The effects of oligoribonucleotides of identical or similar sequences to a
ribosomal protein binding site, specifically S8, on prokaryotic 30S ribosomal assembly
were investigated in Escherichia coli. The oligoribonucleotides were expected to bind to
the target protein S8 and block its incorporation into the 30S subunit, resulting in a nonfunctional...
RNA tumor viruses, such as avian myeloblastosis virus (AMV)
and MC29 virus, cause malignant transformation of the cells they infect.
Transformed cells are able to continually produce progeny virus
particles.
Evidence is given that RNA-dependent RNA polymerase is present
in virus infected cells. RNA-dependent RNA polymerase isolated
from AMV-infected myeloblasts...
Cystic fibrosis is an inherited genetic disease caused by a single gene mutation in the CFTR protein. Affecting 70,000 people worldwide, this disease is debilitating to patients, resulting in persistent lung infections and decreased quality of life. Due to the various mutation types, there is not currently a single drug...
A novel protocol for the study of protein-nucleic acid interactions is presented and demonstrated
to be feasible. The protocol combines photochemical crosslinking techniques and mass
spectrometric methods into a new strategy for identifying protein domains or amino acid residues
that are in close contact with nucleic acid in protein-nucleic acid...
Fluorescence of adenine, guanine, thymine, cytosine, and uracil
has been detected at room temperature in neutral aqueous solution
using a digital signal accumulation technique and right angle fluorescence
detection from solutions with absorbances between 0. 4 and 0. 7.
The quantum yields are respectively 2.6 x 10⁻⁴, 3.0 x 10⁻⁴,...
The binding of eleven polycyclic, aromatic hydrocarbons to the acid
form of poly A has been investigated. The hydrocarbons were chosen to
test a size criterion. Model building showed that some hydrocarbons
could intercalate in the helix and be well protected from contact with
the aqueous solvent. Experimentally it was...
The effects of pH on the torsional flexibility of DNA bound to a
nucleosome core particle were investigated by studying the time-resolved
fluorescence anisotropy decays of ethidium bromide intercalated into the DNA
of the core particle. As the torsional flexibility of DNA is affected by the
presence of an intercalating...
Peptide-phosphorodiamidate morpholino oligomers (PPMOs) are synthetic DNA mimics that bind and silence gene targets. Through designing PPMOs that silence essential or resistance genes in pathogens, these antisense oligomers could be utilized as novel antimicrobials. Towards this end, my thesis employed minimum inhibitory concentration assays (MICs) to identify PPMOs capable of...
Nucleic acid estimates in tissues of marine macrophytes with the fluorochromes Hoechst 33258 and ethidium homodimer were severely impaired by high concentrations of interfering compounds. The isolation and extraction techniques used pointed towards compounds of low molecular weight and of polar characteristics that were insensitive to organic solvent extraction and...
The DNA-binding domains of E. coli uracil-DNA glycosylase (Ung) and human replication protein A (hRPA) were studied using a general protocol developed in our laboratory for probing protein-DNA interactions. The procedure involves purification and mass spectrometric analysis of the nucleopeptide-products of a tryptically digested UV-crosslinked protein-nucleic acid complex. In the...
Messenger RNA (mRNA) therapy has massive potential to treat various genetic disorders by providing effective proteins to the diseased cells and tissues. Its therapeutic applications include protein replacement, genome editing, and immunotherapy. For successful mRNA treatment, mRNA must be delivered to the cytosol of target cells. One of the leading...
The Mx protein family of rainbow trout (Oncorhynchus mykiss) was characterized at the molecular level. With primers derived from a partial sequence of Perch (Perca fluviatis) Mx genomic DNA, a partial rainbow trout Mx genomic DNA sequence was obtained by polymerase chain reaction (PCR) amplification. All salmonid fish investigated contained...
Wildtype sunflower typically produces 12-24% oleic acid (18:1) and 70-82%
linoleic acid (18:2). High-oleic sunflower, by contrast, produces up to 80-94% oleic
acid. The monounsaturated oleic acid has a greater oxidative stability than the
polyunsaturated linoleic acid, predominant in wildtype sunflower, and has greater
nutritional benefits than polyunsaturated and saturated...
Expanding efforts to develop preventive gonorrhea vaccines is critical because of the dire possibility of untreatable gonococcal infections. Reverse vaccinology, which includes genome and proteome mining, has proven very successful in the discovery of vaccine candidates against many pathogenic bacteria. However, progress with this approach for a gonorrhea vaccine remains...
Deoxyribonucleoside triphosphate pools in mammalian mitochondria are highly
asymmetric, and this asymmetry probably contributes toward the elevated
mutation rate for the mitochondrial genome as compared with the nuclear
genome. To understand this asymmetry, we must identify pathways for synthesis
and accumulation of dNTPs within mitochondria. We have identified
ribonucleotide reductase...
Saccharomyces cerevisiae contains three NAD⁺/NADH kinases, one of which is localized in
mitochondria and phosphorylates NADH in preference to NAD⁺. Strand et al (Euk. Cell
2:809 (2003)) reported that a yeast mutation in POS5, which encodes the mitochondrial
NADH kinase, is a mutator, specific for mitochondrial genes. Because of the...
Intrinsically disordered protein (IDP) duplexes composed of two IDP chains cross-linked by bivalent partner proteins form scaffolds for assembly of multiprotein complexes. The N-terminal domain of dynein intermediate chain (N-IC) is one such IDP that forms a bivalent scaffold with multiple dynein light chains including LC8, a hub protein that...
The multistage model of carcinogenesis states that an accumulation of mutations in genes that are important for maintaining cellular homeostasis may lead to cancer. A specific type of mutation observed in particular types of cancer is the CC-->TT mutation found in the TP53 gene of patients with basal or squamous...
Calf diarrhea (scours) is a primary cause of illness and death in young calves. Significant economic losses associated with this disease include morbidity, mortality, and direct cost of treatment. Multiple pathogens are responsible for infectious diarrhea, including, but not limited to, Bovine coronavirus (BCV), bovine Rotavirus A (BRV), and Cryptosporidium...
The most frequent cause of insufficient acid production during
fermentation of dairy products is bacteriophage infection of lactic
starter cultures. Previously, efforts to minimize this problem have
dealt primarily with the host range relationships of the viruses and
susceptibility patterns of the lactic streptococci. This study was
undertaken to characterize...
Mycobacterium abscessus (MAB) is an opportunistic pathogen responsible for infections in immunocompromised or immunosuppressed patients. Even with long-term treatment with multiple antibiotics, MAB pulmonary infections have high rates of treatment failure. Within the host, MAB is phagocytosed by key innate immune cells macrophages. The TLR2 and TLR4 are two critical...
More needs to be done by the private sector to optimize the drug discovery and development pipeline. In addition, significant efforts should also be focused on the understanding of mechanism of diseases, on the characterization of unexplored biochemical pathways and on the validation of new protein targets. Chemical genomics, which...
Certain bioactive food components, including indole-3-carbinol (I3C) and 3,3'-diindolylmethane (DIM) from cruciferous vegetables, have been shown to target cellular pathways regulating carcinogenesis. Previously, our laboratory showed that dietary I3C is an effective transplacental chemopreventive agent in a dibenzo[def,p]chrysene (DBC)-dependent model of murine T-cell lymphoblastic lymphoma. The primary objective of the...
Decline in cardiac pump function is a hallmark of aging where mitochondrial decay is an important underlying cause. Although certainly multifactorial in nature, both dysfunction of the machinery involved in the chemiosmotic process of energy transduction and lower capacity to maintain fatty acid-driven respiration are identified as intrinsic factors of...
This is the 4th edition of a book that was initiated with the annotation of the function of all the genes in the most commonly studied baculovirus, AcMNPV. It has been almost six years since I reviewed this literature. As a measure of the research that has occurred over this...
This is the third edition of a book that was initiated with the annotation of the function of all the genes in the most commonly studied baculovirus, AcMNPV. The second and third editions involved the update of this information and its integration into chapters covering the major processes central to...
Phytophthora infestans, the cause of the devastating late blight disease of potato and tomato, exhibits a clonal reproductive lifestyle in North America. Phenotypes such as fungicide sensitivity and host preference are conserved among individuals within clonal lineages, while substantial phenotypic differences can exist between lineages. Whole P. infestans genomes were...
Respiratory illnesses caused by both viral and bacterial infection are serious issues for global healthcare providers. With the emergence of new, drug-resistant forms of these diseases, innovative drugs and treatment therapies are needed. M tuberculosis is an infection that affects a large portion of the world’s population each year. Aerosol...
This thesis involves analyses of 3₂-helices, a-helices and of the change in residual
entropy of a protein upon chemical modification. Part of the long-term goal of
understanding the formation, function and stability of proteins requires that all
conformations be accurately assigned and structurally understood. By a statistical
analysis of 258...
The potential for nanomedicine to reduce toxicity and improve therapeutic response to radiation is vast. Clinically, this potential has thus far only begun to be realized with the recent success of the radioenhancing nanoparticle, NBTXR3 by Nanobiotix. Radioenhancing nanoparticles, however, represent only a small class of nanomaterials that may be...
Nonalcoholic fatty liver disease (NAFLD) is characterized by the accumulation of excess neutral lipids within the parenchymal cells of the liver. The primary etiology is diet-induced and most often exists as a comorbidity with obesity, metabolic syndrome, and/or Type 2 Diabetes. The development of excess liver lipid can be self-limiting...
Retinoic acid (RA) exerts important effects in the processes of vertebrate development, cellular growth and differentiation, and homeostasis. However, the mechanisms of action of RA in the control of cellular and developmental processes are incompletely understood, as the retinoid target genes have not been fully characterized. The goal of these...
As the volume of genetic sequence data increases due to improved sequencing techniques and increased interest, the computational tools available to analyze the data are becoming inadequate. This thesis seeks to improve a few of the computational methods available to access and analyze data in the genetic sequence databases. The...
Glutathione (GSH) is the predominant low molecular weight thiol antioxidant
in liver tissue. GSH plays an important role in maintaining the intracellular thiol redox
ratio as well as detoxification of electrophiles and xenobiotics. Aging leads to a significant decline (35%; P ≤ 0.05) in hepatocellular GSH levels. Using young (2-4...
The marine psychrophilic bacterium Vibrio marinus MP-4
possessed a maximum temperature and hydrostatic pressure of
20 C and 425 atm for growth. The effects of temperatures of 21 and
25 C and hydrostatic pressures of 200, 400, 500, and 1,000 atm on
protein, RNA and DNA synthesis by V. marinus...
The vascular endothelium is a single cell layer that lines the lumen of the entire vasculature. It is the site of synthesis of nitric oxide (NO), a vasodilatory compound synthesized by endothelial nitric oxide synthase (eNOS). NO causes intracellular calcium sequestration of the vascular smooth muscle cells, relaxing and dilating...
DNA reassociations, measured by optical and double label
membrane filter techniques, were used to assess genetic relationships
among Aeromonas and enterobacteria. Aeromonads from human
sources, food, fresh water, fish, and pet turtles were examined. The
base compositions of three A. salmonicida isolates were 58% G+C
while the base compositions of...
Three samples of the biopolymer hyaluronic acid (HA) were characterized in the following manner: the molecular weights were obtained via multi-angle laser light scattering; the intrinsic viscosities were calculated through dilute solution viscometry, and the rheology of HA solutions was determined with constant rate rotational viscometry and dynamical mechanical testing....
The auxin-resistant diageotropica (dgt) mutant of tomato (Lycopersicon esculentum Mill.) has a pleiotropic phenotype including a lack of lateral roots and reduced gravitropic response, apical dominance, vascular development, and fruit growth. The dgt mutation reduces the auxin sensitivity of only a subset of auxin responses while levels, metabolism, and transport...