Immunology and Infectious Disease Commons^™

Quantitative Dissection Of Clone-Specific Growth Rates In Cultured Malaria Parasites, Heather B. Reilly Ayala, Hongjian Wang, John A. Steuter, Anastasia M. Marx, Michael T. Ferdig

Faculty Publications - Department of Biological & Molecular Science

Measurement of parasite proliferation in cultured red blood cells underpins many facets of malaria research, from drug sensitivity assays to assessing the impact of experimentally altered genes on parasite growth, virulence, and fitness. Pioneering efforts to grow Plasmodium falciparum in cultured red blood cells revolutionized malaria research and spurred the development of semi-high throughput growth assays using radio-labeled hypoxanthine, an essential nucleic acid precursor, as a reporter of whole-cycle proliferation (Trager and Jensen, 1976; Desjardins et al., 1979). Use of hypoxanthine (Hx) and other surrogate readouts of whole-cycle proliferation remains the dominant choice in malaria research. While amenable to high-throughput …

Contrasting Infection Strategies In Generalist And Specialist Wasp Parasitoids Of Drosophila Melanogaster, Todd A. Schlenke, Jorge Morales, Shubha Govind, Andrew G. Clark

Publications and Research

Although host–parasitoid interactions are becoming well characterized at the organismal and cellular levels, much remains to be understood of the molecular bases for the host immune response and the parasitoids’ ability to defeat this immune response. Leptopilina boulardi and L. heterotoma, two closely related, highly infectious natural parasitoids of Drosophila melanogaster, appear to use very different infection strategies at the cellular level. Here, we further characterize cellular level differences in the infection characteristics of these two wasp species using newly derived, virulent inbred strains, and then use whole genome microarrays to compare the transcriptional response of Drosophila to each. While …

Parasites, Proteomics And Performance: Effects Of Gregarine Gut Parasites On Dragonfly Flight Muscle Composition And Function, Rudolf J. Schilder, James H. Marden

School of Biological Sciences: Faculty Publications

In previous work, we found that dragonflies infected with gregarine gut parasites have reduced muscle power output, loss of lipid oxidation in their flight muscles, and a suite of symptoms similar to mammalian metabolic syndrome. Here, we test the hypothesis that changes in muscle protein composition underlie the observed changes in contractile performance. We found that gregarine infection was associated with a 10-fold average reduction in abundance of a ~155·kDa fragment of muscle myosin heavy chain (MHC; ~206·kDa intact size). Insect MHC gene sequences contain evolutionarily conserved amino acid motifs predicted for calpain cleavage, and we found that calpain digestion …

Tissue And Life Stage Distribution Of A Defensin-Like Gene In The Lone Star Tick Amblyomma Americanum, S. Michelle Todd, Daniel E. Sonenshine, Wayne L. Hynes

Biological Sciences Faculty Publications

The transcript sequence of the Amblyomma americanum Linnaeus (Acari: Ixodidae) defensin, termed amercin (amn), was ascertained and a 219-bp amn coding region identified. The gene encodes a 72-amino acid prepropeptide with a putative 37-amino acid mature peptide. This gene shows little similarity to either of the defensins from Amblyomma hebraeum Koch, the only other Amblyomma species for which a defensin has been described. Sequence comparisons with other tick defensins reveal amn to be shorter (6 bp or 2 amino acids) than the Ixodes scapularis Linnaeus (Acari: Ixodidae) and Dermacentor variabilis (Say) (Acari: Ixodidae) defensin sequences. The amercin prepropeptide …

Asymmetric Localizations Of Lin-17/Fz And Mig-5/Dsh Are Involved In The Asymmetric B Cell Division In C. Elegans, Mingfu Wu, Michael A. Herman

School of Biological Sciences: Faculty Publications

LIN-44/Wnt and LIN-17/Frizzled (Fz) function in a planar cell polarity (PCP)-like pathway to regulate the asymmetric B cell division in Caenorhabditis elegans. We observed asymmetric localization of LIN-17/Frizzled (Fz) and MIG-5/Disheveled (Dsh) during the B cell division. LIN-17∷GFP was asymmetrically localized within the B cell prior to and after the B cell division and correlated with B cell polarity. Asymmetric localization of LIN-17∷GFP was dependent upon LIN-44/Wnt and MIG-5/Dsh function. The LIN-17 transmembrane domain and a portion of the cysteine-rich domain (CRD) were required for LIN-17 function and asymmetric distribution to the B cell daughters, while the conserved KTXXXW …

In Vitro Expression And Purification Of Class I Mhc Molecules, Loi Cheng

Honors Scholar Theses

The major histocompatibility complex (MHC) is a gene family responsible for many critical functions of the immune system in most vertebrates. The MHC consists of three classes differentiated by their structure and function, and MHC class I encodes antigen binding proteins as well as chaperone and accessory proteins such as tapasin. The purpose of this project is to reconstitute several human MHC class I molecules in their peptide-filled and peptide-deficient forms, and to purify these proteins for biochemical study. The expressed proteins include wild type and mutant variants of the fusion protein human leukocyte antigen HLA-B*0801-fos, and human beta-2-microglobulin (β2m). …

Variation In The Innate And Acquired Arms Of The Immune System Among Five Shorebird Species, Luisa Mendes, Theunis Piersma, Dennis Hasselquist, Kevin Matson, Robert Ricklefs

Biology Department Faculty Works

To contribute to an understanding of the evolutionary processes that shape variation in immune responses, we compared several components of the innate and acquired arms of the immune system in five related, but ecologically diverse, migratory shorebirds (ruff Philomachus pugnax L., ruddy turnstone Arenaria interpres L., bar-tailed godwit Limosa lapponica L., sanderling Calidris alba Pallas and red knotC. canutus L.). We used a hemolysis-hemagglutination assay in free-living shorebirds to assess two of the innate components (natural antibodies and complement-mediated lysis), and a modified quantitative enzyme-linked immunosorbent assay in birds held in captivity to assess the acquired component (humoral antibodies against …

Glucocorticoid Hormone Levels Increase With Group Size And Parasite Load In Cliff Swallows, Samrrah A. Raouf, Linda C. Smith, Mary Bomberger Brown, John C. Wingfield, Charles R. Brown

School of Biological Sciences: Faculty Publications

Animals often cope with adverse events by releasing glucocorticoid hormones, which in turn promote increased energy assimilation. In captive animals, crowding also leads to increased glucocorticoid activity, probably because of increased levels of social competition. We investigated how group size and ectoparasite infestations affected endogenous levels of the glucocorticoid hormone, corticosterone, in colonial cliff swallows, Petrochelidon pyrrhonota, in southwestern Nebraska, USA. Parasites were removed from some colonies by fumigating nests. Baseline levels of corticosterone in breeding adults varied significantly with whether parasites were present, colony size (measured by total number of active nests at a site), and nesting stage. …

Proceedings Of The Western Australian Dhufish Workshop 2004, Mark Pagano, Terry Fuller

Fisheries Occasional Publications

The Western Australian Dhufish Workshop 2004 was a joint project between Recfishwest and the Western Australian Department of Fisheries.

The workshop incorporated presentations from the Department of Fisheries, Recfishwest, the Western Australian Fishing Industry Council, Murdoch University and Challenger TAFE to showcase current knowledge relating to the iconic dhufish in Western Australia. The perspectives of a recreational fisher, a commercial wetline operator and a metropolitan charter boat operator were also presented.

Through information sharing and discussion the workshop provided a platform to better understand the fundamental issues that underpin current and future management arrangements for this valuable resource. Both commercial …

A Defensin-Like Gene Expressed In The Black-Legged Tick, Ixodes Scapularis (Acari: Ixodidae), Wayne L. Hynes, S. M. Ceraul, S. M. Todd, K. C. Seguin, Daniel E. Sonenshine

Biological Sciences Faculty Publications

The black-legged tick Ixodes scapularis linnaeus (Acari: Ixodidae) is an important vector of microbial pathogens. Knowledge of the tick's innate immune response, particularly defensin and other antimicrobial peptides, is important for understanding how microbes survive in this tick. A defensin gene (slnA) from I. scapularis was obtained by reverse transcription-polymerase chain reaction (RT-PCR) using mRNA extracted from tissues of female ticks. RT-PCR indicated the gene was expressed in the midgut, haemocytes, and fat-body, although no evidence of a peptide was found. Sequencing a cloned cDNA fragment revealed a 225 bp open reading frame encoding a 74 amino acid pre-prodefensin, including …

Contributions To The Mammalogy Of Mongolia, With A Checklist Of The Species Of The Country, David S. Tinnin, Jon L. Dunnum, Jorge A. Salazar-Bravo, Nyamsuren Batsaikhan, M. Scott Burt, Scott Lyell Gardner, Terry L. Yates

Harold W. Manter Laboratory of Parasitology: Faculty Publications

We present accounts for 40 species of mammals collected from 15 localities in the Mongolian People's Republic. Accounts include taxonomic, morphometric, reproductive and ecological information, as well as trap effort and success. In addition, we include a brief history of mammalogical work within Mongolia, a taxonomically updated species list for the country, and a list of institutions with holdings of Mongolian mammals.

Terminal Deoxynucleotidyl Transferase Generation Of Sequence Diversity On Plasmids, Qingbei Zhang

Morehead State Theses and Dissertations

A thesis presented to the faculty of the College of Science and Technology at Morehead State University in partial fulfillment of the requirements for the Degree of Master of Science by Qingbei Zhang on June 19, 2002.

Comparison Of Tnf-A Levels Produced By Macrophages Treated With Brucella Abortus And Escherichia Coli Lps, Tajie Harris

Honors Capstones

Capstone submitted as a graduation requirement for the BSU Honors Program.

Glass Capillary Tube Feeding: A Method For Infecting Nymphal Ixodes Scapularis (Acari: Ixodidae) With The Lyme Disease Spirochete Borrelia Burgdorferi, Anne H. Broadwater, Daniel E. Sonenshine, Wayne L. Hynes, Shane Ceraul, Aravinda M. De Silva

Biological Sciences Faculty Publications

We evaluated an artificial capillary feeding method to infect nymphal Ixodes scapularis (Say) ticks with Borrelia burgdorferi, the causative agent of Lyme disease. Thirty to 70% of the nymphs were infected after feeding for 2.5 h from glass capillary tubes filled with a solution of spirochetes. Capillary infection was stable and persisted in the nymphs for at least 10 d after feeding. Capillary feeding also maintained natural vector competence patterns because I. scapularis ticks acquired infection unlike Dermacentor variablis (Say), which did not become infected. Capillary infected I. scapularis nymphs were capable of transmitting the infection to naive mice …

The Response Of Mice To Infection By The Parasitic Nematode Trichinella: A Comparison Of Trichinella Spiralis And Trichinella Pseudospiralis, Barry Clayton Anderson

Dissertations and Theses

The intracellular parasite Trichinella is a genus in phylum Nematoda that contains six named species including Trichinella spiralis and Trichinella pseudospiralis. These parasites infect a large variety of wild and domestic animals, human beings and a few species of birds. The parasitic strategies and the pathological effects on the host between trichinella spiralis and Trichinella pseudospiralis are quite different.

The purpose of this study is to gain an understanding of the physiological, immunological and pathological differences between these two species of Trichinella using infections in the mouse as a model. In the course of this research I have attempted to …

Contrasts In Tick Innate Immune Responses To Borrelia Burgdorferi Challenge: Immuno-Tolerance In Ixodes Scapularis Versus Immunocompetence In Dermacentor Variabilis (Acari: Ixodidae)., Robert Johns, Jun Ohnishi, Anne Broadwater, Daniel E. Sonenshine, Aravinda M. Desilva, Wayne L. Hynes

Biological Sciences Faculty Publications

The blacklegged tick, Ixodes scapularis Say, transmits the Lyme disease spirochete (Borrelia burgdorferi), whereas the American dog tick, Dermacentor variabilis (Say), is unable to transmit the bacterium. We compared the innate immune response of these ticks against spirochetes directly inoculated into the hemocoel cavity of ticks. In I. scapularis, some Borrelia were found associated with hemocytes, while numerous other spiral-shaped, intact bacteria remained free in the hemolymph. In contrast, in D. variabilis only remnants of the bacteria were evident in the hemolymph, indicating lysis; intact spirochetes were rare. Spirochetes were observed bound to or within the organs …

Identification Of A Defensin From The Hemolymph Of The American Dog Tick, Dermacentor Variabilis, Robert Johns, Daniel E. Sonenshine, Wayne L. Hynes

Biological Sciences Faculty Publications

Hemolymph from partially fed virgin Dermacentor variabilis females was collected following Borrelia burgdorferi challenge and assayed for antimicrobial activity against Bacillus subtilis and B. burgdorferi. A small inducible cationic peptide was identified by SDS-PAGE in the hemolymph of these ticks as early as 1h post challenge. Following purification by a three-step procedure involving sequential SepPak elution, reversed phase high performance liquid chromatography (RP-HPLC) and gel electrophoresis, the yield of the active peptide was approximately 0.1% of the total protein in the hemolymph plasma. The molecular weight, 4.2kDa, was determined by MALDI-TOF mass spectrometry. N-terminal sequencing by the Edman degradation …

Tick Immunity To Microbial Infections: Control Of Representative Bacteria In The Hard Tick Dermacentor Variabilis (Acari: Ixodidae), Robert Johns, Shane Ceraul, Daniel E. Sonenshine, Wayne L. Hynes

Biological Sciences Faculty Publications

No abstract provided.

Integronlike Structures In Campylobacter Spp. Of Human And Animal Origin, Brigid Lucey, D. Crowley, P. Moloney, B. Cryan, M. Daly, F. O'Halloran, E. J. Threlfall, S. Fanning

Department of Biological Sciences Publications

Resistance to antimicrobial agents used to treat severe Campylobacter spp. gastroenteritis is increasing worldwide. We assessed the antimicrobial resistance patterns of Campylobacter spp. isolates of human and animal origin. More than half (n = 32) were resistant to sulphonamide, a feature known to be associated with the presence of integrons. Analysis of these integrons will further our understanding of Campylobacter spp. epidemiology.

Evolutionary Constraints On Population Structure: The Parasites Of Fundulus Zebrinus (Pisces: Cyprinodontidae) In The South Platte River Of Nebraska, John J. Janovy Jr., Scott D. Snyder, Richard E. Clopton

Biology Faculty Publications

Population and community descriptor values (parasites per host, prevalence per parasite species, variance/mean ratios, species density, and diversity indices) for the 7-species parasite community of 61 relatively homogeneous samples of Fundulus zebrinus (Pisces: Cyprinodontidae) in the South Platte River of Nebraska, U.S.A., taken over a 14-yr period, are reported. South Platte River streamflow fluctuates over two orders of magnitude on several time scales-monthly, annually, and over multiple year wet-dry cycles. Relatively homogeneous sampling of a single host species with several parasite species provided a system that allowed assessment of the contribution of evolved parasite life cycles to population structure in …

Behavioral Basis Of Second Intermediate Host Specificity Among Four Species Of Haematoloechus (Digenea: Haematoloechidae), Scott D. Snyder, John J. Janovy Jr.

Biology Faculty Publications

Cercarial behavior patterns were examined in four species of frog lung flukes (Haematoloechus spp.). Cercariae of Haematoloechus complexus, Haematoloechus medioplexus, Haematoloechus longiplexus, and Haematoloechus varioplexus were exposed to three species of experimental arthropods and an inanimate control. The number of cercariae attached to an experimental host at 5 min postexposure was recorded. Haematoloechus longiplexus and H. complexus cercariae attached to experimental hosts at higher rates than cercariae of H. medioplexus and H. varioplexus. Cercariae of H. longiplexus attached to experimental hosts in approximately the same numbers as H. complexus, but H. longiplexus penetrated only damselfly naiads, …

The Effect Of The Mammalian Hormone Estrogen On The Yeast C̲A̲N̲D̲I̲D̲A̲ A̲L̲B̲I̲C̲A̲N̲S̲, Marcia Clifton Finucane

Theses, Dissertations and Capstones

Changes in the balance between commensal microbes and mammalian hosts can cause disease if the scales tip in favor of the microbe. Although usually caused by a defective host immune response, overgrowth of or changes in the microbe may also be responsible for opportunistic infections. Clinical and laboratory investigations have indicated that mammalian hormones, including estrogen, can influence the production of symptomatic disease and the virulence attributes of Candida albicans.

This study demonstrated enhanced growth of some strains of C. albicans when 17β-estradiol was added to the growth medium. Measurement of extracellular protein under the influence of estradiol did …

Second Intermediate Host-Specificity Of Haematoloechus Complexus And Haematoloechus Medioplexus (Digenea: Haematoloechidae), Scott D. Snyder, John J. Janovy

Biology Faculty Publications

Second intermediate host-specificity was examined for two species of the frog lung fluke genus Haematoloechus. Nine species of freshwater arthropods were exposed to cercariae of H. complexus and H. medioplexus. Metacercariae of H. complexus developed in all arthropod species used. Metacercariae of H. medioplexus developed only in anisopteran odonate naiads. This difference in host utilization may have epizootiological implications. The potential development of H. complexus in a greater number of arthropods than H. medioplexus may increase the chances of ingestion of H. complexus by an anuran host. The range of arthropods parasitized by H. complexus indicates that host-specificity …

Cryptococcus Neoformans Serotype Groups Found In Clinical And Environmental Isolates, John Clauson

Masters Theses & Specialist Projects

Cryptococcus neoformans is an encapsulated yeast responsible for severe meningoencephalitis. The importance of epidemiological studies on cryptococcosis has increased since the beginning of the AIDS epidemic. C. neoformans exists in two varieties containing four serotypes, C. neoformans var. neoformans (serotypes A and D) and C. neoformans var. gattii (serotypes B and C). Locally C. neoformans var. neoformans has been associated with pigeon feces during those months having an average temperature of 64.2°F j(17.8°C) and above. Clinical and environmental isolates of C. neoformans obtained from regional hospitals and environmental samplings, respectively, have been grouped into their variety status utilizing canavanine-glycine-bromthymol blue …

Molecular Approaches For Control Of Tomato Bushy Stunt Virus, H. B. Scholthof, M. Borja, Thomas Jack Morris, A. O. Jackson

School of Biological Sciences: Faculty Publications

I N this overview strategies are discussed that are currently being tested in our laboratory in an effort to engineer resistance against tomato bushy stunt virus (TBSV) in plants. TBSV is chosen as a model-system since this is an economically important RNA virus that is well characterized at the molecular level and the virus has some interesting and potentially useful features that can be exploited to produce resistant plants. Moreover, we anticipate that the information derived from our studies with TBSV will be applicable in developing resistance to other plant viruses. Some approaches that we are attempting to apply to …

Evidence Of A Neurogenic Component During Ige-Mediated Inflammation In Murine Skin, Victoria Jean Cavanaugh

Biological Sciences Theses & Dissertations

The direct stimulation of sensory nerves or the intradermal injection of neuropeptides triggers a cutaneous inflammatory response that involves the degranulation of mast cells. These neurogenic inflammatory reactions are physiologically similar to IgE-mediated hypersensitivity responses, which also depend on mast cells. Although non-antigenic stimuli distinguish neurogenic from IgE-mediated inflammation, the similarity of their effector stages suggested that peripheral nerves may also participate in lgE-mediated responses. To examine this hypothesis, IgE responses were elicited in denervated skin. A murine model was developed where the footpad was denervated by surgically removing a 2 mm segment of the sciatic nerve, proximal to the …

Dietary Protein Deficiency And Mycobacterium Bovis Bcg Affect Interleukin-2 Activity In Experimental Pulmonary Tuberculosis, David N. Mcmurray, C. L. Mintzer, R. A. Bartow, Rebecca D. Parr

Faculty Publications

Inbred strain 2 guinea pigs were vaccinated with Mycobacterium bovis BCG or were left unvaccinated. They were maintained for 6 weeks on defined, isocaloric diets containing either 30% (control animals) or 10% (animals receiving low protein) ovalbumin as the sole protein source. Animals were challenged by the respiratory route with a low dose of virulent M. tuberculosis H37Rv and killed 4 weeks later. Protein-malnourished animals were not protected by previous vaccination with BCG. Lymphocytes isolated from various tissues were tested in vitro for proliferative responses to mitogen (concanavalin A) and antigen (purified protein derivative [PPD]), production of interleukin-2 (IL-2), and …

The Effect Of Sodium Chloride On Beta-Hemolytic Streptococci, Bola Fashola

Masters Theses & Specialist Projects

The drug of choice for the treatment of Stieptococcal pharyngitis is penicillin G. However, a common home remedy prescribes the use of salt-water solutions for gargling.

Members of Beta -hemolytic streptococcal groups A, B, and C were isolated from the upper -respiratory tracts of patients diagnosed as having streptococcal pharyngitis. These cultures we:e obtained from HCA Greenview Hospital (Bowling Green, Kentucky) and used to study the effects of sodium chloride on the isolates.

The minimum inhibitory concentration of sodium chloride was determined for each of eight hospital isolates. Croup A streptococci were inhibited at a concentration of 7.2% sodium chloride …

Cellular Immunity In Children With Down Syndrome (Trisomy-21), Roger Lee Noble

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Individuals with Down syndrome (US) suffer from increased incidence of respiratory infections and lymphoblastic leukemia, and a death rate that is particularly high in the first 5 years of life. Relatively few studies have probed immune parameters in young US children. Primary immune defects in DS may be masked by a degree of immune maturity in adults, and hygienic factors may have an effect on immune capability throughout the years. A study of young children can give clearer evidence of the actual primary immune defects in DS.

Blood samples were drawn from 20 DS children under 6 years old and …

Natural Killer Activity In Gardner's Syndrome, Ann Marie Stembridge

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Gardner's syndrome is an autosomal dominant disease presenting multiple colonic polyps with a predisposition for malignant change. In addition to colonic polyp formation by early adolescence, extracolonic lesions appear often prior to polyp formation. One theoretical mechanism for the origin of polyps and malignancies in Gardner's syndrome is a genetic defect in the natural killer cell activity of patients with this disease. Natural killer cells are a subpopulation of lymphocytes that spontaneously lyse tumor cells and virally transformed cells. A study was undertaken to determine the natural killer activity of patients with Gardner's syndrome.

A technique termed chromium release was …