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The Role of Thyroid Hormone on the Development of Endothermy in White Leghorn Chickens [Gallus gallus]
As chickens hatch, there is a rapid change in their physiology and metabolism associated with attaining endothermy. It is thought that thyroid hormones (TH) play a major role in regulating developmental changes at hatching. In birds, TH regulates skeletal muscle growth, which has a direct impact on the chick's ability to thermoregulate via shivering thermogenesis. To better understand the role of TH in the timing of hatching, development of thermogenic capacity, and metabolic rate, we manipulated plasma TH levels in chicken embryos beginning at 85% development (day 17 of a 21 day incubation) with either thyroperoxidase inhibitor methimazole (MMI) or supplemental triiodothyronine (T3). After TH manipulation, we characterized O2 consumption and body temperature in the thermal neutral zone and during gradual cooling. Externally pipped embryos and 1 day post hatch (dph) chicks were cooled from 35 to 15°C. Manipulation of TH altered the timing of hatching, accelerating hatching under hyperthyroid conditions and decelerating hatching with hypothyroid conditions. Cohen's d revealed a large effect size on body temperature (Tb) of EP embryos of hypothyroid animals when compared to euthyroid animals in environmental temperatures of 32°C to 15°C, which was not seen in 1dph animals. Hyperthyroid EP animals were able to maintain metabolic rate over a wider range of ambient temperatures compared to control and hypothyroid animals, but these differences disappeared in 1dph animals. Here, we find that elevating TH levels prior to hatching accelerated hatching and the animal's thermogeneic ability to respond to cooling, but these differences disappear with age.
The Role of Transmembrane Protein 59 in Thrombocyte Function and the Effect of MS-222 on Hemostasis in Zebrafish
Transmembrane protein 59 (tmem59) is a gene that encodes a protein involved in autophagy and apoptosis in human. A previous study in zebrafish showed that tmem59 mRNA was several folds higher in thrombocytes than those found in red blood cells (RBCs). Therefore, we hypothesized that tmem59 has a role in thrombocytes function. We injected a hybrid of control vivo-morpholino (cVMO) and tmem59 specific antisense standard oligonucleotide (tmem59SO) into adult zebrafish to knockdown tmem59.This piggyback knockdown approach resulted in fish that had more bleeding in gill bleeding assay than the control fish. The thrombocytes fromtmem59 knockdown zebrafish aggregated faster with ADP and collagen agonists. Also, the number of blood cells was reduced after the knockdown of tmem59. We also found the effects of MS-222 anesthesia on hemostasis and found that the bleeding was reduced yielding less blood and the blood cell counts increased probably due to vasoconstriction of the blood vessels. In summary, we found tmem59 is a negative regulator of hemostasis and inferred that anesthesia should be avoided in hemostasis studies.
Population Dynamics and Community Structure of Mosquitoes (Diptera: Culicidae) Recorded in Denton, Texas from 2005 to 2015
A population survey was conducted on the mosquito species recorded in Denton, Texas for the years of 2005 to 2015. Data used in this project were obtained from an ongoing, long-term surveillance program led by the City of Denton and conducted through the University of North Texas. Research focused on the population dynamics and community structure of mosquitoes collected within urban areas of Denton, Texas in relation to certain environmental variables. A total of 80,837 female mosquitoes were captured and represented 38 species found under the following genera: Aedes, Anopheles, Coquillettidia, Culex, Culiseta, Mansonia, Orthopodomyia, Psorophora, Toxorhynchites, and Uranotaenia. Culex quinquefasciatus was the most abundant species followed by Aedes vexans. Seasonal patterns of the most abundant species revealed high variability throughout the study. Container breeders were most abundant in August and those that breed in floodwaters were most abundant in the months of May and September. Samples were tested for arbovirus presence through the Texas Department of State Health Services in Austin, Texas and multiple pools tested positive for West Nile virus throughout the study. Stepwise multiple regression and Spearman's rank correlation analyses were performed to examine the relationship between the mosquito community and environmental variables. Data revealed that temperature, precipitation, and dew point were the most important variables influencing the mosquito population in the City of Denton.
In Vitro Exploration of Function Acrolein Toxicity with Cortical Neuronal Networks
Acrolein is produced endogenously after traumatic brain injury (TBI) and is considered a primary mechanism for secondary damage occurring after TBI. We are using frontal cortex networks derived from mouse embryos and grown on microelectrode arrays in vitro to monitor the spontaneous activity of networks and the changes that occur after acrolein application. Networks exposed to acrolein exhibit a biphasic response profile. An initial increase in network activity, followed by a decrease to 100% activity loss in applications ≥ 50 µM. In applications below 50 µM, acrolein was not toxic but generated activity instability with coordinated but irregular population busts lasting for up to 6 days. The increase in activity preceding toxicity may be linked to a decrease in free spermine, a free radical scavenger that modulates Na+, K+, Ca+ channels as well as NMDA, Kainate, and AMPA receptors. Action potential wave shape analysis after 20 and 30 µM acrolein application revealed a concentration-dependent 15-33% increase in peak to peak amplitude within minutes after exposure. For the same concentrations of acrolein (50 µM), the time required to reach 100% activity loss (IT100) was longer in serum-free medium than in medium with 5% serum, in which IT100 values were reduced by a factor of 4. The greater toxicity in the presence of serum may be explained by acrolein adducts on serum proteins. These reaction products have been shown by other labs to be toxic in cell culture. This in vitro system could be used to expand biochemical analyses such as acrolein-induced spermine depletion and may provide an effective platform for investigating cell culture correlates of secondary TBI damage.
"Superbursts": Investigation of Abnormal Paroxysmal Bursting Activity in Nerve Cell Networks In Vitro
Superbursts (SBs) are large, seemingly spontaneous activity fluctuations often encountered in high density neural networks in vitro. Little effort has been put forth to define and analyze SBs which are paroxysmal bursting discharges. Through qualitative and quantitative means, I have described specific occurrences of superbursting activity. A complex of paroxysmal bursting has been termed a "superburst episode," and each individual SB is a "superburst event" which is comprises a fine burst structure. Quantitative calculations (employing overall spike summations and coefficient of variation (CV) calculations), reveal three distinct phases. Phase 1 is a "build up" phase of increasingly strong, coordinated bursting with an average of a 17.6% ± 13.7 increase in activity from reference. Phase 2, the "paroxysmal" phase, is comprised of massive coordinated bursting with high frequency spike content. Individual spike activity increases by 52.9% ± 14.6. Phase 3 is a "recovery phase" of lower coordination and an average of a 50.1% ± 35.6 decrease in spike production from reference. SBs can be induced and terminated by physical manipulation of the medium. Using a peristaltic pump with a flow rate of 0.4ml/min, superbursting activity ceases approximately 28.3 min after the introduction of flow. Alternatively, upon cessation of medium flow superbursting activity reemerges after approximately 8.5 min. Additionally, this study explored other methods capable of inducing superbursting activity using osmotic shocks. The induction and termination of SBs demonstrates that the cell culture environment plays a major role in generating this phenomenon. The observations that high density multi-layer neuronal networks in culture are more likely to enter paroxysmal bursting also supports the hypothesis that enrichment and depletion layers of metabolites and ionic species are involved in such unusual activity. The dynamic similarity of the SB phenomenon with epileptiform discharges make further quantification on the spike pattern level pertinent and important.
Simultaneous Electrophysiological and Morphological Assessment of Impact Damage to Nerve Cell Networks
A ballistic pendulum impulse generator was used to impact networks in primary culture growing on microelectrode arrays. This approach has the advantage of imparting pure tangential acceleration insults (50 to 300 g) with simultaneous morphological and electrophysiological multichannel monitoring for days before and after the impact. Action potential (AP) production, network activity patterns, and cell electrode coupling of individual units using AP waveshape templates were quantified. Network adhesion was maintained after tangential impacts up to 300g with minimal loss of pre-selected active units. Time lapse phase contrast microscopy revealed stable nuclei pre-impact, but post impact nuclear rotation in 95% of observations (n= 30). All recording experiments (n=31) showed a repeatable two-phase spike production response profile: recovery to near reference in 1-2 hrs, followed by a slow activity decay to a stable, level plateau approximately 30-40% below reference. Phase 1 consisted of a complex two-step recovery: rapid activity increase to an average 23.6% (range: 11-34%) below reference, forming a level plateau lasting from 5 to 20 min, followed by a climb to within 20% of reference where a second plateau was established for 1 to 2 hrs. Cross correlation profiles showed changes in firing hierarchy after impact, and in spontaneous network oscillatory activity. Native oscillations were found in the Delta band (2 to 3 Hz), and decreased by approximately 20% after impact. Under network disinhibition with bicuculline, oscillations were slower (0.8-1Hz) and decreased 40% after impact. These data link network performance deficits with microscopically observable subcellular changes.
Phenotypic Analysis of Medicago truncatula NPF1.7 Over-Expressing Plants Grown under Different Nitrate Conditions
Plants have many nitrate transporters; in the model legume Medicago truncatula, MtNPF1.7 is among them. MtNPF1.7 is important for M. truncatula growth and it has been established that MtNPF1.7 is a high affinity nitrate transporter. M. truncatula plants with mutations in MtNPF1.7 gene show defects during plants growth, with striking abnormalities in nodule development and root architecture. Nitrogen fixation is an energy expensive process; when legumes have sufficient bioavailable nitrogen like nitrate available, it suppresses nodulation and nitrogen fixation. Previous preliminary results in our lab showed that plants constitutively expressing MtNPF1.7 have a growth phenotype in the absence of nitrate, but no data was available on how M. truncatula plants constitutively expressing MtNPF1.7 are affected by the presence of nitrate. For my research, I confirmed the preliminary results on the growth of M. truncatula plants overexpressing NPF1.7 and examined these plants' phenotypes when nitrate was not provided in the growth media and when it was provided at two different concentrations. Compared with wild type A17, plants constitutively expressing MtNPF1.7 gene grow larger, have more lateral roots and more nodules when grown in the absence of nitrate and when 0.2 mM KNO3 was provided. At 1 mM KNO3, there are fewer differences between wild type A17 and plants constitutively expressing the MtNPF1.7 gene. Compared with wild type A17, plants constitutively expressing the MtNPF1.7 gene flower earlier, which indicates MtNPF1.7 gene may have a function in plant flowering.
Investigating Human Gut Microbiome in Obesity with Machine Learning Methods
Obesity is a common disease among all ages that has threatened human health and has become a global concern. Gut microbiota can affect human metabolism and thus may modulate obesity. Certain mixes of gut microbiota can protect the host to be healthy or predispose the host to obesity. Modern next-generation sequencing technique allows accessing huge amount of genetic information underlying microbiota and thus provides new insights into the functionality of these micro-organisms and their interactions with the host. Multiple previous studies have demonstrated that the microbiome might contribute to obesity by increasing dietary energy harvest, promoting fat deposition and triggering systemic inflammation. However, these researches are either based on lab cultivation studies or basic statistical analysis. In order to further explore how gut microbiota affect obesity, this thesis utilize a series of machine learning methods to analyze large amount of metagenomics data from human gut microbiome. The publicly available HMP (Human Microbiome Project) metagenomic sequencing data, contain microbiome data for healthy adults, including overweight and obese individuals, were used for this study. HMP gut data were organized based on two different feature definitions: taxonomic information and metabolic reconstruction information. Several widely used classification algorithms: namely Naive Bayes, Random Forest, SVM and elastic net logistic regression were applied to predict healthy or obese status of the subjects based on the cross-validation accuracy. Furthermore, the corresponding feature selection algorithms were used to identify signature features in each dataset that lead to the differences between healthy and obese samples. The results showed that these algorithms perform poorly on taxonomic data than metabolic pathway data though lots of selected taxa are still supported by literature. Among all the combinations between different algorithms and data, elastic net logistic regression has the best cross-validation performance and thus becomes the best model. In this model, several important ...
Neurological Responses to a Glucose Diet in Caenorhabditis elegans
TRPV channels play a role in both mammalian insulin signaling, with TRPV1 expression in pancreatic beta-cells, and in C. elegans insulin-like signaling through expression of OSM-9, OCR-1, and OCR-2 in stress response pathways. In response to a glucose-supplemented diet, C. elegans are know to have sensitivity to anoxic stress, exhibit chemotaxis attraction, and display reduced egg-laying rate. Transcriptome analysis reveals that glucose stimulates nervous system activity with increased transcript levels of genes regulating neurotransmitters. Ciliated sensory neurons are needed for a reduced egg-laying phenotype on a glucose-supplemented diet. Egg-laying rate is not affected when worms graze on glucose-supplemented Delta-PTS OP50 E. coli, which is defective in glucose uptake. This suggests a possible sensory neuron obstruction by exopolysaccharides produced by standard OP50 E. coli on glucose, eliciting a starvation response from the worm and causing reduced egg-laying rate. Glucose chemotaxis is affected in specific TRPV subunit allele mutants: ocr-2(vs29) and osm-9(yz6), serotonin receptor mutants: ser-1(ok345) and mod-1(ok103), and G-alpha protein mutant: gpa-10(pk362). TRPV deletion mutants had no effect on glucose chemotaxis, alluding to the modality role pf TRPV alleles in specific sensory neurons. The role of serotonin in a reduced egg-laying rate with glucose remains unclear.
Delivery of CRISPR/Cas9 into Blood Cells of Zebrafish: Potential for Genome Editing in Somatic Cells
Factor VIII is a clotting factor found on the intrinsic side of the coagulation cascade. A mutation in the factor VIII gene causes the disease Hemophilia A, for which there is no cure. The most common treatment is administration of recombinant factor VIII. However, this can cause an immune response that renders the treatment ineffective in certain hemophilia patients. For this reason a new treatment, or cure, needs to be developed. Gene editing is one solution to correcting the factor VIII mutation. CRISPR/Cas9 mediated gene editing introduces a double stranded break in the genomic DNA. Where this break occurs repair mechanisms cause insertions and deletions, or if a template oligonucleotide can be provided point mutations could be introduced or corrected. However, to accomplish this goal for editing factor VIII mutations, a way to deliver the components of CRISPR/Cas9 into somatic cells is needed. In this study, I confirmed that the CRISPR/Cas9 system was able to create a mutation in the factor VIII gene in zebrafish. I also showed that the components of CRISPR/Cas9 could be piggybacked by vivo morpholino into a variety of blood cells. This study also confirmed that the vivo morpholino did not interfere with the gRNA binding to the DNA, or Cas9 protein inducing the double stranded break.
Generating Molecular Biology Tools to Investigate the Ca2+ Binding Ability of Arabidopsis TON2
The position of the cell division plane in plants is determined by the position of the preprophase band. The pre prophase band (PPB) is a ring of microtubules centered around the nucleus on the inner side of plasma membrane that establishes the cortical division site. The PPB forms at the end of G2 and breaks down at the end of prophase leaving behind protein markers of its position that are collectively called the cortical division site. During cytokinesis the phragmoplast expands towards the cortical division site and mediates the fusion of the new cell plate with the mother cell at that position. Several proteins necessary for PPB formation in plants have been identified, including maize DCD1 and ADD1 and Arabidopsis TON2, which are all type 2A protein phosphatase (PP2A)B" regulatory subunits. DCD1, ADD1, and TON2 localize to the PPB and the cortical division site through metaphase. The PP2A subunits each have two EF-hand domains, which are predicted to bind calcium ions. Since calcium ions are important for some aspects of cell division, we designed a series of constructs to test if TON2 binds calcium. TON2 protein was cloned into expression vectors, pET42a, and expression of TON2 protein was confirmed via Western blotting and immunodetection using a GST antibody. Site directed mutagenesis was used to mutate the TON2 EF-hand domains and mutated cDNAs were also cloned into expression vectors. These were then expressed in bacterial systems. Finally, the GST tagged proteins were purified. In the future, wild-type and mutated proteins TON2 proteins will used in calcium binding assays to determine if TON2 binds calcium.
Evaluating the Role of UV Exposure and Recovery Regimes in PAH Photo-Induced Toxicity
Polyaromatic hydrocarbons (PAHs) are contaminants synthesized through incomplete combustion of carbon based substances. PAHs are known to be photodynamic and toxicity increases exponentially when in contact with ultraviolet radiation (UV). The effect of UV absent recovery periods and potential for latent toxicity during photo-induced toxicity are previously unknown and are not included within the toxicity model. Results of equal interval tests further support the current reciprocity model as a good indicator of PAH photo-induced toxicity. Interval test results also indicate a possible presence of time-dependent toxicity and recovery thresholds and should be included into toxicity risk assessments. Moreover, results of latent effects assays show that latent mortality is a significant response to PAH photo-induced toxicity and should be included into toxicity risk assessments. The present research demonstrates that UV exposure time rate is a significant driving force of PAH photo-induced toxicity.
Respiratory Responses in the Freshwater Snail (Pomacea Bridgesii) are Differently Affected by Temperature, Body Mass,and Oxygen Availability
Pomacea bridgesii is a snail species native to tropical and sub-tropical regions, where it usually faces variability in water, temperature and oxygen level. This study of the effect of temperature on mass-specific oxygen consumption (ṀO2) and its relation to body weight shows that the ṀO2 of juvenile snails in normoxia (18-21 kPa) acclimated at temperature of 25°C ranged from 5 to 58 µMol O2/g/h, with a mean of 41.4 ± 18.3 µMol O2/g/h (n=7). Adult snails in normoxia at 25°C show less variation, ranging from 13 to 23 µMol O2/g/h , with a mean of 24.4± 6.1 µMol O2/g/h (n=12). The Q10 value for juvenile snails was higher in the interval 25-30°C (Q 10=5.74) than in the interval 20-25°C (Q10= 0.286). In adult snails, Q10 was higher in the interval 20-25°C (Q10=3.19). ṀO2 of P. bridgesii in relation to body weight showed a negative linear correlation between metabolic rate and body weight with b values between 0.23 and 0.76. Also, both juvenile and adult snails exhibited weak O2 regulation. In general, the different respiratory characteristics between juvenile and adult snails might be related to the differences of individual life history, which caused them to perform differently in face of temperatures change. Additionally, Pomacean snails species originated in tropical habitats where there is a lack of thermal fluctuation. For this reason, Pomacean snails may be less likely to have evolved effective thermal acclimation capabilities.
Phenotypic Morphological Plasticity Induced by Environmental Salt Stress in the Brine Shrimp, Artemia franciscana
Phenotypic plasticity is the ability of an organism to express different phenotypes in response to biotic or abiotic environmental cues. The ability of an organism to make changes during development to adjust to changes in its environment is a key to survival. Sexually reproducing organisms that have short life cycles and that are easy to raise in the laboratory are more conducive for developmental phenotypic plasticity. Considerable research has already been carried out on the brine shrimp, Artemia franciscana, regarding its morphology due to changing salinities. There is, however, little research considering subsequent generations and how there morphology might be affected by parental experiences. This study has examined: 1) the morphological effects of different rearing regimes of different salinity levels, and 2) the epigenetic transgenerational transfer of these morphological traits in A. franciscana. Measurements included rate of growth (as measured by instar), body size, body length, and other morphological traits. A gradual increase to more hyperosmotic conditions during development produced brine shrimp that were larger in size and also more developmentally advanced. Salinity stress experienced by adults had increased the growth rate in the F1 offspring of A. franciscana. Collectively, these data indicate that Artemia franciscana is a tractable model for investigating phenotypic plasticity. These findings have added to the ever-growing field of developmental phenotypic plasticity while also providing more information on the natural history and adaptive abilities of A. franciscana.
A High-fat Meal Alters Post-prandial mRNA Expression of SIRT1, SIRT4, and SIRT6
Sirtuins (SIRT) regulate the transcription of various genes involved in the development of diet-induced obesity and chronic disease; however, it is unknown how they change acutely following a high-fat meal. The purpose of this study was to determine the effect of a high-fat meal (65% kcals/d; 85% fat recommendation), on SIRT1-7 mRNA expression in blood leukocytes at 1, 3, and 5-h post-prandial. Men and women (N=24) reported to the lab following an overnight fast (>12H). Total RNA was isolated and reverse transcribed prior to using a Taqman qPCR technique with 18S rRNA as a normalizer to determine SIRT1-7 mRNA expression. An additional aliquot of serum was used to measure triglycerides. Data was analyzed using a RM ANOVA with P<0.05. Triglycerides (P<0.001; 124%) peaked at 3-h. SIRT 1 (P=0.004; 70%), and SIRT 6 (P=0.017; 53%) decreased expression at 3-h. SIRT4 (P=0.024) peaked at 5H relative to baseline (70%) and 3-h (68%). To our knowledge, this is the first study to report that consumption of a high-fat meal transiently alters SIRT mRNA expression consistent in a pattern that mirrors changes in serum triglycerides. Decrease in expression of SIRT1 and SIRT6 combined with an increased SIRT4 would be consistent with an increase in metabolic disease risk if maintained on a chronic basis.
The Effect of Post-exercise Ethanol Consumption on the Acute Hormonal Response to Heavy Resistance Exercise in Women
The purpose of this study was to examine the hormonal response to acute ethanol ingestion following a bout of heavy resistance exercise in women. Eight resistance trained women completed two identical acute heavy resistance exercise tasks (AHRET). From 10-20 minutes post-AHRET, participants consumed either a grain ethanol or a placebo beverage. Blood was collected before (PRE) and immediately after the AHRET (IP) and then every 20 minutes for five hours. Blood collected after beverage ingestion was pooled into 3 batches (phases: 20-40 minutes, 60-120 minutes, and 140-300 minutes post-exercise) and analyzed for serum total testosterone (TT), free testosterone (FT), insulin-like growth factor-I (IGF-I), human growth hormone (GH), cortisol (COR), and estradiol (E2) concentrations. Circulating concentrations of TT were significantly greater at P20-40 than at PRE, P60-120, and P140-300. Circulating concentrations of FT were significantly greater at P20-40 than at all other times. Circulating concentrations of GH were significantly greater at IP than at PRE, P60-120, and P140-300. Circulating concentrations of COR were significantly greater at P20-40 than at all other times. Additionally, COR concentrations at P140-300 were significantly lower than at all other times. Circulating concentrations of IGF-1 were significantly greater at P20-40 than at P60-120 and P140-300. Circulating concentrations of E2 were significantly greater at P20-40 than at all other times. In summary, the present study demonstrated an acute modulation of the neuroendocrine milieu following a heavy resistance exercise bout in women. Ethanol ingestion appeared to have no significant effect on the characteristics of acute hormonal augmentation in TT, FT, GH, COR, IGF-1, or E2.
The Role of Cysteinyl Leukotriene Receptor 2 in Thrombocyte Aggregation
Cysteinyl leukotriene receptor 2, a G-protein coupled receptor known to be expressed and functional on human platelets. However, it seems that upon ligand activation the cysteinyl leukotriene receptor 2 activates a variety of signaling pathways in multiple cell types among different species. Previously, a former laboratory member Vrinda Kulkarni found cysteinyl leukotriene receptor 2 to be expressed on the surface of adult zebrafish thrombocytes. In this work I studied the characteristics of aggregation in adult zebrafish thrombocytes with the knockdown of cysteinyl leukotriene receptor 2. I used a newly developed knockdown method to study the function of cysteinyl leukotriene receptor 2. Knockdown of the cysteinyl leukotriene was confirmed using RT-PCR results showed p=.001, reduced sell surface level of expression of the cysteinyl leukotriene receptor 2 results showed that p=.002. I found that the knockdown of cysteinyl leukotriene receptor 2 results in prothrombotic thrombocytes by using flow cytometry p=.0001.
Cytotoxicity and Functional Toxicity of Mefloquine and the Search for Protective Compounds
Mefloquine hydrochloride is an antimalarial agent that has been used for the past 40 years. Numerous reports of neurological side effects have recently led the FDA to issue a strong warning regarding long-term neurological effects. This warning lead to the U.S. Army’s Special Forces and other components to discontinue its use in July of 2013. Despite reported adverse side effects, mefloquine remains in circulation and is recommended to travelers going to specific Asian countries. Mefloquine has been used as a treatment for those already infected with the malaria parasite (blood concentrations ranging from 2.1 to 23 µM), and as prophylaxis (blood concentrations averaging 3.8 µM) (Dow 2003). The purpose of this study was to quantify Mefloquine’s toxicity using spontaneously active nerve cell networks growing on microelectrode arrays in vitro and to identify compounds that alleviate or reduce toxic effects. The current literature on mefloquine toxicity is lacking electrophysiological data. These data will contribute to research on the mechanism of adverse side effects associated with mefloquine use. Sequential titration experiments were performed by adding increasing concentrations of mefloquine solution to cultured neurons. Network responses were quantified and reversibility was examined. In each network, activity decreases were normalized as a percent of reference activity yielding a mean IC50 value of 5.97 ± 0.44 (SD) µM (n=6). After total activity loss, no activity was recovered with two successive medium changes. To test for network response desensitization resulting from sequential applications over 5-6 hr periods, one-point titrations at varying concentrations were conducted with fresh networks. These experiments yielded a single concentration response curve with an IC50 value of 2.97 µM. This represents a statistically significant shift (p < 0.0001) to lower concentrations of mefloquine, demonstrating that sequential applications result in network desensitization. After mefloquine exposures, cells were evaluated for irreversible cytotoxic damage. Over a ...
Neurotoxicity of the Industrial Solvent 4-Methylcyclohexanemethanol: Involvement of the GABA Receptor
A recent chemical spill of 4-Methylcyclohexanemethanol (4-MCHM) in West Virginia left 300,000 people without water. Officials claimed that this compound is not lethally toxic, but potentially harmful if swallowed or inhaled, and can cause eye and skin irritation. Sittig's Handbook of Toxic and Hazardous Chemical Carcinogens reports high exposures from skin contact or inhalation may cause damage to the heart, liver, kidneys, and lungs, and may result in death. However, no quantitative data seem to exist and no references can be found on neurotoxicity. We have investigated the neurotoxicity of 4-MCHM using mammalian nerve cell networks grown on microelectrode arrays. Network spontaneous activity from multiple units (range 48 – 120 per network) were used as the primary readout. Individual units were followed based on spike waveforms digitized at 40 kHz (Plexon MNAP system). Dose response curves show the effective inhibitory concentration at 50 percent decrease (EC50) to average 27.4 microM SD±6.17. However, in the presence of 40 microM bicuculline, a competitive GABAA antagonist, the EC50 shifts to 70.63uM SD ±4.3; implying that early, low concentration exposures to 4-MCHM involve GABA activation. Initial activity loss occurs without active unit loss (defined as 10 or more template threshold crossing per min), indicating functional interference with spike production. Full recovery has not been seen at concentrations above 130 microM, unless the culture was given bicuculline. Direct exposure to 400uM results in immediate, irreversible loss of spike production, followed by necrosis of glia and neurons.
Women Have Higher Skin Temperature on the Back during Treadmill Exercise in a Hot, Humid Environment
A common measurement of body temperature during exercise in a hot, humid environment is mean skin temperature collected from 3-12 sites on the body. However, such an approach fails to demonstrate localized differences in skin temperature that are likely to exist as a function of gender. The purpose of this study was to examine potential differences in skin temperature between men and women at 17 different locations on the body. Young women (21 ± 1 y; n = 11) and men (23 ± 3; n = 10) were recruited to complete a 60-min walk/jog interval protocol in a hot (34 ± 1 °C), humid (64 ± 8%) environment while skin temperature was measured. Data was analyzed using a repeated-measures ANOVA (p < 0.05) and location of interaction effects determined using a Fisher’s least squares difference test. We observed a higher change (p < 0.05) from baseline skin temperatures (ΔTsk) for women in three locations: left upper back (women: avg. ΔTsk = 4.12 ± 0.20 °C; men: avg. ΔTsk = 2.70 ± 0.10 °C), right upper back (women: avg. ΔTsk = 4.19 ± 0.07 °C; men: avg. ΔTsk = 2.92 ± 0.05 °C), and right mid-back (women: avg. ΔTsk = 4.62 ± 0.14 °C; men: avg. ΔTsk =3.55 ± 0.09 °C). Individual time differences between genders occurred after 7- (left upper back) and 15-min (right upper back, right mid-back) of exercise and were maintained until the end of exercise. Women have a greater increase in skin temperature at three locations on the back following the onset of exercise in a hot, humid environment. This report provides important information regarding the implications of women exercising in a hot, humid environment.
Optimization of Novel Culturing and Testing Procedures for Acute Effects on Acartia Tonsa and Tisbe Biminiensis
Copepods comprise an ecologically important role in freshwater and marine ecosystems, which is why they are often considered an important ecotoxicological model organism. The International Organization for Standardization’s (ISO) 14669 protocol is the only guideline for the determination of acute toxicity in three European marine copepod species: Acartia tonsa. The goal of this project was to assess the feasibility of establishing and maintaining cultures of Acartia tonsa, as well as to refine current culturing and egg separation methods. Initial culture methodology proved difficult for consistent production of eggs and collection of nauplii. The development of an airlift system for the separation of eggs from nauplii and adults, based on size, successfully increased the availability of eggs, nauplii and adults. The sensitivity and relative conditions of the copepod species was assessed by running a series of 48h acute toxicity tests with the reference toxicants 3,5-dichlorophenol, 4,4’-methylenebis(2,6-di-tert-butylphenol. The acute 48 hour median lethal dose concentration (LC50), the no observed effect concentration (NOEC), and the lowest observed effect concentration (LOEC) was analyzed for the three reference compounds for of A. tonsa.
In Vitro Investigations of Antibiotic Influences on Nerve Cell Network Responses to Pharmacological Agents
Neuronal networks, derived from mouse embryonic frontal cortex (FC) tissue grown on microelectrode arrays, were used to investigate effects of gentamicin pretreatment on pharmacological response to the L-type calcium channel blocker, verapamil. Gentamicin is a broad spectrum antibiotic used to control bacterial contamination in cell culture. The addition of gentamicin directly to medium affects the pharmacological and morphological properties of the cells in culture. A reproducible dose response curve to verapamil from untreated cultures was established and the mean EC50 was calculated to be 1.5 ± 0.5 μM (n=10). 40 μM bicuculline was added to some cell cultures to stabilize activity and verapamil dose response curves were performed in presence of bicuculline, EC50 1.4 ± 0.1 μM (n=9). Statistical analysis showed no significant difference in verapamil EC50s values obtained in presence of bicuculline and hence the data was combined and a standard verapamil EC50 was calculated as 1.4 ± 0.13 μM (n=19). This EC50 was then used to compare verapamil EC50s obtained from neuronal cell cultures with chronic and acute exposures to gentamicin. FC cultures (21- 38 days old) were found to be stable in presence of 2300 μM gentamicin. The recommended concentration of gentamicin for contamination control is 5uL /1 ml medium (108 μM). At this concentration, the verapamil EC50 shifted from 1.4 ± 0.13 μM to 0.9 ± 0.2 μM. Given the limited data points and only two complete CRCs, statistical comparison was not feasible. However, there is a definite trend that shows sensitization of cells to verapamil in presence of gentamicin. The cultures exposed to 108 μM gentamicin for 5 days after seeding showed loss of adhesion and no data could be collected for pharmacological analysis. To conclude, acute gentamicin exposure of neuronal cell cultures causes increased sensitivity to verapamil and chronic or long term exposure to ...
Acute Effects of the Antibiotic Streptomycin on Neural Network Activity and Pharmacological Responses
The purpose of this study is to find out that if antibiotic streptomycin decreases neuronal network activity or affects the pharmacological responses. The experiments in this study were conducted via MEA (multi-electrode array) technology which records neuronal activity from devices that have multiple small electrodes, serve as neural interfaces connecting neurons to electronic circuitry. The result of this study shows that streptomycin lowered the spike production of neuronal network, and also, sensitization was seen when neuronal network pre-exposed to streptomycin.
Measuring Biomarkers From Dried Blood Spots Utilizing Bead-based Multiplex Technology
Dried blood spots is an alternative method to collect blood samples from research subjects. However, little is known about how hemoglobin and hematocrit affect bead-based multiplex assay performance. The purpose of this study was to determine how bead-based multiplex assays perform when analyzing dried blood spot samples. A series of four experiments outline the study each with a specific purpose. A total of 167 subject samples were collected and 92 different biomarkers were measured. Median fluorescence intensity results show a positive correlation between filtered and non-filtered samples. Utilizing a smaller quantity of sample results in a positive correlation to a larger sample. Removal of hemoglobin from the dried blood spot sample does not increase detection or concentration of biomarkers. Of the 92 different biomarkers measured 56 were detectable in 100-75% of the attempted samples. We conclude that blood biomarkers can be detected using bead-based multiplex assays. In addition, it is possible to utilize a smaller quantity of sample while avoiding the use of the entire sample, and maintaining a correlation to the total sample. While our method of hemoglobin was efficient it also removed the biomarkers we wished to analyze. Thus, an alternative method is necessary to determine if removing hemoglobin increases concentration of biomarkers. More research is necessary to determine if the biomarkers measured in this study can be measured over time or within an experimental model.
Effects of Sertraline Exposure on Fathead Minnow (Pimephales promelas) Steroidogenesis
Sertraline is a selective serotonin reuptake inhibitor (SSRI) that is widely used for the treatment of depression and anxiety. Due to the abundant therapeutic use of sertraline, low levels have been detected in municipal wastewater effluents suggesting that aquatic organisms may be exposed. The purpose of this study was to evaluate the steroidogenic effects of sertraline on larval (FHM) and adult female fathead minnows (FFHM), Pimephales promelas. Larval FHM were exposed to 0.1, 1, and 10 µg/L sertraline for 28 days and analyzed via RT-qPCR for differential expression of 11β-Hydroxysteroid dehydrogenase (11β-HSD), 20β-Hydroxysteroid dehydrogenase (20β-HSD), aromatase (CYP19), and nuclear thyroid receptor alpha (TRα). FFHM were exposed to 3 or 10 µg/L sertraline for 7 days with the brain and ovary excised at exposure termination. Juvenile FHM exposed to 0.1 μg/L sertraline had a significant upregulation of both 20β-HSD and TRα. FFHM exposed to 10 µg/L sertraline had a significant upregulation of 11β-HSD expression in brain tissue, while no steroidogenic changes were observed in the FFHM ovary. Similarly, in FFHM brain tissue, CYP19 and 20β-HSD expression levels were significantly higher in fish exposed to 10µg/L sertraline compared to control. The significance of these findings with respect to survival, growth and reproduction are currently unknown, but represent future research needs.
Monitoring Monocyte Oxldl Phagocytosis As a Cardiovascular Disease Risk Factor Following a High-fat Meal
Macrophage-derived foam cells play a predominant role in the deposition of arterial plaques during the early stages of atherosclerosis. The deposition of arterial plaques is known to be effected by several factors, including a person’s dietary habits. The consumption of a high-fat (>60% of calories from fat) meal is known to elevate serum LDL and triglycerides, which have been previously implicated in the formation pf foam cells. One limitation of current research models is that it is not possible to directly measure foam cells in vivo. Thus, the purpose of the present study was to validate the use of blood derived monocytes as a proxy measure of foam cells. In order to complete this objective, we evaluated monocyte oxLDL phagocytosis capacity following consumption of a high-fat meal. Eight men and women participated in the present study and venous blood samples were collected prior to the meal, 1-h, 3-h, and 5-h post-meal. Monocytes (CD14+/16- and CD14+/16+) were evaluated for adhesion molecule expression (CD11a, CD11b, and CD18), scavenger R (CD36) expression, and oxLDL phagocytosis using an image-based flow cytometry method developed in our laboratory for this purpose. Data was statistically analyzed for significance using a single-factor ANOVA with repeated measures and a p < 0.05. Consumption of a high-fat meal caused an increase significant increase in the proportion of pro-inflammatory monocytes (CD14+/16+) and a decrease in classic monocytes (CD14+/16-), with the greatest difference occurring at 5 h post prandial (p = 0.038). We also found that pro-inflammatory monocyte expression of adhesion molecules and CD36 increased in a manner that would promote in vivo movement of monocytes into the subendothelial space. Finally, over the course of the 5 h postprandial period, the majority of oxLDL uptake occurred in pro-inflammatory compared to classic monocytes. These results suggest that consuming a high-fat meal increases the ...
A Study of Selected Factors Influencing Science Majors Toward Science Careers
"The purpose of this study is to gather information concerning some factors which may have operated to influence science majors in North Texas State College toward science careers."--4.
Microflora in Prepared Foods Dispensed from Eating Establishments in Dallas, Texas
A bacteriological study was made of a number of prepared foods that were bought ready-to-eat, for home consumption, from several food establishments in Dallas, Texas. The purpose of this study was to show whether these foods could be a potential source of food poisoning; whether there are any particular foods that should have greater care than others in order to protect their quality; and whether the bacteriological contamination could have been prevented by strict observance of the city regulations for handling foods.
Responses to Diets High in Phenylalanine Compounds as Genetic Parameters in Mice
The induction of phenylketonuria in mice through the use of excess dietary phenylalanine is an area in which limited research has been done. This study intends to pursue further work in this area, more specifically, to study the effects of excess dietary phenylalanine and the phenylalanine analogue A.P.B.A. (2-amino-3-phenyl butanoic acid) (7) on brain serotonin and brain norepinephrine. In addition, the effects of these two compounds on the incidence of audiogenic seizures will be explored.
Bacteriological Studies of the Campus Drinking Fountains of North Texas State Teachers College Denton, Texas
"In order to gain an adequate idea of the sanitary condition of the drinking fountains on the North Texas State Teachers College campus, it was found necessary to approach these bacteriological studies from a seasonal point of view."--1.
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.
A Study of a Selected Group of Science Related Characteristics of Non-Science Majors
The purpose of this study is to gather information related to the characteristics of two groups of undergraduate non-science majors; namely, those students who once considered science as a career and subsequently changed to a non-science major and those who have never considered a career in science. It is frequently observed that children in the elementary school have an intense interest in science. One of the principal concerns of this study is how and for what reasons has the interest in science changed between elementary school and college? It is also the purpose of this study to gather information related to the attitudes of these college students toward the science courses they have had in high school and toward their science teachers. In attempting to arrive at an answer, the group who once considered a science career and those who have never considered an occupational choice in a field of science will be compared in the following areas: 1. General information such as sex, age, size of high school attended, rank within their graduating classes, college classification and military experiences. 2. Occupations and hobbies of parents. 3. Experiences which may influence attitudes toward science such as elementary school science, high school science courses, high school science teachers, attitudes toward science requirements, high school courses liked most, high school courses liked least, and membership in science clubs. 4. College science courses completed. 5. Extent of the interviewees' present interest in science, Finally, the circumstances surrounding the tentative choice of a science career by those who once considered such a career will be discussed.
Genic Differentiation and Evolution in the Ground Squirrel Subgenus Ictidomys (Spermophilus)
The genetic structure of 26 natural populations of three species (S. tridecemlineatus, S. mexicanus, and S. spilosoma) of the Ictidomys subgenus of ground squirrels was analyzed using chromosomal and electrophoretic techniques. Chromosomal variation was not observed in S. mexicanus, and only slight karyotypic variation was found in the other two species. Chromosomal evidence indicated hybridization between S. tridecemlineatus and S. mexicanus, placing these species within the classical definition of semispecies. Analysis of electrophoretic variation at 29 genetic loci indicated close genetic relationships between these species. Evolution in Ictidomys appears to be linked with Pleistocene events, and speciation appears to have occurred within the last 155,000 years.
Determination of Biotransformation and Biodegradation Rate Constants for Naphthalene, Lindane and Phenol
Biotransformation and biodegradation rate constants were determined for naphthalene, lindane, and phenol in water samples from three different sources. Rate constants produced from monitoring disappearance of the parent chemical (biotransformation) were compared to those obtained from mineralization of the chemical (ultimate biodegradation) by ¹⁴CO₂ evolution as well as acidification of the residual ¹⁴C-labeled compound (primary biodegradation). Rate constants were statistically different for the three chemicals. The water source affected the rate constants. When biomass measurements of the waters were considered and second-order rate constants were derived, there was no statistical evidence that this parameter gave a reliable rate constant statistic that could be useful in predicting the fate of any of naphthalene, lindane, and phenol in these waters.
The Effect of Sodium Salicylate on the Ultrastructure of Trypanosoma lewisi
This study examines ultrastructural changes occurring in seven- and fourteen-day populations of Trypanosorfa lewisi when 60 mg sodium salicylate is administered to the host. These changes were related to the host-immune response. Seven-day trypanosomes showed approximately 10 posterior volutin granules. Seven-day trypanosomes whose host had received salicylate exhibited. 30 to 40 volutin granules, and their posterior tip exhibited volutin granules in high numbers sometimes excluding other cellular elements. Fourteen-day trypanosones showed fewer volutin granules than seven-day, salicylate-treated ones. Salicylate treatment caused no additional ultrastructural alterations. Thus the volutin granules are not linked to the reproduction inhibiting antibody (ablast in) but may be involved in the formation of the trypanocidal antibodies.
The Distribution of Pathogenic Pseudomonas aeruginosa in Sewage
The purpose of this study was to extend our understanding of the ecological relationships of P. aeruginosa by investigating the differences or similarities between the strains of this organism found in sewage and those found as pathogens in human infections. This research was approached by comparing the serological types of P. aeruginosa isolated from sewage contaminated waters in Argentina (South America) to those isolated from sewage contaminated waters in Texas. They were typed with sera obtained using P. aeruginosa isolated from human infections. The data obtained revealed that bacteria isolated from sewage in Texas and from soil and water in Argentina are antigenically similar to those isolated from human infections.
Population Studies of Ulmus Crassifolia in Flood-Plain Forests of Denton County, Texas
The problem with which this investigation was concerned was the comparison of cedar elm populations in different stands along creeks in Denton County, Texas, and the relationship of certain population parameters to various substrates present at stand sites, Parameters investigated eluded average basal area, basal-area density, transect-segment density, intertree distance, lateral distance, frequency, diameter breast-high, diameter breast-high size-class distribution, and immature-tree density. Variations among populations of Ulmus crassifolia Nutt were noted and analyzed in terms of soil particle size and existing community conditions.
The Determination of Uptake and Depuration Rate Kinetics and Bioconcentration Factor of Naphthalene and Lindane in Bluegill Sunfish, Lepomis macrochirus
Bluegill were exposed to 3 and 30 pg/L lindane and 20 and 200 pg/L naphthalene to determine uptake rate constants, K1 depuration rate constants, K2, and bioconcentration factors, BCF. Correlations were determined between lipid normalized and non-lipid normalized BCFs, and between observed Kl, K2 and BCFs and predicted values. The K1 values for both chemicals and concentrations were similar. The K2 values were different (1.04 day~1, 0.46 day 1). Naphthalene was more rapid. BCFs for lindane (315) and naphthalene (98) were different. Lipid normalized BCFs for naphthalene were more variable than non-lipid normalized BCFs. The reverse was observed for lindane BCFs. Predicted K1, K2 , and BCFs were in agreement with observed values.
Induction of Aryl Hydrocarbon Hydroxylase in Ambystoma tigrinum
Aryl hydrocarbon hydroxylase (AHH) was induced 15-fold in Ambystoma tigrinum by intraperitoneal injection of 3-methylcholanthrene in corn oil, or 10-fold by addition of aromatic polycyclic hydrocarbons to the aqueous environment of the neotene animal. The cytochrome P-450-associated microsomal enzyme is similar to the inducible, one-gene, autosomal-dominant system typical in the laboratory mouse and man. Differences in optimal temperature for enzyme induction and activity were noted in organ culture of human and Ambystoma tissues, and ratios of benzpyrene metabolites differed between Ambystoma and Mus. The half life of enzyme activity induced in vivo was related to the excretion of hydrocarbon metabolites.
The Population Dynamics and Trophic Relationships of Seven Species of Fish in a Small Southwestern Pond, with Special Attention Toward Young-of-the-Year Fish
Production rates of seven species of fish, along with food-web interactions, were determined for one year in a pond. Production of fish in the pond amounted to 208.9 q/m2/yr. Over 95% of the production occurred in the summer, with Y of Y fish contributing 83.8%. There were two food-web interactions observed in Y of Y fishes: crappie vs. bass and bullhead vs. sunfish. When Y of Y fish shifted to benthic diets in the spring, the small biomass of benthos available could not sustain the larger biomass of fish. Competition for the available food resulted in decreased production rates for the fish community.
Effects of an Acute Bout of Near-Maximal Intensity Exercise on the Cardiac Enzymes in Human Sera
The Cardiac Profile, a pattern of serum enzyme changes seen within seventy-two hours after an AMI, is diagnostic aid for detecting occurrence of infarcts. The effects of exercise stress on the Cardiac Profile aid clinicians in avoiding diagnostic errors in patients immediately after exercise. Five male volunteers ran from six to ten miles. Serum enzyme levels were monitored serially three days before and five days after stress. Enzyme activity was determined spectrophotometrically and electrophoretically. Significant increases in total CPK and LDH were seen. An LDH 'one-two flip' occurred eight hours after exercise. No MB-CPK was found following the run.
An Investigation of Naturally Occurring Tastes and Odors from Fresh Waters
The Problem of the cause and control of tastes and odors in water supplies has been the subject of a great many investigations during the last half-century; however, many questions in this field remain to be answered. Theories relating to the cause of tastes and odors occurring in water are still rather numerous and, at present, only a few of them are compatible. Although several procedures have been employed for the eradication of tastes and odors from water supplies, none of them are completely successful.
Cigarette Residues Affect Steroidogenesis in Cultured Y-1 Mouse Adrenal Tumor Cells
This study (1) quantitatively compared steroid production in cultured Y-l mouse adrenal tumor cells exposed to Camel and Carlton-smoke derived residues, and (2) localized the effects in the cell. Basal steroid production was increased by Camel residues but not by Carlton, while ACTH stimulation was interfered with by both residues. Camel basal stimulation was comparable to that of cAMP, and was abolished by Cytochalasin D. The stimulation was also comparable to that of cholera toxin, which activates adenyl cyclase. Results indicate that residue components dissolve in the membrane stimulating adenyl cyclase at a point similar to or before that utilized by cholera toxin for its stimulating effect.
The Stoneflies (Plecoptera) of Texas
An illustrated key to the adults and known nymphs of Texas Plecoptera is provided. Species accounts, including geographic distribution within Texas, and biological notes are given. Of the twenty-seven species of Plecoptera known from Texas, six are new state records. TWo species new to science, Isoperla jewetti and Isoperla coushatta are described. Taeniopteryx starki Stewart and Szczytko, Zealeuctra arnoldi Ricker and Ross, and Zealeuctra hitei Ricker and Ross are endemic to the Edwards Plateau area of Texas. Two species, Mesocapnia frisoni (Baumam and Gaufin) and Isoperla jewetti New Species are western in origin. The remaining nineteen species (excluding Anacroneuria) are typically eastern species.
Pelagic Phytoplankton and Physicochemical Correlates for Lake Texoma
An analysis and correlation of phytoplankton communities with physicochemical data from 3 sites in Lake Texoma was conducted to supplement time-series data. Water and phytoplankton were sampled monthly, March, 1976-February, 1977. Simple correlations were run between all physicochemical parameters and phytoplankton standing crop from the 3 sites. Multiple linear regression analyses were used to develop equations predictive of phytoplankton standing crop and chloride concentration. Minerals leached from marine sediments in the Red River chennel contribute to formation of a, halocline which seals the anoxic hypolimnion from the reservoir surface in midsummer. Conductivity decreased west to east, 2980-1800. pmhos/cm. Maximum mean annual phytoplankton standing crop in Red River arm was 36 percent greater than midlake. Eutrophication was evident.
X-irradiation Effects on the Action Potentials of Frog Sciatic Nerves Inhibited by Carbon Monoxide and Ouabain
The response of frog sciatic nerve action potentials to x-irradiation and metabolic (carbon monoxide) or transport (ouabain) inhibition was determined in an attempt to further identify the nature of radiation insult to nervous tissue. Carbon monoxide, ouabain (2 X 10-5 M), and nitrogen anoxia were shown to produce a near linear decline in action potential amplitude. The carbon monoxide and nitrogen inhibitions of activity were reversible in air; the carbon monoxide inhibition was light reversible. Ouabain inhibition was partially reversible by soaking the nerve in aerated Ringer's. Application of 120 kv x-rays (75 Kr at 4.9 Kr/min) to nerves during the linear decline in spike amplitude brought about a marked enhancement (146%) of inhibition by 99% CO/l% 02, nitrogen (136%), and ouabain (265%). All bhanges were shown to be statistically significant by a regression analysis. However, x-irradiation did not appear to alter the air reversibility of carbon monoxide and nitrogen inhibitions nor the reversibility in Ringerts of the ouabain inhibition. Additionally x-irradiation completely blocked light reversal of 98% CO/2% 02 inhibition and produced a decline in activity. A possible interpretation of these results is a compensation for radiation action at this dosage requiring metabolism and ion pump activity.
A Study on the Serological Relationships of Various Fractions of Pseudomonas aeruginosa
The purpose of this research was to determine the relationship of the slime layer antigen(s) to the "101" or LPS antigens and to attempt to evaluate the role of antibodies against the latter in protection against experimental infections in mice with the homologous strain of Pseudomonas aeruginosa. Results from agglutination tests, chromatographic separations, passive protection tests, and characterizations of the antigens by gel double diffusion do not support the concept that LPS is a necessary portion of the immunogenic material. The immunogenicity of LPS can be attributed to co-purification of residual amounts of slime layer antigens on the washed cells from which LPS was extracted.
The Roles of Genic Behavioral and Biochemical Mechanisms in the Adaption of Minnows of the Genus Notropis (Cyprinidae) to Temperature
Electrophoretic variation at twenty gene loci, patterns of behavioral thermoregulation, and genotype-specific malate dehydrogenase kinetics were investigated among populations of the red shiner, Notropis lutrensis, and the blacktail shiner, N. venustus, collected from thermally altered and thermally unaltered portions of their ranges. Genic variation was found to be high among red shiners and low among blacktail shiners. The behavioral response of the blacktail shiner to temperature was fixed among the populations sampled, whereas the response of the red shiner was mutable. Finally, blacktail shiners have incorporated into their genome an Mdh-B allele which functions well at low temperatures; red shiners, displaying high levels of Mdh-B polymorphism, maintain a more complex set of allozymes which function well over a wide range of environmental temperatures. These data are consistent with reported ecotypic distributions of the species in Texas waters; i.e., blacktail shiners occur in cool, thermally static habitats, and red shiners are tolerant of wide temperature ranges.
Evolutionary Genetics of Three Semispecies of Wood Rats--Neotoma Albigula, Neotoma Micropus, and Neotoma Floridana
Electrophoretic variation in 18 proteins encoded by 20 autosomal loci was used to compare the genetic relationships of 19 natural populations representing three species of the subgenus Neotoma. Of the 20 loci examined nine were monomorphic and fixed for the same allele in all populations. No more than seven loci were polymorphic within a single population. Genetic variability was expressed as the proportion of loci heterozygous in the average individual of a population. Heterozygosity in the three species of Neotoma studied averaged 0.078, a value within the range reported for other rodents. Although the levels of heterozygosity seen in Neotoma could not readily be explained, the variation may be attributed to ecological factors. The three species of Neotoma were compared on the basis of genetic similarity and found to form a close taxonomic unit, probably semispecies. Divergence times were obtained for the three species and found to compare well with divergence times obtained from fossil data. In general, the three species have diverged within the last 112,000 years during the Wisconsin glacial period.
The Growth of Azotobacter vinelandii on p-Hydroxybenzoic Acid from Soil Medium
The purpose of this study was to search for the substrates utilized by Azotobacter vinelandii in dialysed soil media. Also, we sought to determine the relationship between these substrates and the growth and morphological variations of A. vinelandii. p-Hydroxybenzoic acid was shown to be used as the carbon and energy source by A. vinelandii in dialysed soil medium. The amount of this compound in the soil dialysed soil medium ranged from 14 to 21 micrograms per gram of soil. In a dialysed soil medium, p-hydroxybenzoic acid induced A. vinelandii to form minute bodies, similar to the filtrable forms reported by Gonzalez and Vela, although no growth of minute bodies was detected.