Cloning and Sequencing of Protein Kinase cDNA from Harbor Seal (Phoca vitulina) Lymphocytes

Protein kinases (PKs) play critical roles in signal transduction and activation of lymphocytes. The identification of PK genes provides a tool for understanding mechanisms of immunotoxic xenobiotics. As part of a larger study investigating persistent organic pollutants in the harbor seal and their possible immunomodulatory actions, we sequenced harbor seal cDNA fragments encoding PKs. The procedure, using degenerate primers based on conserved motifs of human protein tyrosine kinases (PTKs), successfully amplified nine phocid PK gene fragments with high homology to human and rodent orthologs. We identified eight PTKs and one dual (serine/threonine and tyrosine) kinase. Among these were several PKs important in early signaling events through the B- and T-cell receptors (FYN, LYN, ITK and SYK) and a MAP kinase involved in downstream signal transduction. V-FGR, RET and DDR2 were also expressed. Sequential activation of protein kinases ultimately induces gene transcription leading to the proliferation and differentiation of lymphocytes critical to adaptive immunity. PKs are potential targets of bioactive xenobiotics, including persistent organic pollutants of the marine environment; characterization of these molecules in the harbor seal provides a foundation for further research illuminating mechanisms of action of contaminants speculated to contribute to large-scale die-offs of marine mammals via immunosuppression

The utility of twins in developmental cognitive neuroscience research: How twins strengthen the ABCD research design

The ABCD twin study will elucidate the genetic and environmental contributions to a wide range of mental and physical health outcomes in children, including substance use, brain and behavioral development, and their interrelationship. Comparisons within and between monozygotic and dizygotic twin pairs, further powered by multiple assessments, provide information about genetic and environmental contributions to developmental associations, and enable stronger tests of causal hypotheses, than do comparisons involving unrelated children. Thus a sub-study of 800 pairs of same-sex twins was embedded within the overall Adolescent Brain and Cognitive Development (ABCD) design. The ABCD Twin Hub comprises four leading centers for twin research in Minnesota, Colorado, Virginia, and Missouri. Each site is enrolling 200 twin pairs, as well as singletons. The twins are recruited from registries of all twin births in each State during 2006–2008. Singletons at each site are recruited following the same school-based procedures as the rest of the ABCD study. This paper describes the background and rationale for the ABCD twin study, the ascertainment of twin pairs and implementation strategy at each site, and the details of the proposed analytic strategies to quantify genetic and environmental influences and test hypotheses critical to the aims of the ABCD study. Keywords: Twins, Heritability, Environment, Substance use, Brain structure, Brain functio

Associations between diurnal preference, sleep quality and externalizing behaviours: a behavioural genetic analysis

Background - Certain aspects of sleep co-occur with externalizing behaviours in youth, yet little is known about these associations in adults. The present study: (1) examines the associations between diurnal preference (morningness versus eveningness), sleep quality and externalizing behaviours; (2) explores the extent to which genetic and environmental influences are shared between or are unique to these phenotypes; (3) examines the extent to which genetic and environmental influences account for these associations. Method - Questionnaires assessing diurnal preference, sleep quality and externalizing behaviours were completed by 1556 young adult twins and siblings. Results - A preference for eveningness and poor sleep quality were associated with greater externalizing symptoms [r=0.28 (95% CI 0.23–0.33) and 0.34 (95% CI 0.28–0.39), respectively]. A total of 18% of the genetic influences on externalizing behaviours were shared with diurnal preference and sleep quality and an additional 14% were shared with sleep quality alone. Non-shared environmental influences common to the phenotypes were small (2%). The association between diurnal preference and externalizing behaviours was mostly explained by genetic influences [additive genetic influence (A)=80% (95% CI 0.56–1.01)], as was the association between sleep quality and externalizing behaviours [A=81% (95% CI 0.62–0.99)]. Non-shared environmental (E) influences accounted for the remaining variance for both associations [E=20% (95% CI −0.01 to 0.44) and 19% (95% CI 0.01–0.38), respectively]. Conclusions - A preference for eveningness and poor sleep quality are moderately associated with externalizing behaviours in young adults. There is a moderate amount of shared genetic influences between the phenotypes and genetic influences account for a large proportion of the association between sleep and externalizing behaviours. Further research could focus on identifying specific genetic polymorphisms common to both sleep and externalizing behaviours

PAH- and PCB-induced Alterations of Protein Tyrosine Kinase and Cytokine Gene Transcription in Harbor Seal (Phoca Vitulina) PBMC

Mechanisms underlying in vitro immunomodulatory effects of polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were investigated in harbor seal peripheral leukocytes, via real-time PCR. We examined the relative genetic expression of the protein tyrosine kinases (PTKs) Fyn and Itk, which play a critical role in T cell activation, and IL-2, a cytokine of central importance in initiating adaptive immune responses. IL-1, the macrophage-derived pro-inflammatory cytokine of innate immunity, was also included as a measure of macrophage function. Harbor seal PBMC were exposed to the prototypic immunotoxic PAH benzo[a]pyrene (BaP), 3,3',4,4',5,5'-hexachlorobiphenyl (CB-169), a model immunotoxic PCB, or DMSO (vehicle control). Exposure of Con A-stimulated harbor seal PBMC to both BaP and CB-169 produced significantly altered expression in all four targets relative to vehicle controls. The PTKs Fyn and Itk were both up-regulated following exposure to BaP and CB-169. In contrast, transcripts for IL-2 and IL-1 were decreased relative to controls by both treatments. Our findings are consistent with those of previous researchers working with human and rodent systems and support a hypothesis of contaminant-altered lymphocyte function mediated (at least in part) by disruption of T cell receptor (TCR) signaling and cytokine production

Minimising medicine use in organic dairy herds through animal health and welfare planning

Livestock is important in many organic farming systems, and it is an explicit goal to ensure high levels of animal health and welfare (AHW) through good management. This will lead to reduced medicine use and better quality of animal products. In two EU network projects NAHWOA & SAFO it was concluded that this is not guaranteed merely by following organic standards. Both networks recommended implementation of individual animal health plans to stimulate organic farmers to improve AHW. These plans should include a systematic evaluation of AHW and be implemented through dialogue with each farmer in order to identify goals and plan improvements. 15 research institutions in 8 European countries are involved in the proposed project with the main objective to minimise medicine use in organic dairy herds through active and well planned AHW promotion and disease prevention. The project consists of 5 work packages, 4 of which comprise research activities building on current research projects, new applications across borders, exchange of knowledge, results and conclusions between participating countries, and adopting them to widely different contexts. International and national workshops facilitate this exchange. Focus areas are animal health planning, AHW assessment using animal based parameters and development of advisory systems and farmer groups. Epidemiological analyses of the effect on AHW from reduced medicine use and herd improvements are planned in all participating countries

Influence of Behavioral State, Sex, and Season on Resource Selection by Jaguars (Panthera onca): Always on the Prowl?

How a predator uses its landscape to move through its territory and acquire prey is a fundamental question for scientific research. The influence of abiotic and biotic factors on space use of large carnivores has profound implications for their future management and conservation. In the Pantanal, Brazil, jaguars (Panthera onca) are the apex predator, but conflicts with cattle depredations pose a risk to their future conservation. We examined whether behavioral state, sex, and season influenced how jaguars used the landscape in the Pantanal. To accomplish this, we radio‐collared four females and six males; radio‐collared jaguars were monitored for 76 radio‐months with 11,787 GPS locations acquired. We developed resource selection functions (RSFs) examining how female and male jaguars used their landscape during three behavioral states (moving, killing native prey, killing cattle) during two seasons (dry, wet). From the RSF models, we found similar variables and relationships of landscape characteristics that jaguars selected for when moving and when depredating native prey and cattle. While moving, jaguars selected for locations that were either in dense cover or very near dense cover, with higher plant diversity and closer to water than available across the landscape. While null models suggested jaguars opportunistically depredated native prey in the dry season and cattle in the wet season, there was some indication they selected for specific landscape characteristics, mainly dense cover when killing cattle in the dry season and native prey in the wet season. Both sexes killed native prey and cattle within dense cover or close to dense cover as expected of an ambush predator. Particular habitat types were not important as long as there was dense cover for concealment. Additionally, jaguars killed prey closer to water than was available on the landscape. The similar variables across the models showed the importance of dense cover and distance to dense cover during all three behavioral states indicating jaguars in the Pantanal were “always on the prowl.” Understanding the spatial requirements for jaguars during the acquisition of native prey and cattle may lead to improved management strategies to allow for continued coexistence of jaguars in an area of traditional cattle production

Influence of behavioral state, sex, and season on resource selection by jaguars (\u3ci\u3ePanthera onca\u3c/i\u3e): Always on the prowl?

How a predator uses its landscape to move through its territory and acquire prey is a fundamental question for scientific research. The influence of abiotic and biotic factors on space use of large carnivores has profound implications for their future management and conservation. In the Pantanal, Brazil, jaguars (Panthera onca) are the apex predator, but conflicts with cattle depredations pose a risk to their future conservation. We examined whether behavioral state, sex, and season influenced how jaguars used the landscape in the Pantanal. To accomplish this, we radio-collared four females and six males; radiocollared jaguars were monitored for 76 radio-months with 11,787 GPS locations acquired. We developed resource selection functions (RSFs) examining how female and male jaguars used their landscape during three behavioral states (moving, killing native prey, killing cattle) during two seasons (dry, wet). From the RSF models, we found similar variables and relationships of landscape characteristics that jaguars selected for when moving and when depredating native prey and cattle. While moving, jaguars selected for locations that were either in dense cover or very near dense cover, with higher plant diversity and closer to water than available across the landscape. While null models suggested jaguars opportunistically depredated native prey in the dry season and cattle in the wet season, there was some indication they selected for specific landscape characteristics, mainly dense cover when killing cattle in the dry season and native prey in the wet season. Both sexes killed native prey and cattle within dense cover or close to dense cover as expected of an ambush predator. Particular habitat types were not important as long as there was dense cover for concealment. Additionally, jaguars killed prey closer to water than was available on the landscape.The similar variables across the models showed the importance of dense cover and distance to dense cover during all three behavioral states indicating jaguars in the Pantanal were “always on the prowl.” Understanding the spatial requirements for jaguars during the acquisition of native prey and cattle may lead to improved management strategies to allow for continued coexistence of jaguars in an area of traditional cattle production