PRELIMINARY ESTIMATES OF THE DOPAMINE β-HYDROXYLASE CONTENT AND ACTIVITY IN PURIFIED NORADRENERGIC VESICLES1

The dopamine β-hydroxylase (DβH) content and activity of large dense-core noradrenergic vesicles purified from bovine splenic nerve were determined using two assay procedures : enzymic activity expressed in Units per mg protein and homospecific activity based on radioimmunoassay expressed in Units per mg DβH antigen. Approximately two-thirds of the total enzyme activity is latent in these vesicles, even after various treatments designed to compromise vesicle integrity. DβH can be completely unmasked by brief treatment with 0.01-0.05% Triton X-100 and activity increases from 0.20 to 0.64 Units per mg vesicle protein. Calculations based on both assay methods suggested that an average of 7% (range 3-15%) of the total vesicle protein was DβH and that the average vesicle contained about 4 molecules of enzyme (range 2-9 molecules). The estimated homospecific activities indicated an average of 25 and 50% (range 18-72%) of the vesicle enzyme was inactive in the various samples using the two antibodies. The vesicle can synthesize up to 30 molecules of noradrenaline/s per molecule of DβH at near optimal substrate concentration, and 60-270 molecules of norepinephrine/s per vesicle. The assumptions used in the various calculations were critically analyzed and, based on the methods employed, it is tentatively considered to be unlikely that there could be more than 5-12 molecules of DβH per vesicle. The possibility that circulating DβH originates primarily, if not exclusively, from the large dense-core vesicle type is considered and the functional implications of the data support the concept of vesicle reuse during several cycles of exocytosis involving a quantal size equal to a fraction of the vesicle transmitter content.     

Intra- and interspecific chromosome polymorphisms in cultivated Cichorium L. species (Asteraceae)

Endive (Cichorium endivia L.) and chicory (C. intybus L.) both have 2n = 18, but until now, there has been no detailed karyomorphological characterization. The present work evaluated five accessions of each species using FISH with rDNA probes and fluorochrome staining with CMA and DAPI. Both species presented distinct banding patterns after fluorochrome staining: while endive had proximal CMA⁺⁺/DAPI⁻ bands in the short arms of pairs 1, 2 and 3, chicory had proximal CMA-positive bands in chromosomes 1 and 3 and interstitial in the short arm of chromosome 8. Among endive accessions, FISH procedures revealed conserved position and number of 5S and 45S rDNA sites (two and three pairs, respectively), associated with the CMA-positive bands. Notwithstanding, polymorphisms were detected within chicory accessions regarding the number and the distribution of rDNA sites in relation to the most frequent karyotype (two pairs with 45S and one with 5S rDNA). The karyological markers developed allowed karyotypic differentiation between both species, uncovering peculiarities in the number and position of rDNA sites, which suggest chromosome rearrangements, such as translocations in chicory cultivars. The interspecific and intraspecific polymorphisms observed emphasize the potential of karyomorphological evaluations, helping our understanding of the relationships and evolution of the group.     

Distinct Leaf‐trait Syndromes of Evergreen and Deciduous Trees in a Seasonally Dry Tropical Forest

In seasonally dry tropical forests, tree species can be deciduous, remaining without leaves throughout the dry season, or evergreen, retaining their leaves throughout the dry season. Deciduous and evergreen trees specialize in habitats that differ in water availability (hillside and riparian forest, respectively) and in their exposure to herbivore attack (seasonal and continuous, respectively). We asked whether syndromes of leaf traits in deciduous and evergreen trees were consistent with hypothesized abiotic and biotic selective pressures in their respective habitat. We measured seven leaf traits in 19 deciduous and 11 evergreen tree species in a dry tropical forest in Western Mexico, and measured rates of herbivory on 23 of these species. We investigated the covariance of leaf traits in syndromes related to phenology and associated physiology, and to anti‐herbivory defense. We found evidence for syndromes that separated phenological strategies among four traits: toughness, water content, specific leaf area, and carbon:nitrogen (C:N) ratios. We found a trade‐off between two other traits: trichomes and latex. Overall, evergreen species exhibited lower rates of herbivory than deciduous species. Lower rates of herbivory were explained by a syndrome of higher toughness, lower water content, and higher C:N ratios, which are traits representative of evergreen trees. Phenology and trait syndromes did not exhibit significant phylogenetic signal, consistent with the hypothesis of evolutionary convergence among phenologies and associated leaf‐trait syndromes. Our results suggest that deciduous and evergreen trees could respond to differential water availability and herbivory in their respective habitats by converging on distinct leaf‐trait syndromes. Abstract in Spanish is available at http://www.blackwell‐synergy.com/loi/btp .     

Seasonal changes in mixed‐species bird flocks and antipredator information

Animals acquire information produced by other species to reduce uncertainty and avoid predators. Mixed‐species flocks (MSFs) of birds are ubiquitous in forest ecosystems and structured, in part, around interspecific information transfer, with “nuclear” species providing information that other species eavesdrop on. We hypothesized that in a seasonal tropical forest, the amount of information produced by birds about predation would be dynamic and particularly would decrease inside MSFs when the nuclear species leave MSFs to breed. We obtained baseline information on MSF encounter rate and species composition along established sampling routes over 9 months near the Sino‐Vietnamese border. We also conducted three experiments to quantify information produced by different species in response to typical predator encounters, including a moving predator stimulus presented inside of MSFs, and a stationary predator model presented both inside and outside of MSFs. MSFs were much less frequent in the breeding season with fewer individuals of the nuclear species, David's Fulvetta (Alcippe davidi), participating, though the diversity of other species remained stable. Fulvettas were the dominant producer of alarm‐related information both to the moving and stationary stimuli in MSFs and were also among the most active mobbers to stimuli presented outside of MSFs. In the breeding season, they tended to call less to the moving stimulus, and substantially fewer individuals responded to the in‐flock stationary stimulus. Other species increased their own information production at stationary predator stimuli (inside and outside of MSFs) during the breeding season, perhaps due to their increased investment in offspring during this time. Yet even during the breeding season, David's Fulvetta remained the highest producer of information about predators in MSFs. Hence, while we show that information production in MSFs can be somewhat dynamic, we describe a continually asymmetric communication system, in which a nuclear species is important to the whole community.     

Aromatic residue position on the nonpolar face of class a amphipathic helical peptides determines biological activity

The apolipoprotein A-I mimetic peptide 4F (Ac-DWFKAFYDKVAEKFKEAF-NH(2)), with four Phe residues on the nonpolar face of the amphipathic alpha-helix, is strongly anti-inflammatory, whereas two 3F analogs (3F(3) and 3F(14)) are not. To understand how changes in helix nonpolar face structure affect function, two additional 3F analogs, Ac-DKLKAFYDKVFEWAKEAF-NH(2) (3F-1) and Ac-DKWKAVYDKFAEAFKEFL-NH(2) (3F-2), were designed using the same amino acid composition as 3F(3) and 3F(14). The aromatic residues in 3F-1 and 3F-2 are near the polar-nonpolar interface and at the center of the nonpolar face of the helix, respectively. Like 4F, but in contrast to 3F(3) and 3F(14), these peptides effectively inhibited lytic peptide-induced hemolysis, oxidized phospholipid-induced monocyte chemotaxis, and scavenged lipid hydroperoxides from low density lipoprotein. High pressure liquid chromatography retention times and monolayer exclusion pressures indicated that there is no direct correlation of peptide function with lipid affinity. Fluorescence studies suggested that, although the peptides bind phospholipids similarly, the Trp residue in 4F, 3F-1, and 3F-2 is less motionally restricted than in 3F(3) and 3F(14). Based on these results and molecular modeling studies, we propose that the arrangement of aromatic residues in class A amphipathic helical molecules regulates entry of reactive oxygen species into peptide-phospholipid complexes, thereby reducing the extent of monocyte chemotaxis, an important step in atherosclerosis.     

Floral biology,reciprocal herkogamy and breeding system in four Psychotria species (Rubiaceae) in Brazil

Distylous species have two floral morphs with reciprocal positions of sexual whorls, functioning to promote cross‐pollination. Additionally, most distylous species have an incompatibility system linked to the discrete variation of anthers and stigmas. Here we provide new data on heterostyly, reproductive biology and pollination in four Psychotria spp. from the Cerrado Region (Brazil). Psychotria deflexa, P. nitidula and P. trichophoroides fitted the distylous morphological syndrome, whereas P. prunifolia was monomorphic and self‐compatible. Reciprocity varied across species, with the upper whorl of anthers and stigmas being more reciprocal than the lower whorl. Psychotria nitidula has a heteromorphic incompatibility system, whereas P. deflexa and P. trichophoroides showed partial self‐compatibility. Psychotria prunifolia and P. trichophoroides were visited by large bees and exhibited lower reproductive efficacy than P. nitidula and P. deflexa, which are visited by wasps and small bees. Our results provide new evidence that the morphological distylous syndrome does not always co‐occur with heteromorphic incompatibility and additional mechanisms (e.g. disassortative pollen transfer) may operate to maintain the polymorphism and isoplethy of species with different incompatibility system.     

Hawaiian waterbird movement across a developed landscape

A key component for biologists managing mobile species is understanding where and when a species occurs at different locations and scaling management to fit the spatial and temporal patterns of movement. We established an automated radio-telemetry tracking network to document multi-year movement in 2016–2018 of 3 endangered waterbirds among wetlands on Oʻahu, Hawaiʻi, USA: ʻalae ʻula or Hawaiian gallinule (gallinule; Gallinula galeata sandvicensis), ʻalae keʻokeʻo or Hawaiian coot (coot; Fulica alai), and aeʻo or Hawaiian stilt (stilt; Himantopus mexicanus knudseni), each with different ecological requirements. There were marked differences in the movement propensity of the species, with no movement among sites detected in gallinules, 31% of coots moving among wetlands, and very high levels of daily movement in stilts. A network analysis revealed strong evidence for fidelity among individual stilts to specific wetlands, indicating different groups of wetlands supported different birds. There was also strong evidence for patterns in daily and seasonal movement patterns of stilts. Our work indicates the importance of each wetland to the waterbirds they support, as each individual had strong fidelity to a single wetland. In addition, for Hawaiian coots and stilts, which were documented moving among multiple wetlands, a network of wetlands may be key for long-term persistence of these endangered species, and coordinated regional management of waterbirds as a shared resource could provide greater benefits to waterbirds than independent management of each wetland.     

Tremorgenic Mycotoxins from Aspergillus Caespitosus

Two tremorgenic mycotoxins were isolated from Aspergillus caespitosus, and identified as verruculogen and fumitremorgin B. They were produced at the rate of 172 and 325 mg per kg, respectively, on autoclaved cracked field corn.     

Tremorgenic Toxin from Penicillium verruculosum

A new mycotoxin that produces severe tremors and acute toxicity when administered orally or intraperitoneally (ip) to mice and 1-day-old cockerels was obtained from a strain of Penicillium verruculosum Peyronel isolated from peanuts. The ip 50% lethal dose (LD(50)) of this tremorgen was 2.4 mg/kg in mice and 15.2 mg/kg in chickens. Orally administered LD(50) values for the toxin were 126.7 mg/kg in mice and 365.5 mg/kg in chickens. The trivial name "verruculogen" is proposed for this tremorgenic mycotoxin. Physical and chemical characteristics of the mycotoxin are described.