Microbiology Commons^™

A Preclinical Model Of Ionizing Radiation And Tnf-Alpha Gene Therapy For Cancer, Jun Li

Loma Linda University Electronic Theses, Dissertations & Projects

Long term control of high-grade brain tumors is rarely achieved with current therapeutic regimens. This study sought to determine if pretreatment with plasmid DNA expressing Tumor Necrosis Factor-α (TNF-α) could enhance the effect of ionizing radiation in an aggressive, rapidly growing glioma tumor (C6) model. Plasmid DNA is an attractive vector system for delivery of foreign genes into eukaryotic cells due to its advantages including low immunogenicity in vivo, lesser toxicity and relatively low cost. We have constructed a plasmid-based mammalian expression vector (pGL1-TNF) to secrete human TNF-α from cancer cells. Plasmid DNA was successfully transfected into C6 cells …

Azidothymidine And Interferon-Α Induce Apoptosis In Herpesvirus-Associated Lymphomas, Richard K. Lee, Jian-Ping Cai, Vadim Deyev, Parkash Gill, Lisa Cabral, Charles Wood, Ram Agarwal, Wenle Xia, Lawrence Boise, Eckhard Podack, William Harrington Jr.

Nebraska Center for Virology: Faculty Publications

Lymphoproliferative diseases that occur in immunocompromised patients are frequently associated with herpesviruses. These patients often fare poorly after treatment with conventional chemotherapy. We reported previously that patients with AIDS-related Burkitt’s lymphoma (BL) responded to parenteral azidothymidine (AZT) and IFN-α. We found that EBV-positive lymphoma cells derived from these patients cultured with AZT express CD95 and undergo apoptosis. AZT-mediated apoptosis was caspase dependent and occurred despite Fas receptor blockade. In contrast, EBV-negative lymphomas were resistant to AZT-induced apoptosis, as were EBV-positive lymphomas that expressed high levels of bcl-2. Primary effusion lymphoma (PEL) cell lines infected with human herpesvirus type 8 required …

Arthritis And Rosai-Dorfman Disease Of The Skin: A Diagnostic Dilemma, Irshad Nabi Soomro, Kamran Hameed

Department of Pathology and Laboratory Medicine

No abstract provided.

Bioavailability And Trophic Transfer Of Humic-Bound Copper From Bacteria To Zooplankton, Emile M. Lores, Jonathan Pennock

School of Marine Science and Ocean Engineering

ABSTRACT:

The effect of humic acid (HA) on uptake and transfer of Cu by selected marine organisms from the microbial loop was determined. Bacteria grown with and without 15 µg Cu l^-1 and with and without 10 mg Suwannee River Humic Acid (SRHA) l^-1 were fed to Uronema sp. The Uronema were subsequently fed to Acartia tonsa to determine the effect of humic acid on the uptake and transfer of Cu from bacteria to copepods. The presence of 10 mg SRHA l^-1 reduced Cu uptake in A. tonsa by an average of 54% and significantly reduced the …

Lymphotropic Virions Affect Chemokine Receptor-Mediated Neural Signaling And Apoptosis: Implications For Human Immunodeficiency Virus Type 1-Associated Dementia, Jialin Zheng, Anuja Ghorpade, Douglas Niemann, Robin L. Cotter, Michael R. Thylin, Leon Epstein, Jennifer Swartz, Robin B. Shepard, Xiaojuan Liu, Adeline Nukuna, Howard E. Gendelman

Nebraska Center for Virology: Faculty Publications

Chemokine receptors pivotal for human immunodeficiency virus type 1 (HIV-1) infection in lymphocytes and macrophages (CCR3, CCR5, and CXCR4) are expressed on neural cells (microglia, astrocytes, and/or neurons). It is these cells which are damaged during progressive HIV-1 infection of the central nervous system. We theorize that viral coreceptors could effect neural cell damage during HIV-1-associated dementia (HAD) without simultaneously affecting viral replication. To these ends, we studied the ability of diverse viral strains to affect intracellular signaling and apoptosis of neurons, astrocytes, and monocyte-derived macrophages. Inhibition of cyclic AMP, activation of inositol 1,4,5-trisphosphate, and apoptosis were induced by diverse …

A Study On The Effects Of The N-Terminal Amino Acid Sequence On The Activation Of Human T-Cell Leukemia Virus Type 1 Protease, Hidayah Muhammad Kendall

Chemistry & Biochemistry Theses & Dissertations

Human T-cell leukemia virus type 1 (HTL V-1) is dependent upon the enzymatic activity of its protease for maturation. Maturation of the protease is facilitated by cleavage of specific amino acid residues, followed by dimerization. The effects of the amino acid sequence located N-terminally to the cleavage site on the ability of the protease to become active were the focus of the current study. These amino acid sequences were contributed by the plasmid vector into which the protease gene was inserted.

Surface probability analyses (SPAs) of the vectors, as well as for native sequences which produce the mature protease and …

P53 And Pcna Expression In Benign, Atypical And Malignant Meningiomas, R Ahmed, I N. Soomro, S A. Aziz, S H. Hasan

Department of Pathology and Laboratory Medicine

Objective: Alterations: p53 genes are turning out to be the most common genetic alterations in human cancers. Due to long half-life of mutated p53, its detection is possible by immunohistochemistry. Proliferating cell nuclear antigen (PCNA) is expressed by dividing cells, hence has been shown to correlate with prognosis. We have used monoclonal antibodies protein DO-7 (p53) and PC10 (PCNA) to see whether their expression correlates with histological grading in meningethelial tumour.
Material and Methods: a Twenty nine meningiomas (20 benign, 7 atypical and 2 malignant) were selected from the records of our laboratory. p53 and PCNA expression was sought by …

Spinal Cord Compression Caused By Metastatic Epithelial Myoepithelial Carcinoma Of The Parotid Gland, I N. Soomro, A S. Hussainy, K Chishti, M H. Pui, S A. Khan, R Ahmed, S H. Hasan

Department of Pathology and Laboratory Medicine

No abstract provided.

Different Mechanisms Of C-Jun Nh2-Terminal Kinase-1 (Jnk1) Activation By Ultraviolet-B Radiation And By Oxidative Stressors, Mihail S. Iordanov, Bruce E. Magun

CUP Faculty Research

Irradiation of mammalian cells with ultraviolet-B radiation (UV-B) triggers the activation of a group of stress-activated protein kinases known as c-Jun NH₂-terminal kinases (JNKs). UV-B activates JNKs via UV-B-induced ribotoxic stress. Because oxidative stress also activates JNKs, we have addressed the question of whether the ribotoxic and the oxidative stress responses are mechanistically similar. The pro-oxidants sodium arsenite, cadmium chloride, and hydrogen peroxide activated JNK1 with slow kinetics, whereas UV-B potentiated the activity of JNK1 rapidly.N-acetyl cysteine (a scavenger of reactive oxygen intermediates) abolished the ability of all oxidative stressors tested to activate JNK1, but failed …

Evolutionary Endocrinology Of Juvenile Hormone Esterase: Functional Relationship With Wing Polymorphism In The Cricket, Gryllus Firmus, Anthony J. Zera, Yuan Huang

Anthony Zera Publications

The existence, nature, and physiological consequences of genetic variation for juvenile hormone esterase (JHE) activity was studied in the wing-polymorphic cricket, Gryllus firmus. Hemolymph (blood) JHE activity was sixfold lower in nascent short-winged (SW) females, relative to nascent long-winged (LW) females, during the last juvenile stadium (stage). Morph-associated genetic variation for JHE activity had two causes, variation in loci: (1) regulating whole-organism enzyme activity; and (2) controlling the degree to which JHE is secreted into the blood. Reduced JHE activity in nascent SW-selected individuals was associated with reduced in vivo juvenile hormone catabolism. This suggests that variation in JHE …

The Endocrine Genetics Of Wing Polymorphism In Gryllus: Critique Of Recent Studies And State Of The Art, Anthony J. Zera

Anthony Zera Publications

In a series of papers, the most recent of which was published in Evolution, quantitative-genetic experiments were undertaken on reproductive and physiological correlates of wing polymorphism in the sand cricket, Gryllus firmus (Fairbairn 1994; Fairbairn and Yadlowski 1997; Roff et al. 1997). A goal of these studies was to determine the physiological causes underlying: (1) genetic variation for an ecologically important threshold trait (wing polymorphism); and (2) genetic correlations between wing morph and other reproductive and migratory features, such as fecundity and flight muscle histolysis. These authors concluded that genetic variation for the activity of the endocrine regulator, juvenile hormone …

Development Of A Novel Method For Phenol Detection Using High Performance Thin Layer Chromatography, Danette Young Carlton

Theses & Honors Papers

The purpose of this study was to develop a more efficient method to test for phenol in both wastewater and non-wastewater. Phenol is a caustic and toxic contaminant that may be present in our water supplies. Phenol is used in various resins, pharmaceuticals, plastics and disinfectants. Environmental Protection Agency methods are available to test for phenol using gas chromatography along with mass spectrometry or flame ionization detection. Each of these methods can be a long, involved process consisting of extraction, many sample injections, and interference problems. This method developed an easier and less complicated approach of testing for phenol in …

Succinate Dehydrogenase (Sdh) From Bradyrhizobium Japonicum Is Closely Related To Mitochondrial Sdh, David J. Westenberg, Mary Lou Guerinot

Dartmouth Scholarship

The sdhCDAB operon, encoding succinate dehydrogenase, was cloned from the soybean symbiont Bradyrhizobium japonicum. Sdh from B. japonicum is phylogenetically related to Sdh from mitochondria. This is the first example of a mitochondrion-like Sdh functionally expressed in Escherichia coli.

Genetic And Structural Analyses Of Cytoplasmic Filaments Of Wild-Type Treponema Phagedenis And A Flagellar Filament-Deficient Mutant, Jacques Izard, William A. Samsonoff, Mary Beth Kinoshita, Ronald J. Limberger

Department of Food Science and Technology: Faculty Publications

Unique cytoplasmic filaments are found in the treponeme genus of spirochete bacteria. Their function is unknown, but their location underneath the periplasmic flagellar filaments (PFF) suggests a role in motility and/or cell structure. To better understand these unique structures, the gene coding for the cytoplasmic filaments, cfpA, was identified in various treponemal species. Treponema phagedenis cfpA was 2,037 nucleotides long, and the encoded polypeptide showed 78 to 100% amino acid sequence identity with the partial sequence of CfpA from T. denticola, T. vincentii, and T. pallidum subsp. pertenue. Wild-type T. phagedenis and a …

Cullin-3 Targets Cyclin E For Ubiquitination And Controls S Phase In Mammalian Cells, Jeffrey D. Singer, Mark Gurian-West, Bruce Clurman, James M. Roberts

Biology Faculty Publications and Presentations

Cyclin E is an unstable protein that is degraded in a ubiquitin- and proteasome- dependent pathway. Two factors stimulate cyclin E ubiquitination in vivo: when it is free of its CDK partner, and when it is phosphorylated on threonine 380. We pursued the first of these pathways by using a two-hybrid screen to identify proteins that could bind only to free cyclin E. This resulted in the isolation of human Cul-3, a member of the cullin family of E3 ubiquitin–protein ligases. We found that Cul-3 was bound to cyclin E but not to cyclin E-Cdk2 complexes in mammalian cells, and …

A Vibrio Cholerae Lysr Homolog, Aphb, Cooperates With Apha At The Tcpph Promoter To Activate Expression Of The Toxr Virulence Cascade, Gabriela Kovacikova, Karen Skorupski

Dartmouth Scholarship

We describe here a new member of the LysR family of transcriptional regulators, AphB, which is required for activation of the Vibrio cholerae ToxR virulence cascade. AphB activates the transcription of the tcpPH operon in response to environmental stimuli, and this process requires cooperation with a second protein, AphA. The expression of neither aphA or aphB is strongly regulated by environmental stimuli, raising the possibility that the activities of the proteins themselves may be influenced under various conditions. Strains of the El Tor biotype of V. cholerae typically exhibit lower expression of ToxR-regulated virulence genes in vitro than classical strains …

An Extracellular Protein Produced By Staphylococcus Hominis Strain Αm With Antimicrobial Activity Against Mycobacterium Spp, Sandra May Jacobsen

Biological Sciences Theses & Dissertations

The inhibitory activity exhibited by Staphylococcus hominis strain αM toward Mycobacterium species was examined. These studies included determining the conditions that maximized inhibitory agent production, analyzing characteristics of the agent, determining the time of agent production, ascertaining plasmid possession and the role of plasmids in the production of this inhibitory agent, and ascertaining the antimicrobial activity of other strains of S. hominis. On solid and in liquid media, S. hominis strain αM exhibits a unique inhibitory activity toward Mycobacterium species that is not demonstrated by Staphylococcus epidermidis or other S. hominis strains. Maximum agent production and recovery was …

Broad-Spectrum Protection Against Tombusviruses Elicited By Defective Interfering Rnas In Transgenic Plants, Teresa Rubio, Marise Borja, Herman Scholthof, Paul A. Feldstein, Thomas Jack Morris, Andrew O. Jackson

Nebraska Center for Virology: Faculty Publications

We have designed a DNA cassette to transcribe defective interfering (DI) RNAs of tomato bushy stunt virus (TBSV) and have investigated their potential to protect transgenic Nicotiana benthamiana plants from tombusvirus infections. To produce RNAs with authentic 5’ and 3’ termini identical to those of the native B10 DI RNA, the DI RNA sequences were flanked by ribozymes (RzDI). When RzDI RNAs transcribed in vitro were mixed with parental TBSV transcripts and inoculated into protoplasts or plants, they became amplified, reduced the accumulation of the parental RNA, and mediated attenuation of the lethal syndrome characteristic of TBSV infections. Analysis of …

Insertional Inactivation Of Treponema Denticola Tap1 Results In A Nonmotile Mutant With Elongated Flagellar Hooks, Ronald J. Limberger, Linda L. Slivienski, Jacques Izard, William A. Samsonoff

Department of Food Science and Technology: Faculty Publications

The treponemal fla operon is comprised of numerous motility-related genes; however, the initial gene of this operon, tap1, has no known function. A recently developed system to generate specific mutants in Treponema denticola was utilized to determine if Tap1 was essential for motility. T. denticola tap1 and flanking DNA were identified, cloned, and sequenced, and a suicide plasmid that contained tap1 interrupted with an erythromycin resistance cassette (ermF and ermAM) was constructed. Because of potential polar effects from this cassette, a second plasmid that contained tap1 interrupted with a modified erythromycin resistance cassette that lacked the putative ermF transcription …

An Economic Analysis Of The Threat Of An Exotic Disease On Agriculture: The Case Of Sheep Pox In Western Australia, Nhu Che, Tom Kompas

Biosecurity published reports

Sheep pox is an acute disease that causes heavy losses due to high sheep mortality, costly eradication programs and ongoing problems with trade restrictions for many countries around the world. Australia until now has been unaffected, and although the risk of disease entry may be very slight, one case of sheep pox outbreak could cost the Western Australian economy as much as $50 million in damages, mostly from the trade disruption losses ( over the 21-month period of trading bans). The introduction of a Serology Test Program may reduce that amount by $35 million due to a shortening of the …

Functional Analysis Of Human Igfbp-6 In Human Osteoblasts, Tao Yan

Loma Linda University Electronic Theses, Dissertations & Projects

Insulin-like growth factor binding proteins (IGFBPs) are a family of six secreted proteins that bind to and modulate the functions of insulin-like growth factors (IGF-I and IGF-II). As a relatively new member of the IGF system, IGFBP-6 was found to be produced by human osteoblasts and exert significant biological effects on human osteoblasts.

Our previous studies indicated that all-trans retinoic acid (ATRA) caused more than a10 fold increase in IGFBP-6 production and a50-60% reduction of ALP activity in human osteoblasts. Based on these findings, we proposed the hypothesis that ATRA induced human osteoblast differentiation was mediated at least in part …

Use Of Bioinformatics In The Analysis Of Chemotaxis Proteins, Sean Andrew Bulloch

Loma Linda University Electronic Theses, Dissertations & Projects

Bacterial chemotaxis is one of the best-known signal transduction systems. Levels of attractants and repellents are sensed in the surrounding environment by various chemoreceptors. The signal is passed to the excitation pathway via the transfer of a phosphoryl group from the receptor-associated histidine kinase CheA to the response regulator CheY. Phospho-CheY binds to the flagellar motor switching the direction of rotation of the flagella and thus allowing the cell to move towards or away from the attractant or repellent. The phosphatase CheZ removes the phosphoryl group from phospho-CheY restoring default flagellar rotation. Adaptation to stimuli occurs via addition of methyl …

Antitumor Effect Of Il-2, P53, And Bax Naked Gene Transfer In C6 Glioma Cells, Peyman Haghighat

Loma Linda University Electronic Theses, Dissertations & Projects

[Abstract not included]

Acute Effects Of Whole-Body Proton Irradiation On The Immune System Of The C57bl/6 Mouse, Eric H. Kajioka

Loma Linda University Electronic Theses, Dissertations & Projects

The acute effects of proton whole-body irradiation (WBI) on leukocytes, lymphocytes, and hematological parameters in the spleen and blood of C57BI/6 mice were examined and compared to the effects of photon (gamma) WBI derived from a 60cobalt (60Co) source. Adult, female C57BL/6 mice were exposed to a single dose (3 Gy, 0.4 Gy/min dose rate) of either proton WBI at the Bragg peak, proton WBI at the entry plateau, or Co WBI, and sacrifice intervals were at 1,4, 7, and 10 days post- WBI. Flow cytometry analysis of the spleen and peripheral blood showed depression in cell counts for all …

Dna-Dna Checkerboard Hybridization To Identify Bacteria In Human Plaque Samples, Jasan Lee Zimmerman

Loma Linda University Electronic Theses, Dissertations & Projects

Previous research has shown that using DNA probes to identify the presence of specific bacteria in human plaque samples is superior to the commonly used anaerobic cultural procedures. Some periodontal pathogens are either uncultivable or very difficult to grow, such as Prevotella intermedia, P. nigrescens, and Eubacterium brachy. Digoxygenin probes were prepared for 15 oral bacteria. These probes were used with a checkerboard DNA-DNA hybridization scheme to analyze whether human plaque samples contained any of the 15 bacterial species. Our results have shown that this method is successful in identifying bacterial DNA present in the subgingival patient plaque …

Morphological Pattern And Frequency Of Thyroid Tumors, S H. Shah, S Muzaffar, I N. Soomro, Sheema H. Hasan

Department of Pathology and Laboratory Medicine

Objective:
The present study was done to evaluate the frequency of thyroid cancer and to find out the prevalence of histological types of thyroid tumor with respect to age and sex group.
Setting:
This study included consecutive cases of malignant tumors of thyroid gland, which were diagnosed in the Department of Pathology at the Aga Khan University Hospital, Karachi during the period of three years (1995-1997).
Methods:
These cases were evaluated on H & E stained sections from paraffin embedded 10% buffered formalin fixed tissue blocks. Special stains and immunohistochemical analysis were performed whenever required.Results: A total of 8541 malignant …

The Timing And Pattern Of Myogenesis In Hymenochirus Boettgeri, Matthew T. Smetanick, Rafael O. De Sá

Biology Faculty Publications

Differences in the relative timing of homologous developmental events among closely related species, known as heterochronies, may provide valuable clues in understanding evolutionary relationships (McKinney, 1988; McNamara, 1995). Examining the timing of myogenic events is a relatively easy and effective method for finding heterochronic events. For example, whether muscle proteins and myofibrils appear before or after multinucleation can be determined through histological techniques (Kielbowna, 1981). Simple observations of live specimens can pinpoint functional landmarks such as first twitch (spontaneous or due to external stimuli) and first heartbeat.

Engineering Secreted Proteins For Gene Transfer And Dna Vaccination, Jingxue Liu

Loma Linda University Electronic Theses, Dissertations & Projects

In recent years gene therapy has become a promising way of alleviating incurable human ailments, its concept emerging as the ultimate therapy for many infectious and genetic diseases. Two important aspects of the development of successful gene therapy protocols are the ability to monitor gene transfer readily, and the establishment of new protocols for treating specific diseases. In this work, Renilla luciferase and human glutamic acid decarboxylase (GAD) 65 were engineered for secretion to address some aspects of these issues.

Secreted reporter proteins are promising tools to study gene transfer and expression in a non-destructive manner, and bioluminescent proteins are …

Control Of Cell Shape And Elongation: The Mreb Protein Of Escherichia Coli., Jason Gioia

Honors Capstones

No abstract provided.

Multicenter Comparison Of Pcr Assays For Detection Of Human Herpesvirus 8 Dna In Semen, Philip E. Pellett, Thomas J. Spira, Omar Bagasra, Chris Boshoff, Lawrence Corey, Laura De Lellis, Meei-Li Huang, Jung-Chung Lin, Steve Matthews, Paolo Monini, Paola Romessi, Carlos Sosa, Charles Wood, John A. Stewart

Nebraska Center for Virology: Faculty Publications

Reported prevalences of human herpesvirus 8 (HHV-8) (Kaposi’s sarcoma-associated herpesvirus) in semen have ranged widely. This is possibly due to differences in assay sensitivity, geographic or population-based differences in the true presence of the virus in semen, and PCR contamination. This study assessed interlaboratory sensitivity and reproducibility in the analysis of blinded experimental panels, each consisting of 48 specimens and being composed of semen specimens from different healthy artificial-insemination donors (n = 30) and human immunodeficiency virus (HIV)-infected patients (n= 7) plus positive (n = 4) and negative (n = 7) controls. The experimental panels analyzed in each laboratory were …