Microbiology Commons^™

Development And Application Of A Thermistor Current Meter, Carl M. Way, Albert J. Burky, Christine Miller-Way

Biology Faculty Publications

This report provides details for the construction of a hot-bead thermistor current meter which is capable of measuring water velocities on a millimeter spatial scale and for the construction of a compact and accurate calibration system. Hot-bead thermistor current meters can be built with response times of 200 ms capable of measuring velocities between 0.1 and 80 cm s-1. The construction of a sturdy probe for application in lotic systems such as high gradient Hawaiian streams was achieved by the use of heavy-duty acrylic tubing, small stainless steel gas-chromatography tubing, and flexible Tygon spaghetti tubing. An acrylic handle anchors the …

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, …

Molecular And Cellular Analysis Of Chlamydia Trachomatis: Persistence And Reactivation, Kimberly R. Tau

Electronic Theses and Dissertations

Chlamydia trachomatis (CT) is the most prevalent sexually-transmitted infection in the United States. It has been suggested that CT infections can become latent. This has not been substantiated. CT persistence was examined at the molecular and cellular level in vitro and in vivo. Penicillin treatment of CT in vitro results in abnormal inclusions and reduced recovery of infectious CT. Penicillin did not inhibit initial stages of infection, but did downregulate CT rRNA levels after 25 hours post-inoculation (p.i.). DNA amplification was employed to differentiate between a resolved infection and a persistent one. Utilizing a primer pair that amplified a 144 …

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 …

Immunosuppressive Activity Derived From The Uc729-6 Lymphoblastoid And M21-Hpb Melanoma Cell Lines, Charlene J. Repique

Loma Linda University Electronic Theses, Dissertations & Projects

Previous work conducted in our laboratories, using a murine model, suggested that soluble factors secreted by tumor cells suppress lymphocyte responses. To apply this premise to human tumors, we studied the effects of UC729-6 (lymphoblastoid B-cell) and M21-HPB (malignant melanoma) conditioned media (CM) on lymphocyte proliferation assays, as well as on the growth of other cells. The CM was collected at 2-5 day intervals from cultures of UC729-6 and M21- HPB cells in serum-free media. In assays using phytohemagglutinin (PHA) and concanavalin A (ConA), mononuclear peripheral blood cells from healthy human donors showed decreased [³H]-thymidine ([³H]-Tdr) …

A Simple Solid-Phase Electrophoretic Procedure For The Separation Of Plasmid Dna, Linda Ann Simurra

Biological Sciences Theses & Dissertations

A method was developed for extraction of plasmid DNA from bacterial cells embedded in agarose blocks. Cell - containing blocks were treated with various lysing reagents and inserted into the wells of an agarose gel. Upon electrophoresis the plasmid DNA migrated out of the embedding block and into the gel leaving intact chromosomal DNA in the well. This method was tested with various organisms and found to be effective for plasmid isolation. In comparison to "traditional" procedures, this new method is less tedious since chemical separation of plasmids is not required prior to electrophoresis. Also, a higher yield of plasmid …

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 …

Analysis Of Multiple Isoenzyme Expression Among Twenty-Two Species Of Mycoplasma And Acholeplasma, Stephen J. O'Brien, Janice M. Simonson, Marion W. Grabowski, Michael F. Barile

Biology Faculty Articles

Crude extracts of triple-cloned, purified cultures of 22 species of Mycoplasma and Acholeplasma were examined for expression of 21 isozyme systems routinely used to type mammalian cells. Nine previously described enzymes (purine nucleoside phosphorylase, adenylate kinase, dipeptidase, esterase, glyceraldehyde-3-phosphate dehydrogenase, glucose phosphate isomerase, glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and superoxide dismutase) and three enzymes not previously reported in mycoplasma (triose phosphate isomerase, inorganic pyrophosphatase, and acid phosphatase) were detected in some or all of the species examined. These findings provide new information on the enzymatic expressions of these organisms. Three of the isozyme systems (superoxide dismutase, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase) …

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 …

The Biology Of Coprinus Radians, Ozonium, Oidium, And Their Interrelationships, Thomas J. Fahey

All-Inclusive List of Electronic Theses and Dissertations

The pleomorphism of Coprinus radians was studied. The three distinct growth forms--sporophore, oidial stage, and ozonium stage--were examined in relation to ultrastructure, morphology and development. Scanning and transmission electron microscopy are correlated with photomicrographs of developmental stages. Development of the oidia and of the sporophore are of particular interest. The multinucleate oidia develop quite differently from oidia as generally described for Basidiomycete imperfect stages. Development of the oidia of this Coprinus is holoblastic and sympodial. The mushroom primordia are differentiated internally into gill, cap, and stipe hyphae while only l-2mm in diameter. At this stage they look superficially like sclerotial …

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.

Studies On Muropeptide Amino Acids Of Rhizobium Japonicum, Mohammed Alauddin

Masters Theses

The isolation of Rhizobium japonicum cell walls was performed by ultrasonication and the enzymatic digestion with DNase, RNase and pronase separately. Crude cell walls were washed with M NaCl and 5% SDS and were isolated by 20,000 g centrifugation. Muropeptide amino acids were freed by hydrolysis at 121C for 10 minutes in 6N HCl.

Detection of Rhizobium japonicum murein layer amino acids was done by 2-dimensional instant thin layer chromatography on silica gel plates. The solvents found most favorable, were n-butanol: acetic acid: water and n-propanol: 34% ammonium hydroxide. Amino acid spots were developed by spraying ninhydrin on the chromatograms, …

The Role Of D-Gluconic Acid In The Regulation Of The Synthesis Of The Enzymes Of The Entner-Doudoroff Pathway In Pseudomonas Fluorescens, Steven C. Quay

Honors Theses

Induction of Entner-Duodoroff pathway enzymes in Pseudomonas fluorescens was investigated to study the role of gluconate as a possible inducer. Glucose oxidase-deficient mutants were isolated and characterized. One of these mutants, gox-7, was deficient in particulate glucose oxidase; another mutant, gox-17, was deficient in particulate glucose and gluconate oxidase activities. Gluconate, but not glucose, induced synthesis of gluconokinase and 6-phosphogluconate dehydratase in both mutants. High constitute levels of 2-keto-3-deoxy-6-phosphogluconate aldolase were found when both mutants were grown on glucose. Growth of parent and both mutant strains on glycerol also resulted in high levels of Entner-Doudoroff pathway enzymes. It was concluded …

Lysis Of The Rhizobiaceae, Craig Richard Baumrucker

Masters Theses

The following members of the Rhizobiaceae were tested for their lytic response to various treatments: Agrobacterium tumefaciens, Chromobacterium violaceum, Rhizobium phaseoli, Rhizobium trifolii, Rhizobium lupini, and Rhizobium japonicum. Lysis was observed in spent growth media and as a function of saline concentration, pH, buffer composition, disodium (ethylenedinitrilo) tetraacetate (EDTA) concentration, and lysozyme.

Cells were harvested by centrifugation, washed, and resuspended in various treatment solutions. The lytic response was followed spectrophotometrically.

The different responses of the six cultures was applied to a summation of reclassification proposals evolved through modern taxonomic techniques such as DNA homology …

A Balanced Salt Culture Medium For Halteria Grandinella, Yee Wan Mark

All-Inclusive List of Electronic Theses and Dissertations

The animals that the writer worked with liere the unicellular organisms called liaJ,tez:ia grandil.!iilJ.a• They belong to the Pl~lum Protozoa and are placed in the Class Ciliata and the Order Oligotrichia. llftltez'A gDJ}gini•U, like most protozoa, ll ve in waters but they seem to prefer qUiet pools, lakes, and ponds in which aqua i;;!c plants are growing and in which there is much organic matter.

The Chromosomes Of The Yeast Cell, Donald William Penn

Bachelors’ Theses

Saccharomyces cerevisiae has long been a fungus of economic importance in the baking and beverage fields. A large amount of research has been done on this yeast cytologically and biochemically. However, due to its small size and difficulty in staining, the cytology of the yeast cell remains somewhat in question. Chromosomes and division figures have been demonstrated by several workers, but there still remain sharp differences of opinion as to the nature of the nucleus. Recent work on the genetics of yeast has shown evidence of chromosome division similar to that occurring in higher plants and studies of linkage have …

Replicating Single-Cycle Adenovirus Vectors Generate Amplified Influenza Vaccine Responses, Catherine M. Crosby, William E. Matchett, Stephanie S. Anguiano-Zarate, Christopher A. Parks, Eric A. Weaver, Larry R. Pease, Richard J. Webby, Michael A. Barry

Nebraska Center for Virology: Faculty Publications

Head-to-head comparisons of conventional influenza vaccines with adenovirus (Ad) gene-based vaccines demonstrated that these viral vectors can mediate more potent protection against influenza virus infection in animal models. In most cases, Ad vaccines are engineered to be replication-defective (RD-Ad) vectors. In contrast, replication-competent Ad (RC-Ad) vaccines are markedly more potent but risk causing adenovirus diseases in vaccine recipients and health care workers. To harness antigen gene replication but avoid production of infectious virions, we developed “single-cycle” adenovirus (SC-Ad) vectors. Previous work demonstrated that SC-Ads amplify transgene expression 100-fold and produce markedly stronger and more persistent immune responses than RD-Ad vectors …