Structural And Functional Characterization Of An Unusual Camp Responsive Transcription Factor, Cmr, In Tb Complex Mycobacteria, Sridevi Ranganathan

Legacy Theses & Dissertations (2009 - 2024)

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis in humans, is an intracellular pathogen that infects millions of people every year. Mtb can survive inside the host for extended periods of time by sensing and adapting to the host environmental stressors. Transcriptional gene regulation plays a critical role in this adaptation. This dissertation focuses on understanding the structural and functional aspects of one such transcriptional regulatory unit, Cmr (Rv1675c), in Mtb.


Investigating The Role Of Epitranscriptomics In Viral Immunity, Kaycie S. Lawson Jan 2017

Investigating The Role Of Epitranscriptomics In Viral Immunity, Kaycie S. Lawson

Legacy Theses & Dissertations (2009 - 2024)

The characterization of host-virus relationships is essential for understanding the process of viral replication and driving the development of effective viral targeting strategies. In this work, I tested the hypothesis that ribonucleic acid (RNA) post-transcriptional modifications (PTMs) may play a significant role in defining host-virus interactions. The model system consisted of a yeast strain capable of harboring the endogenous L-A virus, which allowed me to evaluate the possible involvement of PTMs in the process of cellular immunity to virus-borne toxicity. A mass spectrometry-based platform was utilized to compare the global PTM profile of wild-type yeast with those of the same …


A Tale Of Two Regulators : Characterization Of The Novel Transcription Factor Abmr And The Small Non-Coding Rna Mcr11 In Mycobacterium Tuberculosis, Roxanne Candice Girardin Jan 2017

A Tale Of Two Regulators : Characterization Of The Novel Transcription Factor Abmr And The Small Non-Coding Rna Mcr11 In Mycobacterium Tuberculosis, Roxanne Candice Girardin

Legacy Theses & Dissertations (2009 - 2024)

Genes of unknown function make up nearly one third of Mtb’s genome (Cole,


Antifungal Genome Mining And Genetics In Filamentous Actinomycete Bacteria Isolated From Local Soils, Jacob Alex, Kevin Lou, Bilal Makhdoom Jan 2017

Antifungal Genome Mining And Genetics In Filamentous Actinomycete Bacteria Isolated From Local Soils, Jacob Alex, Kevin Lou, Bilal Makhdoom

Undergraduate Research Symposium Posters

Actinomycetes are gram positive, filamentous bacteria that produce useful antibiotics, antitumor agents, and agricultural products. A series of enrichments were undertaken to isolate actinomycetes from local soils, varying enrichment media, antibacterials, and soil treatments (including heat and CaCO3). Isolates were characterized by 16S rDNA sequencing, phenotypic and morphological observations, and antibiotic production. The genetic tractability of select isolates was analyzed using a panel of integrating vectors derived from ɸC31, ɸBT1, and OzzyJ phage using intergeneric conjugation. Further, a semi-degenerate multiplex PCR assay to detect ɸBT1 genomic integrants was designed and tested for the first time. Finally, PCR screens were used …


Development And Characterization Of Histidine-Tagged Hpv16 L2 And Ms2-Arginine-Tagged Recombinant Proteins For Downstream Processes, Tahiyat Alothaim Jan 2017

Development And Characterization Of Histidine-Tagged Hpv16 L2 And Ms2-Arginine-Tagged Recombinant Proteins For Downstream Processes, Tahiyat Alothaim

Dissertations, Master's Theses and Master's Reports

Human papillomaviruses (HPVs) are the most common sexually transmitted infections. Persistent infection with HPV can lead to anogenital cancers including head and neck cancers. Three prophylactic vaccines have been approved to prevent against some types of HPV infection. However, the vaccines are HPV-type specific and protect mostly against the HPV types included in the vaccines. To offer broader protection against more HPV types, studies in the field are developing candidate vaccines targeting a conserved minor capsid protein, L2. Nevertheless, reagents for developing and assessing L2 vaccines are limited. For example, antibodies to assess the antigenicity of some L2 epitopes are …


Characterization Of A New Chlorovirus Type With Permissive And Non-Permissive Features On Phylogenetically Related Algal Strains, Cristian F. Quispe, Ahmed Esmael, Olivia Sonderman, Michelle Mcquinn, Irina Agarkova, Mohammed Battah, Garry A. Duncan, David D. Dunigan, Timothy P. L. Smith, Cristina Decastro, Immacolata Speciale, Fangrui Ma, James L. Van Etten Jan 2017

Characterization Of A New Chlorovirus Type With Permissive And Non-Permissive Features On Phylogenetically Related Algal Strains, Cristian F. Quispe, Ahmed Esmael, Olivia Sonderman, Michelle Mcquinn, Irina Agarkova, Mohammed Battah, Garry A. Duncan, David D. Dunigan, Timothy P. L. Smith, Cristina Decastro, Immacolata Speciale, Fangrui Ma, James L. Van Etten

Nebraska Center for Virology: Faculty Publications

A previous report indicated that prototype chlorovirus PBCV-1 replicated in two Chlorella variabilis algal strains, NC64A and Syngen 2-3, that are ex-endosymbionts isolated from the protozoan Paramecium bursaria. Surprisingly, plaque-forming viruses on Syngen 2-3 lawns were often higher than on NC64A lawns from indigenous water samples. These differences led to the discovery of viruses that exclusively replicate in Syngen 2-3 cells, named Only Syngen (OSy) viruses. OSy-NE5, the prototype virus for the proposed new species, had a linear dsDNA genome of 327 kb with 44-nucleotide-long, incompletely base-paired, covalently closed hairpin ends. Each hairpin structure was followed by an identical …


การยับยั้งการเจริญและการผลิตอะฟลาทอกซินของ Aspergillus Flavus ในไซเลจข้าวโพดโดยใช้ยีสต์และแล็กติกแอซิดแบคทีเรียปฏิปักษ์, ทรรศภรณ์ รุ่งชัยวัฒนกุล Jan 2017

การยับยั้งการเจริญและการผลิตอะฟลาทอกซินของ Aspergillus Flavus ในไซเลจข้าวโพดโดยใช้ยีสต์และแล็กติกแอซิดแบคทีเรียปฏิปักษ์, ทรรศภรณ์ รุ่งชัยวัฒนกุล

Chulalongkorn University Theses and Dissertations (Chula ETD)

วัตถุประสงค์ของงานวิจัยนี้เพื่อศึกษาความสามารถของยีสต์และแล็กติกแอซิดแบคทีเรียปฏิปักษ์ในการยับยั้งการเจริญและการผลิตอะฟลาทอกซินรวมของ Aspergillus flavus บนจานอาหารเลี้ยงเชื้อและในไซเลจข้าวโพด และเพื่อหาปริมาณความเข้มข้นและอัตราส่วนของแล็กติกแอซิดแบคทีเรียและยีสต์ผสมที่ให้ผลการยับยั้งสูงสุดในไซเลจข้าวโพด โดยเริ่มจากการทดสอบประสิทธิภาพของยีสต์ (Wickerhamomyces anomalus MSCU 0652 และ Kluyveromyces marxianus MSCU 0655) และแล็กติกแอซิดแบคทีเรีย (Lactobacillus casei AN2 และ Lactobacillus paracasei AN3) ในการยับยั้งการเจริญและการผลิตอะฟลาทอกซินของ A. flavus บนอาหารเลี้ยงเชื้อ พบว่า จุลินทรีย์ปฏิปักษ์ที่ใช้ทดสอบทั้งหมดให้ผลการยับยั้งการเจริญของราค่อนข้างต่ำ (10-20 เปอร์เซ็นต์) อย่างไรก็ตาม ยีสต์ผสมระหว่าง W. anomalus MSCU 0652 และ K. marxianus MSCU 0655 และ L. paracasei AN3 แบบเดี่ยวมีประสิทธิภาพในการลดการผลิตอะฟลาทอกซินรวมสูงสุด (43.25 และ 35.64 เปอร์เซ็นต์ ตามลำดับ) ดังนั้น จึงนำยีสต์ผสมและ L. paracasei AN3 มาทดสอบความสามารถในการยับยั้งการเจริญและการผลิตอะฟลาทอกซินรวมของ A. flavus ในไซเลจข้าวโพดปลอดเชื้อและไซเลจข้าวโพดร่วมกับจุลินทรีย์ประจำถิ่น พบว่า การใช้ L. paracasei AN3 ร่วมกับยีสต์ผสมระหว่าง W. anomalus MSCU 0652 และ K. marxianus MSCU 0655 สามารถยับยั้งการเจริญและลดปริมาณอะฟลาทอกซินรวมของราได้ดีทั้งในไซเลจข้าวโพดปลอดเชื้อและไซเลจข้าวโพดที่หมักร่วมกับจุลินทรีย์ประจำถิ่น จากนั้น เมื่อทดสอบความเข้มข้นและอัตราส่วนของ L. paracasei AN3 ร่วมกับยีสต์ผสมระหว่าง W. anomalus MSCU 0652 และ K. marxianus MSCU 0655 ที่เหมาะสม พบว่า การเติม L. paracasei AN3 ที่ความเข้มข้นมากกว่ายีสต์ผสม (10⁸:10⁶) …


โมโนโคลนอลแอนติบอดีต่อโปรตีน Vp1 ของไวรัสโลหิตจางในไก่ และการระบุเอพิโทป, เอกราช สิทธิเดช Jan 2017

โมโนโคลนอลแอนติบอดีต่อโปรตีน Vp1 ของไวรัสโลหิตจางในไก่ และการระบุเอพิโทป, เอกราช สิทธิเดช

Chulalongkorn University Theses and Dissertations (Chula ETD)

โรคโลหิตจางในไก่ เกิดจากไวรัส Chicken anemia virus (CAV) ไก่ที่ติดไวรัส จะมีภาวะโลหิตจางอย่างรุนแรงตัวแคระแกร็น ต่อมธัยมัสฝ่อ เซลล์ไขกระดูกลดลง และภูมิคุ้มกันบกพร่อง โปรตีน VP1 เป็นโปรตีนโครงสร้างของไวรัส ทำหน้าที่ในการรวมกันเป็นอนุภาคไวรัส มีคุณสมบัติในการเป็นแอนติเจน CAV งานวิจัยก่อนหน้านี้ สามารถผลิตรีคอมบิแนนท์โปรตีน Δ60N_VP1 ของไวรัสโลหิตจางในไก่ จึงเป็นที่มาของวัตถุประสงค์ของงานวิจัยนี้ คือ ผลิตโมโนโคลนอลแอนติบอดีต่อโปรตีน VP1 และ ระบุเอพิโทปต่อโมโนโคลนอลแอนติบอดี โดยนำม้ามที่ได้จากการใช้รีคอมบิแนนท์โปรตีน Δ60N_VP1 กระตุ้นภูมิคุ้มกันในหนูเม้าส์ มาหลอมรวมกับเซลล์ไมอีโลมา พบว่าได้ เซลล์ไฮบริโดมาที่ผลิตโมโนโคลนอลแอนติบอดีจำนวน 5 โคลน ได้แก่ โคลนที่ 1, 2, 3, 7 และ 21 สามารถจำแนกชนิดของแอนติบอดีได้เป็น IgG2a, IgM, IgG2b, IgM และ IgG1 ตามลำดับ การระบุเอพิโทป ทำโดยแบ่งชิ้นส่วนสารพันธุกรรม Δ60N_VP1 เป็น 6 ส่วน ได้แก่ Fragment 1(F1), F2, F3, F4 และ Separating 1(S1) และ S2 ซึ่งแยกมาจากชิ้น F1 จากนั้นเพิ่มปริมาณสารพันธุกรรมด้วยไพรเมอร์จำเพาะ และทำการย้ายชิ้นส่วนแต่ละชิ้นเข้าสู่เวกเตอร์ pET28a นำไปชักนำให้เกิดการแสดงออกใน E. coli Rosetta-gami รีคอมบิแนนท์โปรตีนแต่ละชนิดจะถูกใช้เป็นแอนติเจนเพื่อใช้ระบุเอพิโทปต่อโมโนโคลนอลแอนติบอดี ด้วยวิธี western blotting analysis ผลการทดลองพบโมโนโคลนอลแอนติบอดี ทั้ง 5 โคลน มีความจำเพาะต่อแอนติเจน Δ60N_VP1 และ ไม่เกิดปฏิกิริยาข้ามกับรีคอมบิแนนท์โปรตีนที่มี His-Tag ชนิดอื่น ในการศึกษานี้ใช้ โมโนโคลนอลแอนติบอดีชนิด IgG เท่านั้น ที่จะถูกนำไประบุเอพิโทป ผลการทดลองพบว่าโมโนโคลนอลแอนติบอดีโคลนที่ 1 ทำปฏิกริยาจำเพาะกับโปรตีน …


Biochemical, Structural, And Drug Design Studies Of Norovirus And Zika Virus Proteases, Ben Kuiper Jan 2017

Biochemical, Structural, And Drug Design Studies Of Norovirus And Zika Virus Proteases, Ben Kuiper

Wayne State University Dissertations

Noroviruses, which are the leading cause of acute gastroenteritis, cause an estimated 677 million infections and 213,000 deaths each year worldwide. Noroviruses are classified into seven genogroups (GI-GVII); GI, GII, and GIV have been shown to be infectious in humans. However, GII noroviruses cause the majority of outbreaks (89%). No pharmacologic treatment or vaccine currently exists to treat or prevent norovirus infections.

Recently, the development of a norovirus replicon system, a murine model of norovirus infection, and the development of a biochemical protease assay have allowed for the design and development of norovirus inhibitors. However, the replicon and biochemical assay …


Investigation Into Roseburia Intesinalis For Optimal Growth And Survivability Conditions For Potential Probiotic Use, Lisa Rana Felczak Jan 2017

Investigation Into Roseburia Intesinalis For Optimal Growth And Survivability Conditions For Potential Probiotic Use, Lisa Rana Felczak

Wayne State University Theses

Obesity and diabetes is an epidemic of increasing concern. Certain novel bacteria have been found to be an aid in reversing some of the mechanisms associated. This study focused on investigating the optimal stability conditions for R.intestinalis that can associate with an increase in growth. First, environmental conditions were tested to find the optimal conditions associated with each. Once growth conditions were found, generation time was calculated and various prebiotics were used to investigate a potential for additional growth. Next, common antibiotics were tested to see if the R.intestinalis could still show growth when exposed. Large batches of R.intestinalis were …


Human Cytomegalovirus Cytoplasmic Virion Assembly Complex: Structure In Vivo And Role Of Pul103 In Its Biogenesis, Ma Christina Raye Lim Jan 2017

Human Cytomegalovirus Cytoplasmic Virion Assembly Complex: Structure In Vivo And Role Of Pul103 In Its Biogenesis, Ma Christina Raye Lim

Wayne State University Theses

Human Cytomegalovirus (HCMV) is a linear, double stranded DNA virus that causes severe disease in the immunocompromised, and is one of the common cause of congenital disease. Antivirals that are currently available for treatment target the DNA replication of the virus cycle, and are highly toxic. Finding new drug targets, such as proteins responsible for virion assembly and egress, would help to alleviate the disease burden.

HCMV remodels the host cell to form a structure called the cytoplasmic virion assembly complex (cVAC), a site of virion maturation and egress. The first objective of this work is to study the structure …


The Potential Stimulation Of C. Minuta Growth By Various Prebiotics, And Its Sensitivity To Various Antibiotics, Maria Elena Hakim Jan 2017

The Potential Stimulation Of C. Minuta Growth By Various Prebiotics, And Its Sensitivity To Various Antibiotics, Maria Elena Hakim

Wayne State University Theses

Gut microbiota play a crucial role in maintaining intestinal health and integrity; a feature that is both heritable, as well as affected by the environment and lifestyle. Probiotics are supplements containing live microorganisms that act in a similar manner to gut microflora, and maintain a balance in the latter. C. minuta is a novel bacterium in the gut that was found to be associated with reduction in body weight and adiposity. The aim of this study was to determine the possible effects of different prebiotics on C. minuta growth, and the survival of C. minuta in response to different antibiotics. …


The Neonatal Anti-Viral Response Fails To Control Measles Virus Spread In Neurons Despite Interferon-Gamma Expression And A Th1-Like Cytokine Profile, Priya Ganesan Jan 2017

The Neonatal Anti-Viral Response Fails To Control Measles Virus Spread In Neurons Despite Interferon-Gamma Expression And A Th1-Like Cytokine Profile, Priya Ganesan

Electronic Theses and Dissertations

Neonates are highly susceptible to infections in the central nervous system (CNS) and have a greater risk of viral infections and encephalopathies. Neurotropic viral infections can lead to blindness, hearing loss and neurological deficiencies such as cognitive impairment, epilepsy, and even death in the neonatal and pediatric populations. Viral infections also are hypothesized to indirectly contribute to neurodegenerative and neuropsychiatric diseases such as Schizophrenia and Parkinson’s disease later in life due to early neuronal damage or stress. Many diverse viruses are capable of invading the neonatal CNS including Borna Disease Virus, Coxsackievirus (CV), Herpes simplex viruses (HSV), and measles virus. …


Fractionation And Characterization Of Lignin Streams From Genetically Engineered Switchgrass, Enshi Liu Jan 2017

Fractionation And Characterization Of Lignin Streams From Genetically Engineered Switchgrass, Enshi Liu

Theses and Dissertations--Biosystems and Agricultural Engineering

Development of biomass feedstocks with desirable traits for cost-effective conversion is one of the main focus areas in biofuels research. As suggested by techno-economic analyses, the success of a lignocellulose-based biorefinery largely relies on the utilization of lignin to generate value-added products, i.e. fuels and chemicals. The fate of lignin and its structural/compositional changes during pretreatment have received increasing attention; however, the effect of genetic modification on the fractionation, depolymerization and catalytic upgrading of lignin from genetically engineered plants is not well understood. This study aims to fractionate and characterize the lignin streams from a wild-type and two genetically engineered …


Role Of Intracellular Growth During The Gastrointestinal Stage Of Listeria Monocytogenes Infection, Grant Steven Jones Jan 2017

Role Of Intracellular Growth During The Gastrointestinal Stage Of Listeria Monocytogenes Infection, Grant Steven Jones

Theses and Dissertations--Microbiology, Immunology, and Molecular Genetics

Listeria monocytogenes is a facultative intracellular bacterium that causes foodborne disease in humans. L. monocytogenes invade the gut mucosa and then disseminate, causing systemic infections associated with high mortality rates in immunocompromised individuals. It is unknown how L. monocytogenes traffic to the mesenteric lymph nodes, which represent an important bottleneck for systemic spread. In addition, little is known about the gastrointestinal stage of infection due to the general resistance of mice to oral infection with L. monocytogenes. Our laboratory developed a novel foodborne mouse model of listeriosis utilizing a murinized strain of L. monocytogenes to investigate the gastrointestinal stage …


Evolution Of Equine Arteritis Virus During Persistent Infection In The Reproductive Tract Of The Stallion And The Male Donkey, Bora Nam Jan 2017

Evolution Of Equine Arteritis Virus During Persistent Infection In The Reproductive Tract Of The Stallion And The Male Donkey, Bora Nam

Theses and Dissertations--Veterinary Science

Equine arteritis virus (EAV) establishes persistent infection in the stallion reproductive tract, and the carrier stallion continues to shed virus in semen for weeks to years or lifelong. The objective of this study was to elucidate the intra-host evolution of EAV during persistent infection in stallions. Seven EAV seronegative stallions were experimentally infected with EAV KY84 strain and followed for 726 days post-infection, and sequential clinical samples including semen were collected for virus isolation and next-generation sequencing (NGS). In addition, archived sequential semen samples from two stallions that were naturally infected with EAV KY84 for a long-period (up to 10 …


Structure-Function Investigation Of Proteins Involved In Cellulose Biosynthesis By Escherichia Coli, Thomas Brenner Jan 2017

Structure-Function Investigation Of Proteins Involved In Cellulose Biosynthesis By Escherichia Coli, Thomas Brenner

Theses and Dissertations (Comprehensive)

Bacteria thrive within multicellular communities called biofilms consisting of a self-produced matrix. Biofilm matrices improve bacterial adherence to surfaces while creating a barrier from host immune responses, disinfectants, antibiotics and other environmental factors. Persistent colonization by the widely distributed pathogens, Escherichia coli and Salmonella spp., has been linked to production of biofilms composed of the exopolysaccharide cellulose. Cellulose-containing biofilms are also important to Acetobacter, Sarcina, Rhizobium and Agrobacterium species to form symbiotic and pathogenic interactions. In Enterobacteriaceae, two operons (bcsABZC and bcsEFG) are proposed to encode for proteins that form a cellulose biosynthetic complex that spans the …


An Undergraduate Cell Biology Lab: Western Blotting To Detect Proteins From Drosophila Eye, Neha Gogia, Ankita Sarkar, Amit Singh Jan 2017

An Undergraduate Cell Biology Lab: Western Blotting To Detect Proteins From Drosophila Eye, Neha Gogia, Ankita Sarkar, Amit Singh

Biology Faculty Publications

We have developed an undergraduate laboratory to allow detection and localization of proteins in the compound eye of Drosophila melanogaster, a.k.a fruit fly. This lab was a part of the undergraduate curriculum of the cell biology laboratory course aimed to demonstrate the use of Western Blotting technique to study protein localization in the adult eye of Drosophila. Western blotting, a two-day laboratory exercise, can be used to detect the presence of proteins of interests from total protein isolated from a tissue. The first day involves isolation of proteins from the tissue and SDS-PAGE (sodium dodecyl sulfate-polyacrylamide) gel …


Designing A Luminescence Assay To Measure In Vitro Binding Of The Fibronectin Attachment Protein (Fap) Of Mycobacterium Avium Subspecies Paratuberculosis To Fibronectin, Sirri Ngwa Jan 2017

Designing A Luminescence Assay To Measure In Vitro Binding Of The Fibronectin Attachment Protein (Fap) Of Mycobacterium Avium Subspecies Paratuberculosis To Fibronectin, Sirri Ngwa

All Graduate Theses, Dissertations, and Other Capstone Projects

The attachment and penetration of epithelial cells by Mycobacterium avium subsp. paratuberculosis (MAP) requires a bacterial fibronectin (FN) attachment protein FAP-P, which facilitates binding of FN by MAP. The region of FAP-P that interacts with FN is known, however, the region of FN that is bound by FAP-P is to be determined. Identification of the region of FN to which FAP-P binds would enable the synthesis of a competing peptide that could prevent FN binding by MAP. The first step toward locating this binding site is the design of a reliable FN binding assay. The purpose of this investigation was …


The Development Of New Molecular Tools To Detect Mycobacterium Avium Subspecies Paratuberculosis From A Bovine Faecal Environment By Real-Time Pcr, Marcel De Kruijf Jan 2017

The Development Of New Molecular Tools To Detect Mycobacterium Avium Subspecies Paratuberculosis From A Bovine Faecal Environment By Real-Time Pcr, Marcel De Kruijf

Theses

The slow-growing intracellular pathogen Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne’s disease in domesticated and wild ruminants. Johne’s disease inflicts severe annual economic losses to the dairy industry worldwide. This thesis is divided into five chapters that consist of a literature review' chapter and four experimental chapters focusing on MAP strain genotyping and the development of novel molecular MAP detection tools using real-time PCR. Chapter 1 describes published information regarding MAP characteristics, Johne’s disease, MAP genomes, available MAP detection methods, MAP epidemiology and genotyping.

To efficiently control and monitor MAP infection, molecular t>ping of MAP strains provides extensive knowledge …


Bioelectrohydrogenesis And Inhibition Of Methanogenic Activity In Microbial Electrolysis Cells - A Review, Rengasamy Karthikeyan, Ka Yu Cheng, Ammaiyappan Selvam, Arpita Bose, Jonathan W.C. Wong Jan 2017

Bioelectrohydrogenesis And Inhibition Of Methanogenic Activity In Microbial Electrolysis Cells - A Review, Rengasamy Karthikeyan, Ka Yu Cheng, Ammaiyappan Selvam, Arpita Bose, Jonathan W.C. Wong

Biology Faculty Research

Microbial electrolysis cells (MECs) are a promising technology for biological hydrogen production. Compared to abiotic water electrolysis, a much lower electrical voltage (~0.2V) is required for hydrogen production in MECs. It is also an attractive waste treatment technology as a variety of biodegradable substances can be used as the process feedstock. Underpinning this technology is a recently discovered bioelectrochemical pathway known as "bioelectrohydrogenesis". However, little is known about the mechanism of this pathway, and numerous hurdles are yet to be addressed to maximize hydrogen yield and purity. Here, we review various aspects including reactor configurations, microorganisms, substrates, electrode materials, and …


Frequency Of Colistin And Fosfomycin Resistance In Carbapenem-Resistant Enterobacteriaceae From A Tertiary Care Hospital In Karachi, Salima Qamar, Najma Shaheen, Sadia Shakoor, Joveria Farooqi, Kauser Jabeen, Rumina Hasan Jan 2017

Frequency Of Colistin And Fosfomycin Resistance In Carbapenem-Resistant Enterobacteriaceae From A Tertiary Care Hospital In Karachi, Salima Qamar, Najma Shaheen, Sadia Shakoor, Joveria Farooqi, Kauser Jabeen, Rumina Hasan

Department of Pathology and Laboratory Medicine

Introduction:
Management of infections with carbapenem-resistant Enterobacteriaceae (CRE) is challenging. In recent times, agents such as colistin and fosfomycin have been used in combination with other antibiotics to treat such infections. In this study, we aim to seek frequency of colistin and fosfomycin resistance in CRE from Pakistan.
Methods:
This study was conducted at clinical laboratories, Aga Khan University Hospital. In total, 251 CRE were included in the study. Colistin minimum inhibitory concentrations (MICs) were performed using broth microdilution (BMD) method and VITEK 2 system, whereas fosfomycin susceptibility was performed using Kirby-Bauer method. MIC50 and MIC90 were calculated for colistin …


Rapid Detection Of In Vitro Antituberculous Drug Resistance Among Smear-Positive Respiratory Samples Using Microcolony Detection-Based Direct Drug Susceptibility Testing Method, Irim Iftikhar, Seema Irfan, Joveria Farooqi, Zahida Azizullah, Rumina Hasan Jan 2017

Rapid Detection Of In Vitro Antituberculous Drug Resistance Among Smear-Positive Respiratory Samples Using Microcolony Detection-Based Direct Drug Susceptibility Testing Method, Irim Iftikhar, Seema Irfan, Joveria Farooqi, Zahida Azizullah, Rumina Hasan

Department of Pathology and Laboratory Medicine

Background: With the rise in multidrug-resistant tuberculosis, there is a search for newer techniques that will rapidly detect drug-resistant Mycobacterium tuberculosis. Although molecular techniques can detect resistance, culture is still considered gold standard, especially in resource-limited settings where quick, cheap, and easy techniques are needed. The aim of the study was to evaluate microcolony method thin layer agar (TLA) for quick detection of resistance against the first- and second-line antituberculous drugs in clinical isolates. This was a cross-sectional study performed at Aga Khan University Hospital.
Material and Methods: A total of 87 Z-N stain smear-positive pulmonary samples were received and …


Phenotypic Low-Level Isoniazid Resistance As A Marker To Predict Ethionamide Resistance In Mycobacterium Tuberculosis, Salima Qamar, Joveria Q. Farooqi, Kausar Jabeen, Rumina Hasan Jan 2017

Phenotypic Low-Level Isoniazid Resistance As A Marker To Predict Ethionamide Resistance In Mycobacterium Tuberculosis, Salima Qamar, Joveria Q. Farooqi, Kausar Jabeen, Rumina Hasan

Department of Pathology and Laboratory Medicine


Background:
Tuberculosis is one of the most prevalent diseases in Pakistan. Pakistan has the highest burden of MDR-TB in the Eastern Mediterranean region. Ethionamide is an anti-tuberculous drug frequently used to treat MDR-TB. Its drug susceptibility testing is not easily available in resource limited settings. Since it acts on the same target protein as isoniazid (inhA protein encoded by inhA gene), we sought to find out if phenotypic isoniazid resistance can be a marker of ethionamide resistance.
Materials and Methods: This was a retrospective observational study conducted at the Aga Khan University hospital section of microbiology. Data was retrieved between …