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Studies on Tissue Factor Pathway Inhibitor in Zebrafish
Tissue Factor Pathway Inhibitor (TFPI) is an anticoagulant protein containing three Kunitz domains, K1, K2 and K3. K1 inhibits Factor VIIa, K2 inhibits Factor Xa, and K3 enhances the Factor Xa inhibition by its interaction with Protein S. Since zebrafish is an excellent genetic model, we hypothesized that TFPI regulation could be studied using this model. As a first step, we confirmed the presence of tfpia in zebrafish. Subsequently, we performed knockdown of tfpia, and knockout of tfpia in K3 domain using CRISPR/Cas9. Both the tfpia knockdown and tfpia homozygous deletion mutants showed increased coagulation activities. Our data suggest that zebrafish tfpia is an orthologue for human TFPIα, and silencing it results in a thrombotic phenotype. We then optimized the piggyback knockdown method, where we could simultaneously piggyback 3 or 6 ASOs corresponding to 3 or 6 genes, respectively, using one VMO. These multiple gene knockdowns will increase the efficiency of genome-wide knockdowns. Since there are no studies on chromatin remodeling that control TFPI expression, we hypothesized that the genome-wide knockdowns of the Chromatin Binding and Regulatory Proteins (CBRPs) in zebrafish could help identify novel tfpia gene regulators. We chose 69 CBRPs and subjected them to simultaneous gene knockdowns. Our …
Date:
August 2021
Creator:
Raman, Revathi
System:
The UNT Digital Library
Cardiovascular Fetal Programming in Quail (Colinus virginianus), An Avian Comparative Model
The consequences of early embryonic insults and how they affect subsequent life reflects the emerging concept of "fetal programming". The aim of this project is to study the effects of embryonic insults as they subsequently manifest themselves in adults, with emphasis on the heart and vasculature. My experiments establish that fetal programming operates on the bobwhite quail inducing similar changes as those observed in mammalians and other birds. The quail's fast development provides reliable data in a short period of time than other avian models (e.g. domestic chicken). Data on quail showed a correlation between egg mass and hatchling mass; where small eggs produce small hatchlings but a high mortality made it impractical as a stressor for this study. Hypoxia was used as a stressor during embryonic incubation, where it induced a low hatching weight in quail that was not observable in adult birds. Morphological measurements demonstrated an increased ventricular collagen content and reduced ventricular lumen in birds in adults incubated in hypoxia consistent with hypertension. The hematological analyzes showed few differences indicating organ remodeling instead of hematopoietic compensation. The assessment of vascular reactivity pointed out an impaired endothelium dependent relaxation commonly associated to hypertension in birds and mammals. Fetal …
Date:
December 2016
Creator:
Flores Santin, Josele R.
System:
The UNT Digital Library
Effect of pH on the Persistence and Toxicity of Cyfluthrin to Chironomus Tentans
The effect of pH upon the aquatic toxicity of cyfluthrin was determined in 48 h static acute toxicity tests using 2nd instar Chironomus tentans larvae. Tests were conducted in both dechlorinated tap water and lake water of pH 8.0, 7.2, and 6.6. After 48 h, immobilized and dead larvae were removed and replaced with 2nd instar larvae to assess the persistence of toxicity. Midges were cultured in water adjusted to the pH values used in testing. Toxicity if cyfluthrin varied inversely with pH. An increase in the pH of tap water by 2 units resulted in a 2-decrease in toxicity. Toxicity of cyfluthrin also varied between tap and lake water of the same pH. EC50 values in lake water were approximately 2-3 times lower than those in tap water. Toxicity in the lake water and tap water at every pH tested was also significantly different when regression line elevations were compared. Natural waters amended with cyfluthrin were consistently more toxi to the chironomids than tap water of the same pH. Persistence of cyfluthrin at low pH also influenced chironomid behavior. Recovery of normal behavior generally began within 24 h at pH 8.0 At pH 6.0, recovery did not begin until …
Date:
May 1992
Creator:
Hambleton, Faith Ann (Faith Ann Elizabeth)
System:
The UNT Digital Library
Advanced Techniques in Microbial and Molecular Biology: Laboratory Procedures for a Graduate Level Course
Advanced laboratory techniques for Microbial and Molecular Biology at the graduate level are presented in this thesis. The procedures for the laboratory experiments are set forth in detail. This laboratory is conducted as two parts, each by a different professor. Part 1 covers the experiments conducted by Dr G.A.O. Donovan. These experiments include an introduction, staining procedures, biochemical reactions, mutagenesis experiment, essays,. preparation and analysis of plasmid DNA and various other topics. Part 2 covers the experiments conducted by Dr. Daniel Kunz and includes various topics like media preparation, phenotyping strains, conjugative transfer of plasmids, SDS-PAGE, induction and measurement of enzyme and transposon mutagenesis
Date:
August 1998
Creator:
Kumar, Shalini
System:
The UNT Digital Library
A Water Quality Study of Lake Texoma
A series of four stations for sampling the waters of the Red Rivers and Lake Texoma was established. Water samples from top, middle, and bottom levels of these four stations were taken on monthly intervals during periods of homothermousity and on two-week intervals during periods of thermal stratification. These samples were analysed for the cations sodium, potassium, calcium, and magnesium, and for the anions normal carbonate, bicarbonate, sulfate, chloride, nitrate, water hardness (expressed as calcium carbonate), and phosphates. Determinations for dissolved oxygen, carbon dioxide, pH, temperature, and specific conductance were performed. Using these data, vertical profiles of the lake at these four stations were established. From the study of these data it was concluded that the waters of the Red River improve in quality coincident with impoundment, and that these improvements are sufficient to warrant the use of these waters.
Date:
June 1959
Creator:
Goodman, John Willis
System:
The UNT Digital Library
Toxicological Evaluation for the Ocular Administration of Tolrestat: An Aldose Reductase Inhibitor for the Treatment of Diabetes
Aldose reductase inhibitors (ARI's) have been shown to attenuate or prevent several complications of diabetes in animals. Tolrestat is a potent and unique ARI from Ayerst Laboratories, New York, NY. The efficacy and toxicology of tolrestat via topical ocular administration was examined. Tolrestat effectively enhanced corneal reepithelialization and was efficacious in the prevention of cataracts in the streptozotocin-diabetic rat. Ocular tissues were examined by slit lamp biomicroscopy, scanning and transmission electron microscopy, and light microscopy. Liver and kidney tissue was also examined. The presence of tolrestat was confirmed in urine, feces, and eye specimens, and quantitated in serum. There was no evidence of local or systemic tolrestat induced toxicity. Tolrestat prevented cataract formation at less than one-third the reported oral dose and at approximately one-fiftieth the associated serum concentration in rats. ED-50 and TD-50 calculations indicate that tolrestat is a relatively safe drug by topical ocular administration.
Date:
December 1987
Creator:
Carney, Gerald R.
System:
The UNT Digital Library
Effects of Nutrient Media on Growth and Morphology of Azotobacter Vinelandii
The work described in this thesis was undertaken to study the reasons why Azotobacter vinelandii ATCC 12837 after incubation in Burk's nitrogen-free liquid will not form as many colonies when plated on Difco Tryptic Soy Agar as when planted on Burk's nitrogen-free agar. The difference in growth of A. vinelandii on the two agars was established by performing viable cell-plate counts. The difference in growth was most apparent at 24-hours incubation of the Burk's liquid-media cultures. Phase contrast microscopic observations of Tryptic Soy media cultures of A. vinelandii disclosed the regular formation of fungoid cells at early stages of growth of the bacteria, 18 to 24 hours.
Date:
August 1974
Creator:
Butsch, Robert W.
System:
The UNT Digital Library
Assessment of Radio-Tagged Grass Carp (Ctenopharnygodon idella) Dispersion, Vegetation, and Temperature Preferences in North Lake Reservoir
Twenty-nine (Group One, June 8,1995) grass carp (Ctenopharyngodon idella) and five (Group Two, April 18, 1996) grass carp were radio-tagged to monitor movement patterns and habitat preferences on North Lake, a 335 hectare multi-use reservoir located in Irving, Texas. Overall fish mean Average Daily Movement (ADM) rates were 49.2 meters/day (during Half One, 6/8/95-11/30/95) and 5.3 meters/day (during Half Two, 12/14/95-6/6/96). Aquatic macrophtye distribution data were obtained. Radio-tagged grass carp were located in Hydrilla verticillata infested areas increasingly throughout the study, however, percent frequency of Hydrilla along 15 transects did not decrease. Radio-transmitters were equipped with temperature-sensors (10-35 Celsius range). Results indicated that radio-tagged grass carp showed no avoidance of areas of North Lake with elevated water temperatures. Radio-tagged grass carp dispersed quickly from stocking point, then moved into littoral areas infested with Hydrilla. After an initial movement period, most fish remained in a localized area.
Date:
August 1996
Creator:
Lacewell, Jason (Jason Lawrence)
System:
The UNT Digital Library
A Comparison of Thermogenesis by Selected Substrates on Hypothermic Rat Liver
The thermogenic effects in hypothermia of four substrates--alanine, glycine, ethano, and pyruvate - were studied in seventeen experiments. Albino rats were decapitated, and their livers were removed. The livers were homogenized with phosphate buffer at -5° C. After equilibration in a refrigerated Warburg apparatus at 20° C, the substrates were added and tissue respiration was recorded over three hours. Heat production was calculated from O2 uptake and CO2 production. Results showed that alanine, glycine, and pyrvate yielded 93.19, 89.86, and 89.89 x 10^6 kg-cal compared to a control value of 86.11 x 10^-6 kg-cal. Ethanol provided 110.31 x 10^-6 kg-cal, a value significantly greater than for the other substrates. The substrates studied, especially ethanol, did, therefore increase heat production in an artificially hypothermic environment in homogenized rat livers.
Date:
August 1979
Creator:
Long, James T.
System:
The UNT Digital Library
Spatially Explicit Modeling of West Nile Virus Risk Using Environmental Data
West Nile virus (WNV) is an emerging infectious disease that has widespread implications for public health practitioners across the world. Within a few years of its arrival in the United States the virus had spread across the North American continent. This research focuses on the development of a spatially explicit GIS-based predictive epidemiological model based on suitable environmental factors. We examined eleven commonly mapped environmental factors using both ordinary least squares regression (OLS) and geographically weighted regression (GWR). The GWR model was utilized to ascertain the impact of environmental factors on WNV risk patterns without the confounding effects of spatial non-stationarity that exist between place and health. It identifies the important underlying environmental factors related to suitable mosquito habitat conditions to make meaningful and spatially explicit predictions. Our model represents a multi-criteria decision analysis approach to create disease risk maps under data sparse situations. The best fitting model with an adjusted R2 of 0.71 revealed a strong association between WNV infection risk and a subset of environmental risk factors including road density, stream density, and land surface temperature. This research also postulates that understanding the underlying place characteristics and population composition for the occurrence of WNV infection is important for …
Date:
December 2015
Creator:
Kala, Abhishek K.
System:
The UNT Digital Library
Impact of a Genetically Engineered Probiotic Therapy and IGF-1 Genomics in the PAHenu2 Mouse Model of PKU
Absence of functional phenylalanine hydroxylase results in phenylketonuria (PKU). Viable treatments remain few, expensive and secondary conditions such as osteopenia occur in most PKU patients. Objective 1: Given the recently described roles of gut microbes to aid host digestion, an orally administered genetically engineered probiotic as the delivery vehicle for enzyme replacement therapy was created. The engineered probiotic, pHENOMMenal, produced phenylalanine ammonia lyase with significant production of trans-cinnamate (phenylalanine cleavage product) in vitro and resulted in a reduction of 515 μM in blood phenylalanine when fed to PKU animals for 14 days (from 2307µM ± 264µM to 1792µM ± 261µM, n = 6, P < 0.05). The control probiotic produced no change in blood phenylalanine. Thus, pHENOMMenal treatment in PKU mice demonstrated engineered microbes could compensate for a metabolic deficiency of the host. Objective 2: Evaluate the PAHenu2 mouse model of PKU for a genetic discrepancy causing ocular enlargement and delayed development observed only after the PAHenu2 mutation was crossed to the C57BL/6J mouse. When compared to healthy littermates, ELISA indicated a consistent but insignificant decrease in plasma IGF-1 and an increase in ocular IGF-1 in PKU animals. SNP screening demonstrated a differential inheritance of IGF-1 alleles in healthy and …
Date:
December 2015
Creator:
Durrer, Katherine Elaine
System:
The UNT Digital Library
Genetic Analysis of Medicago truncatula Plants with a Defective MtIRE Gene
Leguminous plants are able to fix nitrogen by establishing a symbiotic relationship with soil dwelling bacteria, called rhizobia. The model plant Medicago truncatula forms a partnership with Sinorhizobium meliloti whereby the plant gains bioavailable nitrogen and in exchange the bacteria gains carbohydrates. This process occurs within nodules, which are structures produced on the roots of the plants within which nitrogen is fixed. M. truncatula incomplete root elongation (MtIRE) was localized to the infection zone, which is zone II of indeterminate nodules. It was shown to encode a signaling kinase so it was anticipated to play a role in nodulation. Mutants of MtIRE in the R108 background, mutagenized with the Tnt1 retrotransposon, were obtained from reverse screen, and were assessed to determine if a disrupted MtIRE gene was the cause of nitrogen fixation defective nodules. Mutant line NF1320, having a mutant phenotype, showed typical Mendelian segregation of 3:1 when backcrossed to R108. Experimental results show that MtIRE gene is not the cause of the mutant phenotype, but was linked to the causative locus. MtIRE co-segregated with the mutant phenotype 83%. Southern blot and the first version of the M. truncatula genome (version 3.5) reported a single MtIRE gene and this was …
Date:
August 2015
Creator:
Alexis, Naudin
System:
The UNT Digital Library
Forward Genetic Characterization of Medicago truncatula Tnt1 Insertion Mutants Defective in Nodule Development and Symbiotic Nitrogen Fixation
Legumes are unique plants because they form special structures “nodules”, via symbiotic relationships with rhizobial bacteria present in the soil. Once rhizobia mature inside nodules, they fix atmospheric nitrogen providing a source of bioavailable nitrogen to the plant. To discover novel genetic components involved in the legume-rhizobia symbiosis by using forward genetic screening, we have isolated Medicago truncatula Tnt1 insertion mutants in the R108 ecotype, which are defective in nodule development and symbiotic nitrogen fixation in response to Sinorhizobium meliloti. Out of three mutants NF11044, NF11217 and NF8324, one of the mutants showed brown nodules and Fix- phenotype that is defective in symbiotic nitrogen fixation. The other two mutants showed white nodules and Fix- phenotype, also indicator of defects in symbiotic nitrogen fixation. To identify the underlying mutation causing the phenotype, we have developed molecular genetic markers by obtaining genomic sequences flanking the Tnt1 insertions by TAIL-PCR and Illumina sequencing. To carry out co-segregation analysis, back-crossed BC1F2 segregating populations were obtained. These are being phenotyped, genotyped and analyzed for co-segregation of the phenotype with the Tnt1 genetic markers. Back-crossing also has the effect of reducing the Tnt1 insertions, which are not linked to the nodulation defective phenotypes. Out of the …
Date:
May 2015
Creator:
Kadel, Khem L.
System:
The UNT Digital Library
A Study of Some Congenital Anomalies of the Teeth
Anomalies of the teeth include a variety of expression such as: anodontia, hyperdontia, peg teeth. opalescent dentine, mottled enamel, and other imperfections of the teeth. This paper is concerned primarily with anodontia in human subjects, although other abnormalities of interest are mentioned. Insufficient datum is the principal difficulty encountered in determining the mechanism and means of transmission in human genetics. Small families, slow breeding, and inability to subject humans to experimentation account, in part, for this difficulty. The lack of reliable data is the second major difficulty in determining the mode of transmission. Often normal or unaffected members of a family are forgotten due to insignificance placed on them--thus an untrue pedigree is the result. The fact that one must rely on information from one or two members of a family lends itself to error. Therefore, an absolutely accurate determination of Mendelian ratios in human genetics would be impossible. This error, however, can be reduced by the gathering and compilation of large numbers of family pedigrees from which calculations can be made and results determined which will reveal the mode of transmission of various hereditary traits. With this view in mind, this paper is presented, first, to add several more …
Date:
August 1955
Creator:
Jones, Alvin Watt
System:
The UNT Digital Library
Relationship of Certain Fungi to Azotobacter in Nitrogen-Free Media
Azotobacter and various fungi were grown together in nitrogen-free media. Maximal fungal growth in the medium used was possible only at the expense of Azotobacter cells and growth was always accompanied by acid production. When the medium reached a pH of 2, the bacterial cells were aggregated on fungal hyphae and the culture fluid appeared to be free of Azotobacter. Aspergillus niger grew well at the expense of viable bacteria and other fungi grew well on heat-killed cells of A. vinelandii. Members of the genus Hormodendrum, although not causing significant decrease in pH, were also able to clear turbid cultures of Azotobacter. However, clearing, which involved the attachment of bacteria to fungal hyphae, was dependent on acid production by the fungi. Bacterial aggregation was followed by hyphal attachment, bacterial inactivation, and finally, bacterial cell lysis.
Date:
August 1976
Creator:
Ray, Manfred G.
System:
The UNT Digital Library
A Multiphasic Study of the Interaction Between the Branchiobdellid Cambarincola Vitrea and its Crayfish Host Procambarus Simulans
The host-parasite interface between the branchiobdellid Cambarincola vitrea Ellis and its crayfish host is investigated along ecological, ultrastructural, serological, and endocrinological lines of evidence. Monthly analysis of a natural branchiobdellid from a vernal habitat demonstrates the annelid to be particularly sensitive to changes involving both the physical habitat as well as the host crayfish.
Date:
August 1973
Creator:
Koepp, Stephen John
System:
The UNT Digital Library
Thyroid Ultrastructural Changes Induced by Hypothermia
Investigations have shown that the hypothalamus and pituitary respond to decreases in body temperature by stimulating the thyroid gland to release T3 and T4 hormones. This study was designed to investigate ultrastructural changes of the thyroid gland induced by hypothermia. The ultrastructural changes produces by stimulation by Thyroid Stimulating Hormone were also examined as an adjunct to the hypothermic stimulation of the gland. There was a significant increase in microvilli on the luminal border of follicle cells along with a remarkable increase in numbers of dense granules. The dense granules also demonstrated a polarity being found near the border of the colloid lumen. The glandular changes induced by Thyroid Stimulating Hormone were very similar to those of hypothermia.
Date:
August 1979
Creator:
Kent, James Simpson
System:
The UNT Digital Library
An Evaluative Study of the Grasslands of the R.J. McMurry Ranch, Denton County, Texas
It is the purpose of this problem to classify the four major pastures of the McMurry ranch using the discussed classification system. The definite measurable qualities characterizing each condition of the system are used extensively in this study. The problem is concerned also with the observation of results of misuse, the present practices that could result in further depletion of portions of the McMurry ranch, and procedures being employed at the present time aimed toward the restoration of these depleted portions.
Date:
June 1951
Creator:
Nolen, Bette Rudd
System:
The UNT Digital Library
Effects of Smoking on Gastric Secretion and Gastric Motility in Man
This thesis is concerned with the use of some of the newer techniques in a study of some of the effects of cigarette smoking on gastric secretion and gastric motility in normal subjects and in patients with active duodenal ulcers.
Date:
June 1956
Creator:
McKenzie, Jess Mack
System:
The UNT Digital Library
A Quantitative and Qualitative Bacterial Analysis of the Sewage Disposal System of Perryton, Texas
This investigation had for its purpose the determination of first, the inefficiency of the Perryton, Texas, sewage disposal plant proper; and second the potential effectiveness of a lagoon now used to complete the sewage purification; and consequently, offset an apparent health hazard. The problem, as pursued, has consisted of first, the collection of sewage samples from the six stations listed in Table 1 over the period of time from September 23, 1949 to May 14, 1950; second, a quantitative analysis of each sample in order to determine the number of bacterial organisms present; third, a qualitative analysis of each sample in order to determine more expecially the presence of members of the genera Aerobacter, Escherischia, Salmonella and Shigella; and fourth, an attempt to evaluate the potentialities of the natural purification factors of the lagoon.
Date:
August 1950
Creator:
Sullivan, Troy G.
System:
The UNT Digital Library
Development of Range Condition Classes for Coleman County, Texas
This paper deals with a study of the rangelands in Coleman County just west south and west of the Western Cross Timbers of Texas. This problem consists of classifying certain rangelands in this area according to whether they are excellent, fair, good, or poor, the chief basis for which is their deterioration.
Date:
June 1951
Creator:
McMinn, Philip Morris
System:
The UNT Digital Library
Investigating the Ability of Pseudomonas aeruginosa pyrE Mutants to Grow and Produce Virulence Factors
Pseudomonas aeruginosa are medically important bacteria that are notorious for causing nosocomial infections. To gain more knowledge into understanding how this organism operates, it was decided to explore the pyrimidine biosynthetic pathway. Pyrimidine synthesis, being one half of the DNA structure, makes it a very important pathway to the organism’s survivability. With previous studies being done on various genes in the pathway, pyrE has not been studied to the fullest extent. To study the function of pyrE, a site directed mutagenesis was done to completely knock out pyrE, which encodes the protein orotate phosphoribosyl transferase that is responsible for converting orotate into orotate monophosphate (OMP). A mutation in this step leads to accumulation and secretion of orotate into the medium. Analyzing virulence factors produced by the mutant and comparing to the wild type, some intriguing features of the mutant were discovered. One of the findings was the over expression of virulence factors pyoverdin and pyocyanin. Pyocyanin over expression, based on the results of this study, is due to the accumulation of orotate while over production of pyoverdin is due to the accumulation of dihydroorotate. The other virulence factors studied were motility assays, exoproducts, and growth analysis. All virulence factor production …
Date:
December 2014
Creator:
Niazy, Abdurahman
System:
The UNT Digital Library
The Impact of Developmental Stress on Cardiovascular Physiology of Two Archosaur Species: American Alligator (Alligator mississippiensis) and Domestic Chicken (Gallus gallus)
Crocodilians and birds comprise sister taxa of archosaurs, the development of these vertebrates occurs within an egg case that leaves developing embryos susceptible to fluctuations in the nesting environment. Studies suggest that sub-optimal conditions alter morphological growth and cardiovascular physiology. Regulation of the cardiovascular system is immature in the subjects studied, and embryos may rely on humoral rather than neural control of the cardiovascular system. The primary focus of this dissertation was to assess regulatory mechanisms responsible for maintenance of arterial pressure and heart rate. Dehydration stress had marked effects on embryo growth, and altered baseline cardiovascular parameters, while leaving the response to humoral regulator, angiotensin II (Ang II), unaffected. However, dehydrated alligator embryos developed cholinergic tone on heart rate. Hypoxic incubated chicken embryos were reduced in embryo mass, and altered response to humoral regulatory components Ang I and adenosine in addition identifying a novel regulatory component of the cardiovascular response to acute hypoxia. Collectively, these studies add to the existing knowledge of cardiovascular physiology in embryonic archosaurs and suggest that some components of cardiovascular regulation are plastic following developmental stress.
Date:
December 2014
Creator:
Tate, Kevin B.
System:
The UNT Digital Library
The Effects of Common Electrolytes on Growth and Development of Selected Species of Aquatic Actinomycetes
The role that aquatic actinomycetes play, in the production of tastes and odors in water supplies has been investigated since 1948. The ability of these organisms to produce by-products in lakes and streams, which renders the water unpalatable, is of considerable public health importance. It is desirable that the waterworks industry has as much information as possible concerning the factors that contribute to the growth of these organisms. Since it appears that the aquatic actinomycetes may be isolated from most fresh-water sources, the problem of diversified environments and nutritional requirements offers an excellent field of investigation. The fresh-waters of the world contain variable quantities of electrolytes that may determine in part the biological activity of these organisms. The unsolved questions in this instance are concerned with the electrolytes present and their quantitative effects on the growth and development of these forms.
Date:
August 1959
Creator:
Sissom, Stanley L.
System:
The UNT Digital Library