Molecular Genetics Commons^™

Reversal Of Age-Associated Decline In Immune Response To Pnu-Imune Vaccine By Supplementation With The Steroid Hormone Dehydroepiandrosterone, Manju Garg, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Recently, we reported that murine antibody responses to the 23-valent pneumococcal polysaccharide (Pnu-Imune) vaccine declined with age. Here we present data to support the concept that age-associated immune defects are not only due to intrinsic defects in immune cells but are also due to extrinsic factors emanating from the neuroendocrine system. We found that supplementation with dehydroepiandrosterone, a steroid hormone known to be reduced in the aged, corrects the immune deficiency of aged mice and significantly enhanced their splenic immune responses to the Pnu-Imune vaccine.

A Family Showing No Evidence Of Linkage Between The Ataxia Telangiectasia Gene And Chromosome, D. Hernandez, C. M. Mcconville, Michael W. Stacy, C. G. Woods, M. M. Brown, P. Shutt, G. Rysiecki, A. M. R. Taylor

Bioelectrics Publications

We have studied an inbred family in which two cousins presented with the same clinical features of ataxia telangiectasia (AT). Both patients are still ambulatory at ages 25 and 20. Cellular features of both patients are typical of AT and include increased radiosensitivity and an increased level of spontaneously occurring chromosome aberrations in peripheral blood lymphocytes. Linkage studies and haplotype analysis show no clear evidence that the gene for AT in this family is on chromosome 11q22-23. As previously reported AT families from complementation groups AB, C, and D have all shown linkage to this region of 11q22-23. Our study …

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 …

The Expression And Regulation Of The Hsdk Genes Of Escherichia Coli K-12, Ainu Parkash

Loma Linda University Electronic Theses, Dissertations & Projects

The DNA restriction and modification (R-M) genes in bacteria play a defensive role in the organism, offering protection to the host from invaders such as bacteriophages. The expression of these genes in E. coli K-12 had not previously been studied due to the complexity of the EcoK R-M enzymes. The use of lacZ gene fusion techniques allowed studies of the expression of the hsdK genes for the first time. Operon fusions of Mu(/ac) to the hsdK genes show that the two hsdK promoters, p _res and p _mod, are weak (50 Miller units) and that …

The Cγ Subunit Is A Unique Isozyme Of The Camp-Dependent Protein Kinase, Stephen J. Beebe, Paul Salomonsky, Tore Jahnsen, Yixin Li

Bioelectrics Publications

There are at least three isozymes (Cα, Cβ, and Cγ) of the mammalian catalytic (C) subunit of cAMP-dependent protein kinase (PKA) (Beebe, S., Oyen, O., Sandberg, M., Froysa, A., Hansson, V., and Jahnsen, T. (1990) Mol. Endocrinol. 4, 465-475). To compare the Cγ and Cα isozymes, the respective cDNAs were expressed in permanently transformed Kin-8 PKA-deficient Y1 adrenal cells using the mouse metallothionein promoter. The recombinant C subunits were characterized as immunoreactive, zinc-inducible, cAMP-dependent kinase activities. In contrast to Cα, histone was a better substrate than Leu-Arg-Arg-Ala-Ser-Leu-Gly (Kemptide) for Cγ. Furthermore, Cγ histone kinase activity was not inhibited by the …

Ecdysterone Regulatory Elements Function As Both Transcriptional Activators And Repressors., Leonard Dobens, Karen Rudolph, Edward M. Berger

Dartmouth Scholarship

A synthetic, 23-bp ecdysterone regulatory element (EcRE), derived from the upstream region of the Drosophila melanogaster hsp27 gene, was inserted adjacent to the herpes simplex virus thymidine kinase promoter fused to a bacterial gene for chloramphenicol acetyltransferase (CAT). Hybrid constructs were transfected into Drosophila S3 cells and assayed for ecdysterone-inducible CAT expression. In the absence of ecdysterone a tandem pair of EcREs repressed the high constitutive level of CAT activity found after transfection with the parent reporter plasmid alone. After hormone addition very high levels of CAT activity were observed. Insertion of the EcRE pair 3' of the CAT gene …

Characterization Of Hsd::Mudx128 Operon Fusion Mutant Of Escherichia Coli K-12, Marjorie Ann T. Reyno

Loma Linda University Electronic Theses, Dissertations & Projects

The study of the regulation of gene expression in complex genes can be facilitated by the use of operon fusions which place a well characterized lacZ gene under the control of a promoter of interest. An hsd::MudX128 operon fusion mutant of E. coli K-12 isolated by Prakash (1986) was observed to have a high β-galactosidase activity (1000 units), ten to twenty times higher than other hsd::MudX mutants. This β-galactosidase activity was suppressed to a very low level (25 units) by the introduction of an F’ plasmid, F'101, which carries the 98 min to 2 min region of the …

Freshwater Fishes Of Shipstern Nature Reserve, Caspar F. A. Bijleveld

Physiology Faculty Research Publications

A survey of fishes in and around Shipstern Nature Reserve in Northern Belize, Central America, was carried out over a period of eight months, from February to September 1990. Its purpose was to establish a preliminary list of the fishes of Shipstern Nature Reserve, including descriptions, distribution and collecting data and additionally to describe some of the habitats in which the fishes were found. This study focuses on fishes considered to be freshwater species. These occurred for the greater part in brackish habitats. Nevertheless, a few fish considered as marine species were caught in the waters of the reserve and …

Molecular Genetics Of The Caenorhabditis Elegans Heterochronic Gene Lin-14, Gary Ruvkun, Victor Ambros, Alan Coulson, Robert Waterston, John Sulston, H. Horvitz

Victor R. Ambros

We describe a general strategy for the genetic mapping in parallel of multiple restriction fragment length polymorphism (RFLP) loci. This approach allows the systematic identification for cloning of physical genetic loci within about 100 kb of any gene in Caenorhabditis elegans. We have used this strategy of parallel RFLP mapping to clone the heterochronic gene lin-14, which controls the timing and sequence of many C. elegans postembryonic developmental events. We found that of about 400 polymorphic loci in the C. elegans genome associated with the Tc1 family of repetitive elements, six are within 0.3 map unit of lin-14. The three …

Caribbean Island Zoogeography: A New Approach Using Mitochondrial Dna To Study Neotropical Bats, Carleton J. Phillips, Dorothy E. Pumo, Hugh H. Genoways, Phillip E. Ray

University of Nebraska State Museum: Mammalogy Papers

Genetic analysis of animal mitochondrial DNA is a new and valuable addition to the battery of techniques available to zoogeographers. This paper describes characteristics of mitochondrial DNA (mtDNA) that make it applicable for the study of island zoogeography.

Some traditional zoogeographic questions are examined using mtDNA from the Neotropical fruit bat, Artibeus jamaicensis. The specific questions are: 1) To what extent are island populations isolated (that is, does interbreeding occur between the insular subspecies)? 2) Can a single founding female account for the mitochondrial genomes on specific islands in the Antilles? 3) Is there a correlation between the genomic …

Genetic Transformation In Salmonella Typhimurium, Siao Ping Tasi

Loma Linda University Electronic Theses, Dissertations & Projects

Plasmid transformation is an essential step in many gene cloning experiments. In Salmonella tynhimurium, mutants of rough strain have shown improved transformation frequency previously. In this work a stable restriction-minus (r^-) and modification-plus (m⁺) strain and its derivatives were constructed. Effect of restriction barrier on transformation of plasmid pBR322 was tested. Several factors which may affect transformation frequency have been systematically examined to determine the optimum conditions for transformation in S. tvphimurium. Efforts were made to simplify the procedure.

Using pBR322 plasmid as an experimental model, the r^- strain demonstrated 10⁵ to …

The Characterization Of Ribosomal Rna Gene Chromatin From Physarum Polycephalum, Sally A. Amero, Vicky L. Montoya, Wendy L. Murdoch, Roy C. Ogle, John L. Keating, Robert M. Grainger

Medical Diagnostics & Translational Sciences Faculty Publications

We have isolated ribosomal RNA gene (rDNA) chromatin from Physarum polycephalum using a nucleolar isolation procedure that minimizes protein loss from chromatin and, subsequently, either agarose gel electrophoresis or metrizamide gradient centrifugation to purify this chromatin fraction (Amero, S. A., Ogle, R. C., Keating, J. L., Montoya, V. L., Murdoch, W. L., and Grainger, R. M. (1988) J. Biol. Chem. 263, 10725-10733). Metrizamide-purified rDNA chromatin obtained from nucleoli isolated according to the new procedure has a core histone/DNA ratio of 0.77:1. The major core histone classes comigrate electrophoretically with their nuclear counterparts on Triton-acid-urea/sodium dodecyl sulfate two-dimensional gels, although they …

Mitochondrial Dna Polymorphism In Three Antillean Island Populations Of The Fruit Bat, Artibeus Jamaicensis, Dorothy E. Pumo, Everett Z. Goldin, Beth Elliot, Carleton J. Phillips, Hugh H. Genoways

University of Nebraska State Museum: Mammalogy Papers

The Neotropical fruit bat, Artibeus jamaicensis, occurs throughout Latin America and on many islands in the Caribbean. Populations from Jamaica (in the Greater Antilles) to Barbados (in the Lesser Antilles) have been classified as a subspecies (A. j. jamaicensisi separate from that on the Lesser Antillean island of St. Vincent (A. j. schwartzi). Mitochondrial DNA (mtDNA) was isolated from 54 individuals collected on these islands, analyzed by digestion with restriction endonucleases, and the restriction sites were mapped. Three different mtDNA genotypes (16,000 ± 200 bp) were identified: J-1 (16 animals from Jamaica, one from St. Vincent, …

Partial Characterization Of Lpss From Rhizobium Fredii Usda 205 Before And After Nod Gene Induction, Bradley Lawrence Reuhs

Masters Theses

The lipopolysaccharides (LPS) from Rhizobium fredii USDA 205 and its nod- mutant, HC 205, which has been cured of the 192-MDa symbiotic plasmid, were isolated by the phenol-water extraction method. The LPS from the wild-type strain which was grown in association with host-plant root extract or a flavone analog to a constituent of the root extract, which is required for the induction of at least a portion of the nodulation genes, was also isolated. The conditions required for the induction of the nod genes were determined by work with a nod- mutant of R. fredii USDA 201 which …

Karyotypic Analysis Of Five Rodents And A Marsupial From Belize, Central America, David W. Burton, John W. Bickham, Hugh H. Genoways, Timothy J. Mccarthy

University of Nebraska State Museum: Mammalogy Papers

Karyotypes and chromosomal banding patterns of six species of small mammals from Belize are presented and discussed in light of relevant data in the literature. Individuals of Heteromys desmarestianus had FN values of 72, which differs from previous reports. G-bands were obtained and compared to fluorescent bands (DAPI) and with the location of NOR sites. The karyotypes of Peromyscus mexicanus, Sigmodon hispidus, Oryzomys couesi, and Ototylomys phyllotis are identical to the karyotypes for these species described in the literature. Active NORs were located on four pairs of chromosomes in O. phyllotis and fluorescent bands obtained with the …

The Effects Of Strain Differences On The Binding Of Concavalian A To Mouse Sperm, Lisa A. Mckay

Honors Theses

The current study, lectins were used as genetic probes to determine the phenotype of the sperm from different strains of mice. Lectins are proteins of non-immune origin, which bind to specific carbohydrates (McCoy, 1986). Being that virtually all cell membranes contain carbohydrate components, lectins have been used to identify all types of cells. In this study, a biotinylated lectin Concavalian A (Con A) derived from Canavalia ensiformis, was used and binding was detected using an avidin-biotin peroxidase (ABP) complex technique. The object of the current research was to 1) determine if mouse strain differences have an effect on lectin binding, …

A Comparison Of The Acidic Extracellular Polysaccharides From Rhizobium Trifolii Mutants Affected In Trifolium Repens Root Hair Curling, Brian Andrew Hanley

Masters Theses

The compositions and sugar linkages of the acidic extracellular polysaccharide (EPS) from several Rhizobium trifolii mutants were compared. These mutants were derived from a wild type Rhizobium trifolii, ANU843, and are unable to cause normal root hair curling, Hac-, or nodulate clover, Nod-. Three of the mutants contain a Tn5 insertion in a 5.3 kb BglII fragment of the symbiotic plasmid (pSym). A fourth mutant is missing the pSym, which includes much of the 5.3 kb BglII fragment, both of these deletions result in Hac-Nod- phenotypes. When the 5.3 kb BglII fragment is cloned (pRT150) and reinserted into the …

Seasonal Responses Of Filtration Rates To Temperature, Oxygen Availability, And Particle Concentration Of The Freshwater Clam, Musculium Partumeium (Say), Albert J. Burky, Richard B. Benjamin, Denis G. Conover, Jennifer Detrick

Biology Faculty Publications

Seasonal filtration rates of a pond population of the freshwater pisidiid clam, Musculium par­tumeium (Say), were assessed by measuring the clearance of 2.02 µ.m latex beads from suspension and expressed in terms of FR = a(AFDW}b where FR = filtration rate (ml H20 · hr- 1 ) , AFDW = mg ash-free dry weight of a whole clam and "a" and "b" are constants. The a-values ( = FR of a 1-mg AFDW clam} vary seasonally with the highest rates corresponding to periods of maximum growth and reproduction in the spring and fall. Changes in "b" reflect the influence of …

Explorations, Vol. 1, No. 1, R. D. Blake, Katherine Carter, David Mackinnon Ebitz, Carole J. Bombard, Stephen A. Norton

Explorations — A Journal of Research

Welcome to the first issue of Explorations, A Journal of Research at the University of Maine at Orono.

Join us as we explore a representative selection of the pure and applied research of our faculty. At UMO, we believe that research can capture the imagination and invigorate the mind, as well as contribute directly to the quality of life of the citizens we serve.

In this first issue, we have selected four areas of research that span the disciplines of biological and environmental sciences and the arts. This is but a small part of the research conducted by the faculty …

Interpopulation Variation In Calcareous And Proteinaceous Shell Components In The Stream Limpet, Ferrissia Rivularis, Carl M. Way, Daniel J. Hornbach, Albert J. Burky

Biology Faculty Publications

Natural populations of the North American stream limpet, F. rivularis, were studied in upstate New York [USA], in a set of localities whose waters have a 15-fold range of dissolved Ca (4.6-67.6 mg/liter) and also range from oligotrophy to eutrophy. Shell component analyses (CaCO3, total organic C and total N) are reported as component mass-fractions (mg/g or .mu.g/g dry weight) and as values for a standard limpet shell of 35 mm aperture length (AL). More than 2-fold differences occur between populations in all 3 components, with relatively little variation occurring within each population. Expressed per standard limpet, CaCO3 values for …

Seasonal Metabolism Of The Sphaeriid Clam, Musculium Partumeium (Say), From A Permanent And A Temporary Pond, Carl M. Way, Daniel J. Hornbach, Albert J. Burky

Biology Faculty Publications

Seasonal metabolic rates were examined for the sphaeriid clam, Musculium partumeium (Say) from a permanent and a temporary pond. For the fall-born generation from the permanent pond and for the single generation from the temporary pond, metabolic rates peak during periods of greatest growth and reproduction. Metabolic rates were consistently higher for the permanent pond population. These rates for Musculium partumeium extend the available information on the family Sphaeriidae to the genus Musculium.

Growth Of Pisidium Casertanum (Poli) In West Central Ohio, Albert J. Burky, Daniel J. Hornbach, Carl M. Way

Biology Faculty Publications

Clarke (1973) states that Pisidium casertanum (Poli) is perhaps the most common species of Pisidium and is truly cosmopolitan, occurring throughout Eurasia, Africa, Australia, and South, Central and North America. Populations of these clams are reported in habitats ranging from ephemeral ponds to benthic zones of deep lakes. Our study describes Manuscript received 29 May 1979 and in revised form 25 July 1980 (#79-34). 2 Present address: Department of Biology, University of Virginia, Charlottesville, VA 22901. 3 Present address: Department of Zoology, Miami University, Oxford, OH 45056. 41 aspects of the life-cycle of P. casertanum in a spring-fed stream with …

Buoyancy Changes As Related To Respiratory Behavior In An Amphibious Snail, Pomacea Urceus (Müller), From Venezuela, Kathleen A. Burky, Albert J. Burky

Biology Faculty Publications

The ampullariid Pomacea urceus uses its ctenidium and lung in respiration. The snails reach the surface with their siphon and ventilate their lung by withdrawing their head-foot in a pumping action. The lung gas also serves to change the buoyancy of the snails. After a ventilation a snail does not necessarily return to the same buoyancy level. They may achieve overall specific gravities of <1 to > 1 after a ventilation. They may remain submerged (s.g. > 1) or float (s.g.

Molecular Weight Determinations Of Some Salmonella Potsdam Bacteriophage Dna's, Winston H. Richards

Loma Linda University Electronic Theses, Dissertations & Projects

Two group A phages, P9c/2 and P4/2, were studied. Their density, sedimentation velocity, and biochemical properties were compared. Their densities were determined by density gradient centrifugation in 38.3% cesium chloride. Sedimentation velocity was used to determine their molecular weights. To accomplish this, the DNA of P9c/2 and P4/2 was labeled with tritium and T₂ DNA, labeled with ³²P, was used as a standard. The DNA of P9c/2 and P4/2 was separately mixed with T₂ DNA and centrifuged in a 0-15% sucrose gradient. The molecular weights were calculated using the formula D₂/D₁= (M_{2 …}

Biochemical Properties Of Salmonella Potsdam Bacteriophage P4/2, Clyde L. Roggenkamp

Loma Linda University Electronic Theses, Dissertations & Projects

Over 90% recovery of phage P4/2 was realized with differential centrifugation concentration and purification, whereas only about 16% was recovered from the aqueous two-phase polymer - Genetron - ECTEOLA-cellulose column system. Purity of preparations was greatest with the density gradient centrifugation (A₂₆₀ = 2.66 and A₂₈₀ = 1.80 per 10¹² plaque-forming units). With the chemical system of concentration and purification followed by an ECTEOLA anion exchange run, the A₂₆₀ was 6.6 and the A₂₈₀ was 4.3.

Three tests were employed to determine the configuration of P4/2 DNA: (1) a formalin denaturation test, (2) a thermal …