A CE assay for the detection of agonist-stimulated adenylyl cyclase activity

A CE assay was developed for the detection of adenylyl cyclase (AC) activity stimulated at the AC and G protein-coupled receptor (GPCR) level. In the assay, cell membranes overexpressing GPCR and/or AC were incubated with modulators and substrate ATP to produce cAMP in a dose-dependent manner. In both the CE-UV and a radiochemical assay, the addition of forskolin (FSK) resulted in a two- to three-fold maximum increase in AC activity with EC 50 s of 4.2 14± 140.7 and 2.4 14± 140.7 14ΜM, respectively, demonstrating that similar results were obtained by both assays. GPCR activation was also detected using cell membranes overexpressing AC and the Β 2 -adrenergic receptor (Β 2 AR) fused to the stimulatory G protein. Terbutaline (Β 2 AR agonist) increased the basal rate of cAMP formation 1.7 14± 140.1-fold resulting in an EC 50 of 62 14± 1410 14nM. The assay's ability to detect antagonists is demonstrated by the expected right-shifted EC 50 of terbutaline by the Β 2 AR antagonist propranolol. The CE-UV assay offers advantages over the traditional radioactivity assay in terms of safety and labor.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/56060/1/1913_ftp.pd

Mating system variation in neotropical black mangrove, Avicennia germinans, at three spatial scales towards an expanding northern distributional limit

Climate-driven range expansion of ecosystem-defining foundation species can have wide-reaching ecological consequences. Expansion may also result in mating system changes in these foundation species because of the ecological characteristics of range margins, such as greater conspecific isolation and reduced pollinator availability. It is important to understand how mating systems may change during expansion due to their direct influence on intraspecific genetic and demographic dynamics. Here, we used 12 microsatellite loci to genotype progeny arrays of the neotropical black mangrove (Avicennia germinans) at six collection sites (n = 23 maternal trees; 1,612 genotyped propagules) along a latitudinal gradient towards a northern distributional limit on the Atlantic coast of Florida, USA (27.56–30.01oN), where mangroves have expanded into salt marsh over the past several decades. We assessed mating system variation at three spatial scales. First, at the species-distribution level, published outcrossing rates for tropical conspecifics were more than two times higher than those for subtropical Florida A. germinans, consistent with reductions in pollinator diversity and in mangrove abundance with latitude. Second, at the population level, Florida outcrossing rates did not systematically decline towards the northern range limit, but instead, a more open pollen-dispersal neighbourhood at the transition from mangrove to salt marsh dominance may elevate outcrossing until conspecific abundances become too low towards the range limit. Third, at the individual level, outcrossing increased as conspecific cover increased at the Florida range margin, consistent with density-dependent plastic shifts in mating system. These findings suggest that ecological structure influences the A. germinans mating system at varying spatial scales. Further research needs to evaluate the effect of A. germinans mating system variation on the survival and fitness of offspring and on the extent of population-level local adaptation at expanding distributional limits

An expert assessment on playspace designs and thermal environments in a Canadian context

Playgrounds are a hub for child play and concerns that may impact children\u27s play there may hinder their health and well-being. Extreme temperatures can increase risks in children of sunstroke, burns from playground surfaces, and exposure to ultraviolet radiation. Despite health risks from extreme heat to children, existing playground design standards around the world, including in Canada, make little-to-no mention of how to design playgrounds for thermal comfort, particularly in summer. To help fill this gap in the Canadian context, several organizations collaborated to develop guidance for thermally comfortable playgrounds in Canada. As part of this project, an online survey was administered to 55 experts with diverse professional backgrounds, largely from Canada and the United States, to determine how thermal comfort is viewed in playground design and safety. Survey results showed agreement among experts that thermal comfort receives low or no priority in playground design but should be prioritized or considered alongside other safety factors in relevant playground safety guidelines and standards. The results of this survey not only helped inform the 2020 publication of a Thermal Comfort annex to the CSA Group\u27s Children\u27s playgrounds and equipment standard (CAN/CSA Z614) but could also help inform future research and practice globally

Geology and palaeontology of the Hindon Maar Complex: A Miocene terrestrial fossil Lagerstätte in southern New Zealand

Highlights • Hindon Maar Complex is a new mid-Miocene Fossil-Lagerstätte in New Zealand. • Anoxia in maar lakes allowed exquisite preservation of plant and animal fossils. • The biota is from a lake and Nothofagus/podocarp/mixed broadleaf forest ecosystem. • Fossils record high diversity at humid, warm Southern Hemisphere mid-latitudes. Abstract This paper highlights the geology, biodiversity and palaeoecology of the Hindon Maar Complex, the second Miocene Konservat-Lagerstätte to be described from New Zealand. The Lagerstätte comprises four partly eroded maar-diatreme volcanoes, with three craters filled by biogenic and highly fossiliferous lacustrine sediments. The exceptionally well-preserved and diverse biota from the site is derived from a mid-latitude Southern Hemisphere lake-forest palaeoecosystem, including many fossil taxa not previously reported from the Southern Hemisphere. The most common macrofossils are leaves of Nothofagus, but the flora also includes conifers, cycads, monocots (such as Ripogonum and palms), together with Lauraceae, Myrtaceae and Araliaceae leaves and flowers. The small maar lakes were surrounded by Nothofagus/podocarp/mixed broadleaf forest growing under humid, warm temperate to subtropical conditions. The fossil fauna comprises insects in the orders Odonata, Hemiptera, Thysanoptera, Coleoptera, Diptera, Hymenoptera and Trichoptera, and the fish assemblage includes a non-migratory species of the Southern Hemisphere Galaxias (Galaxiidae) and a significant new record of the freshwater eel Anguilla (Anguillidae). The fossil assemblage also includes the first pre-Quaternary bird feathers from New Zealand and abundant coprolites derived from fish and volant birds, presumably waterfowl. Palynomorph analysis and a 40Ar/39Ar age of 14.6 Ma obtained from basanite associated with the maar complex indicate that the Hindon Maar Complex is of mid-Miocene age (Langhian; New Zealand local stage: Lillburnian). It thus provides a new and unique perspective on Neogene terrestrial biodiversity and biogeography in the Australasian region, around the end of the mid-Miocene thermal optimum and prior to late Miocene–Pleistocene climate cooling episodes when many warm-temperate and subtropical forest components became extinct in New Zealand

Miocene Fossils Reveal Ancient Roots for New Zealand’s Endemic Mystacina (Chiroptera) and Its Rainforest Habitat

This is an open access article distributed under the terms of the Creative Commons Attribution License , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.The New Zealand endemic bat family Mystacinidae comprises just two Recent species referred to a single genus, Mystacina. The family was once more diverse and widespread, with an additional six extinct taxa recorded from Australia and New Zealand. Here, a new mystacinid is described from the early Miocene (19–16 Ma) St Bathans Fauna of Central Otago, South Island, New Zealand. It is the first pre-Pleistocene record of the modern genus and it extends the evolutionary history of Mystacina back at least 16 million years. Extant Mystacina species occupy old-growth rainforest and are semi-terrestrial with an exceptionally broad omnivorous diet. The majority of the plants inhabited, pollinated, dispersed or eaten by modern Mystacina were well-established in southern New Zealand in the early Miocene, based on the fossil record from sites at or near where the bat fossils are found. Similarly, many of the arthropod prey of living Mystacina are recorded as fossils in the same area. Although none of the Miocene plant and arthropod species is extant, most are closely related to modern taxa, demonstrating potentially long-standing ecological associations with Mystacina

Hurricanes overcome migration lag and shape intraspecific genetic variation beyond a poleward mangrove range limit

Expansion of many tree species lags behind climate‐change projections. Extreme storms can rapidly overcome this lag, especially for coastal species, but how will storm‐driven expansion shape intraspecific genetic variation? Do storms provide recruits only from the nearest sources, or from more distant sources? Answers to these questions have ecological and evolutionary implications, but empirical evidence is absent from the literature. In 2017, Hurricane Irma provided an opportunity to address this knowledge gap at the northern range limit of the neotropical black mangrove (Avicennia germinans ) on the Atlantic coast of Florida, USA. We observed massive post‐hurricane increases in beach‐stranded A. germinans propagules at, and past, this species’ present‐day range margin when compared to a previously‐surveyed, non‐hurricane year. Yet, propagule dispersal does not guarantee subsequent establishment and reproductive success (i.e., effective dispersal). We also evaluated prior effective dispersal along this coastline with isolated A. germinans trees identified beyond the most northern established population. We used 12 nuclear microsatellite loci to genotype 896 hurricane‐driven drift propagules from nine sites and 10 isolated trees from four sites, determined their sources of origin, and estimated dispersal distances. Almost all drift propagules and all isolated trees came from the nearest sources. This research suggests that hurricanes are a prerequisite for poleward range expansion of a coastal tree species and that storms can shape the expanding gene pool by providing almost exclusively range‐margin genotypes. These insights and empirical estimates of hurricane‐driven dispersal distances should improve our ability to forecast distributional shifts of coastal species

The Vehicle, Spring 1985

Vol. 26, No. 2 Table of Contents Beyond the FieldsKeila Tooleypage 3 Lonely Sculptor Accustomed to Living AloneMichelle Mitchellpage 4 Mona LisaBob Zordanipage 4 Poet Born in Pearl HarborAngelique Jenningspage 5 IntroductionsGraham Lewispage 6 Living InsideJennifer Soulepage 9 PictureKathy Greypage 10 Salvadore Dali in a Wheelchair on TVAngelique Jenningspage 11 Sonata in E FlatBecky Lawsonpage 12 Myopia and Wild KingdomMichelle Mitchellpage 12 On Becoming a GrandmotherKeila Tooleypage 13 A VisionJennifer D. Pringlepage 14 The Covered BridgeDebbie Woodleypage 14 Jacob\u27s LifeJoan Sebastianpage 15 ForgotGraham Lewispage 15 A Dozen and One TrainsongsAngelique Jenningspage 16 Women\u27s PlaceJennifer Soulepage 19 Night SailingKim Dumentatpage 20 She Isn\u27t There WhenMichelle Mitchellpage 20 A Case for the Common ColdMaggie Kennedypage 21 the cityTammy Batespage 22 The RattlesnakeEric S. McGeepage 22 New PictureKeila Tooleypage 23 Lewis and SinGraham Lewispage 24 Funny BarbecueBob Zordanipage 26 In a DreamF. Link Rapierpage 26 The Winter\u27s ColdJennifer Soulepage 27 Diary EntryTammy Batespage 27 Minor God and Patron Saint of Rabbits SpeaksAngelique Jenningspage 28 A MomentBrett Wilhelmpage 29 The Bishop SeatF. Link Rapierpage 30 The Thought of Being Rid of MyselfKeila Tooleypage 33 I Saw A ChildBea Cessnapage 33 Complacent gourmetGary Burrowspage 34 Night DreamsJennifer Soulepage 35 Changing ImagesAmy Callpage 35 Olsen Rug Co. Waterfall & ParkMaggie Kennedypage 36 Edge of the WildF. Link Rapierpage 37 DragonS. Hillpage 37 Harvests of CornBob Zordanipage 38 The Club JeromeGary Burrowspage 39 Tarzan And The CabPatrick Peterspage 39 The Rain That Never CameLynanne Feilenpage 40 Wonderment of the Far CrescentF. Link Rapierpage 40https://thekeep.eiu.edu/vehicle/1047/thumbnail.jp

Priorities for HIV and chronic pain research results from a survey of individuals with lived experience

The Global Task Force on Chronic Pain in HIV published seven research priorities in the field of HIV-associated chronic pain in 2019: (1) causes; (2) management; (3) treatment individualization and integration with addiction treatment; (4) mental and social health factors; (5) prevalence; (6) treatment cost effectiveness; and (7) prevention. The current study used a web-based survey to determine whether the research topics were aligned with the priorities of adults with lived experiences of HIV and chronic pain. We also collected information about respondents' own pain and treatment experiences. We received 311 survey responses from mostly US-based respondents. Most respondents reported longstanding, moderate to severe, multisite pain, commonly accompanied by symptoms of anxiety and/or depression. The median number of pain treatments tried was 10 (IQR = 8, 13), with medications and exercise being the most common modalities, and opioids being viewed as the most helpful. Over 80% of respondents considered all research topics either "extremely important" or "very important". Research topic #2, which focused on optimizing management of pain in people with HIV, was accorded the greatest importance by respondents. These findings suggest good alignment between the priorities of researchers and US-based people with lived experience of HIV-associated chronic pain.</p

The Vehicle, Spring 1984

Vol. 25, No. 2 Table of Contents The OpeningKathy Fordpage 3 PaintThomas B. Waltrippage 3 PhotographLisa Fresepage 4 City as Castigating AnimalJerry McAnultypage 5 LimitsMichelle Mitchellpage 6 In SilenceKeila Tooleypage 6 PhotographLisa Fresepage 7 Indian CreekJerry McAnultypage 8 The Right Idea about RainMichelle Mitchellpage 8 Van Gogh\u27s Young Male PeasantKeila Tooleypage 9 AbigailMaggie Kennedypage 9 CleaningJennifer K. Soulepage 10 Heat WaveJerry McAnultypage 11 VigilJanet Gracepage 11 Hatred of ClownsMichelle Mitchellpage 12 Trees in the YardKeila Tooleypage 13 Lost on Back RoadsToby Mirabellipage 13 The Death of My Father\u27s HandsGina Grillopage 14 Soft LandingMaggie Kennedypage 14 Incident in the Generic AisleMaggie Kennedypage 15 Happy Well-adjusted Individuals, ConsideringJennifer K. Soulepage 16 Old ManGina Grillopage 17 It Doesn\u27t MatterTed Boonepage 18 A Love Poem for Charles DonaldCathy Moepage 18 David Willey at Whitman WharfSue Oiler Millerpage 19 I Am the ArtsBrenda Renee Drummerpage 20 Mothers and DaughtersJennifer K. Soulepage 22 ScratchesMichelle Mitchellpage 26 SedativeStacey Flanniganpage 26 Calloused HandJanet Gracepage 27 Cool CatStacey Flanniganpage 27 For My History Teacher, Miss WilhelmCathy Moepage 28 Short Poem for Chris\u27s EyesBob Zordanipage 28 MissingMichelle Mitchellpage 29 Over ThereGraham Lewispage 30 Strange BedfellowsMaggie Kennedypage 31 Retired PlumberMichelle Mitchellpage 32 Rumors of War DeadKathy Fordpage 33 Night LightsKeila Tooleypage 33 Fish on the Train TracksJerry McAnultypage 34 DaddyKeily Tooleypage 34 A Message to the MessengerAmy Eadespage 35 Dancing AloneKathy Bowerpage 36 HandsMaggie Kennedypage 36 The Key to the Ladies\u27s RoomMichelle Mitchellpage 37 Crashing the BlueStacey Flanniganpage 37 The Cradle will FallMichelle Mitchellpate 38 One IslandKeila Tooleypage 38 Black Walnut PaintSue Oiler Millerpage 39 SerendipityGraham Lewispage 40 Cover ArtLisa Fresehttps://thekeep.eiu.edu/vehicle/1045/thumbnail.jp

Human Tyrosine Hydroxylase Natural Allelic Variation: Influence on Autonomic Function and Hypertension

The catecholamine biosynthetic pathway consists of several enzymatic steps in series, beginning with the amino acids phenylalanine and tyrosine, and eventuating in the catecholamines norepinephrine (noradrenaline) and epinephrine (adrenaline). Since the enzyme tyrosine hydroxylase (TH; tyrosine 3-mono-oxygenase; EC 1.14.16.2; chromosome 11p15.5) is generally considered to be rate-limiting in this pathway, probed as to whether common genetic variation at the TH gene occurred, and whether such variants contributed to inter-individual alterations in autonomic function, either biochemical or physiological. We began with sequencing a tetranucleotide (TCAT) repeat in the first intron, and found that the two most common versions, (TCAT)6 and (TCAT)10i, predicted heritable autonomic traits in twin pairs. We then conducted systematic polymorphism discovery across the ~8 kbp locus, and discovered numerous variants, principally non-coding. The proximal promoter block contained four common variants, and its haplotypes and SNPs (especially C-824T, rs10770141) predicted catecholamine secretion, environmental stress-induced BP increments, and hypertension. Finally, we found that two of the common promoter variants, C-824T (rs10770141) and A-581G (rs10770140), were functional in that they differentially affected transcriptional activity of the isolated promoter, disrupted recognition motifs for specific transcription factor binding, altered the promoter responses to the co-transfected (exogenous) factors, and bound the endogenous factors in the chromatin fraction of the nucleus. We concluded that common variation in the proximal TH promoter is functional, giving rise to changes in autonomic function and consequently cardiovascular risk