Variance in the production of homovanillic acid and 3-methoxy-4-hydroxyphenethyleneglycol by the awake primate brain

Previous experimental results, using a new technique whereby the production rates of the neurotransmitter metabolites homovanillic acid (HVA) and 3-methoxy-4-hydroxyphenethyleneglycol (MHPG) by the awake primate brain are determined, have shown a wide variance in metabolite production among both animal and human subjects. These data suggested that either individual subjects differ in the activity of brain dopamine (DA) or norepinephrine (NE) neurons and/or that the activities of these neurons fluctuate over time. For these reasons a series of experiments were performed in which measures of HVA and MHPG production were obtained at three time points in the same animal (monkeys) over a three hour period. It was found that the group mean values for the production of HVA and MHPG by brain were similar for each of the three time points. However, it was also found that marked variations in HVA and MHPG production occur within a single animal over a three hour period. The coefficients of variation for individual animals for HVA ranged from 9.3 to 31.9% and for MHPG from 10.1 to 62.3%. These variations were not correlated with grossly observable changes in behavioral states. Using an analysis of variance it was found that the variance in MHPG production was significantly greater than that for HVA (F = 6.2, p < 0.05) suggesting that brain NE systems are more liable and/or show greater change than do brain DA systems. These data are interpreted as indicating that in the awake, resting primate brain fluctuations in the activities of DA and NE neurons occur, i.e. there is not a steady, invariant production of metabolites but rather they are produced in pulses of varying lengths. This interpretation of the data is generally consistent with electrophysiological studies which indicate that catecholamine neurons fire in bursts which are then followed by silent periods. Finally, in terms of practical application of the V-A difference technique, these data indicate that replicable group mean estimates of brain HVA and MHPG production can be obtained by averaging values from a single time point whereas accurate information about an individual animal will require multiple samplings.Recent reports from this laboratory have described a method whereby a direct measure of the rates of production of neurotransmitter metabolites such as homovanillic acid (HVA), 3-methoxy-4-hydroxyphenethyleneglycol (MHPG), and 5-hydroxyindoleacetic acid (5-HIAA) by the awake primate brain can be determined (1, 2, 3, 4). Since the quantities of HVA, MHPG, and probably 5-HIAA in the brain vary as a function of the activity of dopamine (DA), norepinephrine (NE), and serotonin (5-HT) neurons (1, 5, 6, 7, 8), it is likely that these measures of neurotransmitter metabolite production reflect the functional state of brain DA, NE, and 5-HT neuronal systems. The experimental results thus far obtained with this technique have shown a wide variance in the rates of neurotransmitter metabolite production across both animal and human subjects even though the subjects were not in clearly different behavioral or emotional states (1, 2, 4, 9). These data suggested that either individual subjects differ markedly in the activities of brain DA, NE, and 5-HT neurotransmitter systems and/or that the activity of these systems fluctuates markedly over time. For these reasons, experiments were undertaken in which repeated measures of HVA and MHPG production by brain within the same animal were determined over a three hour period. The results of these experiments, which are reported here, indicate that there are marked changes in brain metabolite production which occur within animals. The implications of these findings for our understanding of the functioning of brain neurotransmitter systems and for the practical applications of this technique are discussed.     

Population Structure of Clinical and Environmental Vibrio parahaemolyticus from the Pacific Northwest Coast of the United States

Vibrio parahaemolyticus is a common marine bacterium and a leading cause of seafood-borne bacterial gastroenteritis worldwide. Although this bacterium has been the subject of much research, the population structure of cold-water populations remains largely undescribed. We present a broad phylogenetic analysis of clinical and environmental V. parahaemolyticus originating largely from the Pacific Northwest coast of the United States. Repetitive extragenic palindromic PCR (REP-PCR) separated 167 isolates into 39 groups and subsequent multilocus sequence typing (MLST) separated a subset of 77 isolates into 24 sequence types. The Pacific Northwest population exhibited a semi-clonal structure attributed to an environmental clade (ST3, N = 17 isolates) clonally related to the pandemic O3:K6 complex and a clinical clade (ST36, N = 20 isolates) genetically related to a regionally endemic O4:K12 complex. Further, the identification of at least five additional clinical sequence types (i.e., ST43, 50, 65, 135 and 417) demonstrates that V. parahaemolyticus gastroenteritis in the Pacific Northwest is polyphyletic in nature. Recombination was evident as a significant source of genetic diversity and in particular, the recA and dtdS alleles showed strong support for frequent recombination. Although pandemic-related illnesses were not documented during the study, the environmental occurrence of the pandemic clone may present a significant threat to human health and warrants continued monitoring. It is evident that V. parahaemolyticus population structure in the Pacific Northwest is semi-clonal and it would appear that multiple sequence types are contributing to the burden of disease in this region.     

Pathway to domestic natural rubber production: a cradle-to-grave life cycle assessment of the first guayule automobile tire manufactured in the United States

The International Journal of Life Cycle Assessment - Guayule (Parthenium argentatum) is a perennial shrub that can be cultivated in the Southwestern US. It produces natural rubber that could be a...     

High-frequency generation of conditional mutations affecting Drosophila melanogaster development and life span.

Genome sequencing reveals that a large percentage of Drosophila genes have homologs in humans, including many human disease genes. The goal of this research was to develop methods to efficiently test Drosophila genes for functions in vivo. An important challenge is the fact that many genes function at more than one point during development and during the life cycle. Conditional expression systems such as promoters regulated by tetracycline (or its derivative doxycycline) are often ideal for testing gene functions. However, generation of transgenic animals for each gene of interest is impractical. Placing the doxycycline-inducible ("tet-on") promoter directed out of the end of the P transposable element produced a mobile, doxycycline-inducible promoter element, named PdL. PdL was mobilized to 228 locations in the genome and was found to generate conditional (doxycycline-dependent), dominant mutations at high frequency. The temporal control of gene overexpression allowed generation of mutant phenotypes specific to different stages of the life cycle, including metamorphosis and aging. Mutations characterized included inserts in the alpha-mannosidase II (dGMII), ash1, and pumilio genes. Novel phenotypes were identified for each gene, including specific developmental defects and increased or decreased life span. The PdL system should facilitate testing of a large fraction of Drosophila genes for overexpression and misexpression phenotypes at specific developmental and life cycle stages.     

A Regional Retrospective Assessment of the Potential Stressors Causing the Decline of the Cherry Point Pacific Herring Run

The Pacific herring stock that spawns at Cherry Point, northwest of Bellingham, WA, has undergone a dramatic decline in the last 20 years. The population decline corresponds with a collapse of the age structure. The Cherry Point area contains three deep water shipping piers, two refineries, an aluminum smelter, and urban development. The Cherry Point Aquatic Reserve was formed initially to protect the spawning habitat of the Cherry Point Pacific herring run. We conducted a retrospective assessment using the relative risk model (RRM) to investigate the causes of the current decline of the Cherry Point run. The RRM combines aspects of the weight-of-evidence (WoE) approach and other methods of establishing causality into a framework that deals with multiple stressors, uncertainty, and spatial scale.

An analysis of the Cherry Point Pacific herring age structure and population dynamics indicates that the loss of reproductive potential of the older age class fish was the population characteristic that led to the decline of the run. Exploitation, habitat alteration and climate change are the risk factors that contribute to the decline of the Cherry Point Pacific herring. The retrospective assessment identified the cyclic nature of climate change, as expressed by the warmer sea surface temperatures associated with a warm Pacific Decadal Oscillation (PDO), as the primary factor altering the dynamics of the Pacific herring. Other factors are ranked accordingly along with the associated uncertainty. Criteria for selecting alternative endpoints for managing the Cherry Point Aquatic Reserve are also provided.

The strengths of the retrospective RRM include its ability to combine a WoE and causality criteria with a multitude of stressors at a regional scale. The difficulties include how to deal with differences in the magnitude of effects, and expressing the uncertainty as distributions.     

Probing the nature of H2 activation in catalytic asymmetric hydrogenation

Despite many years of intensive study, the natures of turnover-limiting and enantio-determining steps in catalytic asymmetric hydrogenation of prochiral enamides are poorly understood. An intriguing set of studies involving isotopic labeling distributions in catalytic enamide hydrogenation reactions were reported by Brown and Parker (Organometallics, 1 (1982) 950–956) more than a decade ago. In this paper we report the results of studies re-examining the application of isotopic probes to the catalytic hydrogenation enamides. These results provide some insights into the nature of the H₂ activation step in enamide hydrogenation.     

The influence of arbuscular mycorrhizae and light on Wisconsin (USA) sand savanna understories 2. Plant competition

Wisconsin (USA) oak savannas are endangered plant communities that have remarkably high plant species diversity. To investigate factors underlying this richness, we experimentally investigated the potentially interacting effects of light gradients and arbuscular mycorrhizal fungi (AMF) on plant competition in the greenhouse, using a fully randomized block design. We used four plant species, soil, and AMF from a remnant sand savanna, under two light and five AMF treatments. Plants were grown four per pot under two competition treatments (either one or four species per pot) for 20 weeks. Using ANOVA, we found that all species showed significant treatment effects on total and shoot biomass, primarily due to differences in competition and light, less to AMF. However, effects were the opposite of predictions. Putatively mycorrhizal plants showed neutral to negative responses to AMF, and a nonmycorrhizal species outcompeted AMF species in infected pots. We concluded that our experimental setup of small pots, sandy soil, and long growing period had induced parasitism by the AMF on susceptible hosts. This unexpected result is consistent with field data from the sand savanna, and may help explain how nonmycorrhizal plants can compete successfully with AMF species in established, species-rich communities.