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Separate Metabolic Pathways Leading To Dna Fragmentation And Apoptotic Nuclear Chromatin Condensation, D. Sun, Shibo Jiang, Li-Mou Zheng, David M. Ojcius, John Ding-E. Young

All Dugoni School of Dentistry Faculty Articles

Apoptosis is the predominant form of cell death observed in a variety of physiological and pathological conditions such as cancer involution, insect metamorphosis, the development of the immune and nervous systems, and embryogenesis. The typical nuclear changes taking place in apoptotic cells include extensive condensation of chromatin and internucleosomal DNA fragmentation into units of 200 base pairs. However, the mechanisms responsible for both chromatin condensation and DNA fragmentation have yet to be elucidated. In this study, micrococcal nuclease and the divalent cations, Ca2+ and Mg2+, were applied to isolated nuclei in an attempt to reconstitute in vitro the digestion of …

Morphometric Analysis Of Enteric Lesions In C3h/Hen Mice Inoculated With Serpulina Hyodysenteriae Serotypes 2 And 4 With Or Without Oral Streptomycin Pretreatment, Jagannatha V. Mysore, Gerald Duhamel

School of Veterinary and Biomedical Sciences: Faculty Publications

The segmental distribution and sequential progression and the role of the indigenous bacterial flora in the development of enteric lesions associated with Serpulina hyodysenteriae infection in laboratory mice have not been defined. We examined the distribution and sequential morphometric changes in the large intestine of mice orally inoculated with S. hyodysenteriae serotypes 2 and 4. To determine the role of colonization resistance conferred by the indigenous bacterial flora, 40 female C3H/HeN mice were administered water alone or water containing 5 mg/mL streptomycin sulfate ad libitum for seven days prior to orogastric inoculation either with S. hyodysenteriae or sterile trypticase soy …

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 …

A Molecular Basis For Erythromycin Sensitivity And Resistance In Escherichia Coli, Harold S. Chittum

Electronic Theses and Dissertations

The effect of erythromycin on the 50S ribosomal subunit during cell growth has been extensively investigated. Sucrose density gradient analysis of ribosomes formed in the presence and absence of the drug revealed a 50S specific assembly defect is partially responsible for erythromycin's inhibitory effects on wild type cells. Examination of two erythromycin-resistant mutants of E. coli (N281 and N282) revealed that mutant N281 (L22 mutant) but not N282 (L4 mutant) was assembly defective in the presence of the drug, although only at much higher drug concentrations (300 ug/ml vs. 75 ug/ml for wild type cells). The altered genes from each …

Probing Protein-Protein Interactions Among Proteins Of A Nonaggregated Fatty Acid Synthetase From Euglena Gracilis Variety Bacillaris, Sande G. Williams

Electronic Theses and Dissertations

Enoyl-acyl carrier protein (ACP) reductase from chloroplast nonaggregated fatty acid synthetase (FAS) of Euglena gracilis variety bacillaris was purified to a single band on a denaturing polyacrylamide gel. The enzyme was partially characterized with respect to substrate specificity, reduced nucleotide requirement, and the effect of ACP and Ca$\sp{++}$ on enzyme activity. Antibodies against the purified protein were raised in hens and isolated from eggs. ACP was purified from Euglena in yields of about 1mg/100g (wet weight) of cells. Antibodies were raised against the purified protein. ACP antibodies inhibited the Euglena chloroplast FAS using Euglena or E. coli ACP as a …

Investigation Of The Structural Heterogeneity Of Lipooligosaccharides From Pathogenic Haemophilus And Neisseria Species And Of R-Type Lipopolysaccharides From Salmonella Typhimurium By Electrospray Mass Spectrometry, B W. Gibson, William Melaugh, Nancy J. Phillips, M A. Apicella, A A. Campagnari, J M. Griffiss

Chemistry Faculty Publications

Heterogeneity in the lipooligosaccharides (LOS) of pathogenic Haemophilus and Neisseria species is evident from the multiplicity of components observed with electrophoretic analyses. Knowledge of the precise structures that make up these diverse LOS molecules is clearly the key to reaching an understanding of pathogenic processes such as phase variation and molecular mimicry. Except for a few cases, little is known about the specific structural features of LOS that underlie phase variation and molecular mimicry, partly because of the inherent difficulties in the structural elucidation of these complex glycolipids. In the lipopolysaccharides (LPS) from Salmonella typhimurium and Escherichia coli, rough, or …

Detection Of Serum Antibody Responses In Cattle With Natural Or Experimental Neospora Infections, Patricia Conrad, Karen Sverlow, Mark Anderson, Joan Rowe, Robert Bondurant, Gwen Tuter, Richard Breitmeyer, California Department Of Food And Agriculture, Animal Health Branch, Mark Thurmond, Alex Ardans, J. P. Dubey, Gerarld Duhamel, Bradd Barr

School of Veterinary and Biomedical Sciences: Faculty Publications

Parasite-specific antibody responses were detected using an indirect fluorescent antibody (IFA) test in cattle that were naturally or experimentally infected with Neospora parasites. The test was developed using Neospora tachyzoites isolated from an aborted bovine fetus and grown in bovine cell cultures (isolate BPA1). In all cases, infections were confirmed by the identification of Neospora tachyzoites and/or bradyzoite cysts in fetal or calf tissues using an immunoperoxidase test procedure. Fifty-five naturally infected cows that aborted Neospora-infected fetuses had titers of 320-5,120 at the time of abortion. The titer of 6 cows that were serologically monitored over a prolonged period decreased …

Purification And Ligand Binding Of A Soluble Class I Mhc Molecule Consisting Of The First Three Domains Of H-2kd Fused To B2-Microglobulin Expressed In The Baculovirus/Insect Cell System, Francois Godeau, Immanuel F. Luescher, David M. Ojcius, Cecile Saucier, Estelle Mottez, Lucien Cabanie, Philippe Kourilsky

All Dugoni School of Dentistry Faculty Articles

A recombinant baculovirus encoding a single-chain murine major histocompatibility complex class I molecule in which the first three domains of H-2Kd are fused to beta 2-microglobulin (beta 2-m) via a 15-amino acid linker has been isolated and used to infect lepidopteran cells. A soluble, 391-amino acid single-chain H-2Kd (SC-Kd) molecule of 48 kDa was synthesized and glycosylated in insect cells and could be purified in the absence of detergents by affinity chromatography using the anti-H-2Kd monoclonal antibody SF1.1.1.1. We tested the ability of SC-Kd to bind antigenic peptides using a direct binding assay based on photoaffinity labeling. The photoreactive derivative …

Lysosomal Enzyme Secretion In Tumor Cells And The Purification And Partial Characterization Of A 67 Kda Cathepsin B-Like Enzyme Secreted By B16 Melanoma Cells, Charles Achkar

Dissertations

No abstract provided.

Purification And Characterization Of The Prostate Cancer-Associated Antigen: 7e11-C5, Qi Feng

Theses and Dissertations in Biomedical Sciences

A murine monoclonal antibody (MAb), 7E11-C5, recognizes a prostate organ-specific antigen. Under reducing conditions, the 7E11-C5 antigen exhibited major 100-Kd and minor 70-Kd bands, suggesting that the antigen comprises two peptide chains. The antigen was detected in pooled normal human seminal plasma, xenograft tissue extract of a prostate adenocarcinoma cell line, LNCaP, and tissue extracts of benign prostatic hyperplasia (BPH) and prostate carcinomas, but was not identified in any of the non-prostate tissue extracts tested. Antibodies of the three major prostate-associated antigens (prostatic acid phosphatase (PAP), prostate specific antigen (PSA) and prostate secretory protein (PSP)) were unable to block the …

Cytolytic And Ion Channel Forming Properties Of The N-Terminus Of Lymphocyte Perforin, David M. Ojcius, Pedro M. Persechini, Li-Mou Zheng, Paulo C. Notaroberto, Sandro C. Adeodato, John Ding-E. Young

All Dugoni School of Dentistry Faculty Articles

Perforin lyses cells by binding to the target cell membrane, where it polymerizes into large nonspecific pores. It is shown here that the first 34 amino acids of the N-terminal region of either human or murine perforin are soluble in aqueous medium and spontaneously insert into membranes. The N-terminal peptides lyse liposomes and nucleated cells, and they form ion channels in planar bilayers, some of which are comparable to those previously described for perforin. The lytic activity of the N-terminal domains does not require calcium, is independent of the lipid headgroup composition, and can be inhibited by heparin. Tumor cells …

Cells That Express All Five Proteins Of Vesicular Stomatitis Virus From Cloned Cdnas Support Replication, Assembly, And Budding Of Defective Interfering Particles, Asit K. Pattnaik, Gail W. Wertz

School of Veterinary and Biomedical Sciences: Faculty Publications

An alternative approach to structurefunction analysis of vesicular stomatitis virus (VSV) gene products and their interactions with one another during each phase of the viral life cycle is described. We showed previously by using the vaccinia viruslT7 RNA polymerase expression system that when cells expressing the nucleocapsid protein (N), the phosphoprotein (NS), and the large polymerase protein (L) of VSV were superinfected with defective interfering (DI) particles, rapid and efflicient replication and amplification of DI particle RNA occurred. Here, we demonstrate that all five VSV proteins can be expressed simultaneously when cells are cotransfected with plasmids containing the matrix protein …

Monoclonal Antibodies To The Fusion Protein Of Bovine Respiratory Syncytial Virus, Kent M. Mulkey, Gary A. Anderson

School of Veterinary and Biomedical Sciences: Faculty Publications

Five monoclonal antibodies specific for bovine respiratory syncytial virus were characterized by Western immunoblotting, radioimmunoprecipitation, and epitope mapping assays. The monoclonal antibodies were found to be specific for the fusion protein, and there were at least two antigen binding sites, one of which was neutralizing.

Pharmacokinetics, Immune Response, And Biodistribution Of Iodine-131-Labeled Chimeric Mouse/Human Igg1, K 17-1a Monoclonal Antibody, Ruby Meredith, Albert Lobuglio, Walter Plott, Roger Orr, Ivan Brezovich, Charles Russell, Elizabeth Harvey, Michael Yester, Mansoor Saleh

Haematology and Oncology, East Africa

Pharmacokinetics,immunogenicity,and biodistributionof a 131I-Iabeledmouse/human chimenc monoclonal antibody (C17-1A) was studiedin six metastaticcoloncancerpatients. Pharmacokinetics obtalned from serum radioactivity or chi mera concentration were identical after 5 mCi of 131I(>17-1A withmeanalphahalf-livesof 17.6±2.3 and19.7±2.9 and meanbeta half-livesof 100.9±16.1 and 106.4±14.1 hr, respectively. HPLC analysis documented the monomeric chi rneiic17-lA withoutevidenceof immunecomplexesorfree 1311 None of the patients developed antibody after ‘31I-chi merle17-1A exposure.Radiolocalizationoccurredin known areas of disease >4 cm in all patients. The half-life of total body radioactivity was 58 ±7 hr by whole-body counts and 64 ±13 hr by urine measurements. Whole-body and bone marrow dose estimates ranged from 0.75-1 .03 and …

Replication And Amplification Of Defective Interfering Particle Rn As Of Vesicular Stomatitis Virus In Cells Expressing Viral Proteins From Vectors Containing Cloned Cdnas, Asit K. Pattnaik, Gail W. Wertz

School of Veterinary and Biomedical Sciences: Faculty Publications

Replication and amplification of RNA genomes of defective interfering (DI) particles of vesicular stomatitis virus (VSV) depend on the expression of viral proteins and have untD now been attained only in ceUs coinfected with helper VSV. In the work described in this report, we used a recombinant vaccinia virus-T7 RNA polymerase expression system to synthesize individual VSV proteins in cells transfected with plasmid DNAs that contain cDNA copies of the VSV genes downstream of the T7 RNA polymerase promoter. In this way, we were able to examine the ability of VSV proteins, individually and in combination, to support DI particle …

Efficacy Of A Cell Extract From Actinobacillus (Haemophilus) Pleuropneumoniae Serotype 1 Against Disease In Swine, P. J. Fedorka-Cray, M.J. Huether, D.L. Stine, G. A. Anderson

School of Veterinary and Biomedical Sciences: Faculty Publications

We partially characterized a cell extract (CE) from Actinobacillus pleuropneumoniae serotype 1 and used the CE to test the efficacy of secreted proteins against disease. Secreted products from 4-h culture supernatants were precipitated with 20% polyethylene glycol. Analysis of the CE indicated the presence of protein, endotoxin, and carbohydrate. Hemolytic activity to bovine erythrocytes and cytotoxic activity to porcine mononuclear leukocytes was also demonstrated. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the CE from a 4-h culture showed a major band at 110 kilodaltons (kDa), while a CE of a 26-h culture indicated the presence of a number of additional …

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 …

Improved Selective Medium For The Isolation Of Treponema Hyodysenteriae, Robert A. Kunkle, J. M. Kinyon

School of Veterinary and Biomedical Sciences: Faculty Publications

An agar medium with improved selection for Treponema hyodysenteriae was developed. Cultures of T. hyodysenteriae and T. innocens, feces from 11 clinically normal pigs, and colonic contents from 6 pigs with gross lesions consistent with swine dysentery were diluted in phosphate-buffered saline and plated on Trypticase soy agar (BBL Microbiology Systems, Cockeysville, Md.) with 5% citrated bovine blood (TSA), TSA with 400 ,ug of spectinomycin per ml (TSA-S400), TSA-S400 with 25 ,ug each of colistin and vancomycin per ml, and TSA with 5% pig feces extract and five antimicrobial agents (spiramycin, rifampin, vancomycin, colistin, and spectinomycin) (BJ). Viable numbers …

An Immunochemical Analysis Of Alcoholic Hyaline And Its Relationship To Keratin Intermediate Filaments, David L. Drexler

Loma Linda University Electronic Theses, Dissertations & Projects

Alcoholic Hyaline (AH) and keratin intermediate filaments possess ultrastructural, biochemical and immunochemical similarities. This study compares human AH, human stratum corneum keratin (HSCK) and normal human liver cytokeratin (HCK) by quantitative immunochemical means, using polyclonal as well as monoclonal antibodies. Enzyme-linked immunosorbent assays (ELISAs) were employed, as well as "immunoblotting" and immunocytochemical techniques. HCK was isolated using Triton X-100 and "high-salt” buffers and exhibited seven major polypeptide bands on SDS-PAGE (41-58 kd). Antisera were made toward HCK and the two major HCK bands. Other immunochemicals included antisera toward AH, the three major AH bands, and HSCK, as well as 3 …

Formation Of Influenza Virus Particles Lacking Hemagglutinin On The Viral Envelope, Asit K. Pattnaik, Donald J. Brown, Debi P. Nayak

School of Veterinary and Biomedical Sciences: Faculty Publications

We investigated the intraceUular block in the transport of hemagglutinin (HA) and the role of HA in virus particle formation by using temperature-sensitive (Is) mutants (1s134 and 1s61S) of inOuenza virus AlWSN/33. We found that at the nonpermissive temperature (39.5°C), the exit of ts HA from the rough endoplasmic reticulum to the Golgi complex was blocked and that no additional block was apparent in either the exit from the Golgi complex or post-Golgi complex transport. When MDBK ceUs were infected with these mutant viruses, they produced noninfectious virus particles at 39.5°C. The efficiency of particle formation at 39.5°C was essentiaUy …

Lectins, El-Nasir Lalani, M Wells

Department of Pathology and Laboratory Medicine

No abstract provided.

Identification Of Four Complementary Rna Species In Akabane Virus-Infected Cells, Asit K. Pattnaik, G. Abraham

School of Veterinary and Biomedical Sciences: Faculty Publications

The analysis of RNA extracted from purified Akabane virus demonstrated the presence of three size classes of single-stranded RNAs with sedimentation coefficients of 31S (large, L), 26S (medium, M), and 13S (small, S). Molecular weights of these RNA species were estimated to be 2.15 x 10⁶,1.5 X 10⁶, and 0.48 x 10⁶ for the L, M, and S RNAs, respectively. Hybridization analysis involving viral genomic RNA and RNA from virus-infected cells resulted in the identification of four virus-specific cRNA species in infected cells. These cRNAs were found to be nonpolyadenylated by their inability to bind …

Electron Spin Resonance Studies Of Membrane Fluidity Of Normal And Malignant Human Neural Tissues, Robert P. Wersto

Dissertations

No abstract provided.

Role Of Membrane-Associated Components Of Mycoplasma Arthritidis On The Biosynthesis Of Humoral Antibodies And Arthritogenic Properties In Experimental Animals, Lois M. Bergquist

Loma Linda University Electronic Theses, Dissertations & Projects

The virulence of animal-passaged Mycoplasma arthritidis (158) was demonstrated to be enhanced in rats by growth in oleic acid-enriched broth. The virulence of a laboratory-maintained culture of the same strain was unaffected by similar growth conditions. No differences either in the immune response as measured by complement-fixing (CF) antibodies or in the histogenesis of the disease were detected when rats were injected with animal-passaged M. arthritidis grown either in the presence or in the absence of oleic acid. All tests for metabolism-inhibiting (MI) antibodies in the rat were negative.

A Mycoplasma-associated immunosuppression of the primary and of the secondary hemagglutinin …

The Comparative Histochemistry Of The Infective Oncospheres Of Oochoristica Osheroffi Meggitt, 1934, And Oochoristica Scelopori Voge And Fox, 1950, (Cyclophyllidea: Linstowiidae) With Notes On The Origins Of The Egg Membranes, Gary R. Coulter

Loma Linda University Electronic Theses, Dissertations & Projects

Infective oncospheres of Oochorlstica osheroffi and Oochoristica scelopori were compared using fifteen histochemical tests. Vital stains were also used.

Carnoy’s-fixed gravid proglottids were vacuum-embedded in a paraffin-piccolyte mixture and cut transversely at 6 microns. Sections were tested for the presence of phosphatases, proteins, sclerotin precursors, nucleic acids, carbohydrates and acid mucopolysaccharides.

Oncospheres were histochemlcally very similar. Heavy concentrations of mucoproteins and glycoproteins were detected in the oncospheral coat of O. scelopori. Acid phosphatase was found in the embryophore of O. osheroffi, but could not be demonstrated in O. scelopori. No evidence of a sclerotized egg capsule was …

Cell Free Amino Acid Incorporating Systems From Mesophilic And Thermophilic Bacteria, Nisson Schechter

Masters Theses

No abstract provided.

Viral Infections Of Domestic Animals, George A. Young

School of Veterinary and Biomedical Sciences: Faculty Publications

There is a strong tendency in current research on mammalian virus diseases to concentrate on virus activity at a cellular or subcellular level. This trend is summarized adequately by Enders in relation to viral replication as follows: ','The large and ever-increasing volume of published experimental work on viral replication strikingly reveals the central position of this phenomenon in contemporary virological research. There are good reasons, both biological and practical, underlying this intensive effort to understand, i n detail, how a virus particle, without energy transforming apparatus of its own, manages to utilize the metabolic equipment of the cell to produce …

Worm Eggs: Cost You Money, E.F. Frolik, E.W. Janike

School of Veterinary and Biomedical Sciences: Faculty Publications

Contents

One Million Eggs Daily.......... 3

Damaging Trip......... 4

Worm Remedies......... 5

Modern Worm Remedies......... 6

A Control Program.......... 8

The Corn-Stalk Disease In Cattle, Frank S. Billings

School of Veterinary and Biomedical Sciences: Faculty Publications

MORPHO-BIOLOGICAL CORRESPONDENCE BETWEEN TWO OR MORE MICRO- ETIOLOGICAL ORGANISMS NOT SUFFICIENT GROUNDS FOR PRONOUNCING THE DISEASES WITH WHICH THEY ARE CONNECTED IDENTICAL. The details of this discussion will be found in my report on the swine-plague. It is necessary, however, to touch upon the essential points here also. As was there shown, Hueppe asserts that the European diseases previously mentioned as being caused by a member of this group of belted, ovoid germs, viz., the “ Huhne Cholera, Kaninchen Septikasmie, und Wild Seuche ’ ’ are all one and the same disease, because their micro-etiological organisms have the same form, …

Keratitis Contagiosa In Cattle, Frank S. Billings

School of Veterinary and Biomedical Sciences: Faculty Publications

This is not a new disease by any means, so far as the United States are concerned, nevertheless I have been unable to find any description of it in the literature at my command. While new to myself until the past Summer, there have been quite a number of reports of its existence, and complaints about it, from farmers and breeders of cattle in some of the live-stock journals of our western States. Under these circumstances, it would seem that a description of its clinical phenomena and gross pathological lesions may not be without scientific interest to the opthalmologist, and …