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A discrete acyl carrier protein (ACP) bearing a photolabile nonhydrolysable carba(dethia) malonyl pantetheine cofactor was chemoenzymatically prepared and utilised for the trapping of biosynthetic polyketide intermediates following light activation. From the in vitro assembly of the polyketides SEK4 and SEK4b, by the type II actinorhodin “minimal” polyketide synthase (PKS), a range of putative ACP‐bound diketides, tetraketides, pentaketides and hexaketides were identified and characterised by FT‐ICR‐MS, providing direct insights on active site accessibility and substrate processing for this enzyme class., A novel photoactivatable acyl carrier protein‐based tool provides novel insights on substrate iterative processing for a model type II iterative polyketide synthase.

Simple Summary Basal cell carcinoma is the most common human cancer worldwide. The molecular basis of BCC involves an interplay of inherited genetic susceptibility and somatic mutations, commonly induced by exposure to UV radiation. In this review, we outline the currently known germline and somatic mutations implicated in the pathogenesis of BCC with particular attention paid toward affected molecular pathways. We also discuss polymorphisms and associated phenotypic traits in addition to active areas of BCC research. We finally provide a brief overview of existing non-surgical treatments and emerging targeted therapeutics for BCC such as Hedgehog pathway inhibitors, immune modulators, and histone deacetylase inhibitors. Abstract Basal cell carcinoma (BCC) is a significant public health concern, with more than 3 million cases occurring each year in the United States, and with an increasing incidence. The molecular basis of BCC is complex, involving an interplay of inherited genetic susceptibility, including single nucleotide polymorphisms and genetic syndromes, and sporadic somatic mutations, often induced by carcinogenic exposure to UV radiation. This review outlines the currently known germline and somatic mutations implicated in the pathogenesis of BCC, including the key molecular pathways affected by these mutations, which drive oncogenesis. With advances in next generation sequencing and our understanding of the molecular genetics of BCC, established and emerging targeted therapeutics are offering new avenues for the non-surgical treatment of BCC. These agents, including Hedgehog pathway inhibitors, immune modulators, and histone deacetylase inhibitors, will also be discussed.

More than 100 monoclonal antibodies (mAbs) are in industrial and clinical development to treat myriad diseases. Accurate quantification of mAbs in complex media, derived from industrial and patient samples, is vital to determine production efficiency or pharmacokinetic properties. To date, mAb quantification requires time and labor-intensive assays. Herein, we report a novel dual-affinity ratiometric quenching (DARQ) assay, which combines selective biorecognition and quenching of fluorescence signals for rapid and sensitive quantification of therapeutic monoclonal antibodies (mAbs). The reported assay relies on the affinity complexation of the target mAb by the corresponding antigens and Protein L (PrL, which targets the Fab region of the antibody), respectively, labeled with fluorescein and rhodamine. Within the affinity complex, the mAb acts as a scaffold framing the labeled affinity tags (PrL and antigen) in a molecular proximity that results in ratiometric quenching of their fluorescence emission. Notably, the decrease in fluorescence emission intensity is linearly dependent upon mAb concentration in solution. Control experiments conducted with one affinity tag only, two tags labeled with equal fluorophores, or two tags labeled with fluorophores of discrete absorbance and emission bands exhibited significantly reduced effect. The assay was evaluated in noncompetitive (pure mAb) and competitive conditions (mAb in a Chinese Hamster Ovary (CHO) cell culture harvest). The "DARQ" assay is highly reproducible (coefficient of variation ∼0.8-0.7%) and rapid (5 min), and its sensitivity (∼0.2-0.5 ng·mL-1), limit of detection (75-119 ng·mL-1), and dynamic range (300-1600 ng·mL-1) are independent of the presence of CHO host cell proteins.

For over a century there have been continual efforts to incorporate nature into urban planning. These efforts – known as urban reconciliation– aim to manage and create habitats that support biodiversity within cities. Given that species select habitat at different spatial scales, understanding the scale at which urban species respond to their environment is critical to the success of urban reconciliation efforts. We assessed species-habitat relationships for common bat species at local (50-m), medium (500-m), and broad (1-km) spatial scales in the Chicago metropolitan area and predicted bat activity across the greater Chicago region. We found that habitat characteristics across all measured scales were important predictors of silver-haired bat (Lasionycteris noctivagans) and eastern red bat (Lasiurus borealis) activity, and big brown bat (Eptesicus fuscus) activity was significantly lower at urban sites compared to rural sites. We also found that open vegetation had a negative effect on silver-haired bat activity at the local scale but a positive effect at the medium-sized scale, indicating potential shifts in the relative importance of some habitat characteristics at varying scales. These results demonstrate that local-scale effects may be constrained by broader spatial patterns. Our findings highlight the importance of considering scale in urban reconciliation efforts and our landscape predictions provide information that can help prioritize urban conservation work.

Species differences between domestic cats (Felis catus) and dogs (Canis familiaris) has led to differences in their ability to digest, absorb and metabolize carbohydrates through poorly characterized mechanisms. The current study aimed to first examine biopsied small intestine, pancreas, liver and skeletal muscle from laboratory beagles and domestic cats for mRNA expression of key enzymes involved in starch digestion (amylase), glucose transport (sodium-dependent SGLTs and -independent glucose transporters, GLUT) and glucose metabolism (hexokinase and glucokinase). Cats had lower mRNA expression of most genes examined in almost all tissues compared to dogs (p < 0.05). Next, postprandial glucose, insulin, methylglyoxal (a toxic glucose metabolite) and d-lactate (metabolite of methylglyoxal) after single feedings of different starch sources were tested in fasted dogs and cats. After feeding pure glucose, peak postprandial blood glucose and methylglyoxal were surprisingly similar between dogs and cats, except cats had a longer time to peak and a greater area under the curve consistent with lower glycolytic enzyme expression. After feeding starches or whole diets to dogs, postprandial glycemic response, glycemic index, insulin, methylglyoxal and d-lactate followed reported glycemic index trends in humans. In contrast, cats showed very low to negligible postprandial glycemic responses and low insulin after feeding different starch sources, but not whole diets, with no relationship to methylglyoxal or d-lactate. Thus, the concept of glycemic index appears valid in dogs, but not cats. Differences in amylase, glucose transporters, and glycolytic enzymes are consistent with species differences in starch and glucose handling between cats and dogs.

Responses to chemical and physical stressors are commonly expected among organisms residing near the Athabasca oil sands. Physiological effects have been observed in fishes during field studies; but further effects associated with development are not clear or consistent among species. For instance, data from a fish fence in 2009 show declines in the relative abundances of some species, including Arctic grayling ( Thymallus arcticus). In contrast, increases were seen in white sucker ( Catostomus commersoni). This divergence suggests incomplete understanding of the status of fishes residing near the oil sands. However, an important challenge limiting understanding is the lack of reliable baseline or reference data. To overcome this challenge, we used iterative normal ranges and a historical data set (electrofishing surveys done from 1987 to 2014) to determine if changes have occurred in fishes captured in the lower Athabasca River. These analyses revealed clear increases in the lengths of white sucker and walleye ( Sander vitreus) and their relative abundances during the spawning season. The occurrence of these changes may be associated with overwintering location, but reduced fishing pressure in Lake Athabasca, eutrophication, or a cumulative effect may explain the form of changes detected in this study.

Aggressive behavior is common in many species and is often adaptive because it enables individuals to gain access to limited resources. However, aggression is also highly plastic and the degree of plasticity could be influenced by factors such as resource limitation and the social environment. In this study, we examined how the effects of social experience and resource limitation could persist to affect future aggressive interactions. Using naturally inbred strains of Drosophila melanogaster that differ in aggressiveness, we manipulated the level of available resources by varying fly density (two treatments: high and low per capita resources) and group composition by varying strain frequency (five treatments: homogeneous strains, or mixed at 1:3, 1:1 or 3:1 ratios of the more aggressive to less-aggressive strain). For each treatment group, we measured aggression before and after flies were placed through a 4-day period of fixed resources. There was no consistent effect of resource competition on aggression. Instead, changes in aggression depended on resource availability in combination with group composition. In homogeneous groups made up of only one strain, all males became more aggressive following the fixed-resource period, regardless of fly density. In mixed-strain treatments at high density, we observed plastic shifts in aggression of males from both strains, but the direction of plastic responses depended on social composition. Our results show that aggression may not only be influenced by the intensity of previous competitive experiences caused by resource limitation, but also through social effects caused by the composition of the group. Lay Summary An animal's expression of aggression can change over time, partly due to previous experiences with other individuals, and can persist across time and space. We created environments where fruit flies competed for food, competitors were either aggressive or non-aggressive, and they were mixed at different ratios. We tested subsequent changes in aggression as a result of these competitions. Male aggression was affected by their competitive experience, but the impact depended on the group makeup during competition.

Identical twin pairs of dairy cows (one twin being placed with 4 calves for multiple suckling while the other was machine milked) were used to study the hormonal changes associated with anoestrus. Experiments were conducted to examine the effect of multiple suckling on the interval to first oestrus post partum, concentrations of LH and progesterone post partum, and the LH response to LH-RH or oestradiol-17 beta. The effects of hormone therapy on anoestrus were also examined. Multiple suckled cows had a 5-9-week longer interval to first oestrus post partum than did their non-suckling twins in all trials. There were no significant differences between the suckling and non-suckling cows in basal plasma levels of LH. The suckling cows showed a delay of 2 and 6 weeks post partum in their response to LH-RH and oestradiol-17 beta respectively when compared with their non-suckling twins. Treatment with a progesterone-releasing intravaginal device and bromocriptine to lower prolactin levels did not reduce the interval to the first post-partum oestrus while treatment with the progesterone device and PMSG reduced the interval to both first oestrus and conception. These findings suggest that suckling affects the response of the hypothalamic-pituitary axis to both LH-RH and oestradiol in the post-partum cow.

This study investigates the association of PD-L1 expression and immune cell infiltrates and their impact on clinical outcome, in addition to their overlap with microsatellite instability (MSI), HER2 and ATM molecular subgroups of gastric cancer (GC). PD-L1 membrane expression on tumour cells (TC) and infiltrating immune cells (IC), CD3 + T-lymphocytes, CD8+ cytotoxic T-cells, ATM and HER2 were assessed by immunohistochemistry (IHC) in the ACRG (Asian Cancer Research Group) GC cohort (N = 380). EBV status was determined using in situ hybridization and MSI status was performed using PCR and MLH1 IHC. The PD-L1 segment was associated with increased T-cell infiltrates, while the MSI-high segment was enriched for PD-L1, CD3, and CD8. Multivariate analysis confirmed PD-L1 positivity, high CD3 and high CD8 as independent prognostic factors for both disease-free survival and overall survival (all p

Corticosteroid-binding globulin (CBG) transports glucocorticoids in blood and is a serine protease inhibitor family member. Human CBG has a reactive center loop (RCL) which, when cleaved by neutrophil elastase (NE), disrupts its steroid-binding activity. Measurements of CBG levels are typically based on steroid-binding capacity or immunoassays. Discrepancies in ELISAs using monoclonal antibodies that discriminate between intact vs RCL-cleaved CBG have been interpreted as evidence that CBG with a cleaved RCL and low affinity for cortisol exists in the circulation. We examined the biochemical properties of plasma CBG in samples with discordant ELISA measurements and sought to identify RCL-cleaved CBG in human blood samples. Plasma CBG-binding capacity and ELISA values were consistent in arterial and venous blood draining skeletal muscle, liver and brain, as well as from a tissue (adipose) expected to contain activated neutrophils in obese individuals. Moreover, RCL-cleaved CBG was undetectable in plasma from critically ill patients, irrespective of whether their ELISA measurements were concordant or discordant. We found no evidence of RCL-cleaved CBG in plasma using a heat-dependent polymerization assay, and CBG that resists immunoprecipitation with a monoclonal antibody designed to specifically recognize an intact RCL, bound steroids with a high affinity. In addition, mass spectrometry confirmed the absence of NE-cleaved CBG in plasma in which ELISA values were highly discordant. Human CBG with a NE-cleaved RCL and low affinity for steroids is absent in blood samples, and CBG ELISA discrepancies likely reflect structural differences that alter epitopes recognized by specific monoclonal antibodies.

FGFR1 and FGFR2 are amplified in many tumor types, yet what determines response to FGFR inhibition in amplified cancers is unknown. In a translational clinical trial, we show that gastric cancers with high-level clonal FGFR2 amplification have a high response rate to the selective FGFR inhibitor AZD4547, whereas cancers with subclonal or low-level amplification did not respond. Using cell lines and patient-derived xenograft models, we show that high-level FGFR2 amplification initiates a distinct oncogene addiction phenotype, characterized by FGFR2-mediated transactivation of alternative receptor kinases, bringing PI3K/mTOR signaling under FGFR control. Signaling in low-level FGFR1-amplified cancers is more restricted to MAPK signaling, limiting sensitivity to FGFR inhibition. Finally, we show that circulating tumor DNA screening can identify high-level clonally amplified cancers. Our data provide a mechanistic understanding of the distinct pattern of oncogene addiction seen in highly amplified cancers and demonstrate the importance of clonality in predicting response to targeted therapy. Significance: Robust single-agent response to FGFR inhibition is seen only in high-level FGFR-amplified cancers, with copy-number level dictating response to FGFR inhibition in vitro, in vivo, and in the clinic. High-level amplification of FGFR2 is relatively rare in gastric and breast cancers, and we show that screening for amplification in circulating tumor DNA may present a viable strategy to screen patients. Cancer Discov; 6(8); 838–51. ©2016 AACR. This article is highlighted in the In This Issue feature, p. 803

Human-wildlife conflict is increasing as urbanization expands and wildlife species adjust to living near people. Translocation is often used to manage human-wildlife conflict because it is considered to be humane, yet fates of translocated animals are largely unknown. As an urban adapter, woodchucks (Marmota monax) are a common source of human-wildlife conflict due to their burrowing, foraging, and scent-marking behavior. We examined survival and movements of 27 nuisance woodchucks captured by a nuisance wildlife operator in the Chicago metropolitan area, radiomarked with internal transmitters, and translocated to exurban release sites mimicking typical practices. We also captured and radiomarked 16 resident woodchucks from the release landscape for comparison. Translocated woodchucks moved farther than residents immediately post release with no evidence of homing and most left the release site. Annual survival did not differ between translocated and resident woodchucks. However, survival was extremely low (0.18) compared to previous estimates for woodchucks, primarily due to high predation by coyotes (Canis latrans). Translocation should be used only when other nonlethal methods are ineffective (e.g., exclusion, removing food sources, selecting unpalatable plants for gardens). When necessary, the practice of translocation could be improved by reducing predation risk for translocated animals, either by selecting release sites with low predation risk, or by using soft-release methods, such as acclimation enclosures or artificial burrows.

Baseline variations in trout-perch energy use (growth, gonad size) and energy storage (condition, liver size) were characterized between 2009 and 2015 in 8 reaches of the Athabasca River (AB, Canada), including 2 reaches upstream of the city of Fort McMurray (AB, Canada) and 6 reaches downstream of Fort McMurray among existing oil sands operations. Generalized linear models, used to account for background variation, indicated that fork length, gonad size, and liver size decreased, whereas body weight increased, in relation to river discharge, for both male and female trout-perch. Air temperature was positively correlated with liver size and negatively correlated with gonad size and body weight for females, but only positively correlated with gonad weight for males. These linear models explained approximately 20 to 25% of the variation in adjusted body size, and upward of 80% of the variation in adjusted body weight, liver weight, and gonad weight. Residuals from linear models were used to estimate normal ranges of variation for each of the fish population performance measures. Combined, the models and normal ranges can be used to assess subsequent monitoring data, providing potential triggers for follow-up monitoring activities. Environ Toxicol Chem 2019;38:0-0. Environ Toxicol Chem 2019;38:1890-1901. © 2019 SETAC.

This article reviews the biochemical and physiological observations underpinning the synergism between ghrelin and ghrelin agonists with exercise, especially progressive resistance training that has been shown to increase muscle mass. The synergy of ghrelin agonists and physical exercise could be beneficial in conditions where muscle wasting is present, such as that found in patients with advanced cancer. The principal mechanism that controls muscle anabolism following the activation of the ghrelin receptor in the central nervous system involves the release of growth hormone/insulin-like growth factor-1 (GH/IGF-1). GH/IGF-1 axis has a dual pathway of action on muscle growth: (a) a direct action on muscle, bone and fat tissue and (b) an indirect action via the production of both muscle-restricted mIGF-1 and anti-cachectic cytokines. Progressive resistance training is a potent inducer of the secretion the muscle-restricted IGF-1 (mIGF-1) that enhances protein synthesis, increases lean body mass and eventually leads to the improvement of muscle strength. Thus, the combination of ghrelin administration with progressive resistance training may serve to circumvent ghrelin resistance and further reduce muscle wasting, which are commonly associated with cachexia.

Context and Objective: 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) catalyses regeneration of cortisol in liver, adipose tissue, and skeletal muscle, making a substantial contribution to circulating cortisol as demonstrated in humans by combining stable isotope tracer infusion with arteriovenous sampling. In the brain, 11βHSD1 is a potential therapeutic target implicated in age-associated cognitive dysfunction. We aimed to quantify brain 11βHSD1 activity, both to assess its contribution to systemic cortisol/cortisone turnover and to develop a tool for measuring 11βHSD1 in dementia and following administration of 11βHSD1 inhibitors. Design, Setting, and Participants: With ethical approval and informed consent, 8 healthy men aged 38.1 years (sd 16.5) underwent an ECG-gated phase-contrast magnetic resonance scan to quantify internal jugular vein blood flow and were infused with 1,2 [2H]2-cortisone and 9,11,12,12 [2H]4-cortisol for 3 h before samples were obtained from the internal jugular vein and an arterialized hand vein. Steroids were quantified by liquid chromatography-tandem mass spectrometry. Main Outcome Measures and Results: Steady state tracer enrichments were achieved and systemic indices of cortisol/cortisone interconversion were consistent with previous studies in healthy men. However, there was no measurable release or production of cortisol, 9,12,12 [2H]3-cortisol or cortisone into the internal jugular vein. Conclusions: Although cerebral 11βHSD1 reductase activity may be greater in cognitively impaired patients, in healthy men any contribution of 11βHSD1 in the brain to systemic cortisol/cortisone turnover is negligible. The influence of 11βHSD1 in the brain is likely confined to subregions, notably the hippocampus. Alternative approaches are required to quantify pharmacodynamics effects of 11βHSD1 inhibitors in the human brain.

Aggression can be beneficial in competitive environments if aggressive individuals are more likely to access resources than non-aggressive individuals. However, variation in aggressive behaviour persists within populations, suggesting that high levels of aggression might not always be favoured. The goal of this study was to experimentally assess the effects of population density and phenotypic frequency on selection on aggression in a competitive environment. We compared survival of two strains of Drosophila melanogaster that differ in aggression across three density treatments and five frequency treatments (single strain groups, equal numbers of each strain and strains mixed at 3:1 and 1:3 ratios) during a period of limited resources. While there was no difference in survival across single-strain treatments, survival was strongly density dependent, with declining survival as density increased. Furthermore, at medium and high densities, there was evidence of negative frequency-dependent selection, where rare strains experienced greater survival than common strains. However, there was no evidence of negative frequency-dependent selection at low density. Our results indicate that the benefits of aggression during periods of limited resources can depend on the interaction between the phenotypic composition of populations and population density, both of which are mechanisms that could maintain variation in aggressive behaviours within natural populations.

International audience; Collective decision processes remain a common management approach in most organizations. In such processes, it seems important to offer participants the opportunity to confront the differences in their points of view. To this end, cognitive and technical tools are required that facilitate the sharing of individuals’ reasoning and preferences, but at the same time allow them to keep some information and attitudes to themselves. The aim of our study is to assess whether, in the multi-criteria approach to problem structuring, decision-makers can be comfortable using shared criteria in addition to private criteria. For this purpose, an exploratory experiment with student subjects was conducted using the Group Decision Support System, GRUS.

This paper presents results of analyses of body burdens of metals and hydrocarbons, and taste tests for taint, in Iceland scallop and American plaice performed as part of the Environmental Effects Monitoring (EEM) program for the Terra Nova offshore oil development (Grand Banks of Newfoundland, Canada). Scallop and plaice were collected in a Study Area located within approximately 1 km of drill centres at Terra Nova and in a Reference Area located approximately 20 km from the development. Samples were collected in 1997 to establish a baseline, and from 2000 to 2010, during drilling periods. Scallop adductor muscle tissue was contaminated with >C10–C21 aliphatic hydrocarbons resembling the drilling fluid in the synthetic drilling mud (SBM) used at Terra Nova in 2000, 2002 and 2004, but contamination of adductor muscle was not noted in 2006, 2008 and 2010. The maximum concentration in muscle was 28 mg/kg wet weight, noted in 2002. Scallop viscera was contaminated with hydrocarbons resembling drilling fluid in SBMs near drill centres in all EEM years except 2010. Viscera contamination with >C10–C21 hydrocarbons gradually decreased from a maximum of 150 mg/kg in 2000, to a maximum of 27 mg/kg in 2008; all values were below the laboratory reporting detection limit of 15 mg/kg in 2010. Therefore, evidence from both muscle and viscera indicates a decrease in tissue hydrocarbon contamination in recent years. Barium, another major constituent in drilling muds, has not been noted in scallop adductor muscles at concentrations above the reporting detection limit, but barium was detected in viscera in baseline and EEM years. The maximum concentration of barium in viscera during baseline sampling was 8 mg/kg. The maximum concentration in EEM years (29 mg/kg) was noted in 2000. The maximum concentration in 2010 was 25 mg/kg. The concentration of metals other than barium in scallop tissues was similar between the Terra Nova Study Area and the Reference Area. Hydrocarbons resembling the fluid in SBMs were noted in one American plaice liver sample collected near drill centres in 2000. Otherwise, there has been no evidence of project-related metals or hydrocarbon contamination in plaice liver or fillet samples. There has been no evidence of taint (off-taste) for scallop adductor muscle and plaice fillet tissue in baseline or EEM years. Combined with a parallel study on fish bioindicators at Terra Nova that showed that fish health at Terra Nova was similar to that at the Reference Area ( Mathieu et al., 2011 ), these results indicate little to no detectable biological effects on Iceland scallop and American plaice as a result of Terra Nova activities.

An environmental effects monitoring (EEM) program was developed by Suncor (formerly Petro-Canada) in 1997/98 to assess effects of the Terra Nova offshore oil and gas development on the receiving environment. The Terra Nova Field is located on the Grand Banks approximately 350 km southeast of Newfoundland (Canada), at approximately 100 m water depth. The EEM program was developed with guidance from experts in government, academia and elsewhere, and with input from the public. The EEM program proposed by Suncor was accepted by Canadian regulatory agencies and the program was implemented in 2000, 2001, 2002, 2004, 2006, 2008 and 2010, with pre-development sampling in 1997. The program continues to be implemented every two years. EEM includes an assessment of alterations in sediment quality through examination of changes in sediment chemistry, particle size, toxicity and benthic invertebrate community structure. A second component of the program examines potential effects on two species of commercial fishing interest: Iceland scallop (Chlamys islandica) and American plaice (Hippoglossoides platessoides). Chemical body burden for these two species is examined and taste tests are performed to assess the presence of taint in edible tissues. Effects on American plaice bioindicators are also examined. A final component of the program assesses potential effects of the Terra Nova development on water quality and examines water column chemistry, chlorophyll concentration and physical properties. The papers presented in this collection focus on effects of drill cuttings and drilling muds on the seafloor environment and, as such, report results on sediment quality and bioaccumulation of drilling mud components in Iceland scallop and American plaice. This paper provides information on drilling discharges, an overview of the physical oceanography at the Terra Nova Field, and an overview of the field program designed to assess environmental effects of drilling at Terra Nova.

This paper describes effects of drilling with water and synthetic-based drilling muds on benthic macro-invertebrates over 10 years at the Terra Nova offshore oil development. As such, the paper provides insight on the effects of relatively new synthetic-based drilling muds (SBMs), and makes an important contribution to our understanding of the long-term chronic effects of drilling on benthic communities. The Terra Nova Field is located approximately 350 km offshore on the Grand Banks of Newfoundland (Canada). Sediment and invertebrate samples were collected in 1997 (baseline) prior to drilling, and subsequently in 2000, 2001, 2002, 2004, 2006, 2008 and 2010. Approximately 50 stations were sampled in each year at distances of less than 1 to approximately 20 km from drill centres. Summary benthic invertebrate community measures examined were total abundance, biomass, richness, diversity and multivariate measures of community composition based on non-Metric Dimensional Scaling (nMDS). Decreases in abundance, biomass and richness were noted at one station located nearest (0.14 km) to a drill centre in some environmental effects monitoring (EEM) years. These decreases coincided with higher levels of tracers of drill muds in sediments (barium and >C 10 –C 21 hydrocarbons). Abundances of selected individual taxa were also examined to help interpret responses when project-related effects on summary measures occurred. Enrichment effects on some tolerant taxa (e.g., the polychaete family Phyllodocidae and the bivalve family Tellinidae) and decreased abundances of sensitive taxa (e.g., the polychaete families Orbiniidae and Paraonidae) were detected to within approximately 1–2 km from discharge source. Lagged responses three to five years after drilling started were noted for Phyllodocidae and Tellinidae, suggesting chronic or indirect effects. Overall, results of benthic community analyses at Terra Nova indicate that effects on summary measures of community composition were spatially limited but, as seen elsewhere, some taxa were more sensitive to drilling discharges.

3,3-Di-methyl-azetidin-2-ones were identified as potent and selective 11β-HSD1 inhibitors against the human and mouse forms of the enzyme. Structure guided optimisation of LLE was conducted, utilising a key polar interaction and identifying stereochemical preference for the 4S isomer. Metabolic stability was improved to afford oral exposure, providing tool compounds suitable for pre-clinical evaluation.

Following a cluster of haematology patients with carbapenem-resistant Klebsiella pneumoniae (CRKP) septicaemia, we initiated screening for rectal carriage of CRKP and multidrug-resistant K. pneumoniae (MDRKP) in this patient group. Haematology inpatients submit a rectal swab once weekly. When plated onto chromogenic Brilliance™ UTI Agar (Oxoid), and incubated overnight with a 10 µg ertapenem disc (Oxoid), K. pneumoniae is identified and semi-automated antibiotic susceptibility testing is performed using the Vitek 2 analyser (Biomerieux). When no zone of inhibition occurs, immediate intervention through patient isolation and enhanced environmental cleaning can be instigated to control further spread while empirical antibiotic prescribing is adapted to take account of identified resistances. Over 2 years, six patients with CRKP and 20 patients with MDRKP were identified. These isolates were resistant to first-line empirical treatment choices for neutropenic sepsis and presented a clinical risk of treatment failure for sepsis post cytotoxic chemotherapy. We describe how this rectal screening methodology was developed and how the results influenced appropriate antibiotic prescribing, patient placement in single rooms and the cleaning of the ward environment to prevent person-to-person transmission of MDRKP and CRKP.

Among social species, iterative interactions may lead to social preferences among group-mates and often are associ- ated with increased mating opportunities or improved indirect fitness benefits. Although preferential associations have been documented in multiple species, this phenomenon has never been empirically studied in bats—the second largest order of mammals, where many of the 1200+ species live in groups of tens to hundreds of individuals. Given the current understanding of the social behaviour exhibited in this species, we explored the possibility that adult female big brown bats (Eptesicus fuscus (Beauvois, 1796)) show preferential association between group-mates using a pairwise choice design with individuals from a captive colony. Focal individuals were placed in a Y-maze and were given free choice of two familiar conspecifics. We measured the time focal individuals spent in close proximity to each conspecific. Our results indicate that some bats exhibit preferential association between group-mates, as multiple individuals spent significantly more time in close proximity to one conspecific versus another, despite randomizing the position of stimulus bats between trials. Given the frequent and long-term associations between group members of this species, social preferences could play a significant role in the outcomes of their long-term fitness. Resume : Chez les especes sociales, les interactions iteratives peuvent mener ades preferences sociales entre membres d'un meme groupe et sont souvent associees aune augmentation du nombre de possibilites d'accouplement ou ade meilleurs avantages indirects sur le plan de l'aptitude. Si des associations preferentielles ont ete documentees chez de nombreuses especes, ce phenomene n'a jamais ete etudie de maniere empirique chez les chauves-souris, le deuxieme ordre de mammiferes en importance, dont bon nombre des plus de 1200 especes vivent en groupes de dizaines de milliers d'individus. Etant donne la comprehension actuelle du comportement social de cette espece, nous avons explore la possibilite que les femelles adultes de la serotine brune (Eptesicus fuscus (Beauvois, 1796)) fassent preuve d'association preferentielle au sein de leur groupe, en appliquant ades individus d'une colonie captive un schema de choix par paire. Les individus vises etaient places dans un labyrinthe en Y et avaient le choix entre deux congeneres qui leur etaient familiers. Nous avons mesure le temps que les individus vises passaient en etroite proximite avec chaque congenere. Nos resultats indiquent que certaines chauves-souris font preuve d'association preferentielle avec des membres de leur groupe, puisque de multiples individus passaient significativement plus de temps en etroite proximite avec un congenere plutot qu'un autre, malgre le fait que, entre les essais, les chauves-souris stimuli etaient reparties aleatoirement. Etant donne les associations frequentes et along terme de membres du meme groupe chez cette espece, les preferences sociales pourraient jouer un important role dans la determination de leur aptitude a long terme. (Traduit par la Redaction) Mots-cles : associations sociales, Eptesicus fuscus, bais social, labyrinthe en Y.

Historical eruptions from Mt. Ruapehu (New Zealand) have been small (

Behavioural plasticity is a critical component of natural selection leading to evolution. However, a surge of studies in the last two decades has discovered a distinct limit to behavioural plasticity, commonly referred to as behaviour types and behavioural syndromes. We set out to understand the relationships across behavioural categories in wild-caught adult, female big brown bats and how they compare between social and solitary behaviours. Using bats sampled from four different maternity colonies, we ran a series of behavioural assays to create a behavioural profile for each individual. The behavioural profile encompassed exploratory, learning, competitive and aggressive categories. We found that Big brown bats exhibit a mean profile relatively unique to other well-documented species, where aggression was linked to increased competitive ability but not to boldness. Our results indicate that the solitary and socially directed behaviours of individuals are not necessarily related and that behaviours pertaining to social interactions are linked most closely to learning abilities. Furthermore, we found evidence that poor body condition may be a predictor of increased social interactions and that behaviours exhibited in the presence of conspecifics are unrelated to those exhibited in solitude. These findings indicate importance of social affiliations on individual behaviours in this species and their uniqueness relative to other well-studied taxa.

The discovery of novel pathogenic mechanisms engaged during bacterial infections requires the evolution of advanced techniques. Here, we evaluate the dual polarity matrix norharmane (NRM) to improve detection of bacterial lipid A (endotoxin), from host and vector tissues infected withFrancisella novicida (Fn). We evaluated NRM for improved detection and characterization of a wide range of lipids in both positive and negative polarities, including lipid A and phospholipids across a range of matrix-assisted laser desorption-ionization-coupled applications. NRM matrix improved the limit of detection (LOD) for monophosphoryl lipid A (MPLA) down to picogram level representing a 10-fold improvement of LOD versus 2,5-dihydroxybenzoic acid and 100-fold improvement of LOD versus 9-aminoacridine (9-AA). Improved LOD for lipid A subsequently facilitated detection of theFn lipid A major ion (m/z 1665) from extracts of infected mouse spleen and the temperature-modifiedFn lipid A atm/z 1637 from infectedDermacentor variabilis ticks. Finally, we simultaneously mapped bacterial phospholipid signatures within anFn-infected spleen along with an exclusively host-derived inositol-based phospholipid (m/z 933) demonstrating coprofiling of the host-pathogen interaction. Expanded use of NRM matrix in other infection models and endotoxin-targeting imaging experiments will improve our understanding of the lipid interactions at the host-pathogen interface., This study presents a versatile tool for host-pathogen interaction studies by mass spectrometry imaging; we emphasize the improved detection of pathogen-specific lipids (endotoxin) achievable only with norharmane (NRM), an underutilized matrix substance facilitating simultaneous detection of host and pathogen lipids.

The challenge of developing effective pharmacodynamic biomarkers for preclinical and clinical testing of FGFR signaling inhibition is significant. Assays that rely on the measurement of phospho-protein epitopes can be limited by the availability of effective antibody detection reagents. Transcript profiling enables accurate quantification of many biomarkers and provides a broader representation of pathway modulation. To identify dynamic transcript biomarkers of FGFR signaling inhibition by AZD4547, a potent inhibitor of FGF receptors 1, 2, and 3, a gene expression profiling study was performed in FGFR2-amplified, drug-sensitive tumor cell lines. Consistent with known signaling pathways activated by FGFR, we identified transcript biomarkers downstream of the RAS-MAPK and PI3K/AKT pathways. Using different tumor cell lines in vitro and xenografts in vivo, we confirmed that some of these transcript biomarkers (DUSP6, ETV5, YPEL2) were modulated downstream of oncogenic FGFR1, 2, 3, whereas others showed selective modulation only by FGFR2 signaling (EGR1). These transcripts showed consistent time-dependent modulation, corresponding to the plasma exposure of AZD4547 and inhibition of phosphorylation of the downstream signaling molecules FRS2 or ERK. Combination of FGFR and AKT inhibition in an FGFR2-mutated endometrial cancer xenograft model enhanced modulation of transcript biomarkers from the PI3K/AKT pathway and tumor growth inhibition. These biomarkers were detected on the clinically validated nanoString platform. Taken together, these data identified novel dynamic transcript biomarkers of FGFR inhibition that were validated in a number of in vivo models, and which are more robustly modulated by FGFR inhibition than some conventional downstream signaling protein biomarkers. Mol Cancer Ther; 15(11); 2802–13. ©2016 AACR.

One of the “Five Freedoms” proposed by the Farm Animal Welfare Council in Great Britain is that animals should be free to express normal behaviour. For various domesticated species, the study of still-existing wild ancestors has been one approach in the definition of normal. However, for domesticated cattle, no such wild ancestor exists, so the best that can be done is to study domesticated cattle in environments with little human interference. While several studies have been made of the behaviour of cattle under such conditions, no critical review of the literature has been published. This is the purpose of this paper. While it is surprising that no such review has been undertaken, given its usefulness in the study of the welfare of cattle, what is more surprising is that only 22 studies, published during the period 1927–2009, were considered for inclusion in the review. From the review, it can be concluded that cattle have quite an extensive repertoire of behaviour, comprising 40 identifiable categories and that, of all of the behaviours in their repertoire, grazing is the most common followed, generally, by ruminating and resting. In general, these three categories take up 90–95% of an animal's day. The review also showed that most grazing is performed during the hours of daylight, with very little grazing is observed at night. However, there were exceptions where considerable grazing was observed at night. Furthermore, the greater part of rumination occurred while animals were lying rather than standing and, although the information was scant, ruminating and resting tended to occur at night rather than during the day. Finally, cattle generally follow a diurnal rhythm characterised by peaks of grazing activity associated with sunrise and sunset.

We demonstrate that after implementation of recommendations for universal infant hepatitis B vaccination, HBV infection prevalence among children of foreign-born Asian parents in Georgia declined dramatically; horizontal transmission of infection within households has occurred infrequently; and the vast majority of infants and children have received the recommended hepatitis B vaccinations. These results provide evidence of the success of the hepatitis B infant vaccination program and highlight its potential impact on reducing chronic HBV infection morbidity and mortality among U.S. populations at high risk.

Sticklebacks are used as a model system in behavioral and evolutionary research, and therefore it is important to have a complete understanding of their biology. Despite this, the presence of acoustic signals has never been explored. This study examines acoustic cues in brook sticklebacks during courtship, spawning, egg guarding, and fry guarding. Although some fish produce sounds during spine flaring, results of this experiment showed that brook sticklebacks do not. Females did produce incidental sounds while depositing eggs in the nest. However, these were low intensity and seem unlikely to be heard over background noise.

Summary Data from 838 Finnish Landrace or Finnish Landrace crossbred sheep showed a highly significant correlation between phenotypes of the C blood group system and erythrocyte amino acid transport variants. Erythrocytes with normal amino acid transport properties (GSH high, Ly- type) were Cb-positive or Cb-negative. Erythrocytes with the amino acid transport lesion (GSH low, Ly +) were never Cb-negative. Sheep erythrocytes homozygous for Cbshowed stronger lysis reactions with anti-Cb than heterozygous cells. Ly + sheep were nearly always homozygous for Cb, whereas most Ly- sheep were heterozygous or Cb-negative. Inheritance studies provided strong evidence that this association is due to close genetic linkage.

To consider cattle welfare, time budget of behaviors of young cattle in pen and pasture conditions were compared. Behavioral observations of 103 steers in seven pens and 1136 steers in six ranges were performed during daylight over 3 days in each rearing condition. The pen condition had Japanese Black × Holstein cross, and the pasture conditions had assorted breeds of Angus, Murray Grey, Shorthorn, Hereford, Santa Gertrudis and their crosses aged 5–15 months. The ranges varied in vegetation from extensive native pasture to intensive improved pasture. The daytime proportion of behaviors and activity patterns were compared between rearing conditions. The proportion of walking was much lower in the pen condition (1.0 ± 0.2%) compared to all pasture conditions (at least 9.4 ± 4.0%; all P

[The galatheoid, Eumunida picta, primarily has been reported from deep sea corals and methane seeps; we examined its association with the deep-sea coral, Lophelia pertusa in a non-seep habitat. We used a remotely operated vehicle (ROV) to quantify invertebrate epifauna on six shipwrecks in the Gulf of Mexico in depths of 150 to 1950 m during summer, 2004. Video transects were conducted from over the shipwrecks to 333 m away from the wrecks. Eumunida picta specimens were observed on one of the six wrecks, from 530 to 560 m depth. When present, E. picta were primarily in association with Lophelia pertusa. Eumunida picta were associated with L. pertusa in >80% of the observations and were usually within 1 m of the coral when not in direct contact. The coral may be providing an optimal feeding location for the galatheoid, refuge from predation, or some combination of those. Le Galatheoidea Eumunida picta a ete tout d'abord decouvert dans les coraux profonds et les suintements de methane; nous avons examine son association avec le corail profond, Lophelia pertusa dans un habitat non suintant. Nous avons utilise un systeme teleopere a grande profondeur (ROV, Remotely Operated Vehicle) pour quantifier l'epifaune invertebree sur six epaves de bateaux dans le golfe du Mexique a des profondeurs de 150 a 1950 m au cours de l'ete 2004. Des transects videos ont ete realises depuis les epaves jusqu'a 333 m autour de celles-ci. Des specimens d'Eumunida picta ont ete observes sur l'une des six epaves, a une profondeur de 530 a 560 m. Quand ils etaient presents, les specimens de E. picta etaient associes au Lophelia pertusa. Les Eumunida picta etaient associes a L. pertusa dans plus de 80% des observations et etaient habituellement presents dans une zone de 1 m autour des coraux quand ils n'etaient pas en contact direct. Les coraux pourraient procurer a ce Galatheoidea un excellent site pour se nourrir, un refuge face a la predation ou une combinaison des deux., The galatheoid, Eumunida picta, primarily has been reported from deep sea corals and methane seeps; we examined its association with the deep-sea coral, Lophelia pertusa in a non-seep habitat. We used a remotely operated vehicle (ROV) to quantify invertebrate epifauna on six shipwrecks in the Gulf of Mexico in depths of 150 to 1950 m during summer, 2004. Video transects were conducted from over the shipwrecks to 333 m away from the wrecks. Eumunida picta specimens were observed on one of the six wrecks, from 530 to 560 m depth. When present, E. picta were primarily in association with Lophelia pertusa. Eumunida picta were associated with L. pertusa in >80% of the observations and were usually within 1 m of the coral when not in direct contact. The coral may be providing an optimal feeding location for the galatheoid, refuge from predation, or some combination of those. Le Galatheoidea Eumunida picta a ete tout d'abord decouvert dans les coraux profonds et les suintements de methane; nous avons examine son association avec le corail profond, Lophelia pertusa dans un habitat non suintant. Nous avons utilise un systeme teleopere a grande profondeur (ROV, Remotely Operated Vehicle) pour quantifier l'epifaune invertebree sur six epaves de bateaux dans le golfe du Mexique a des profondeurs de 150 a 1950 m au cours de l'ete 2004. Des transects videos ont ete realises depuis les epaves jusqu'a 333 m autour de celles-ci. Des specimens d'Eumunida picta ont ete observes sur l'une des six epaves, a une profondeur de 530 a 560 m. Quand ils etaient presents, les specimens de E. picta etaient associes au Lophelia pertusa. Les Eumunida picta etaient associes a L. pertusa dans plus de 80% des observations et etaient habituellement presents dans une zone de 1 m autour des coraux quand ils n'etaient pas en contact direct. Les coraux pourraient procurer a ce Galatheoidea un excellent site pour se nourrir, un refuge face a la predation ou une combinaison des deux.]

This study observed the behavioral characteristics of 122 steers in eight pens and 1,136 steers at six pastures. Nonhuman animals kept in pens performed less nutritive oral behaviors and more nonnutritive oral behaviors than animals kept at pasture. Although these could not be described as stereotypies, they did represent a replacement of nutritive oral behaviors by nonnutritive oral behaviors, rather than simply an increase in resting time. This could be indicative of a level of oral frustration. At pasture, there was a greater proportion of oral behaviors in animals with low pasture availability as compared to high availability, but this was an increase in nutritive oral behaviors rather than nonnutritive oral behaviors. Factors other than oral frustration--for example, rumen fill--probably drove this increase.

Two experiments were conducted to determine the effect of weaning in small yards, with or without a feed bunk training procedure, on the subsequent behaviour and performance of Bos taurus steers in a feedlot. A reduction in the incidence of bovine respiratory disease (BRD) was the primary objective. In each experiment, about 200 male beef calves (Angus × Hereford and Hereford) were separated from their mothers at 7–9 months of age and allocated to one of three matched weaning treatment groups. The treatments were (1) yard weaning with hay or silage, (2) yard weaning with hay or silage plus a novel handling procedure to train the animals to be able to find a grain ration in a trough, and (3) paddock weaning without supplement or handling according to common industry practice in southeastern Australia. Paddock weaning is the practice of abrupt separation of cows and calves followed by return to separate pasture paddocks, whereas yard weaning involves abrupt separation but the calves remain in the yards for several days. Experimental vaccines against the major BRD pathogens were given to half of each group 1–2 months prior to entry into a commercial feedlot. The yard weaned and yard trained cattle had a significantly greater weight gain in the first month and over the 90-day feeding period than the paddock weaned control groups. There was no difference between the groups in pre-feedlot weight gain. The yard trained groups showed greater feeding activity during the first few days in the feedlot, but were not significantly different in weight gain from yard weaned. The vaccination treatment also significantly improved the weight gain in the first month and over 80 days. The combination of yard weaning and vaccination produced the highest weight gains overall. There was consistently less morbidity in the yard weaned groups compared to paddock weaned controls. The morbidity in yard trained groups was intermediate between these two. Weaning in small yards and the appropriate use of effective BRD vaccines 1–2 months before feedlot entry are recommended for B. taurus feeder steers in southeastern Australia to minimise sickness and improve productivity in the feedlot. Associated benefits are reduced risks of antibiotic residues and of animal welfare problems.

Commercial dairy cows milked in a parlor system are packed close together in the holding area before milking. The present study examined the relationships of ‘looking up’ behavior with some other behaviors and the productivity of 1116–1153 cows from five farms. The individual identities of the cows looking up in the holding area were recorded at 5 min intervals during six intermittent afternoon milking sessions. Entrance into the milking parlor and the numbers of steps and kicks by cows while the milking person was attaching the milking cups, were recorded in six milking sessions. Flight responses in the pasture after milking were recorded over four days intermittently. The frequency of ‘looking up’ behavior weakly, but significantly correlated with flight starting distance (r = 0.10, P

The pyruvate dehydrogenase multienzyme complex catalyses the oxidative decarboxylation of pyruvate, which is an important regulatory step in oxidative metabolism. Phosphorylation of the E1 (pyruvate decarboxylase) subunit on one of three specific serine residues results in loss of enzyme activity. Four dedicated PDHK (pyruvate dehydrogenase kinase) isoenzymes have been identified, each of which display a distinct tissue-specific expression profile, and have differential regulatory properties. Thus PDHK play a key role in controlling the balance between glucose and lipid oxidation according to substrate supply. Increasing glucose oxidation by inhibiting PDHK may be an effective mechanism to increase glucose utilization; additionally, increasing pyruvate oxidation may further contribute to lowering of glucose level by decreasing the supply of gluconeogenic substrates. A number of PDHK inhibitors are now available to enable this mechanism to be evaluated as a therapy for diabetes. The isoenzyme selectivity profile of AZD7545 and related compounds will be described and evidence for their non-ATP-competitive mode of action presented. These compounds increase PDH activity in vivo, and when dosed chronically, improve glycaemic control in Zucker rats. Furthermore, glucose lowering has been demonstrated in the hyperglycaemic Zucker diabetic fatty rat. This result supports the hypothesis that inhibition of PDHK may be an effective therapy for Type II diabetes.

Methylene chloride (dichloromethane) is used in a variety of industrial applications. To date, there has been no formal assessment of immunotoxicity attributed to methylene chloride. Studies were undertaken to examine whether methylene chloride has any potential to influence the integrity of immune function. For this purpose, Sprague-Dawley rats of both genders were exposed by inhalation to a single high dose (5000 ppm) of methylene chloride for 6 h/d, 5 d/wk for 28 d. This was considered the relevant route of administration, as not only is inhalation a primary route for human exposure to methylene chloride, but, also, the chemical is absorbed rapidly via the lungs. Under these conditions of exposure, methylene chloride failed to influence absolute or relative thymus weights in either gender and produced a significant reduction in relative, but not absolute, spleen weight in female rats only. Immunocompetence was measured as a function of the ability of treated animals to mount immunoglobulin M (IgM) antibody responses to sheep red blood cells (SRBC) as determined by enzyme-linked immunosorbent assay (ELISA). Exposure to methylene chloride did not affect antibody production. Evidence indicates that under these conditions of exposure, methylene chloride did not compromise immune function.

AZD4785 is a potent and selective high affinity 2’-4’ constrained ethyl residues (cEt) containing therapeutic antisense oligonucleotide (ASO) targeting KRAS. In a patient-derived explant model of non-small cell lung cancer (NSCLC) carrying a KRAS G12C mutation systemic dosing of AZD4785 resulted in tumor regression. To determine the level of KRAS mRNA knockdown achieved by AZD4785 at a dose which causes regression (250 mg/kg/wk), tumor samples were collected and analyzed by RT-PCR. Mean levels of KRAS mRNA knockdown relative to the PBS control group were determined to be 61% and 78% from studies dosed for 2 and 4 weeks, respectively. However, by RT-PCR there was significant variability in KRAS mRNA levels within the PBS control group, and level of knockdown varied with different housekeeper normalization genes. In early clinical studies it will be essential to measure KRAS expression in tumor samples as a pharmacodynamic marker for AZD4785, hence we sought to develop a more robust assay compatible with FFPE clinical samples with low RNA yields. KRAS mRNA levels were quantified by the nanoString platform, using the panCancer pathways 770 gene codeset. This assay generated robust data from RNA extracted from 2 x 5µm sections of FFPE material from each sample, despite very limited yields of RNA. AZD4785 treatment resulted in comparable levels of KRAS knockdown to the RT-PCR assay, but assay variability was greatly reduced, from stdev >50% (RT-PCR) to 5% (nanoString) for PBS control group samples. Using the nanoString assay, mean KRAS mRNA knockdown was 52% and 68% (p50μm apart within four NSCLC tissues and was found to be ≤21%. Taken together, these data validate the use of the KRAS nanoString assay for use in clinical FFPE tissues, and demonstrate the level of knockdown of KRAS mRNA to be >50% at a dose of AZD4785 that causes regression in a preclinical PDX model. Citation Format: Claire Rooney, Sarah Ross, Anna Staniszewska, Sabine Lennarz, Elaine Kilgour, Alexey S. Revenko, A. Robert Macleod, Andrew Pierce, Paul D. Lyne, J. Carl Barrett, Elizabeth A. Harrington. Development of a pharmacodynamic biomarker assay for AZD4785, an antisense oligonucleotide targeting KRAS [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5095. doi:10.1158/1538-7445.AM2017-5095

We determined differences in the behavior of the progeny of two major sire lines of Japanese Black cattle by recording the behavior of 35 and 70 half-sib steers of sires from fast (FG) and slow (SG) growing lines, respectively. Two sire lines of steers were mixed and allocated to nine pens with 11-12 animals per pen. The proportion of steers lying was significantly (P < 0.001) higher in the SG line (43.4 ± 5.7% compared to 40.3 ± 6.0%). The proportion of time spent eating concentrate feed (FG: 12.1 ± 2.3%; SG: 11.4 ± 2.1%), drinking (FG: 0.8 ± 1.1%; SG: 0.4 ± 0.6%), licking the feed trough (FG: 0.4 ± 0.6%; SG: 0.2 ± 0.4%) and performing tongue-playing (FG: 3.1 ± 4.6%; SG: 1.0 ± 1.9%) was significantly higher in FG, whereas the proportion of time spent resting (FG: 41.5 ± 12.8%; SG: 43.7 ± 10.9%) and performing self-licking (FG: 1.7 ± 1.4%; SG: 2.1 ± 1.3%) was higher in SG (all P < 0.05). These results show progeny of the FG sire engaged in more active behaviors compared to the progeny of the SG sire line.

An in vitro model of the rat nasal cavity has been used to compare the responses of nasal tissues in vitro, using loss of intracellular ATP and potassium as indices of toxicity, with the pathological changes occurring following in vivo exposure to four test compounds. Turbinates were incubated in vitro with the test compounds for 4 h, for 24 h or for 4 h followed by 20 h in fresh medium. Titanium dioxide caused little or no loss of ATP in either olfactory epithelium (OE) or respiratory epithelium (RE). Sodium carbonate decreased olfactory, but not respiratory ATP, while acetic acid and 3-methylindole markedly decreased ATP in both tissues. Intracellular potassium concentrations were generally affected to a lesser degree. In vivo, no morphological changes were observed in the nasal cavity following inhalation exposure to either titanium dioxide or sodium carbonate. Inhalation of acetic acid resulted in a very focal lesion in the RE of the dorsal meatus of level 1, while administration of 3-methylindole by intraperitoneal injection caused severe degeneration of OE. In further experiments olfactory turbinates were exposed to a range of concentrations (0–100 mM) of sodium carbonate, acetic acid and 3-methylindole for 4 h and ATP concentrations determined. Concentration-dependent decreases in ATP were observed for sodium carbonate and 3-methylindole, with EC 50 values estimated as 2.57 and 0.91 mM, respectively. Acetic acid only decreased ATP significantly at the 100-mM concentration. In summary, this in vitro model has predicted the nasal toxicity of several compounds, including both direct-acting agents (sodium carbonate, acetic acid) and one requiring metabolic activation (3-methylindole). However, the lack of airflow-dependent dosimetry, results in some lack of discrimination between the different regions of the nasal cavity and may make this model overly sensitive.

The nearshore benthic communities on the Canadian side of Lake Ontario were surveyed in 1991 at depths of 5 and 20 m, and compared to data obtained in 1981 at the same depths and locations. The comparison was made to determine if the nearshore community reflected known reductions in nutrient concentrations over the 10-year period. The study also was designed to determine the extent to which natural environmental factors such as depth, substrate, fetch, and the likelihood of upwelling alter responses of benthic communities to improvements in water quality. In general, both 5 and 20-m communities reflected the reductions in nutrient concentrations and were indicative of the improved conditions. In 1981 at 5 m, worms were the most dominant group, accounting for 79% of the total fauna. Chironomidae (9%), Gastropoda (4%), Amphipoda (3%), and Sphaeriidae (2%) were subdominant. In 1991 at 5 m, the dominance of worms in the total fauna was reduced to about 15%, while chironomids increased to 34%, amphipods to 27%, gastropods to 18%, and sphaeriid clams to 4%. Similar to 5 m, the fauna at 20 m in 1981 were dominated by worms (65%), while chironomids (10%), amphipods (15%), and sphaeriids (8%) were less dominant. In 1991 at 20 m, the fauna remained dominated by worms but their proportion of the total numbers declined to 48%. At the same time, the proportion of chironomids increased to 20%, amphipods decreased to 7%, gastropods increased to 3%, and sphaeriids increased to 21%. With this data set, a variety of indices of benthic community composition were calculated based on lowest-practical-level identifications. Relationships between changes in these indices and habitat features were explored. Upwelling and substrate type modified the response of the benthic community at 20 m, and are thus important factors to consider in future designs examining whole lake changes in benthic community composition. In contrast, changes in benthic community indices over time at 5 m were unrelated to the type of substrate, fetch or upwelling. As a result, future surveys of 5-m benthos require data from only one substrate type and do not require stratification of study designs by fetch or upwelling. Zebra mussels were found along the western shoreline of Lake Ontario in this 1991 survey in low abundances (averaged 730 indiv/m2), as well as sporadically along the northern shore. At these low abundances, zebra mussels had no significant effects on changes in benthic community composition.

In this study we first established, by immunoblotting with specific antibodies, the temporal changes in cellular levels of protein kinase C (PKC) isoforms during differentiation of 3T3-F442A pre-adipocytes. Both pre-adipocyte and adipocyte 3T3-F442A cells were found to express PKC-alpha, -gamma, -delta, -epsilon, -zeta and -mu. However we were unable to detect PKC-beta, -eta or -theta. The same PKC isoform expression profile was found in rat adipocytes. The alpha, delta and gamma isoforms displayed similar temporal patterns of expression during differentiation of 3T3-F442A cells; all increased rapidly, peaking at day 2 of differentiation. Subsequently, the expression of these isoforms decreased, resulting in lower levels in fully differentiated adipocytes than in pre-adipocytes. The expression of PKC-epsilon increased steadily during differentiation, resulting in markedly elevated levels in adipocytes. Although expression of PKC-mu increased during differentiation, this was attributable to prolonged confluence rather than to the differentiation process itself. No change was observed in PKC-zeta levels during adipocyte development. Anti-sense oligodeoxynucleotides (ODNs) were used to deplete selectively the individual PKC subtypes. Each of the ODNs used effectively depleted the specific isoforms to undetectable levels and did not affect expression of the other PKC subtypes. This approach indicated that pre-adipocyte differentiation is not dependent upon PKC-zeta but that PKC-alpha,-delta and -mu each exert an inhibitory influence upon differentiation. Use of anti-sense ODNs to deplete PKC-epsilon and -gamma revealed that pre-adipocyte differentiation is dependent upon each of these isoforms. However, PKC-gamma, but not PKC-epsilon, appeared to be necessary for the clonal expansion of differentiating cells, suggesting that PKC-epsilon is required at a later phase in the differentiation process, when its expression is elevated, for the attainment and maintenance of the adipocyte phenotype.

In order to further assess the usefulness of behaviour as a possible selection criterion for the improvement of lamb-rearing ability, the arena behaviour of rams and ewes of a flock selected for rearing ability (Fertility Flock) was compared to that of rams and ewes of an unselected flock (Random Flock). Arena behaviour was measured in an approach/avoidance test wherein animals were placed alone in a narrow arena for 10 min. During this period, their attraction to a group of sheep in a pen at one end of the arena relative to their avoidance of a human being sitting in the arena in front of this pen was assessed by quantifying five behaviours; minimum, mean and maximum distance from the human, total distance moved in the arena and number of bleats. These behavioural measurements were made at 6, 12 and 20 months of age. Of the five behaviours measured, total distance travelled consistently differed between the two flocks, Fertility Flock animals travelling, on average, only 60% as far as the Random Flock animals. Number of bleats also differed between the two flocks, but only at 6 and 12 months of age. Importantly, these differences appeared in both ewes and rams. The study also showed that these behaviours were all moderately to highly repeatable. These results further support the possible usefulness of arena behaviour as a means of genetically improving the lamb-rearing ability of ewes and indicate that rapid genetic progress may be possible since these behaviours can be measured early in the life of animals and in animals of both sexes.

We have examined the effects of cyclic AMP on the differentiation of 3T3-F442A preadipocytes. High concentrations of intracellular cyclic AMP potently inhibited differentiation whereas low concentrations of intracellular cyclic AMP, induced by a number of different agents, promoted differentiation. To analyse these effects of cyclic AMP more closely, we developed a two-phase protocol for the differentiation of 3T3-F442A cells. Growth hormone (GH) was necessary to prime confluent cells during the first phase, following which, the addition of insulin and other adipogenic agents then promoted terminal differentiation. Cyclic AMP potentiated the priming action of GH but exerted an inhibitory effect on terminal differentiation when added to cells which had previously been primed with GH showing that the effects of cyclic AMP on preadipocyte differentiation are stage-dependent. We analysed the stimulatory effects of cyclic AMP during GH priming and found that cyclic AMP induced phosphorylation of the cyclic AMP response element (CRE) binding protein CREB and activated transcription of a CRE-linked reporter gene. Furthermore, GH also stimulated CREB phosphorylation and activation and this effect was potentiated by cyclic AMP. These results suggest a mechanism for the synergistic priming of preadipocytes for terminal differentiation by cyclic AMP and GH via the activation of differentiation genes containing CREs.

Nasal toxicology is an important, expanding area of research, which has, in the past, centered on in vivo studies. There is now a need for in vitro methodology to study this target organ, and the aim of this work was to develop such a model using isolated rat nasal turbinates. Initially, optimal culture conditions for olfactory epithelium (OE)and respiratory epithelium (RE) were determined using lactate dehydrogenase (LDH) release as an indicator of viability. The optimal culture conditions were found to be incubation at 31 C under an atmosphere of 95%O:5%CO2, in six-well plates containing 2 mL of William's E medium supplemented with glutamine (2 mM) and gentamycin (100 mug/mL), with shaking at 60 strokes/min (amplitude 3.5 cm). Under these conditions, LDH release from both tissues was 30-40% by 24 h. The model was further characterized in terms of a rangeof viability parameters reflecting different aspects of cellular function. Initial values for intracellular potassium, ATP, and nonprotein sulfhydryl le...