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Gyrb Mutations In Coumermycin A1-Resistant Borrelia Burgdorferi., D. Scott Samuels, Richard T. Marconi, Wai Mun Huang, Claude F. Garon

Biological Sciences Faculty Publications

We have isolated and characterized mutants of Borrelia burgdorferi that are resistant to the antibiotic coumermycin A1, which targets the B subunit of DNA gyrase. Mutants had either 100- or 300-fold higher resistance to coumermycin A1 than wild-type B. burgdorferi. In each case, a single point mutation in the gyrB gene converted Arg-133 to Gly or Ile. Mutations in the homologous Arg residue of Escherichia coli DNA gyrase are also associated with resistance to coumarin antimicrobial agents.

Phylogenetic Analysis Of Segment 10 From African Horsesickness Virus And Cognate Genes From Other Orbiviruses, Rafael O. De Sá, Marla Zellner, Marvin J. Grubman

Biology Faculty Publications

Utilizing the reverse transcriptase-polymerase chain reaction (RT-PCR) procedure, we have synthesized full-length copies of segment 10 from African horsesickness virus (AHSV) serotypes 1,4 and 8. The genes were cloned, sequenced and compared with the sequence of the cognate gene from AHSV serotypes 3 and 9. Sequences were analyzed to assess evolutionary relationships among serotypes using cladistics. Based on this analysis the data support a close relationship between serotypes 4 and 9 and between serotypes 1 and 8 and a closer relationship of serotype 3 to the 4 and 9 group.

The Effect Of High Intensity Exercise Training Verses Low Intensity Exercise Training On Fractionated Plasma High-Density Lipoprotein Cholesterol, Teresa L. Spate

Masters Theses

Exercise training is thought to result in an elevation of high-density lipoprotein (HDL) and the cardioprotective subfraction HDL{dollar}\sb2{dollar}. Twenty-five healthy women (39 {dollar}\pm{dollar} 7.8 yrs; 70 {dollar}\pm{dollar} 14 kg) walked two miles three times per week for 12 weeks to examine the effect of exercise intensity on the HDL profile. The L.I. group (N = 12) walked at 60% of the heart rate reserve (HRR) and the H.I. group (N = 13) walked at 80% HRR both maintaining the prescribed distance and frequency. A 22% increase in total HDL (from 32 {dollar}\pm{dollar} 6 to 39 {dollar}\pm{dollar} 8 mg/dl) and a …

Susceptibility Of Eumeces Species Skinks To A Southeastern Isolate Of The Lyme Disease Spirochete (Borrelia Burgdorferi), Gregory A. Cummins

College of Graduate Studies: Theses & Dissertations (1964–2006)

The SI-4 strain of Borrelia burgdorferi obtained from field collected Ixodes scapularis on Sapelo Island, Georgia, was infective to seven of eight cotton mice (Peromyscus gossypinus) by needle inoculation. I. scapularis larvae were then fed on them, and nymphs molting from these larvae were then fed on laboratory mice, Mus musculus, in an attempt to transmit the spirochete. One of three mice was successfully infected; confirmation of the presence of B. burgdorferi was made using BSK II culture and the polymerase chain reaction (PGR). A subsequent attempt was made to determine if SI-4 was infective for Eumeces …

Complete Nucleotide Sequence Of Psti-L Fragment Of Tanapox Virus Genomic Dna, Sarah J. Neering

Masters Theses

Tanapox virus (TPV), of the genus Yatapoxvirus, is a human poxvirus that was originally isolated from the Tana River Valley in Kenya in 1962. TPV produces a mild disease characterized by a short febrile illness associated with one or more nodular skin lesions. Although there have been limited studies on TPV, other poxviruses such as vaccinia virus, have been extensively characterized and their genomic DNAs sequenced. In order to examine the molecular features of TPV, we have cloned approximately 17% of the genomic DNA. TPV DNA was digested with Pstl restriction endonuclease and cloned into pUC19. The Pst …

Identification Of A Protein In Several Borrelia Species Which Is Related To Ospc Of The Lyme Disease Spirochetes., Richard T. Marconi, D. Scott Samuels, Tom G. Schwan, Claude F. Garon

Biological Sciences Faculty Publications

Using oligonucleotide probes which have previously been shown to be specific for the ospC gene found in the Lyme disease spirochete species Borrelia burgdorferi, B. garinii, and group VS461, we detected an ospC homolog in other Borrelia species including B. coriaceae, B. hermsii, B. anserina, B. turicatae, and B. parkeri. In contrast to the Lyme disease spirochetes, which carry the ospC gene on a 26-kb circular plasmid, we mapped the gene in other Borrelia species to linear plasmids which varied in size among the isolates tested. Some isolates carry multiple copies of the gene residing on linear plasmids of different …

Protein Inhibitors Of Calcium Salt Crystal Growth In Saliva, Bile And Pancreatic Juice, J. M. Verdier, B. Dussol, Y. Berland, J. C. Dagorn

Scanning Microscopy

The control of the formation of crystals in biological fluids is one of the most exciting field of research involving both organic and biochemical areas. Many organisms have evolved mechanisms which minimize or avoid the effects of nucleation and crystal growth formation. One of the most important mechanism is the interaction of specific proteins, called inhibitors, with crystals which alters their habits and leads to their elimination. This article, focused on saliva, pancreatic juice and bile, reviews our present knowledge on the structure-function relationships existing between these proteins and their ability to inhibit the growth of different calcium salt crystals.

The Syringomycins: Finding Their Role In Nature, Jon Takemoto

Faculty Honor Lectures

About ten years ago, I began looking for a research problem in the field of plant-microbe interactions. As the name implies, this is the study of how plants and microorganisms associate-either for their mutual benefit or at the expense of one partner. An example is symbiotic nitrogen fixation between leguminous plants and root-colonizing bacteria. Both benefit-the plant gains nitrogen in a useful form and the bacteria gain nutrients and protection. A better understanding of a plant-microbe interaction system could be used to benefit agriculture. For example, boosting nitrogen fixation would mean less petroleum-derived nitrogen fertilizers. These interactive biological systems and …

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 …

Transcriptional Analyses And Mapping Of The Ospc Gene In Lyme Disease Spirochetes, Richard T. Marconi, D. Scott Samuels, Claude F. Garon

Biological Sciences Faculty Publications

In Lyme disease spirochetes, the ospC gene encodes a 22.7-kDa protein referred to as either the pC or the OspC protein. Using a variety of electrophoretic approaches followed by Southern blotting and probing with oligonucleotide probes, we mapped the ospC gene to a circular 26-kb plasmid. The ospC gene represents the first gene to be mapped to a circular plasmid in Lyme disease spirochetes. The occurrence of this gene in isolates belonging to each of the three Lyme disease-associated species, Borrelia burgdorferi, Borrelia garinii, and the VS461 group, was evaluated. The ospC gene was found to occur in all 21 …

Coumermycin A1 Inhibits Growth And Induces Relaxation Of Supercoiled Plasmids In Borrelia Burgdorferi, The Lyme Disease Agent, D. Scott Samuels, Claude F. Garon

Biological Sciences Faculty Publications

Coumermycin A1 is an inhibitor of DNA gyrase, an enzyme that catalyzes supercoiling of DNA and is required for bacterial DNA replication. We have investigated the activity of this coumarin antibiotic on Borrelia burgdorferi, a spirochete and the causative agent of Lyme disease. B. burgdorferi was more susceptible than many other eubacteria to coumermycin as well as novobiocin, another coumarin antibiotic; this contrasted with its relative resistance to the DNA gyrase inhibitors nalidixic acid, oxolinic acid, and ciprofloxacin. Coumermycin at 0.2 micrograms/ml inhibited the growth of B. burgdorferi B31 in BSK II medium. A 100-fold-lower concentration induced the relaxation of …

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 …

Partial Characterization Of The Major Lipooligosaccharide From A Strain Of Haemophilus Ducreyi, The Causative Agent Of Chancroid, A Genital Ulcer Disease, William Melaugh, Nancy J. Phillips, A A. Campagnari, R Karalus, B W. Gibson

Chemistry Faculty Publications

The first preliminary structure of a surface lipooligosaccharide from Haemophilus ducreyi has been determined. The major oligosaccharide was released by mild acid hydrolysis and analyzed by liquid secondary ion and tandem mass spectrometry. The mass spectral data combined with composition and methylation analysis yielded the most probable structure; Gal1----4GlcNAc1----3Gal1----4Hep1----6Glc1----( Hep1----2Hep1----)3,4Hep1---- KDO, where the reducing terminal 3-deoxy-D-manno-octulosonic acid (or KDO) exists in an anhydro form. This anhydro species results from the elimination of a phosphate from C-4 of KDO during mild acid hydrolysis. The core heptose trisaccharide consists of L-glycero-D-manno-heptose, but analysis of the peracetylated sugars indicated that the 1,4-linked heptose …

The Microvasculature Of Human Oral Mucosa Using Vascular Corrosion Casts And India Ink Injection I. Tongue Papillae, Q. X. Yu, W. Ran, K. M. Pang, H. P. Philipsen, J. Theilade, X. H. Chen, Y. C. Mok

Scanning Microscopy

The microvasculature of human tongue papillae originating from 9 males and 6 females, aged 0.5 to 2 years was studied by scanning electron microscopy (SEM) of vascular corrosion casts and by light microscopy (LM) of India ink injected specimens. All papillae showed a microvasculature characterized by primary, secondary and tertiary capillary loops. In the filiform papillae the loops were generally arranged in a corolla-like pattern with the tertiary loops demonstrating a hair-pin shape. The fungiform papillae showed basically a similar architectural pattern although the loops were somewhat more compact and complex in structure. A small, shallow depression of the tertiary …

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 …

Site-Directed Mutagenesis Of The Putative Processing Site Of Recombinant Human Immunodeficiency Virus Reverse Transcriptase, Susan M. Poppe

Masters Theses

The human immunodeficiency virus (HIV) is the etiological agent of the acquired immune deficiency syndrome (AIDS). This retrovirus, encodes a RNA dependent DNA polymerase, reverse transcriptase (RT), which is essential to virus replication. Biochemical analysis indicated that RT is composed of 2 polypeptides of 66,000 and 51,000 daltons (p66 and p51) which combine to form a heterodimer. This heterodimer is thought to arise from the post-translational processing where p66 is made and either autocatalytic or proteolytic cleavage events result in a heterogeneous p66/p51. Site-directed mutagenesis was used at the proposed processing site to probe HIV-RT processing.

Three prokaryotic expression vectors …

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 …

Mounting Sex Pheromone: A Novel Pheromone Responsible For Mate Recognition In The Ixodidae, Jgc Hamilton

Theses and Dissertations in Biomedical Sciences

Cholesterol oleate was demonstrated to be the cuticular contact sex pheromone of Dermacentor variabilis. This pheromone which has been termed the Mounting Sex Pheromone (MSP) was also demonstrated to be present on the surface of D. andersoni, Amblyomma maculatum and A. americanum. This contact sex pheromone enables males excited and attracted by 2,6-dichlorophenol to identify the female as a potential mating partner. The MSP is the second in the series of three sex pheromones guiding the hierarchy of behavioral responses which constitute tick courtship behavior. Tests with D. variabilis and D. andersoni showed that this pheromone …

Export And Intercellular Transfer Of Dna Via Membrane Blebs Of Neisseria-Gonorrhoeae, David W. Dorward, Claude F. Garon, Ralph C. Judd

Biological Sciences Faculty Publications

Naturally elaborated membrane bleb material is frequently observed in cultures of Neisseria gonorrhoeae. This material was purified and analyzed for protein, lipopolysaccharide, and nucleic acid content. The electrophoretic protein profiles of two bleb-rich fractions, called BI and BII, were distinct, with only BII containing lipopolysaccharide and outer membrane proteins I and III. Both fractions contained RNA, circular DNA, and linear DNA. Exogenous pancreatic DNase I appeared to hydrolyze all bleb-associated DNA in fraction BI and the linear DNA in fraction BII. The circular DNA molecules associated with fraction BII resisted digestion. Electron microscopy of the bleb fractions verified their DNA …

Effects Of Methylene Chloride On Immune Function In Mice And The In Vitro Effect Of Methylene Chloride In Immunologic Assays, Man-Ping Wang

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

A number of toxicities associated with methylene chloride have been found in both human subjects and mice. However, relatively few studies have probed immunotoxicities of methylene chloride. In order to examine possible immunotoxicities or immunomodulating effects of methylene chloride, several tests of cellular immune function were performed using both human in in vitro studies and a mouse model.

Body weights and specific organ weights of thymus, spleen, liver, and kidney were normal in CD-1 mice given various concentrations of methylenechloride. However, a significantly reduced mitogenic response to phytohemagglutinin (PHA} and reduced interleukin-2 (IL-2} production was found in these methylene-chloride-treated mice. …

In Vitro Identification Of The Effect Of Serotonin On Lymphocyte Dna Synthesis And Natural Killer Cell Activity, Kim Kartchener Kane

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The purpose of this study was to identify the effects of the neurotransmitter, serotonin (SE), on the immune function of peripheral blood mononuclear cells (PBMC) of normal, healthy subjects. This was done as a preliminary investigation to studies on the association of SE with immune changes in autistic subjects.

The PBMC isolated from normal male subjects were treated with various concentrations of SE for 48 hrs. Their incubation in SE at a concentration of 10^-3 M induced about a 35% decrease in DNA synthesis. However, incubation of the cells in lower concentrations (10^-4 to 10^-10) of …

Protein I: Structure, Function, And Genetics, Ralph C. Judd

Biological Sciences Faculty Publications

No abstract provided.

Ua66/6/2 Bionews From The Hill, Wku Biology

WKU Administration Documents

Newsletter created by and about the WKU Biology department highlighting students, faculty and alumni activities.

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 …

Characterization Of Protein-I From Serum-Sensitive And Serum-Resistance Transformants Of Neisseria-Gonorrhoeae, Ralph C. Judd, Milton Tam, Keith Joiner

Biological Sciences Faculty Publications

No abstract provided.

Evidence For N-Terminal Exposure Of The Protein Ia Subclass Of Neisseria-Gonorrhoeae Protein-I, Ralph C. Judd

Biological Sciences Faculty Publications

The JS3 and FA638 strains of Neisseria gonorrhoeae bear a protein IA subclass of protein I (P.I). The purified P.Is of surface-labeled strains JS3 and FA638 were cleaved with the N-terminal degradation enzyme leucine amino peptidase (LAP), and the resultant fragments were separated in sodium dodecyl sulfate-polyacrylamide gels. Autoradiography demonstrated that the surface radiolabel was absent in a LAP-generated P.I peptide that was about 1,900 daltons lower in apparent molecular mass than the native P.I in both strains. Moreover, the 4G5 monoclonal epitope, known to be located on the surface of the organism, was also absent in the LAP-generated P.I …

Intrauterine Device (Iud) Associated Pathology: A Review Of Pathogenic Mechanisms, Waldemar A. Schmidt, Karmen L. Schmidt

Scanning Electron Microscopy

This paper summarizes our studies of IUD-related disease with those previously published by others. Our data are based upon 51 IUDs and 42 index cases of IUD-related disease demonstrating specific processes. Gross, dissecting microscope, scanning electron microscope and X-ray microanalysis examinations were made of selected IUDs and associated tissues. Tissue associated with the IUDs revealed inflammation in 59.4%, calcific material in 6.3% and no abnormality in 34.4%. JUD-associated tissue responses were accompanied by changes of the IUD; these changes involved deposition of substances upon the IUD surface and degradation of the JUD itself. Disintegration of the IUD, its string or …

Viruses In Water, Nasrin Mohammadpour Daryaee

All Graduate Plan B and other Reports, Spring 1920 to Spring 2023

The use of water by man, plants and animals is universal. Without it there can be no life. Every living thing requires water. Man can live nearly two months without food, but can live only three or four days without water. In our homes, whether in the city or in the country, water is essential for cleanliness and health. Water can be considered as the principal raw material and the lowest cost raw material from which most of our farm produce is made. It is essential for the growth of crops and animals and is a very important factor in …

Purification Of Anthrax Toxin Protective Antigen Component And Characterization Of Its Binding Interaction With Bovine Kidney Cells, Daniel Dalton Martin

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Protective antigen component of B. anthracis toxin was produced and purified to the >99% level. Toxin was purified from culture supernatant utilizing concentration and liquid chromatography techniques. Purity was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

The purified protective antigen retained biological and antigenic activity as evidenced respectively by lethality in Fischer 344 rats when injected in combination with lethal factor, and by positive results on the Ouchterlony double diffussion assay.

Radioiodinated protective antigen was used both in the in vivo and the in vitro experiments.

In vivo distribution of labelled protective antigen was determined in Fischer 344 rats. Assay …

Conserved Chromosomal Positions Of Dual Domains Of The Ets Protooncogene In Cats, Mice, And Humans, Dennis K. Watson, Mary J. Mcwilliams-Smith, Christine Kozak, Roger Reeves, John Gearheart, Michael F. Nunn, William Nash, John R. Fowle Iii, Peter Duesberg, Takis S. Papas, Stephen J. O'Brien

Biology Faculty Articles

The mammalian protooncogene homologue of the avian v-ets sequence from the E26 retrovirus consists of two sequentially distinct domains located on different chromosomes. Using somatic cell hybrid panels, we have mapped the mammalian homologue of the 5' v-ets-domain to chromosome 11 (ETS1) in man, to chromosome 9 (Ets-1) in mouse, and to chromosome D1 (ETS1) in the domestic cat. The mammalian homologue of the 3' v-ets domain was similarly mapped to human chromosome 21 (ETS2), to mouse chromosome 16 (Ets-2), and to feline chromosome C2 (ETS2). …