UNT Libraries - 19 Matching Results

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Advanced Molecular and Microbial Techniques: a Complete Laboratory Notebook

Description: The purpose of this project is to produce a complete and thorough notebook that may be used to supplement laboratory coursework. Its intent is to be used primarily by the students to aid them in understanding background information and the proper laboratory procedures involved in various types of experiments. The laboratory notebook is a summation of all the experiments and procedures used in the six-credit hour Advanced Microbial and Molecular Biology (BIOL 5160) course offered during the summer semester at the University of North Texas. This class is a team taught effort by Professors O'Donovan and Kunz. The course is constructed as an intensive practice exercise to teach the student about gene mutations, biosynthetic pathways, preparation and analysis of plasmid DNA, and many other topics included in the notebook.
Date: May 1998
Creator: Brito-Rodriquez, Carmen Lydia

Analysis of the Trypanosoma brucei Genome and Identification and Characterization of a Gene Family Encoding Putative EF-Hand Calcium-Binding Proteins

Description: The flagellum of Trypanosoma brucei contains a family of antigenically related EF-hand calcium-binding proteins which are called the calflagins. Genomic Southern blots indicated that multiple copies of calflagin genes occur in T brucei. All of the copies were contained in a single 23 kb Xhol-Xhol fragment. Genomic fragments of 2.5 and 1.7 kb were cloned that encoded calflagin sequences. Two new members of the calflagin family were found from genomic clone sequences. The deduced amino acid sequences of the genomic clones showed the calflagin genes were arranged tandemly along the genomic fragments and were similar to previously described calflagins. The calflagin genes were related by two unrelated 3' flanking sequences. An open reading frame that was unrelated to any calflagin was found at the 5' end of the 2.5 kb genomic fragment. Each encoded protein (~24,000u) contained three EF-hand calcium-binding motifs and one degenerate EF-hand motif. In general, variability among the T. brucei calflagins is greater than related proteins in T. lewisii and T. cruzi. This variability results from amino acid substitutions at the amino and carboxy termini, and duplication of internal segments.
Date: May 1998
Creator: DeFord, James H. (James Henry), 1956-

Biochemical Identification of Molecular Components Required for Cyanide Assimilation in Pseudomonas fluorescens NCIMB 11764

Description: Utilization of cyanide as a nutritional nitrogen source in P. fluorescens NCIMB 11764 was shown to involve a novel metabolic mechanism involving nonenzymatic neutralization outside of cells prior to further enzymatic oxidation within. Several cyanide degrading enzymes were produced by NCIMB 11764 in response to growth or exposure to cyanide, but only one of these cyanide, oxygenase (CNO), was shown to be physiologically required for assimilation of cyanide as a growth substrate.
Date: May 1998
Creator: Chen, Jui-Lin

Biogeographic Relationships of Pocket Gophers (Geomys breviceps and Geomys bursarius) in the Southeastern Portion of Their Ranges

Description: This research utilized population genetic analyses (protein starch-gel electrophoresis and DNA sequencing of the cytochrome b mtDNA gene), host-parasite specificity (lice coevolution), remote sensing of satellite data, and geographic information systems (GIS) to characterize newly discovered populations of pocket gophers (genus: Geomys) in Arkansas. These populations are isolated and occur in seemingly unsuitable habitat in the Ozark Mountains of Arkansas. Analyses of electrophoretic and ectoparasite data suggested the populations in the Ozark Mountains represented isolates allied to Geomys bursarius, a species not known to occur in Arkansas. Comparison of mitochondrial DNA sequence data of the cytochrome b gene with that of other taxa and morphometric analyses confirmed that these populations are most closely allied to G. bursarius occurring to the north in Missouri. Moreover, these mtDNA sequence analyses indicated a degree of differentiation typical of that between other subspecies of pocket gophers. Therefore, these populations represent a distinct genetic entity in an intermediate stage of speciation and should be designated as a new subspecies, Geomys bursarius ozarkensis. Molecular clock analysis revealed a time of lineage divergence for this new subspecies as approximately 511,000 YBP. Due to the isolated nature and limited distribution of this subspecies, an evaluation of critical habitat needs was initiated. Remote sensing and GIS technologies were used to identify and describe suitable habitat Computerized classification of satellite imagery of suitable vegetation, integrated with ancillary digital information on soil associations, roads, and water systems, revealed that human activity had played a positive role in the establishment and dispersal of pocket gophers in this area. This research represents an initial combination of classical systematic tools with remote sensing and GIS to investigate biogeographic patterns and evolution. This project establishes a framework for using an interdisciplinary approach to studying organisms with limited distributions, determining evolutionary status, and providing recommendations for ...
Date: August 1998
Creator: Elrod, Douglas Allen

Dalbergia and Albizia: Plantlet Production via Tissue Culture, Karyological Evaluation, and Seed Anatomy with Scanning Electron Microscopy

Description: A publication by the National Academy of Sciences, USA (1979) outlined some of the research need for a great variety of economically important woody species whose remaining genetic resources need urgently to be collected and conserved. A viable regeneration system was established via tissue and cell suspension culture for Albizia falcataria and A. lebbeck, two important wood yielding leguminous tree species. The culture medium was standardized after several trials to obtain callus from the leaflet explants of these two tree species. The optimum use of casein hydrolysate (w/v) and coconut milk (v/v) in addition to 6-Benzylaminopurine and Indole-3-butyric acid could induce morphogenesis and somatic embryogenesis in the cultured tissue. This reports the first observation on somatic embryogenesis ofA. lebbeck using leaflets as the explants. Scanning Electron Microscopy and histological studies were done on the different stages plant development following standard techniques. Embryogenesis in suspension culture followed regeneration of plantlets in A. lebbeck. In A.falcaaria the regenerative process followed via organogenesis from the shoot buds developed on the leaf explants. After hardening the regenerated plants were transferred to the greenhouse. Some of the trees grew more than 25 feet tall within a few months outside the greenhouse. Karyotype of the three leguminous trees Albizia lebbeck, A. falcataria, and Dalbergia sissoo was analyzed. In D. sissoo, various chromosomal anomalies were observed in the cultured tissue. The abnormality indices and ploidy level varied with the age and the frequency of the subculture. In the aged culture the regenerative potential declined but was reinstated to some extent with the addition of two complex growth factors, coconut milk and casein hydrolysate. Seed anatomy of 26 species of 4 leguminous genera was studied with SEM. The main distinguishing anatomical features observed in the seed sections were uniseriate or multiseriate epidermis, epidermal projections, and number of rows ...
Date: December 1998
Creator: Ghosh, Nabarun

Development and Application of an Assessment Protocol for Watershed Based Biomonitoring

Description: With numerous bioassessment methodologies available, a regional protocol needs to be developed to ensure that results are comparable. A regional assessment protocol was developed that includes collecting five benthic macroinvertebrate samples, identifying organisms to genus, and calculating the following metrics: Number of Taxa, Total Number of Individuals, Simpson's Diversity Index, Shannon's Diversity Index, Percent Contribution of Dominant Taxa, Hilsenhoffs Biotic Index, and Percent Contribution of Dipterans. Once the protocol was developed, it was used to assess the Bayou Chico tributaries and watershed. All three tributaries had been significantly impacted by human activity as had the watershed as a whole. This study indicates that a regional protocol could be developed and is appropriate for biomonitoring at the watershed scale.
Date: May 1998
Creator: Schwartz, Joseph Howard

Development of an In Vitro Protoplast Culture System for Albizia Lebek (L.) Benth., an Economically Important Leguminous Tree

Description: An in vitro system of generating protoplasts from their callus cultures was established. The friable callus was more productive in terms of producing protoplasts than the green compact callus. The concentration of the various cell wall degrading enzymes had an effect on the viability of the protoplasts in the medium. The protoplast system developed from the experiments was stable and could be used for the transformation experiments of Albizia lebek and for other plant improvement practices.
Date: August 1998
Creator: Sinha, Debleena

Documentation of Biodiversity Impacts (Including Cumulative Biodiversity Impacts) in Environmental Impact Statements

Description: In the United States, biodiversity impact assessment has historically received little attention. Responding in 1993, the Council on Environmental Quality (CEQ) released guidelines on incorporating biodiversity into environmental impact assessment under the National Environmental Policy Act of 1969. The objectives of the study here were to identify the level of documentation of biodiversity impact assessment in sample Environmental Impact Statements (EISs); identify whether in the years following the release of 1993 CEQ guidelines any significant changes have taken place in assessment of biodiversity; identify deficiencies, and if the need exists, formulate appropriate recommendations and approaches for addressing biodiversity in EISs. The study involved a systematic review of 30 EISs published since the release of CEQ guidelines, and five EISs published prior to it. The review involved answering a series of standard questions, which attempted to ascertain the level of biodiversity impacts included in each impact statement. Trends in approaches to biodiversity impact assessment were investigated and deficiencies summarized. The analysis resulted in a series of recommendations for improving the manner in which biodiversity impact assessment can be approached.
Date: August 1998
Creator: Bhatia, Sarika

Ecological Association Between the Red-Cockaded Woodpecker and Southern Pine Beetle in the Homochitto National Forest: a Geographic Information System Approach

Description: Since the introduction of management practices by the Forest Service to stabilize red-cockaded woodpecker (RCW) populations, the number of cavity trees killed by southern pine beetles (SPB) has increased. A model of the landscape ecology of RCW and SPB in the Homochitto National Forest was created using data collected from the Forest Service and Global Atmospherics. The conclusions of the study were that the RCW and SPB utilize the same type of habitat and the stand hazard maps are an accurate means of determining the locations of SPB infestations. The functional heterogeneity maps created for the SPB and RCW would be useful predictors of future occurrences of either species if complete data were obtained.
Date: December 1998
Creator: Skordinski, Karen R. (Karen Renee)

Forensic DNA Extraction Strategies for PCR Analysis

Description: There is a transition nationwide on the analysis of forensic evidentiary stains containing biological material from traditional serology to Polymerase Chain Reaction (PCR) methodologies. The increased sensitivity of PCR, the limited number of alleles at each locus, and the necessity of producing unambiguous data for entry into the FBI's Combined DNA Index System make this study of extraction procedures of utmost importance. A "single tube" extraction procedure for blood stains collected onto FTA™ paper and a modified differential nonorganic extraction method from spermatozoa containing mixed stains were analyzed and compared. The extraction success was evaluated by amplification and typing of the amplified fragment length polymorphism, D1S80. These modifications of the nonorganic method utilized gave an improved separation of the spermatozoa-containing mixed stains.
Date: May 1998
Creator: Van Winkle, Carolyn

Identification and Characterization of the Pyrimidine Biosynthetic Operon in Streptomyces griseus

Description: To further understand the ATCase/DHOase bifunctional complex formed in Streptomyces, the genes encoding these and other pyrimidine enzymes were identified and characterized. Polymerase chain reaction (PCR) was utilized in this effort. Primers were constructed by selecting conserved regions of pyrimidine genes from known gene and protein sequences of a wide variety of organisms. These sequences were then optimized to Streptomyces codon usage. PCR products were obtained from internal sites within pyrimidine genes and also from primer combinations of different genes. The size, orientation, and partial sequence of the resulting products shows that Streptomyces has a gene organization of pyrR followed by pyrB, pyrC, carA, carB, and pyrF in an operon similar to that found in other Gram-positive bacteria.
Date: May 1998
Creator: Hooten, Jody J. (Jody Jeran)

Medial Medulla Networks in Culture: a Multichannel Electrophysiologic and Pharmacological Study

Description: Spontaneously active primary cultures obtained from dissociated embryonic medial medulla tissue were grown on microelectrode arrays for investigating burst patterns and pharmacological responses of respiratory-related neurons. Multichannel burst rates and spike production were used as primary variables for analysis. Pacemaker-like neurons were identified by continued spiking under low Ca++/high Mg++conditions. The number of pacemakers increased with time under synaptic blocking medium. Sensitivity to CO2 levels was found in some neurons. Acetylcholine changed activity in a complex fashion. Curare, atropine and gallamine modified ACh effects. Eserine alone was ineffective, but potentiated ACh-induced responses. Norepinephrine caused channel-specific increases or decreases, whereas dopamine and serotonin had little effect at 30 μM. GABA and glycine stopped most spiking at 70 μM. Developmental changes in glycine sensitivity (increasing with age) were also observed. It is concluded that pacemaker and chemosensitive neurons develop in medial medulla cultures, and that these cultures are pharmacologically histiotypic.
Date: August 1998
Creator: Keefer, Edward W. (Edward Wesley)

Physical and Functional Characterization of the xy1XYZ Region From TOL Plasmid pDK1 and its Associated Downstream Regulatory Elements

Description: The nucleotide sequence for the pDKl TOL plasmid region encoding toluate-1,2-dioxygenase (Xy1XYZ, TO) was determined. TO is the first enzyme in the meta-cleavage operon, responsible for the conversion of toluates and benzoates to their carboxy-substituted diols. DNA sequence analysis revealed the presence of three open reading frames (ORF). The three ORFs correspond to xylX (1353 bp), xylY (486 bp) and xylZ (1008 bp), encoding predicted protein products of 51370 Da, 19368 Da and 36256 Da, respectively.
Date: August 1998
Creator: Hares, Douglas R. (Douglas Ryan)

Responses of Cultured Neuronal Networks to the Cannabinoid Mimetic Anandamide

Description: The effects of cannabinoid agonists on spontaneous neuronal network activity were characterized in murine spinal cord and auditory cortical cultures with multichannel extracellular recording using photoetched electrode arrays. Different cultures responded reproducibly with global decreases of spiking and bursting to anandamide and methanandamide, but each agonist showed unique minor effects on network activity. The two tissues responded in a tissue-specific manner. Spontaneous activity in spinal tissue was terminated by 1 μM anandamide and 6.1 μM methanandamide. Cortical activity ceased at 3.5 μM and 2.8 μM respectively. Irreversible cessation of activity was observed beyond 8 μM for both tissues and test substances. Palmitoylethanolamide, demonstrated that CB2 receptors were not present or not responsive. However, the data strongly suggested the presence of CB1 receptors.
Date: May 1998
Creator: Morefield, Samantha I. (Samantha Irene)

Role of α-Keto Acids In Cyanide Detoxification and Assimilation by Pseudomonas Bacteria

Description: Cyanide was rapidly removed when added to culture supernatants of seven different Pseudomonas. The ability to remove cyanide was correlated with the accumulation of α-keto acids (pyruvate and α-ketoglutarate). These compounds react with cyanide forming less toxic cyanohydrins, thus conferring a mechanism for bacterial cyanide tolerance. When added to growth media the α-keto acids were shown also to serve as effective cyanide antagonists. While all bacteria tested accumulated α-keto acids, only those capable of utilizing cyanide as a nutritional nitrogen source were able to metabolize cyanohydrins. In P. fluorescens NCIMB 11764, the same enzyme (cyanide oxygenase) shown previously to be involved in cyanide metabolism appears responsible for cyanohydrin transformation. Keto acid excretion is believed to represent a new mechanism of bacterial cyanide detoxification with further enzymatic metabolism of the cyanohydrins helping to explain how cyanide can satisfy the nitrogen requirement in cyanide-utilizing bacteria.
Date: December 1998
Creator: Pan, Guangliang

Underwater Optical Properties of Lake Texoma (Oklahoma-Texas) Using Secchi Disk, Submarine Photometer, and High-Resolution Spectroscopy

Description: The underwater optical climate of Lake Texoma was measured at eleven fixed stations from August 1996 to August 1997. Secchi transparency and submarine photometry characterized seasonal and spatial values of secchi depth (SD), vertical attenuation coefficient (η''), and depth of euphotic zone (Zeu). Indices of Zeu:SD and η'' × SD were compared with universally applied values derived from inland and coastal waters. Turbidity explained 76% of the variation (p = 0.0001) of η'' among water quality parameters, including chlorophyll-α. Using a spectroradiometer, spectral signatures of chlorophyll-α and turbidity were located. Stations with low turbidity exhibited a distinct green reflectance peak around 590-610 nanometers, indicating presence of chlorophyll-α. Stations with high turbidity exhibited a reflectance peak shift towards the red spectrum, making it difficult to detect the chlorophyll signature. Derivative analysis of the reflectance signal at 590-610, and 720-780 nanometers allowed discrimination of this chlorophyll signature from those of turbidity (0.66 ≤ r^2 ≤ 0.99).
Date: August 1998
Creator: Rolbiecki, David A. (David Alan)