Digital Commons Network^™

How To Make A Blastocyst., A J Watson, G M Kidder, G A Schultz

Obstetrics & Gynaecology Publications

Several of the new reproductive technologies have been cultivated from our current understanding of the genetic programming and cellular processes that are involved in the major morphogenetic events of mammalian preimplantation development. Research directed at characterizing the patterns of gene expression during early development has shown that the embryo is initially under maternal control and later superseded by new transcriptional activity provided by the activation of the embryonic genome. Several embryonic transcripts encoding: (i) growth factors, (ii) cell junctions, (iii) plasma membrane ion transporters, and (iv) cell adhesion molecules have been identified as contributing directly to the progression of the …

On The Mechanism Of Parathyroid Hormone Stimulation Of Calcium Uptake By Mouse Distal Convoluted Tubule Cells, F A. Gesek, A. Friedman

Dartmouth Scholarship

PTH stimulates transcellular Ca2+ absorption in renal distal convoluted tubules. The effect of PTH on membrane voltage, the ionic basis of the change in voltage, and the relations between voltage and calcium entry were determined on immortalized mouse distal convoluted tubule cells. PTH (10(-8) M) significantly increased 45Ca2+ uptake from basal levels of 2.81 +/- 0.16 to 3.88 +/- 0.19 nmol min-1 mg protein-1. PTH-induced 45Ca2+ uptake was abolished by the dihydropyridine antagonist, nifedipine (10(-5) M). PTH did not affect 22Na+ uptake. Intracellular calcium activity ([Ca2+]i) was measured in cells loaded with fura-2. Control [Ca2+]i averaged 112 +/- 21 nM. …

Mechanism Of Calcium Transport Stimulated By Chlorothiazide In Mouse Distal Convoluted Tubule Cells., Frank A. Gesek, Peter A. Friedman

Dartmouth Scholarship

Thiazide diuretics inhibit Na+ and stimulate Ca2+ absorption in renal distal convoluted tubules. Experiments were performed on immortalized mouse distal convoluted tubule (MDCT) cells to determine the mechanism underlying the dissociation of sodium from calcium transport and the stimulation of calcium absorption induced by thiazide diuretics. Control rates of 22Na+ uptake averaged 272 +/- 35 nmol min-1 mg protein-1 and were inhibited 40% by chlorothiazide (CTZ, 10(-4) M). Control rates of 36Cl- uptake averaged 340 +/- 50 nmol min-1 mg protein-1 and were inhibited 50% by CTZ. CTZ stimulated 45Ca2+ uptake by 45% from resting levels of 2.86 +/- 0.26 …

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 …

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.

Expression Of A Human Cartilage Procollagen Gene (Col2a1) In Mouse 3t3 Cells., Leena Ala-Kokko, James Hyland, Carol Smith, Kari I. Kivirikko, Sergio A. Jimenez, Darwin J. Prockop

Department of Medicine Faculty Papers

Expression in a recombinant system has been difficult to obtain for any of the major fibrillar collagens that require processing by eight or more post-translational enzymes. Here, two DNA constructs were designed so that the promoter region of the gene for the pro-alpha 1(I) chain of human type I procollagen drove expression of the human type II procollagen gene in mouse NIH 3T3 cells, a culture line that normally synthesizes type I procollagen but not any cartilage-specific protein such as type II procollagen. Both constructs were expressed as both mRNA and protein. In clones expressing the construct at high levels, …

Innervation And Vascular Pharmacodynamics Of The Mouse Spleen, Frank D. Reilly

Scanning Microscopy

Neurohistochemical and in vivo and electron microscopic methods demonstrated α-and ,β-adrenergic receptors and adrenergic innervation in arterioles and "arterial" capillaries of the mouse spleen. Such innervation and receptors in venules and channels within the red pulp were sparse. Cholinergic innervation and receptors were judged to be absent in the microvasculature. Histamine elicited arteriolar dilation which was blocked by metiamide suggesting the presence of H₂ receptors. However , following blockade of H₂ receptors, histamine produced arteriolar constriction. Serotonin elicited only venular constriction. Lactic acid caused arteriolar constriction; bradykinin and prostaglandins (PG) E₂ and PGF_2α triggered arteriolar constriction, …

Nucleotide Sequence Of The Epsilon-Subunit Of The Mouse Muscle Nicotinic Acetylcholine Receptor, P D. Gardner

Dartmouth Scholarship

The two predominant types of nicotinic acetylcholine receptors expressed in mammalian muscle differ with respect to a variety of electrophysiological and biochemical properties. A developmental, innervation-dependent switch in the subunit structure of the receptor, in which a y subunit is replaced by an E subunit, is thought to account, in large part, for these differences(1). Because of the interest in the regulatory mechanisms underlying this switch, much attention has focused on these two subunits. Here I report the nucleotide and deduced amino acid sequences of a clone coding for the mouse muscle E subunit isolated from a cDNA library constructed …

Primary Antibody Responses To Thymus-Independent Antigens In The Lungs And Hilar Lymph Nodes Of Mice, S. Niranjan Goud, Alan M. Kaplan, Subbarao Bondada

Sanders-Brown Center on Aging Faculty Publications

B lymphocytes from the pulmonary lymphoid tissues were stimulated with a variety of thymus-independent (TI) antigens by intratracheal (i.t.) immunization. Immune responses in the lungs and hilar lymph nodes (HLN), which are part of the localized lymphoid tissue, as well as in the spleen, the systemic lymphoid organ, were studied. Thus, primary i.t. immunization of mice with the TI-1 antigen trinitrophenyl-lipopolysaccharide (TNP-LPS) elicited both antigen-specific and polyclonal plaque-forming cell responses from HLN, lung, and splenic B lymphocytes. These responses appeared as early as 3 days after immunization and declined by day 7. Similar immunization with another TI-1 antigen, TNP-Brucella …

Mechanism Of Escape Of Endogenous Murine Leukemia Virus Emv-14 From Recognition By Anti-Akr/Gross Virus Cytolytic T Lymphocytes., Hillary D. White, Michael D. Robbins, William R. Green

Dartmouth Scholarship

It was previously shown that spleen cells from endogenous ecotropic murine leukemia virus emv-14+ AKXL-5 mice fail to stimulate an anti-AKR/Gross virus cytolytic T-lymphocyte (CTL) response in a mixed lymphocyte culture with primed C57BL/6 responder spleen cells, whereas spleen cells from AKXL strains carrying the very similar emv-11 provirus do stimulate a response (Green and Graziano, Immunogenetics 23:106-110, 1986). We wished to determine whether the lack of response with AKXL-5 spleen cells was at the level of recognition between effector cell and target cell and whether the relevant mutation was within the emv-14 provirus. It is shown here that EMV-negative …

Murine Gamma Interferon Fails To Inhibit Toxoplasma Gondii Growth In Murine Fibroblasts., Joseph D. Schwartzman, Steven L. Gonias, E R. Pfefferkorn

Dartmouth Scholarship

Although treatment of human macrophages or fibroblasts with human gamma interferon results in the inhibition of intracellular Toxoplasma gondii, murine gamma interferon stimulated only murine macrophages, not murine fibroblasts, to inhibit T. gondii. This species difference may be important in understanding the control of acute and chronic toxoplasmosis.

Immunity To Larval Brugia Malayi In Balb/C Mice: Protective Immunity And Inhibition Of Larval Development., D. Abraham, R. B. Grieve, J. M. Holy, B. M. Christensen

Department of Microbiology and Immunology Faculty Papers

The objective of this study was to analyze the immune response of mice to the larval stages of Brugia malayi. Male BALB/c mice were inoculated with 3 doses of irradiated third-stage larvae (L-3) of B. malayi and were subsequently challenged with L-3 implanted ip within diffusion chambers. After 3 weeks, larvae were recovered to determine their viability, length, and stage of development. A significant reduction in parasite survival was observed in immunized mice. Furthermore, larvae recovered from immunized mice were significantly shorter than larvae recovered from control mice. All larvae recovered from immunized mice were L-3, whereas 96% of larvae …

Physical Linkage Of The Genes For Platelet Membrane Glycoproteins Iib And Iiia., Paul F. Bray, Greg Barsh, Jean-Philippe Rosa, Xue Ya Luo, Ellen Magenis, Marc A. Shuman

Cardeza Foundation for Hematologic Research

The fibrinogen receptor on human platelets is a prototypic member of the integrin family and is composed of subunit glycoproteins IIb (gpIIb) and IIIa (gpIIIa) in a 1:1 stoichiometric ratio. We have isolated cDNA clones for gpIIb and gpIIIa and localized both genes to chromosome 17. In the current study, several approaches were used to localize and map the genes for gpIIb and gpIIIa. A preliminary evaluation of subchromosomal localization was performed by using a panel of mouse-human somatic cell hybrids that contain different amounts of the long arm of human chromosome 17. Southern hybridization to the DNA of these …

K+ Efflux In Nih Mouse 3t3 Cells And Transformed Derivatives: Dependence On Extracellular Ca2+ And Phorbol Esters., Martin Lubin

Dartmouth Scholarship

In culture medium deficient in Ca2+, NIH mouse 3T3 cells lose K+, gain Na+, and stop growing. A marked increase in the rate of K+ efflux accounts for this loss; Na+, K+-ATPase pump activity increases but does not fully compensate for enhanced K+ efflux. Phorbol esters and cycloheximide inhibit K+ loss in Ca2+-deficient medium. Phorbol esters inhibit K+ efflux from human fibroblasts as well, even at physiological levels of Ca2+. Two cell lines derived from NIH-3T3, one transformed by a simian virus 40 deletion mutant, the other by the polyoma virus oncogene encoding the middle-sized tumor antigen, retain K+ and …

Studies On Encephalomyocarditis Virus-Induced Diabetes In Mice, John Henry Huffman

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

The D-variant of encephalomyocarditis (EMC-D) virus was given to SJL/J mice by the artificial, intraperitoneal (ip) route of infection, or by the natural routes of infection per os (po) or intranasal (i.n.), in comparable concentrations. The po route of infection was found to be ineffective. Mice given virus by either the ip or i.n. routes of infection became diabetic. Mice were more resistant to the infectious and diabetogenic properties of the virus when given by the i.n. than when given by the ip route. Glycosuria in mice given virus i.n. usually lagged one day behind that in mice given virus …

A Comparative Study On In Vitro Invasion Rates By Melanoma Cells In The Human Amniotic Basement Membrane Model Versus In Vivo Tumor Nodule Formation In C57bl6 Mice, B. Persky, G. W. Chmielewski

Scanning Microscopy

Invasion by murine B16-F10 melanoma cells was studied using the human amniotic basement membrane (HABM) assay. B16-F10 cells were collected after a single passage through the amnion and grown to near confluency. The cycle of plating, passaging, collecting, and culturing B16-F10 cells was repeated five times. The invasion rate for B16-F10 cells remained relatively unchanged after six passages through the amnion. Injection of first-passage B16-F10 cells into C57BL6 mice resulted in 29 lung tumors per animal whereas sixth-passage cells resulted in 300+ lung tumors. While there exists no correlation of the number of cells penetrating the amnion with colonization number, …

The Homeo Domain Of A Murine Protein Binds 5' To Its Own Homeo Box., Abraham Fainsod, Leonard D. Bogarad, Tarmo Ruusala, Martin Lubin

Dartmouth Scholarship

Nuclear protein extracts from day 12.5 mouse embryos were used to study protein binding to DNA sequences 5' of the Hox 1.5 homeo box. Embryos of this developmental stage are known to express this gene. DNA binding protein blotting and retardation gel techniques show that murine embryonic nuclear proteins specifically bind a 753-base pair (bp) DNA fragment from the region upstream of the Hox 1.5 homeo box. A fusion protein containing the Hox 1.5 homeo domain constructed in lambda gt11 also binds the same 753-bp DNA fragment. Specific binding of the fusion protein to the upstream DNA fragment shows that …

Manganese Toxicity In Mice: Evaluation Of Neruochemical And Immunologial Alteration, Benjamart Srisuchart

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Neurochemical and immunological effects of manganese chloride (MnCl₂) were evaluated using young male CD-1 mice as the animal model. Intraperitoneal (i.p.) injection of MnCl₂ (1, 3 and 10 mg/kg) daily for 4 weeks did not produce mortality or overt signs of toxicity in any of the treated animals. Manganese treatment increased the thymidine uptake of cultured splenic cells and the selective lymphoproliferative response to T-cell mitogens, phytohaemagglutinin and concanavalin A, in all the treated groups. No difference in the response to B-cell mitogen, lipopolysaccharide was observed. A dose of 10 mg/kg MnCl₂ increased the mixed lymphocyte …

Declining Sensitivity To Interleukin 3 Of Murine Multipotential Hemopoietic Progenitors During Their Development: Application To A Culture System That Favors Blast Cell Colony Formation, Kenichi Koike, James N. Ihle, Makio Ogawa

MUSC Faculty Journal Articles

When spleen cells of 5-fluorouracil (5-FU)-treated mice were cultured in the presence of interleukin 3 (IL-3), most colonies revealed IL-3 concentration-dependent colony formation except for mast cell colonies and blast cell colonies. While most colonies were smaller in lower concentrations of IL-3, the size of the blast cell colonies were similar between high and low IL-3 groups. These data suggested that blast cell colony development requires less IL-3 than the development of multilineage colonies from blast cell colonies. This notion was supported by experiments in which IL-3 was added twice to cultures of spleen cells of 5-FU-treated mice. When low …

A Microinjected Monoclonal Antibody Against Human Dna Polymerase-Alpha Inhibits Dna Replication In Human, Hamster, And Mouse Cell Lines., L Kaczmarek, M R Miller, R A Hammond, W E Mercer

Faculty & Staff Scholarship

No abstract provided.

Identification Of Sequences In The Herpes Simplex Virus Thymidine Kinase Gene Required For Efficient Processing And Polyadenylation., Charles N. Cole, Terryl P. Stacy

Dartmouth Scholarship

The herpes simplex virus (HSV) type 1 thymidine kinase gene (tk) was resected from its 3' end with BAL 31 exonuclease. Two sets of plasmids were isolated that lacked information distal to the two copies of the hexanucleotide 5'-AATAAA-3' located at the 3' end of the HSV tk gene. The presence of a simian virus 40 origin of DNA replication in each plasmid facilitated analysis of patterns of transcription in transfected Cos-1 monkey cells. Transcription analyses were performed with an S1 nuclease protection assay. Efficient processing and polyadenylation at the normal site still occurred when all sequences more than 44 …

Ec85-219 1985 Nebraska Swine Report, Roger J. Kittok, James E. Kinder, Rodger K. Johnson, Donald G. Levis, R. K. Christenson, Casey B. Frye, Chris R. Calkins, Roger W. Mandigo, Robert M. Timm, Daryl D. Fisher, Roy Carlson, E.R. Peo Jr., William R. Schneider, Martin L. Wiernusz, Alex Hogg, R. D. Fritschen, Lee I. Chiba, William Ahlschwede, M. C. Brumm, D. Carlson, G.W. Jesse, H.F. Mayes, G.M. Zinn, J.A. Deshazer, Jerry D. Plessing, Dennis D. Schulte, Marcus J. Milanuk, Gerald Bodman, Laverne E. Stetson, Jack L. Schinstock

University of Nebraska-Lincoln Extension: Historical Materials

This 1985 Nebraska Swine Report was prepared by the staff in Animal Science and cooperating departments for use in the Extension and Teaching programs at the University of Nebraska-Lincoln. Authors from the following areas contributed to this publication: Swine Nutrition, swine diseases, pathology, economics, engineering, swine breeding, meats, agronomy, and diagnostic laboratory. It covers the following areas: breeding, disease control, feeding, nutrition, economics, housing and meats.

Inhibition Of Gluconeogenesis And Glycogenolysis By 2,5-Anhydro-D-Mannitol, R L Hanson, R S Ho, J J Wiseberg, R Simpson, E S Younathan, J B Blair

Faculty & Staff Scholarship

No abstract provided.

Monoclonal Antibodies To Novel Myeloid Antigens Reveal Human Neutrophil Heterogeneity., Edward D. Ball, Robert F. Graziano, Li Shen, Michael W. Fanger

Dartmouth Scholarship

Three cytotoxic murine monoclonal antibodies that recognize myeloid-specific antigens have been produced by immunization with normal human neutrophils or myeloblasts from a patient with acute myelomonocytic leukemia. Two of these, PMN 6 and PMN 29, are specific for neutrophils; the third monoclonal antibody, AML-2-23, is reactive with the majority of normal monocytes as well as a subpopulation of mature neutrophils. Although neutrophils from all individuals tested expressed these antigens, cytofluorographic analysis revealed that the percentage of cells bearing the PMN 6 and AML-2-23 antigens varied among individuals. Significant additional heterogeneity in the density of each antigen among antigen-bearing cells was …

Evaluation Of Allergic Hypersensitivity To 2,4-D, Malathion, And Captafol In Mice, Janette R. Cushman

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

2,4-D-specific IgE antibodies were detected in serum of BALB/c mice using the rat passive cutaneous anaphylaxis (PCA) test following the second intraperitoneal immunization with 1, 10, or 100 ug of 2,4-D0keyhole limpet hemocyanin (KLH) conjugate administered with aluminum hydroxide adjuvant. The groups that received 1 ug of 2,4-D conjugate had the highest antibody titers (geometric mean of 60). A paper radioallergosorbent test (PRAST) was developed for determination of 2,4-D-specific IgE. The PRAST was equally as sensitive as and showed a positive correlation with the PCA assay.

The anhydride of the diacid metabolite of malathion (MMA) coupled to KLH elicited MMA-specific …

Toxicologic Effects Of Extracts Of Cestrum Parqui L'Héritier In Mice, Tomás A. López

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

Cestrum parqui L'Héritier is a Solanaceous weed of wide distribution throughout the world. It causes poisoning of animals, particularly cattle and sheep, and constitutes a serious problem for animal production in some countries.

The present study was undertaken to develop a biological test for the detection of toxicity of extracts obtained from the plant. This test will be utilized to assess the toxicity of extracts obtained from the plant in future studies.

The plant is known to induce poisoning in laboratory animals, but there are no precise references on doses to be utilized and of appropriate routes of dosing. Oral …

Cellular Action Of Vasopressin In Medullary Tubules Of Mice With Hereditary Nephrogenic Diabetes Insipidus, Brian A. Jackson, Richard M. Edwards, Heinz Valtin, Thomas P. Dousa

Dartmouth Scholarship

Our previous studies (1974. J. Clin. Invest.54: 753-762.) suggested that impaired metabolism of cyclic AMP (cAMP) may be involved in the renal unresponsiveness to vasopressin (VP) in mice with hereditary nephrogenic diabetes insipidus (NDI). To localize such a defect to specific segments of the nephron, we studied the activities of VP-sensitive adenylate cyclase, cAMP phosphodiesterase (cAMP-PDIE), as well as accumulation of cAMP in medullary collecting tubules (MCT) and in medullary thick ascending limbs of Henle's loop (MAL) microdissected from control mice with normal concentrating ability and from mice with hereditary NDI.

Adenylate cyclase activity stimulated by VP or by …

The Effects Of Tri-O-Tolyl Phosphate (Totp) On The Immune System Of Mice, Craig R. Brinkerhoff

All Graduate Theses and Dissertations, Spring 1920 to Summer 2023

TOTP is a prevalent industrial and environmental contaminant which has been shown to be a neurotoxic agent. This study was undertaken to investigate the effects of this compound on the immune system. Several techniques were employed to test the humoral as well as cellular effects. Male Swiss Webster mice were administered by gastric gavage with three doses of TOTP (5, 50, and 500 mg/kg) and one dose TMTP (50 mg/kg) in corn oil once a week for 13 weeks. Control animals were given corn oil alone.

Lymphocyte transformation was determined on cultures of splenic cells obtained from animals sacrificed at …

Ec80-219 1980 Nebraska Swine Report, Roger W. Mandigo, Louise M. Dalton, Dennis G. Olson, Clayton L. Kelling, Roy Carlson, E. R. Peo Jr., T.J. Janssen, F. Caporaso, Donald G. Levis, Michael Brumm, David P. Shelton, D.M. Danielson, William Ahlschwede, Robert M. Timm, Bobby D. Moser, Elbert C. Dickey, Gerald Bodman, Austin Lewis

University of Nebraska-Lincoln Extension: Historical Materials

This 1980 Nebraska Swine Report was prepared by the staff in Animal Science and cooperating departments for use in the Extension and Teaching programs at the University of Nebraska-Lincoln. Authors from the following areas contributed to this publication: Swine Nutrition, swine diseases, pathology, economics, engineering, swine breeding, meats, agronomy, and diagnostic laboratory. It covers the following areas: breeding, disease control, feeding, nutrition, economics, housing and meats.

Host-Parasite Interaction During Tularemia In The Mouse, Todd Allen Damrow

Graduate Student Theses, Dissertations, & Professional Papers

No abstract provided.