Medicine and Health Sciences Commons^™

Mechanisms Counteracting Swelling In Brain Cells During Hyponatremia, Herminia Pasantes-Morales, Rodrigo Franco, Benito Ordaz, Lenin D. Ochoa

School of Veterinary and Biomedical Sciences: Faculty Publications

Water gain in the brain consequent to hyponatremia is counteracted by mechanisms that initially include a compensatory displacement of liquid from the interstitial space to cerebrospinal fluid and systemic circulation and subsequently an active reduction in cell water accomplished by extrusion of intracellular osmolytes to reach osmotic equilibrium. Potassium (K⁺), chloride (Cl^–), amino acids, polyalcohols, and methylamines all contribute to volume regulation, with a major contribution of ions at the early phase and of organic osmolytes at the late phase of the regulatory process. Experimental models in vitro show that osmolyte fluxes occur via leak pathways …

Evidence For Two Mechanisms Of Amino Acid Osmolyte Release From Hippocampal Slices, R. Franco, M. E. Torres-Marquez, H. Pasantes-Morales

School of Veterinary and Biomedical Sciences: Faculty Publications

A 30% decrease in osmolarity stimulated ³H-taurine, ³H-GABA and glutamate (followed as ³H-D-aspartate) efflux from rat hippocampal slices. ³H-taurine efflux was activated rapidly but inactivated slowly. It was decreased markedly by 100 μM 5-nitro-(3-phenylpropylamino) benzoic acid (NPPB) and 600 μM niflumic acid and inhibited strongly by tyrphostins AG18, AG879 and AG112 (25–100 μM), suggesting a tyrosine kinase-mediated mechanism. Hyposmolarity activated the mitogen-activated protein kinases (MAPK) extracellular-signal-related kinase-1/2 (ERK1/ERK2) and p38, but blockade of this reaction did not affect ³H-taurine efflux. Hyposmosis also activated phosphatidyl-inositide 3-kinase (PI3K) and its prevention by wortmannin (100 nM) essentially abolished …

Amino Acid Osmolytes In Regulatory Volume Decrease And Isovolumetric Regulation In Brain Cells: Contribution And Mechanisms, Herminia Pasantes-Morales, Rodrigo Franco, M. Eugenia Torres-Marquez, Karla Hernandez-Fonseca, Arturo Ortega

School of Veterinary and Biomedical Sciences: Faculty Publications

Brain adaptation to hyposmolarity is accomplished by loss of both electrolytes and organic osmolytes, including amino acids, polyalcohols and methylamines. In brain in vivo, the organic osmolytes account for about 35% of the total solute loss. This review focus on the role of amino acids in cell volume regulation, in conditions of sudden hyposmosis, when cells respond by active regulatory volume decrease (RVD) or after gradual exposure to hyposmotic solutions, a condition where cell volume remains unchanged, named isovolumetric regulation (IVR). The amino acid efflux pathway during RVD is passive and is similar in many respects to the volume-activated anion …

Efflux Of Osmolyte Amino Acids During Isovolumic Regulation In Hippocampal Slices, Rodrigo Franco, Octavio Quesada, Herminia Pasantes-Morales

School of Veterinary and Biomedical Sciences: Faculty Publications

The efflux of potassium (K⁺) and amino acids from hippocampal slices was measured after sudden exposure to 10% (270 mOsm), 25% (225 mOsm) or 50% (150 mOsm) hyposmotic solutions or after gradual decrease (22.5 mOsm/min) in external osmolarity. In slices suddenly exposed to 50% hyposmotic solutions, swelling was followed by partial (74%) cell volume recovery, suggesting regulatory volume decrease (RVD). With gradual hyposmotic changes, no increase in cell water content was observed even when the solution at the end of the experiment was 50% hyposmotic, showing the occurrence of isovolumic regulation (IVR). The gradual decrease in osmolarity elicited …

Isovolumic Regulation In Nervous Tissue: A Novel Mechanism Of Cell Volume Regulation, O. Quesada, R. Franco, K. Hernandez-Fonseca, K. Tuz

School of Veterinary and Biomedical Sciences: Faculty Publications

Cell volume regulation is a property present in most animal cell lineages that allows them to recover their original volume after events of swelling or shrinkage. Such events can be caused by changes in external osmolarity or to osmotic gradients generated during normal cell functioning.4,6 The mechanism of cell volume regulation involves transmembrane fluxes of osmotically active solutes in the necessary direction to counteract the net gain or loss of intracellular water.9 The process through which cells recover their normal volume after swelling is named Regulatory Volume Decrease (RVD). This consists of the efflux of inorganic osmolytes, such as K …

Optimal Replication Activity Of Vesicular Stomatitis Virus Rna Polymerase Requires Phosphorylation Of A Residue(S) At Carboxy-Terminal Domain Ii Of Its Accessory Subunit, Phosphoprotein P, Leroy N. Hwang, Nathan Englund, Tapas Das, Amiya K. Florida, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

The phosphoprotein, P, of vesicular stomatitis virus (VSV) is a key subunit of the viral RNA-dependent RNA polymerase complex. The protein is phosphorylated at multiple sites in two different domains. We recently showed that specific serine and threonine residues within the amino-terminal acidic domain I of P protein must be phosphorylated for in vivo transcription activity, but not for replication activity, of the polymerase complex. To examine the role of phosphorylation of the carboxy-terminal domain II residues of the P protein in transcription and replication, we have used a panel of mutant P proteins in which the phosphate acceptor sites …

Nucleotide Sequences For Detection Of Serpulina Hyodysenteriae, Gerald E. Duhamel, Robert Elder

School of Veterinary and Biomedical Sciences: Faculty Publications

The invention provides a method for detecting the presence of Serpulina hyOdysenteriae in a biological Sample, an oligonucleotide primer and an S. hyodysenteriae-specific oligonucleotide probe useful in that method, and an article of manufacture that contains the primers and/or probe. Also provided are an about 2.3-kb DNA fragment derived from genomic DNA of S. hyodysenteriae and encoding for an about 56 kDa polypeptide, a recombinant expression vector containing the DNA fragment, the 56 kDa polypeptide and a monoclonal antibody reactive with the peptide, and a method of assaying for antibodies reactive with the 56 kDa peptide.

In Situ Hybridization For The Detection And Localization Of Swine Chlamydia Trachomatis, C. Chae, D.-S. Cheon, D. Kwon, O. Kim, B. Kim, J. Suh, D. G. Rogers, K. D. E. Everett, A. A. Anderson

School of Veterinary and Biomedical Sciences: Faculty Publications

Gnotobiotic piglets were inoculated intralaryngeally with swine Chlamydia trachomatis strain R33 or orally with swine C. trachmatis strain R27. Archived formalin-fixed, paraffin-embedded tissues from piglets euthanatized 4–7 days postinoculation were examined by in situ hybridization for C. trachomatis nucleic acid using a nonradioactive digoxigenin-labeled DNA probes that targeted specific ribosomal RNA or omp1 mRNA molecules of the swine C. trachomatis strains. Positive hybridization signals were detected in bronchial epithelial cells, bronchiolar epithelial cells, pneumocytes, alveolar and interstitial macrophages, and jejunal and ileal enterocytes. Chlamydia-infected cells had a strong signal that was confined to the intracytoplasmic inclusions. Positive hybridization signals were …

Plasmd Bearing A Cdna Copy Of The Genome Of Bovine Viral Diarrhea Virus, Chimeric Derivatives Thereof, And Method Of Producing An Infectious Bovine Wral Darrheavirus Using Sad Plasmid, Ruben O. Donis, Ventzislav B. Vassilev

School of Veterinary and Biomedical Sciences: Faculty Publications

A plasmid bearing a cDNA copy of the genome of bovine viral diarrhea virus (BVDV), chimeric derivatives of the plasmid and a method of producing an infectious bovine viral diarrhea virus using the plasmid are disclosed. The invention relates to a plasmid DNA molecule that replicates easily in E. coli and contains a sufficient portion of the genome of BVDV, cloned as cDNA, to be a suitable template to produce RNA in vitro which, upon transfection into bovine cells, gives rise to infectious BVDV. The BVDV created by the process of the invention can be engineered for use as a …

Polyadenylation Of Vesicular Stomatitis Virus Mrna Dictates Efficient Transcription Termination At The Intercistronic Gene Junctions, Leroy N. Hwang, Nathan Englund, Asit K. Pattnaik

School of Veterinary and Biomedical Sciences: Faculty Publications

The intercistronic gene junctions of vesicular stomatitis virus (VSV) contain conserved sequence elements that are important for polyadenylation and transcription termination of upstream transcript as well as reinitiation of transcription of downstream transcript. To examine the role of the putative polyadenylation signal 3' AUACU₇5' at the gene junctions in polyadenylation and transcription termination, we constructed plasmids encoding antigenomic minireplicons containing one or two transcription units. In plasmid-transfected cells, analyses of the bicistronic minireplicon containing the wild-type or mutant intercistronic gene junctions for the ability to direct synthesis of polyadenylated upstream, downstream, and readthrough mRNAs showed that the AUACU …

Phosphorylation Within The Amino-Terminal Acidic Domain I Of The Phosphoprotein Of Vesicular Stomatitis Virus Is Required For Transcription But Not For Replication, Asit K. Pattnaik, Leroy Hwang, Tong Le, Nathan Englund, Manjula Mathur, Tapas Das, Amiya Banerjee

School of Veterinary and Biomedical Sciences: Faculty Publications

Phosphorylation by casein kinase II at three specific residues (S-60, T-62, and S-64) within the acidic domain I of the P protein of Indiana serotype vesicular stomatitis virus has been shown to be critical for in vitro transcription activity of the viral RNA polymerase (P-L) complex. To examine the role of phosphorylation of P protein in transcription as well as replication in vivo, we used a panel of mutant P proteins in which the phosphate acceptor sites in domain I were substituted with alanines or other amino acids. Analyses of the alanine-substituted mutant P proteins for the ability to support …

Bovine Cell Line Resistant To In Vtro Infection By Bovine Viral Darrheavirus And All Other Known Pestiviruses, Ruben O. Donis, Eduardo F. Flores

School of Veterinary and Biomedical Sciences: Faculty Publications

Bovine cell line resistant to infection by the pestiviruses Bovine Viral Diarrhea Virus (BVDV), Hog Cholera Virus (HCV) and Border Disease Virus (BDV) and all progeny and mutants thereof. A bovine cell line (CRIB) that is resistant to infection by cytopathic and non-cytopathic BVDV and by other Pestiviruses due to a stable, recessive genetic defect which blocks infection at the level of viral entry.

Heterozygosity Mapping Of Partially Congenic Lines: Mapping Of A Semidominant Neurological Mutation, Wheels ( Whl), On Mouse Chromosome 4, Patrick M. Nolan, Patricia J. Sollars, Barbara A. Bohne, Warren J. Ewens, Gary E. Pickard, Maja Bucan

School of Veterinary and Biomedical Sciences: Faculty Publications

We identified a semidominant, chemically induced, mouse use mutation with a complex array of abnormal behaviors including bidirectional circling and hyperactivity, abnormal circadian rhythmicity and abnormal responses to light. In this report, we genetically and phenotypically characterized the circling/ waltzing component of the abnormal behavior. We mapped the locus controlling this trait by heterozygosity mapping of partially congenic lines carrying the mutagenized chromosome outcrossed to different inbred strains for three generations. Analysis of 68 PCR-based markers in 13 affected individuals indicated that the mutant locus, named Wheels (Whl), resides in the subcentromeric portion of mouse chromosome 4. The …

Restoration Of Circadian Behavior By Anterior Hypothalamic Heterografts, Patricia J. Sollars, Daniel P. Kimble, Gary E. Pickard

School of Veterinary and Biomedical Sciences: Faculty Publications

The suprachiasmatic nucleus (SCN) of the anterior hypothalamus (AH) is a circadian oscillator and an important component of the mammalian circadian system. To determine whether the SCN is the dominant circadian pacemaker responsible for generating a species-typical characteristic of circadian rhythms [i.e., period length (T)], neural transplantation was conducted using fetal AH donors of different species and SCN-lesioned (SCNx) hosts. The circadian behavior of each of the three donor species is clearly distinguishable by its species-typical T. The extent of SCN pacemaker autonomy was assessed by noting whether the period of the restored circadian rhythm following heterograft transplantation was characteristic …

Certain Canine Weakly Β-Hemolytic Intestinal Spirochetes Are Phenotypically And Genotypically Related To Spirochetes Associated With Human And Porcine Intestinal Spirochetosis, Gerald E. Duhamel, Nagaraja Muniappa, Michelle R. Mathiesen, J. L. Johnson, J. Toth, R. O. Elder, A. R. Doster

School of Veterinary and Biomedical Sciences: Faculty Publications

Four canine weakly β-hemolytic intestinal spirochetes associated with intestinal spirochetosis (IS-associated WBHIS) were compared with IS-associated human and porcine WBHIS and the type species for Serpulina hyodysenteriae and S. innocens by using phenotypic and genotypic parameters. The IS-associated canine, human, and porcine WBHIS belonged to a phyletic group distinct from but related to previously described Serpulina type species.

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 …

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 …

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.

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 …

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 …

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 …

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 …

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 …

Inoculation—Vaccination, Frank S. Billings

School of Veterinary and Biomedical Sciences: Faculty Publications

When one comes to discuss this question with the members of the profession as he casually meets them, he is often surprised to find how uncertain and clouded the prevailing ideas are upon the subject. In fact, the majority of physicians do not seem to have any clear conception of the act connected with the words “ inoculation ” and “ vaccination.”