Differential regulation of dynein and kinesin motor proteins by tau

Dynein and kinesin motor proteins transport cellular cargoes toward opposite ends of microtubule tracks. In neurons, microtubules are abundantly decorated with microtubule-associated proteins (MAPs) such as tau. Motor proteins thus encounter MAPs frequently along their path. To determine the effects of tau on dynein and kinesin motility, we conducted single-molecule studies of motor proteins moving along tau-decorated microtubules. Dynein tended to reverse direction, whereas kinesin tended to detach at patches of bound tau. Kinesin was inhibited at about a tenth of the tau concentration that inhibited dynein, and the microtubule-binding domain of tau was sufficient to inhibit motor activity. The differential modulation of dynein and kinesin motility suggests that MAPs can spatially regulate the balance of microtubule-dependent axonal transport.     

Parent volatiles in comet 9P/Tempel 1: before and after impact

We quantified eight parent volatiles (H2O, C2H6, HCN, CO, CH3OH, H2CO, C2H2, and CH4) in the Jupiter-family comet Tempel 1 using high-dispersion infrared spectroscopy in the wavelength range 2.8 to 5.0 micrometers. The abundance ratio for ethane was significantly higher after impact, whereas those for methanol and hydrogen cyanide were unchanged. The abundance ratios in the ejecta are similar to those for most Oort cloud comets, but methanol and acetylene are lower in Tempel 1 by a factor of about 2. These results suggest that the volatile ices in Tempel 1 and in most Oort cloud comets originated in a common region of the protoplanetary disk.     

Research in plant genetic resources 1978–2003: General index of the second 25 volumes 'Kulturpflanze'/'GRACE'

A general index of the second 25 volumes of 'Kulturpflanze' 1978–1990/'GRACE' (Genetic Resources and Crop Evolution) 1992–2003 is provided. Its predecessor covered the first 25 volumes of the periodical 'Die Kulturpflanze' and appeared in 1977. The index can be considered as a unique source for research in plant genetic resources.     

Probabilistic patterns of inundation and biogeomorphic changes due to sea-level rise along the northeastern U.S. Atlantic coast

Context

Coastal landscapes evolve in response to sea-level rise (SLR) through a variety of geologic processes and ecological feedbacks. When the SLR rate surpasses the rate at which these processes build elevation and drive lateral migration, inundation is likely.

Objectives

To examine the role of land cover diversity and composition in landscape response to SLR across the northeastern United States.

Methods

Using an existing probabilistic framework, we quantify the probability of inundation, a measure of vulnerability, under different SLR scenarios on the coastal landscape. Resistant areas—wherein a dynamic response is anticipated—are defined as unlikely (p < 0.33) to inundate. Results are assessed regionally for different land cover types and at 26 sites representing varying levels of land cover diversity.

Results

Modeling results suggest that by the 2050s, 44% of low-lying, habitable land in the region is unlikely to inundate, further declining to 36% by the 2080s. In addition to a decrease in SLR resistance with time, these results show an increasing uncertainty that the coastal landscape will continue to evolve in response to SLR as it has in the past. We also find that resistance to SLR is correlated with land cover composition, wherein sites containing land cover types adaptable to SLR impacts show greater potential to undergo biogeomorphic state shifts rather than inundating with time.

Conclusions

Our findings support other studies that have highlighted the importance of ecological composition and diversity in stabilizing the physical landscape and suggest that flexible planning strategies, such as adaptive management, are particularly well suited for SLR preparation in diverse coastal settings.

     

Weak connectivity and population cohesiveness in rudderfish Kyphosus sandwicensis (Teleostei: Kyphosidae) inhabiting remote oceanic islands

1. Population connectivity has a fundamental role in metapopulation dynamics, with important implications in conservation. Easter Island (EI) and Salas y Gómez Island (SG) in the Pacific Ocean are ideal for the study of population connectivity because they are separated by 415 km and isolated from other islands in the Pacific Ocean by >2,000 km.
2. Considering that dispersal processes could play a critical role in the persistence of its populations, the connectivity pattern of the rudderfish Kyphosus sandwicensis was evaluated between EI and SG using both a population genetics and a biophysical modelling approach.
3. The variability in the control region of the mitochondrial DNA did not show a significant phylogeographical pattern, and the variability in 16 microsatellite loci suggested that individuals of K. sandwicensis located at EI and SG belong to the same genetic population. However, historical migration showed that 0.2% of the recruits at EI come from SG and that 0.15% at SG come from EI per year.
4. Using simulated larval release during September and a larval development of 30 days in the plankton, biophysical modelling did not detect migration between the islands. Furthermore, self-recruitment shows interannual variation ranging from 5 to 10% of the total released larvae.
5. Whereas the genetic data showed a lack of population genetic structure but low connectivity of K. sandwicensis between EI and SG, the biophysical modelling showed null movement of particles between the islands. Stochastic movement of larvae or adults could explain the pattern observed, with rafting as an example. These low-frequency and stochastic movements may be important in maintaining the cohesiveness between EI and SG.

     

Tracing trophic pathways through the marine ecosystem of Rapa Nui (Easter Island)

1. The structure of food webs provides important insight into biodiversity, organic matter (OM) pathways, and ecosystem functioning.
2. Stable isotope analysis (δ¹³C and δ¹⁵N) was used to characterize the trophic structure and the main OM pathways supporting food webs in the Rapa Nui coastal marine ecosystem.
3. The trophic position of consumers and isotopic niche metrics were estimated for different assemblages (i.e. mesozooplankton, emergent zooplankton, reef invertebrates, reef fishes, pelagic fishes, and seabirds). Furthermore, the relative importance of different OM sources (i.e. macroalgae, zooxanthellate corals, and particulate OM [POM]) was assessed for heterotrophic consumers using Bayesian mixing model (MixSIAR).
4. Results show a clear pattern of ¹³C and ¹⁵N enrichment from small-sized pelagic and benthic invertebrates, to reef and pelagic fishes, and seabirds. Most invertebrates were classified as primary consumers, reef fishes as secondary consumers and pelagic predators and seabirds as tertiary and quaternary consumers.
5. Isotopic niche metrics indicate a low trophic diversity for pelagic assemblages (mesozooplankton and pelagic fishes), in contrast to reef fauna (invertebrates and fishes), whose higher trophic diversity suggest the exploitation of a wider range of trophic resources. Overlapping of standard ellipses areas between reef invertebrates and reef fishes indicates that both assemblages could be sharing trophic resources.
6. Mixing models results indicate that POM is the main trophic pathway for mesozooplankton, macroalgae (Rhodophyta) for emergent zooplankton, and a mix of coral-derived OM and Rhodophyta for coral reef assemblages such as macrobenthos and reef invertebrates. In contrast, POM contribution was notably more important for some pelagic fishes and seabirds from upper trophic levels.
7. This study provides key elements for conservation efforts on coral reefs, management planning and full-implementation of the recently created Rapa Nui Multiple Use Marine Protected Area.

     

Insulin binding to isolated hepatocytes of Atlantic salmon and rainbow trout

The questions addressed in this study were: 1) whether insulin added to the incubation medium can down-regulate ¹²⁵I insulin binding to isolated hepatocytes of Atlantic salmon (Salmo salar) and rainbow trout (Oncorhynchus mykiss); 2) whether quantitative assessment of insulin processing can be made on isolated fish liver cells; 3) how ambient temperatures can affect insulin binding, and down-regulation of insulin receptors. After isolation and a short (up to 4h) “metabolic recovery period”, liver cells were used either directly in ¹²⁵I insulin binding assay or first preincubated for 18h at 4°C or for 3h at 15°C, with or without mammalian or salmon insulin in concentrations ranging from 1 to 1000 nM. Preincubation at 15°C, decreased binding capacity (number of binding sites per liver cell) in all five independent hepatocyte preparations treated with 1000 nM insulin and in four out of five preparations treated with 100 nM insulin. At 4°C insulin binding sites were down-regulated in less than 50% of all hepatocyte preparations and only in the presence of 1000 nM insulin. Differential quantitive assessment was made of a) intact free insulin; b) insulin degraded; c) intact insulin bound to the cell membrane; d) internalized but degraded insulin, and e) intact insulin internalized by liver cells. Hepatocytes preincubated with 100 – 1000 nM insulin at 15°C bound and internalized less ¹²⁵I insulin. We hypothesize that in vivo, at water temperatures of 15°C and higher, extreme physiological levels of plasma insulin may regulate the numbers of insulin receptors in the salmonid liver. In contrast, in fish inhabiting cold waters the regulation of insulin receptors by circulating plasma insulin seems to be of little physiological importance. Presented in part at the Western Regional Conference on Comparative Endrocrinology, Tempe, Arizona, U.S.A., 1991 and at the Meeting of Italian Society of Experimental Biology, Sorrento, Italy, 1991. Supported by grants from NSF of the USA#DCB 8915935 to E.M.P., NSERC of Canada OPGA 6944 to T.W.M., North Atlantic Treaty Organization (NATO) grant #0926/87 to C.O. and E.M.P., and CYCIT grant of Spain to J.G.     

Insulin-like growth factor I (IGF-I) mRNA expression in coho salmon,Oncorhynchus kisutch: Tissue distribution and effects of growth hormone/prolactin family proteins

To examine the hormonal and nutritional regulation of insulin-like growth factor I (IGF-I) mRNA expression, a sequence-specific solution hybridization/RNase protection assay for coho salmon IGF-I mRNA was developed. This assay is both rapid and sensitive and has low inter- (less than 15%) and intra-assay variations (less than 5%). Using this assay, the tissue distribution of IGF-I mRNA and effects of growth hormone (GH), prolactin (PRL) and somatolactin (SL) on hepatic IGF-I mRNA expression in coho salmon were examined in vivo. Liver had the highest IGF-I mRNA level of 16 pg/μg DNA. Significant amounts of IGF-I mRNA were also found in all other tissues examined (intestine 4.1, kidney 3.8, gill arch 2.4, brain 2.4, ovary 2.3, muscle 2.1, spleen 1.7 and fat 1.1 pg/μg DNA). Injection of coho salmon GH at doses of 0.1 and 1 μg/g body weight significantly increased the hepatic IGF-I mRNA levels in a dose-dependent manner. Injection of coho salmon SL, a recently discovered member of the GH/PRL family, stimulated the IGF-I mRNA expression at the higher dose (1 μg/g), whereas coho salmon PRL had no effect at either dose. Concentration-dependent stimulation by coho salmon GH was also obtained in vitro in primary culture of salmon hepatocytes in concentrations ranging from 0.01 to 1 μg/ml. These results indicate that IGF-I mRNA expression occurs in a variety of tissues in coho salmon, and that at least the hepatic expression is under the regulation of GH and possibly other hormones. The sequence-specific assay established in the present study can be used for accurate quantitation of IGF-I mRNA in salmonid species, and can contribute to a better understanding of the physiology of IGF-I in salmonids.

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Résumé Afin d'étudier les régulations homronales et nutritionnelles de l'expression des ARNm de l'IGF-I (insulin-like growth factor I), un dosage spécifique par hybridation en solution des ARNm d'IGF-I de saumon coho et protégé des RNases, a été développé. Ce dosage, à la fois rapide et sensible, présente un faible coefficient de variation inter- (< 15%) et intra- (< 5%) dosage. L'étude de la distribution tissulaire des ARNm de l'IGF-I et des effets de l'hormone de croissance (GH), de la prolactine (Prl) et de la somatolactine (SI) sur l'expression hépatique des ARNm de l'IGF-I, a été entreprise in vivo chez le saumon coho en utilisant ce dosage. Le foie présente les plus grandes quantités d'ARNm d'IGF-I (16 pg/μg d'ADN). Des quantités significatives d'ARNm d'IGF-I ont été également détectées dans tous les autres tissus étudiés (intestin 4,1; rein 3,8; branchie 2,4; ovaire 2,3; muscle 2,1; rate 1,7 et graisse 1,1 pg/μg d'ADN). L'injection à des saumons coho, de GH à des doses de 0,1 et 1 μg/g de poids vif, augmente significativement et de manière dose dépendante les niveaux hépatiques d'ARNm d'IGF-I. L'injection de SI de saumon coho, un membre récemment découvert de la famille GH/Prl, stimule avec la plus haute dose utilisée, l'expression des ARNm d'IGF-I alors que la Prl n'a aucun effet. La GH augmente de manière dose dépendante (0,01–1 μg/ml) l'expression in vitro des ARNm d'IGF-I par des ARNm d'IGF-I par des hépatocytes de saumon coho en culture. Ces résultats indiquent que, chez le saumon coho, l'expression des ARNm d'IGF-I est présente dans le nombreaux tissus et que, l'expression hépatique est, au moins en partie, régulée par la GH et peut-être par d'autres hormones. Le dosage par séquence spécifique mise au point dans le présent travail, peut-être utilisé pour la quantification précise des ARNm, d'IGF-I de salmonidés et devrait permettre une meilleure connaissance de la physiologie de L'IGF-I chez les salmonidés.

     

Subsoil compaction by heavy sugarbeet harvesters in southern Sweden: III. Risk assessment using a soil water model

Due to its persistence, subsoil compaction should be avoided, which can be done by setting stress limits depending on the strength of the soil. Such limits must take into account soil moisture status at the time of traffic. The objective of the work presented here was to measure soil water changes during the growing period, use the data to calibrate a soil water model and simulate the soil susceptibility to compaction using meteorological data for a 25-year period. Measurements of soil water content were made in sugarbeet (Beta vulgaris L.) from sowing until harvest in 1997 on two sites classified as Eutric Cambisols in southern Sweden. Sampling was carried out at 2-week intervals in 0.1 m layers down to 1 m depth, together with measurements of root growth and crop development. Precompression stress of the soil at 0.3, 0.5 and 0.7 m depth was determined from uniaxial compression tests at water tensions of 6, 30, 60 and 150 kPa and adjusted as a logarithmic function of the soil water tension. Soil water content was simulated by the SOIL model for the years 1963–1988. Risk calculations were made for a wheel load of 8 t and a ground pressure of 220 kPa, corresponding to a fully loaded six-row sugarbeet harvester. Subsoil compaction was expected to occur when the major principal stress was higher than the precompression stress. The subsoil water content was very low in late summer, but increased during the autumn. At the end of August, there was practically no plant available water down to 1 m depth. There was in general good agreement between measured and simulated values of soil water content for the subsoil, but not for the topsoil. In the 25-year simulations, the compaction risk at 50 cm depth was estimated to increase from around 25% to nearly 100% between September and late November, which is the period when the sugarbeet are harvested. The types of simulation presented here may be a very useful tool for practical agriculture as well as for society, in giving recommendations as to how subsoil compaction should be avoided.     

Reliable detection and identification of genetically modified maize, soybean, and canola by multiplex PCR analysis

Multiplex PCR procedures were developed for simultaneously detecting multiple target sequences in genetically modified (GM) soybean (Roundup Ready), maize (event 176, Bt11, Mon810, T14/25), and canola (GT73, HCN92/28, MS8/RF3, Oxy 235). Internal control targets (invertase gene in corn, lectin and beta-actin genes in soybean, and cruciferin gene in canola) were included as appropriate to assess the efficiency of all reactions, thereby eliminating any false negatives. Primer combinations that allowed the identification of specific lines were used. In one system of identification, simultaneous amplification profiling (SAP), rather than target specific detection, was used for the identification of four GM maize lines. SAP is simple and has the potential to identify both approved and nonapproved GM lines. The template concentration was identified as a critical factor affecting efficient multiplex PCRs. In canola, 75 ng of DNA template was more effective than 50 ng of DNA for the simultaneous amplification of all targets in a reaction volume of 25 microL. Reliable identification of GM canola was achieved at a DNA concentration of 3 ng/microL, and at 0.1% for GM soybean, indicating high levels of sensitivity. Nonspecific amplification was utilized in this study as a tool for specific and reliable identification of one line of GM maize. The primer cry1A 4-3' (antisense primer) recognizes two sites on the DNA template extracted from GM transgenic maize containing event 176 (European corn borer resistant), resulting in the amplification of products of 152 bp (expected) and 485 bp (unexpected). The latter fragment was sequenced and confirmed to be Cry1A specific. The systems described herein represent simple, accurate, and sensitive GMO detection methods in which only one reaction is necessary to detect multiple GM target sequences that can be reliably used for the identification of specific lines of GMOs.