In translational research, disease models in preclinical studies are used as media for discovery of drugs or novel therapeutics. Development of in vitro models for various neurological diseases that enable efficient pharmacological or toxicological screening has been ongoing but challenging. Recognizing the potential benefit of in vitro disease models, dysfunctions in the cortical neuronal networks were induced to mimic the functional pathology of neurological symptoms using microelectrode arrays. Two different disease states – tinnitusand excitotoxicity – were investigated and discussed. In this model, pentylenetetrazol-induced increase in spontaneous firing rate and synchrony in the auditory cortical networks was used as correlate of tinnitus. Potential tinnitus treatment drugs from several different classes – including the novel class of potassium channel openers – were screened and quantified. The potentialtherapeutic values of these drugs were also discussed as the basis for drug repurposing. Functional excitotoxicity was induced by cisplatin (a cancer drug that causes neurological sideeffects) and glutamate (the major excitatory neurotransmitter). As proof-of-principle that the model may contribute to expediting the development of therapeutics, cisplatin excitotoxicity wasprevented by the antioxidant D-methionine, while glutamate excitotoxicity was prevented by ceftriaxone (a modulator of a glutamate reuptake transporter). In the latter part of the study, with …

The presence of pharmaceuticals in the environment is becoming an increasing regulatory and scientific concern. Thus, the metabolic profile and bioconcentration potential of diazepam, a model benzodiazepine, were examined, as well as effects on the endocrine system in channel catfish. Through the use of specific and non-specific cytochrome P450 (CYP450) inhibitors, it was determined that CYP3A-like enzymes may play a role in the biotransformation of diazepam into temazepam; however, the isoform(s) required for the formation of other metabolites is still unknown. Overall, only around 7-8% of diazepam is biotransformed into two known metabolites. Due to the lack of inherent metabolism of diazepam in channel catfish, further analysis was conducted to determine the tissue-specific bioconcentration potential of diazepam in catfish. Various tissues were analyzed for the presence of diazepam as well as metabolites and bioconcentration factors (BCF) were calculated, which were all well below regulatory threshold values (> 2000). Additionally, modulation of the endocrine system by diazepam was examined by measuring steroid hormone concentrations and analyzing mRNA expression of selected steroidogenic enzymes and receptors. Two steroidogenic enzymes were modulated following diazepam exposure, indicating potential endocrine disrupting properties of diazepam. Together, these data suggest that diazepam exhibits low metabolic transformation rates in …

Glyphosate based herbicides are among the most widely used herbicides in the world. The purpose of this study was to determine developmental toxicity of glyphosate, the active ingredient in the common herbicide Roundup, on developing chicken embryos. Few studies have examined toxic effects of glyphosate alone versus the full compound formulations of Roundup, which include adjuvants and surfactants. Adjutants and surfactants are added to aid in solubility and absorption of glyphosate. In this study chicken embryos were exposed at the air cell on embryonic day 6 to 19.8 or 9.9 mg / Kg egg mass of glyphosate in Roundup or glyphosate only. Chickens treated with 19.8 and 9.9 mg / Kg glyphosate in Roundup showed significant reduction in survivability compared to glyphosate alone treatments and controls. On embryonic day 18, embryos were sacrificed for evaluation of developmental toxicity using wet embryo mass, dry embryo mass, and yolk mass as indicators. Morphology measurements were taken on liver mass, heart mass, tibiotarsus length and beak length. Embryos treated with 19.8 mg / Kg glyphosate and 9.9 mg / Kg glyphosate in Roundup showed significant reductions in wet and dry embryo mass and yolk mass. Tibiotarsus length in 9.9 mg / Kg glyphosate …

The lumbar motor activity in isolated spinal cords of 72 postnatal Balb/C mice aged 2, 5, 10 and 21 days (PN2-21) was electroneurographically recorded (ENG) via bilateral ventral roots following treatment with three different concentrations (25, 100 and 200 pM) of the neurotransmitter, 5-hydroxytryptamine (5-HT), i.e., serotonin, to determine its effects on spinal pattern generation.

The proposed biology curriculum for non-majors has one main objective, namely to improve scientific literacy among college students. The National Science Education Standards defines scientific literacy as "the knowledge and understanding of scientific concepts and processes required for personal decision making, participation in civic and cultural affairs, and economic productivity". The suggested strategies to accomplish this goal are to limit the number of topics covered, introduce relevant scientific terminology, emphasize general biological concepts and themes, and hone critical thinking and problem-solving skills. Activities such as group projects, written and oral assignments, and class discussions are effective tools to assess student ability to communicate scientifically. It is also important for students to make connections between the course subject matter and how it affects real life events.

This research evaluated the potential of activation and attachment, as sequential companion biomarkers of phagocytosis by earthworm, Lumbricus terrestris, immunoactive coelomocytes for use in immunotoxicology. The potential was assessed by exposing earthworms to sublethal concentrations of CuSO4 and Arochlor 1254®, chemicals used as reference or standard immunotoxicants.

A 2 x 2 factorial design investigated effects of sediment nitrogen and water potassium levels on autofragment production. Reduced nitrogen levels significantly increased autofragment production whereas potassium levels did not significantly alter production. Up to 50% of autofragment production abscised from parent plants grown under low nitrogen conditions compared to 12% or less under high nitrogen conditions.

Recent studies regarding research on oral microorganisms and the oral diseases are presented. The normal flora of the mouth and the oral environment are first described. Dental plaque and dental caries are primary causes of oral disease. Streptococcus mutans is the major contributor in the initiation and progression of dental caries. Lactobacillus, Actinomyces, and Veillonella are other genera of bacteria linked to dental caries. Periodontitis and gingivitis are periodontal diseases that are caused by oral microorganisms. New research has indicated that various antimicrobial agents and techniques to eliminate or lessen the severity of periodontal diseases. Premature delivery of low birth weight babies in pregnant women has been strongly linked to periodontal disease. Present and future microbiological tests are available to easily determine the causative organisms for most oral diseases that help in diagnosis and treatment of a particular disease.

A 30-mer oligonucleotide probe encoding the "A box" and anticodon loop regions of a human glycine tRNA gene was used to isolate a 581bp DNA fragment from a bovine genomic DNA library. Although the cross-hybridizing segment of DNA was found not to encode any tRNA gene or pseudogene, a region with homology to the "C-element" of the "BOV-tA" type Alulike artiodactyl retroposons was identified. This cross-hybridization was determined to be the result of conserved RNA polymerase III promoter elements in the probe portion of the tRNA gene and these repetitive elements. A microsatellite repeat (TC) was also found associated with this element. Future screening for bovine tRNA genes will require the use of a) longer probes and higher stringency hybridization conditions or b) the simultaneous screening with probes from the 5' and 3' ends of the gene which avoid the conserved Pol III promoter boxes.

Bacterial aspartate transcarbamoylases (ATCase's) are divided into three classes that correspond to taxonomic relationships within the bacteria. The opportunistic pathogen Moraxeila catarrhalis has undergone several reclassifications based on traditional microbiological criteria. The previously uncharacterized ATCase from M. catarrhalis was purified to homogeneity and its chemical properties characterized. The ATCase from M. catarrhalis is a class C ATCase with an apparent molecular mass of 480-520 kDa. The M. catarrhalis ATCase is a dodecomer composed of six 35 kDa polypeptides and six 45 kDa polypeptides. The enzyme has an unusually high pH optimum of greater than pH 10. The enzyme exhibited hyperbolic kinetic with a Km for aspartate of 2 mM. A single, separate 78 kDa dihydroorotase from M. catarrhalis was identified and it was not associated with ATCase. These data support the reclassification of M. catarrhalis out of the Neisseriaceae family.

Aspartate transcarbamoylase (ATCase) of Streptomyces was characterized and its interaction with other pyrimidine enzymes explored.

Several biomarkers (cell viability and phagocytosis) based on earthworm (Lumbricus terrestris) immune cells (coelomocytes), together with whole-worm mortality (LC/LD50's), were used to assess a bioremediation attempt to reduce pentachlorophenol (PCP) toxicity in a former wood processing hazardous waste site (HWS).

A total of 192 largemouth bass were caught at Lake Ray Roberts, Texas (1995) to investigate five controversial bass angling techniques relative to hooking mortality. The bait types were Texas-rigged scented and non-scented plastic worms, Carolina-rigged scented and non-scented plastic worms, and live golden shiners. Overall hooking mortality was 21.87% and mortality was dependent upon bait type. Highest mortality resulted from the Texas-rigged scented lures, while the lowest mortality was generated by live golden shiners. A creel survey indicated that few anglers were having success with the investigated baits. Factors that had a confirmed effect on hooking mortality were hooking location and water temperature. Hooking mortality was not excessive compared to other similar studies.

N-acylation of phosphatidylethanolamine (PE) with free fatty acids catalyzed by N-acyl phosphatidylethanolamine (NAPE) synthase was reported in cotyledons of 24-h-old cotton seedlings. Here I report subcellular localization of this enzyme. Differential centrifugation, sucrose density gradient fractionation,aqueous two-phase partitioning and electron microscopy techniques were utilized to elucidate subcellular site(s) of NAPE synthase. Marker enzymes were used to locate organelles in subcellular fractions. Differential centrifugation indicated that NAPE synthase is present in more than one organelle and it is a membrane bound enzyme. Sucrose density gradient fractionations indicated that NAPE synthase is present in membranes derived from endoplasmic reticulum (ER),Golgi and possibly plasma membrane (PM) but not mitochondria, glyoxysomes or plastids. Aqueous two-phase partitioning experiments with cotton and spinach tissues supported these results but Goigi appeared to be the major site of NAPE synthesis. Electron microscopy of subcellular fractions was used to examine isolated fractions to provide visual confirmation of our biochemical results. Collectively, these results indicate that NAPE is synthesized in plant ER, Golgi and possibly PM.

The clcD gene encoding dienelactone hydrolase (DLH) is part of the clc gene cluster for the utilization of the B-ketoadipate pathway intermediate chlorocatechol. The roles that individual amino acids residues play in catalysis and binding of the enzyme were investigated. Using PCR a 1.9 kbp clcD fragment was amplified and subcloned yielding a 821 bp BamHi to ZscoRI subclone in the plasmid pUC19.

Wetland wastewater treatment offers low-cost, energy efficient alternatives to conventional wastewater technologies. In this study, an artificial wetland was constructed at the City of Denton, Texas Pecan Creek Water Reclamation Plant to facilitate diazinon removal from treated effluent.

An in vitro model of Rapid Acceleration Impact (RAI) Injury was used to study the effects of multiple impact (220 g/impact, 3-5 sec intervals) trauma on cultures of murine CNS cells. Investigations with spinal cord cultures showed that 1) multiple impacts delivered tangential to the plane of cell growth caused neuronal death (12% after 3 impacts to 46% after 10 impacts); 2) multiple impacts delivered normal to the plane of cell growth were much less effective (8% dead after 10 impacts); 3) most neuronal death occurred within 15 minutes after injury 4) morphological changes observed included increased nuclear prominence and somal swelling; and 5) pretreatment with ketamine (0.1mM) reduced cell death from 51 to 14% and reduced somal swelling. Identical studies performed on cortical cultures revealed minimal differences between the two tissues in their response to multiple tangential impacts.

The objective of this research was to determine if physical and microbial sediment characterizations could be used to explain benthic macroinvertebrate distribution in two branches of Lake Ray Roberts, Texas.

Salvage of pyrimidine nucleosides and bases by S. griseus and the regulation of aspartate transcarbamoylase (ATCase) were studied. The velocity-substrate curve for S. griseus ATCase was hyperbolic for both aspartate and carbamoylphosphate. The enzyme activity was diminished in the presence of ATP, CTP, or UTP. The synthesis of ATCase was repressed in cells grown in the presence of exogenous uracil. The specific activity of cells grown with uracil was 43 percent of that for cells grown in minimal medium only. Maximal ATCase and dihydroorotase activities were found in the same column fraction after size-exclusion chromatography, suggesting that both activities could reside in the same polypeptide. The pyrimidine salvage enzymes cytosine deaminase and uridine phosphorylase were identified in S. griseus using HPLC reversed-phase chromatography.

The physiological characteristics and organization of cat primary somatosensory cortex (SI) were studied in electrophysiological and anatomical experiments. In single cell recording experiments, quantitatively controlled mechanical stimuli were used to examine the responses of SI cortical neurons to the velocity component of skin or hair displacement. The firing frequency of most rapidly adapting neurons increased as stimulus velocity was increased. Rapidly adapting neurons were classified based on their response patterns to constant-velocity ramp stimuli. Neurons in these classes differed significantly in sensitivity to stimulus velocity and amplitude, adaptation rate, and spontaneous firing rate. The results suggest that frequency coding of stimulus displacement velocity could be performed by individual SI rapidly adapting neurons, and that the classes of rapidly adapting neurons may play different roles in sensation of tactile stimuli. Tract-tracing experiments were used to investigate the ipsilateral corticocortical connections of areas 3b and 2 in SI. Different patterns of connections were found for these areas: area 2 projects to areas 3b, 1, 3a, 5a, 4 and second somatosensory cortex (SII), and area 3b projects to areas 2, 1, 3a and SII. To further compare the organization of these areas, the thalamic input to the forepaw representation within each area was …

Chronic ventricular sympathectomy elicits changes in the coronary circulation, myocardial oxygen consumption and size of infarction resulting fromcoronary occlusion. These changes indicate a change occurring in the basic metabolism of the heart in response to the removal of its sympathetic nervous input. This hypothesis was tested using two groups of dogs, a shamoperated control and a ventricular sympathectomized group. The sympathectomy procedure was an intrapericardial surgical technique which selectively removes ventricular sympathetic input. Four weeks after surgery, left ventricular tissue samples were obtained and rapidly frozen to -80°C. Selected metabolic variables were then compared between the two groups.

General objectives of my study were to describe the reproductive ecology of gray gulls in the large Lealtad colony, with emphasis on demographic parameters and physiological adaptations of eggs and chicks, which would complete some original objectives established in the early 1980's by Guerra and Fitzpatrick. Specifically, my study focused on describing, then comparing with other desert and non-desert nesting larids, interactive effects of ambient physical conditions and nest-site predation on eggs and chicks.

This investigation studied the physiological means by which myocardium can survive and function properly when oxygen supply is limited and cannot initially match oxygen demand. The effects of isoproterenol (ISO) stimulations during low coronary perfusion pressure or hypoxemia on myocardial oxygen demand, work, and oxygen utilization efficiency were investigated.

Products of hydrolytic polymerization of Cr(III) hydroxide were investigated in Milli-Q water and in natural water matrices. Products were first fractionated on Sephadex column using eluents of increasing strength. Ion chromatography (IC) with UV detection at 436 nm was then used to separate the ionic species.