Life Sciences Commons^™

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 …

Book Review: The Baculovirus Expression System: A Laboratory Guide (1992) King, L. A. & Possee, R. D., David D. Dunigan

Nebraska Center for Virology: Faculty Publications

The power of molecular biology is unleashed with the ability to clone and sequence genes, and then express these genes in heterologous systems. This sets the stage for the full analysis of proteins that are otherwise difficult to isolate and/or purify, especially when present at very low copy number per cell or when isolated from relatively precious materials. Overexpression of protein is now possible in a number of systems including prokaryotes (e.g., E. coli) and various eukaryotes (yeast, insects, and plants). The issue then becomes, which system (1) most closely reflects the homologous expression with respect to posttranslational modifications, …

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 …

Book Review, Gianna Julian-Arnold

RISK: Health, Safety & Environment (1990-2002)

Review of the following: BIOMEDICAL POLITICS. (Kathi E. Hanna, ed., National Academy Press 1991) [352 pp.] Preface and acknowledgements, Carl W. Gottschalk, Chair, Institute of Medicine Committee to Study Biomedical Decision Making. Appendices, biographical notes on authors and commentators, index, notes, references. LC 91- 18394, ISBN 0-309-04486-3. [Cloth $29.95. Export $36.00. 2101 Constitution Ave., NW, Washington DC 20418.]

Old Remedies In The Biotechnology Age: Moore V. Regents, Michelle J. Burke, Victoria M. Schmidt

RISK: Health, Safety & Environment (1990-2002)

The California Supreme Court, by excluding a potential remedy, has eliminated many concerns generated by a lower court decision resolving rights to the cell line derived from John Moore's spleen. Yet, the Supreme Court gave little attention to remedies that remain. The authors argue that a constructive trust, a remedy of considerable vintage and great flexibility, could be applied to achieve a just result in such circumstances without affecting the biotechnology world at large.

Immune Responses Of Systemic And Mucosal Lymphoid Organs To Pnu-Imune Vaccine As A Function Of Age And The Efficacy Of Monophosphoryl Lipid A As An Adjuvant, Manju Garg, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

A murine model system was established to study immune responses to the Pnu-Imune vaccine, which is made up of 23 different pneumococcal capsular polysaccharides. In this animal model, antibody-forming cell responses to 21 of 23 individual polysaccharides in the vaccine were detected. The Pnu-Imune vaccine elicited good antibody responses from the spleens and mesenteric lymph nodes (MLN) of young mice, whereas a variety of other peripheral lymph nodes were unresponsive. The immunoglobulin M plaque-forming cell (PFC) response in the spleen to the Pnu-Imune vaccine (given intraperitoneally or subcutaneously) decreased dramatically with increasing age. However, the spleen and MLN differed in …

Book Review, Bradley J. Olson

RISK: Health, Safety & Environment (1990-2002)

Review of the following: THE CODE OF CODES: SCIENTIFIC AND SOCIAL ISSUES IN THE HUMAN GENOME PROJECT. (Daniel J. Kevles & Leroy Hood, eds., Harvard University Press 1992) [397 pp.] Contributors, figures, index, notes, preface, selected bibliography, tables. LC 91-38477, ISBN 0- 674-13645-4. [Cloth $29.95. 79 Garden Street, Cambridge MA 02138.]

Transformation Of A Continuous Rat Embryo Fibroblast Cell Line Requires Three Separate Domains Of Simian Virus 40 Large T Antigen., Jiyue Zhu, Philip W. Rice, Lisa Gorsch, Marina Abate, Charles N. Cole

Dartmouth Scholarship

Mouse C3H 10T1/2 cells and the established rat embryo fibroblast cell line REF-52 are two cell lines widely used in studies of viral transformation. Studies have shown that transformation of 10T1/2 cells requires only the amino-terminal 121 amino acids of simian virus 40 (SV40) large T antigen, while transformation of REF-52 cells requires considerably more of large T antigen, extending from near the N terminus to beyond residue 600. The ability of a large set of linker insertion, small deletion, and point mutants of SV40 T antigen to transform these two cell lines and to bind p105Rb was determined. Transformation …

Cytolytic T Lymphocytes Specific For Tumors And Infected Cells From Mice With A Retrovirus-Induced Immunodeficiency Syndrome., Jennifer G. Erbe, Kathy A. Green, Karen M. Crassi, Herbert C. Morse, W R. Green

Dartmouth Scholarship

LP-BM5 retrovirus complex-infected C57BL/6 mice develop immunodeficiency, somewhat analogous to AIDS, termed murine AIDS (MAIDS). After secondary stimulation with syngeneic B-cell lymphomas from LP-BM5-infected mice, C57BL/6 mice produced vigorous CD8+ cytotoxic T lymphocytes specific for MAIDS-associated tumors. An anti-LP-BM5 specificity was suggested because spleen and lymph node cells from LP-BM5-infected mice served as target cells in competition assays, and cells from LP-BM5, but not ecotropic, virus-infected mice functioned as secondary in vitro stimulators to generate cytotoxic T lymphocytes to MAIDS tumors.

Characterization Of A Novel Prostate Tumor-Associated Antigen, Grayson B. Lipford

Theses and Dissertations in Biomedical Sciences

The murine monoclonal antibodies (MAb) TURP-27 and HNK-1 have been shown to detect antigens that are heavily expressed by benign prostatic hyperplasia (BPH) and carcinoma of the prostate (CaP). Western blot analysis of prostate extracts, showed that MAb TURP-27 and MAb HNK-1 bound glycoproteins of 180, 140, 120, 100, 90 and 69 kDa. Studies have shown that the HNK-1 carbohydrate epitope may be involved in cell adhesion and that it is a component of several characterized adhesion proteins. TURP-27 was found to bind at least three of these adhesion proteins: neural cell adhesion molecules (N-CAM), myelin-associated glycoprotein (MAG) and a …

The Creation Of Transgenic Animal “Models” For Human Genetic Disease, Bernard Rollin

Biomedicine and Animal Models in Research Collection

Transgenic animals will be created to study human genetic disease as soon as the technological capability exists to do so. Extant laws permit such animals to be created. The mindset of the research community makes it inevitable. It is also clear that such diseases can cause enormous amounts of pain and suffering. Responsible researchers need to explore all possible avenues for controlling such pain and suffering. Thus far the research community has not engaged this issue vis a vis animals. The development of methodologies for controlling pain and suffering is likely to be exportable to numerous areas of animal research, …

Priming Of Peripheral Lymph Node B Cells With Tnp-Ficoll: Role Of Lymphokines In B Cell Differentiation, S. Niranjan Goud, Alan M. Kaplan, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

We have previously shown that peripheral lymph node (PLN) B lymphocytes of adult DBA/ 25 mice failed to make an antibody responseto type 2 antigen TNP-Ficoll, but exhibited a good antibody response to type 1 antigen TNP-BruceNa abortus. In the present study we wanted to find out whether the unresponsivenessof PLN B cells to TNP-Ficoll is due to defectsin the early activation and proliferation stage or in the final differentiation stageof B cells. Therefore, we have used a two-step protocol of in vivo immunization of mice with TNP-Ficoll and the subsequent in vitro challenge with TNP-Brucella abortus and studied the …

Multiple Alleles And Estimation Of Genetic Parameters: Computational Equations Showing Involvement Of All Alleles, R Chakraborty

Faculty, Staff and Student Publications

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year‐on‐year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non‐vesicular extracellular particles, characterisation and functional …

Altered Expression Of Adenovirus 12 Dna-Binding Protein But Not Dna Polymerase During Abortive Infection Of Hamster Cells, Lynne A. Lucher, Benjawan Khuntirat, Jiansheng Zhao, Peter C. Angeletti

Nebraska Center for Virology: Faculty Publications

Replication of human adenovirus type 12 DNA is blocked in abortively infected baby hamster kidney cells. The activity and accumulation of adenovirus 12 DNA polymerase is equivalent in infected hamster and human cell extracts. However, the accumulation of adenovirus type 12 DNA-binding protein is approximately 120-fold lower in extracts from infected hamster cells when compared to infected permissive human cells. This difference in accumulation is not because of replication of viral DNA during productive infection, since this difference is observed in the presence of hydroxyurea. The DNA-binding protein from infected hamster cells retains the ability to bind denatured DNA-cellulose. An …

The Ability Of Simian Virus 40 Large T Antigen To Immortalize Primary Mouse Embryo Fibroblasts Cosegregates With Its Ability To Bind To P53., Jiyue Y. Zhu, Marina Abate, Philip W. Rice, Charles N. Cole

Dartmouth Scholarship

The large T antigen encoded by simian virus 40 (SV40) plays essential roles in the infection of permissive cells, leading to production of progeny virions, and in the infection of nonpermissive cells, leading to malignant transformation. Primary mouse embryo fibroblasts (MEFs) are nonpermissive for SV40, and infection by wild-type SV40 leads to immortalization and transformation of a small percentage of infected cells. We examined the ability of an extensive set of mutants whose lesions affect SV40 large T antigen to immortalize MEFs. We found that immortalization activity was retained by all mutants whose lesions are located upstream of codon 346. …

Statistical Interpretation Of Dna Typing Data, R Chakraborty

Faculty, Staff and Student Publications

No abstract provided.

Inclusion Of Data On Relatives For Estimation Of Allele Frequencies, R Chakraborty

Faculty, Staff and Student Publications

No abstract provided.

Mapping The Transcriptional Transactivation Function Of Simian Virus 40 Large T Antigen., Jiyue Y. Zhu, Philip W. Rice, Michele Chamberlain, Charles N. Cole

Dartmouth Scholarship

T antigen is able to transactivate gene expression from the simian virus 40 (SV40) late promoter and from several other viral and cellular promoters. Neither the mechanisms of transactivation by T antigen nor the regions of T antigen required for this activity have been determined. To address the latter point, we have measured the ability of a set of SV40 large T antigen mutants to stimulate gene expression in CV-1 monkey kidney cells from the SV40 late promoter and Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter. Transactivation, although reduced, was retained by an N-terminal 138-amino-acid fragment of T …

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 …

Strain-Specific Neutralizing Determinant In The Transmembrane Protein Of Simian Immunodeficiency Virus, Toshiaki Kodama, Dawn P. Wooley, Daniel P. Silva, Fulvia Dimarzo Veronese, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Monoclonal antibody SF8/5E11, which recognizes the transmembrane protein (TMP) of simian immunodeficiency virus of macaque monkeys (SIVmac), displayed strict strain specificity. It reacted with cloned and uncloned SIVmac251 but not with cloned SIVmac142 and SIVmac239 on immunoblots. This monoclonal antibody neutralized infection by cloned, cell-free SIVmac251 and inhibited formation of syncytia by cloned SIVmac251-infected cells; these activities were specific to cloned SIVmac251 and did not occur with the other viruses. Site-specific mutagenesis was used to show that TMP amino acids 106 to 110 (Asp-Trp-Asn-Asn-Asp) determined the strain specificity of the monoclonal antibody. This strain-specific neutralizing determinant is located within a …

Selection Of Genetic Variants Of Simian Immunodeficiency Virus In Persistently Infected Rhesus Monkeys, Dawn P. Wooley, Ronald C. Desrosiers

Neuroscience, Cell Biology & Physiology Faculty Publications

Genetic and antigenic variation may be one means by which lentiviruses that cause AIDS avoid elimination by host immune responses. Genetic variation in the envelope gene (env) was studied by comparing the nucleotide sequences of 27 clones obtained from two rhesus monkeys infected with molecularly cloned simian immunodeficiency virus. All 27 clones differed from each other and differed from the input clone in the gp120 (SU) portion of the envelope gene. Nucleotide substitutions were shown to accumulate with time at an average rate of 8.5 per 1,000 per year in SU. Surprisingly, the majority of nucleotide substitutions (81%) resulted in …

Catecholamines And The Regulation Of Uterine Contractions In The Pregnant Rhesus Macaque, Cahleen Mae Mcnutt

Loma Linda University Electronic Theses, Dissertations & Projects

This study was designed to test the hypothesis that: 1) maternal and/or amniotic fluid catecholamines or prostaglandins display nocturnal activity rhythms similar to those of the myometrial contractility in the pregnant rhesus macaque and, 2) catecholamines stimulate prostaglandin production by fetal membranes. Paired maternal arterial blood and amniotic fluid samples were collected at 0900 h (AM) and 2100 h (PM) from chronically catheterized rhesus macaques until delivery and analyzed for prostaglandin metabolites (PGFM and PGEM-II) and catecholamines (norepinephrine, epinephrine, and dopamine). Uterine activity data was continuously recorded. A rhythm in uterine contractility was observed, with peak activity between 1900 and …

Molecular Mechanism Of The Stimulation Of Alkaline Phosphatase Activity In Human Bone Cells By 1,25(Oh)2 D3, Eru Kyeyune-Nyombi

Loma Linda University Electronic Theses, Dissertations & Projects

To facilitate this study an in vitro human model system was established that exhibited many aspects of normal osteoblasts. The human osteosarcoma cell line (TE85 cells) expressed a skeletal alkaline phosphatase activity (an accepted bone cell differentiation marker) which was stimulated by 1,25(OH)₂D₃ (a potent differentiating agent), under serum-free conditions in a dose-dependent, time-dependent, and cell density-dependent manner. Cytochemical analysis of the stimulation of ALP activity by 1,25(OH)₂D₃ showed that 1,25(OH)₂D₃ increased the number of TE85 cells that expressed detectable ALP activity, suggesting that 1,25(OH)₂D₃ promoted the process …

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 …

Physiology Of Murine B Lymphocytes Ii. Life-Spans Of Mitogen And Thymus-Independent Antigen (Type 2) Reactive B Lymphocytes From Aged Mice, Venkatachalam Udhayakumar, S. Niranjan Goud, Richard J. Cross, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

We showed earlier that life spans of murine B lymphocytes could be estimated by measuring the functional reactivities of normal B cells upon transfer into x-linked immunodeficient (xid) mice, which do not respond to anti-mouse IgM (anti-μ) antibodies and thymic-independent type -2 (TI-2) antigens. Here the same approach was adopted, to evaluate the life spans of B-lymphocytes from aged mice. Spleen cells from normal young and aged mice were transferred into young or aged xid recipients and the decay kinetics were followed by measuring the proliferative response to anti-μ and PFC response to TNP-Ficoll, a prototype TI-2 antigen. The results …

The Effects Of Glutathione On Human Keratinocyte, Manisha N. Trivedi

Theses and Dissertations in Biomedical Sciences

Cultured human keratinocytes were grown in vitro to assess the effects of glutathione, a thiol, on human keratinocyte proliferation. Single keratinocytes were harvested from newborn (greater than 5 days old) foreskin and were grown in culture flasks such that experiments could be performed in approximately 2 to 3 weeks. In order to evaluate the effects of glutathione (GSH) on keratinocyte growth, experiments were performed to determine the growth curve, plating efficiency, and incorporation of radiolabelled thymidine. Growth studies, performed with different concentrations of glutathione, revealed that 0.065 μM was the most effective concentration when compared to control cell cultures, and …

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.

Proto-Oncogene Regulation By Growth Factors In Bone Cells, Harold Lyndon Merriman

Loma Linda University Electronic Theses, Dissertations & Projects

Previous studies have shown that bone cells in culture produce a number of growth factors that are important in bone regulation. Chick and mouse primary calvarial cultures consist of a mixture of cell types that make it difficult to interpret the results from these model systems. In contrast, MC3T3-E1 cells are a clonal mouse osteoblast-like cell line. Since MC3T3-E1 cells consist of a single population of calvarial cells, they make an ideal system in which to study the autocrine effects of bone growth factors.

Based on this work, MC3T3-E1 cells are now known to produce IGF-I, TGF-beta and IGF-II in …

The Growth Of Simian Virus 40 (Sv40) Host Range/Adenovirus Helper Function Mutants In An African Green Monkey Cell Line That Constitutively Expresses The Sv40 Agnoprotein., Terryl P. Stacy, Michele Chamberlain, Susan Carswell, Charles N. Cole

Dartmouth Scholarship

The simian virus 40 T-antigen carboxy-terminal mutants, dlA2459 and dlA2475, are cell line and temperature dependent for growth and plaque formation in monkey kidney cells. Although these mutants did form plaques on BSC-1 cells at 37 degrees C, they were about fivefold less efficient for plaque formation than wild-type simian virus 40. These mutants did not grow in CV-1 cells and did not synthesize agnoprotein in those cells. CV-1 cells which constitutively express the agnoprotein were permissive for mutant plaque formation. However, late mRNAs, virion proteins, and progeny virion yields did not accumulate to wild-type levels during mutant infection of …

Effect Of Dietary Aluminum Sulfate On Calcium And Phosphorus Metabolism Of Broiler Chicks, Akmed S. Hussein, Austin H. Cantor, Thomas H. Johnson, Robert A. Yokel

Animal and Food Sciences Faculty Publications

The effect of dietary aluminum sulfate on Ca and P metabolism was studied using 1-day-oldmale broiler chicks. In Experiment 1, practical diets providing .90% Ca plus .45% available P (P_av), .90% Ca plus .78% P_av, 1.80% Ca plus .45% P_av, or 1.80% Ca plus .90% P_av were fed with 0 or .392% Al as aluminum sulfate for 21 days. The control diet (.90% Caplus .45% P_av) without added A1 was fed to all chicks during Days 22 to 49. In general, Al significantly (Pi), tibia breaking strength, tibia weight, percentage of …