Analysis of protoplasts and somatic embryogenesis in Medicago truncatula

This thesis examined protoplast proliferation and somatic embryogenesis, by comparing a highly with a poorly embryogenic Medicago truncatula line through microscopic, proteomic and in situ hybridization analysis. Proteome analysis of M. truncatula was used to identify proteins involved in protoplast proliferation and the initiation of somatic embryogenesis. Furthermore, an in situ hybridization study was done to compare the expression of genes known to be involved in zygotic embryogenesis with the expression during somatic embryogenesis. A large number of proteins were up-, and down-regulated during the first 5 days of protoplast culture indicating that cellular reorganization took place. An up-regUlation of PR 1 O-like proteins and flavonoid synthesis proteins and a down-regulation of energy metabolism proteins were observed, indicating an initiation of a stress response. The observed stress response in protoplasts was down-regulated before the first cell divisions at 5-7 d. A stress-inducing bioassay on protoplasts showed that the ability of protoplasts to overcome stress and to proliferate under stress conditions depended on the level of stress and density of the protoplast culture, whereby more stress or a lower culture density resulted in higher levels of cell death. Proteomic analysis of the initiation of somatic embryogenesis showed that similar metabolic pathways were involved in the initiation of somatic embryogenesis and protoplast proliferation. By using a highly embryogenic (2HA) line, and a poorly embryogenic (A 17) line of M. truncatu!a, it was shown that particular proteins were specifically accumulated during the initiation of somatic embryogenesis. A high accumulation of a peroxidase was observed only in At7 tissue at the time of initiation of somatic embryogenesis and might be the reason why the initiation of somatic embryogenesis is inhibited in A 17 tissue. The specific accumulation of flavonoid synthesis proteins might also indicate that flavonoids are involved during the initiation of somatic embryogenesis. In situ hybridization with probes to genes known to be involved in zygotic embryogenesis, showed that M. truncatula somatic and Arabidopsis thaliana zygotic embryogenesis both followed similar developmental pathways. However, a few genes showed distinct patterns of gene expression in M. truncatula somatic embryos.ARC Centre of Excellence for Integrative Legume Research for funding and scholarshi

Culture Matters: French-German Conflicts on European Central Bank Independence

With the Maastricht treaty, the members of the Eurozone agreed on the establishment of a very independent European Central Bank, as well as making their National Central Banks far more independent. However, over the years French political leaders systematically brought forward proposals undermining the ECB's independence, to the dismay of Germany. A pattern that surfaced again during the current sovereign debt crisis and has complicated finding a timely and unified answer to the problems. The article conducts tests of various factors expected to influence the preference for central bank independence. It shows that economic explanations are unable to account for the persistent differences amongst European member-states on this issue. Instead, cultural differences in attitudes, especially a nation's score on the dimension of Power Distance - its acceptance of centralisation of power in a small set of political leaders or institutions - does show a correlation with the different levels of internalisation of the Central Bank independence norm.Central Bank Independence; Culture; European Central Bank; Franco- German relations

Decadal and multi-decadal variability of Labrador Sea Water in the north-western North Atlantic Ocean derived from tracer distributions: Heat budget, ventilation, and advection

Time series of profiles of potential temperature, salinity, dissolved oxygen, and planetary potential vorticity at intermediate depths in the Labrador Sea, the Irminger Sea, and the Iceland Basin have been constructed by combining the hydrographic sections crossing the sub-arctic gyre of the North Atlantic Ocean from the coast of Labrador to Europe, occupied nearly annually since 1990, and historic hydrographic data from the preceding years since 1950. The temperature data of the last 60 years mainly reflect a multi-decadal variability, with a characteristic time scale of about 50 years. With the use of a highly simplified heat budget model it was shown that this long-term temperature variability in the Labrador Sea mainly reflects the long-term variation of the net heat flux to the atmosphere. However, the analysis of the data on dissolved oxygen and planetary potential vorticity show that convective ventilation events, during which successive classes of Labrador Sea Water (LSW) are formed, occurring on decadal or shorter time scales. These convective ventilation events have performed the role of vertical mixing in the heat budget model, homogenising the properties of the intermediate layers (e.g. temperature) for significant periods of time. Both the long-term and the near-decadal temperature signals at a pressure of 1500 dbar are connected with successive deep LSW classes, emphasising the leading role of Labrador Sea convection in running the variability of the intermediate depth layers of the North Atlantic. These signals are advected to the neighbouring Irminger Sea and Iceland Basin. Advection time scales, estimated from the 60 year time series, are slightly shorter or of the same order as most earlier estimates, which were mainly based on the feature tracking of the spreading of the LSW(94) class formed in the period 1989-1994 in the Labrador Sea

Extreme variability in Irminger Sea winter heat loss revealed by ocean observatories initiative mooring and the ERA5 reanalysis

Author Posting. © American Geophysical Union, 2018. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geophysical Research Letters 46(1), (2018): 293-302, doi:10.1029/2018GL080956.Ground‐breaking measurements from the ocean observatories initiative Irminger Sea surface mooring (60°N, 39°30′W) are presented that provide the first in situ characterization of multiwinter surface heat exchange at a high latitude North Atlantic site. They reveal strong variability (December 2014 net heat loss nearly 50% greater than December 2015) due primarily to variations in frequency of intense short timescale (1–3 days) forcing. Combining the observations with the new high resolution European Centre for Medium Range Weather Forecasts Reanalysis 5 (ERA5) atmospheric reanalysis, the main source of multiwinter variability is shown to be changes in the frequency of Greenland tip jets (present on 15 days in December 2014 and 3 days in December 2015) that can result in hourly mean heat loss exceeding 800 W/m2. Furthermore, a new picture for atmospheric mode influence on Irminger Sea heat loss is developed whereby strongly positive North Atlantic Oscillation conditions favor increased losses only when not outweighed by the East Atlantic Pattern.We are grateful to Meric Srokosz and the two reviewers for helpful comments on this work. S. J. acknowledges the U.K. Natural Environment Research Council ACSIS programme funding (Ref. NE/N018044/1). M. O. acknowledges support from EU Horizon 2020 projects AtlantOS (grant 633211) and Blue Action (grant 727852). G. W. K. M. acknowledges support from the Natural Sciences and Engineering Research Council of Canada. Support for the Irminger Sea array of the ocean observatories initiative (OOI) came from the U.S. National Science Foundation. Thanks to the WHOI team and ships' officers and crew for the field deployments and to Nan Galbraith for processing the data and computing the air‐sea fluxes. Support for this processing, and making available and sharing the OOI data, came from the National Science Foundation under a Collaborative Research: Science Across Virtual Institutes grant (82164000) to R. A. W. Data used are available from the following sites: NOAA Climate Prediction Center NAO and EAP indices ftp://ftp.cpc.ncep.noaa.gov/wd52dg/data/indices/tele_index.nh, ECMWF Reanalysis 5 (ERA5) https://www.ecmwf.int/en/forecasts/datasets/archive‐datasets/reanalysis/datasets/era5, and ocean observatories initiative Irminger Mooring https://ooinet.oceanobservatories.org/.2019-06-1

Deep convection in the Irminger Sea observed with a dense mooring array

Deep convection is a key process in the Atlantic Meridional Overturning Circulation, but because it acts at small scales, it remains poorly resolved by climate models. The occurrence of deep convection depends on weak initial stratification and strong surface buoyancy forcing, conditions that are satisfied in only a few ocean basins. In 2014, one of the Ocean Observatories Initiative (OOI) global arrays was installed close to the Central Irminger Sea (CIS) and the Long-term Ocean Circulation Observations (LOCO) moorings in the central Irminger Sea. These programs’ six moorings are located in the center of an area of deep convection and are distributed within a 50 km radius, thus offering detailed insight into spatial differences during the strong convection events that occurred during the winters of 2014/2015 and 2015/2016. Deep mixed layers, down to approximately 1,600 m, formed during both winters. The properties of the convectively renewed water mass at each mooring converge to a common temperature and salinity before restratification sets in at the end of winter. The largest differences in onset (or timing) of convection and restratification are seen between the northernmost and southernmost moorings. High-resolution atmospheric reanalysis data show there is higher atmospheric forcing at the northernmost mooring due to a more favorable position with respect to the Greenland tip jet. Nevertheless, earlier onset, and more continuous cooling and deepening of mixed layers, occurs at the southernmost mooring, while convection at the northern mooring is frequently interrupted by warm events. We propose that these warm events are associated with eddies and filaments originating from the Irminger Current off the coast of Greenland and that convection further south benefits from cold inflow from the southwest

Lagrangian perspective on the origins of Denmark Strait Overflow

Author Posting. © American Meteorological Society, 2020. This article is posted here by permission of American Meteorological Society for personal use, not for redistribution. The definitive version was published in Journal of Physical Oceanography 50(8), (2020): 2393-2414, doi:10.1175/JPO-D-19-0210.1.The Denmark Strait Overflow (DSO) is an important contributor to the lower limb of the Atlantic meridional overturning circulation (AMOC). Determining DSO formation and its pathways is not only important for local oceanography but also critical to estimating the state and variability of the AMOC. Despite prior attempts to understand the DSO sources, its upstream pathways and circulation remain uncertain due to short-term (3–5 days) variability. This makes it challenging to study the DSO from observations. Given this complexity, this study maps the upstream pathways and along-pathway changes in its water properties, using Lagrangian backtracking of the DSO sources in a realistic numerical ocean simulation. The Lagrangian pathways confirm that several branches contribute to the DSO from the north such as the East Greenland Current (EGC), the separated EGC (sEGC), and the North Icelandic Jet (NIJ). Moreover, the model results reveal additional pathways from south of Iceland, which supplied over 16% of the DSO annually and over 25% of the DSO during winter of 2008, when the NAO index was positive. The southern contribution is about 34% by the end of March. The southern pathways mark a more direct route from the near-surface subpolar North Atlantic to the North Atlantic Deep Water (NADW), and needs to be explored further, with in situ observations.This work was financially supported by the U.S. National Science Foundation under Grant Numbers OAC-1835640, OCE-1633124, OCE-1433448, and OCE-1259210

Survival time and prognostic factors in canine leishmaniosis in a non-endemic country treated with a two-phase protocol including initial allopurinol monotherapy

Background: Leishmania infantum is an intracellular protozoan parasite which is endemic in countries of the Mediterranean Basin. Leishmaniosis is increasingly diagnosed in non-endemic areas due to the relocation of dogs from endemic areas and the travel of dogs to and from these areas. The prognosis of leishmaniosis in these dogs may differ from that of those in endemic areas. The aims of this study were (1) to determine the Kaplan–Meier estimated survival time for dogs with leishmaniosis in the Netherlands (a non-endemic country), (2) to determine if clinicopathological variables at the time of diagnosis predicted the survival of these dogs, and (3) to evaluate the effect of a two-phase therapy protocol of allopurinol monotherapy followed by meglumine antimoniate and/or miltefosine in the case of incomplete remission or relapse. Methods: The database of the Department of Clinical Sciences of Companion Animals of the Faculty of Veterinary Medicine, Utrecht University was investigated for leishmaniosis patients. Patient records were reviewed for signalment and clinicopathological data at the time of diagnosis. Only treatment-naive patients were included. Follow-up was performed during the study by phone contact and included treatment received and date and cause of death. Univariate analysis was performed using the Cox proportional hazards regression model. Results: The estimated median Kaplan–Meier survival time was 6.4 years. In the univariate analysis, increases in monocyte, plasma urea and creatinine concentrations, and urine protein to creatinine ratio were all significantly associated with decreased survival time. The majority of patients only received allopurinol monotherapy. Conclusions: Canine leishmaniosis patients in our study population in the Netherlands, which is non-endemic for the disease, had an estimated Kaplan–Meier median survival time of 6.4 years, which is comparable to the outcome of other reported therapy protocols. Increased plasma urea and creatinine concentrations and monocyte concentration were statistically associated with an increased risk of death. We conclude that initial allopurinol monotherapy for 3 months should be effective in more than half of canine leishmaniosis cases, provided there is adequate follow-up, and that meglumine antimoniate or miltefosine therapy should be started as the second phase of the protocol in cases where remission is incomplete or there is a relapse. Graphical Abstract: [Figure not available: see fulltext.

Rotterdam Aphasia Therapy Study (RATS) - 3: " The efficacy of intensive cognitive-linguistic therapy in the acute stage of aphasia"; design of a randomised controlled trial

Background: Aphasia is a severely disabling condition occurring in 20 to 25% of stroke patients. Most patients with aphasia due to stroke receive speech and language therapy. Methodologically sound randomised controlled trials investigating the effect of specific interventions for patients with aphasia following stroke are scarce.

Systematic Review and Meta-Analysis of Preterm Birth and Later Systolic Blood Pressure

Lower birth weight because of fetal growth restriction is associated with higher blood pressure later in life, but the extent to which preterm birth ( <37 completed weeks' gestation) or very low birth weight ( <1500 g) predicts higher blood pressure is less clear. We performed a systematic review of 27 observational studies that compared the resting or ambulatory systolic blood pressure or diagnosis of hypertension among children, adolescents, and adults born preterm or very low birth weight with those born at term. We performed a meta-analysis with the subset of 10 studies that reported the resting systolic blood pressure difference in millimeters of mercury with 95% CIs or SEs. We assessed methodologic quality with a modified Newcastle-Ottawa Scale. The 10 studies were composed of 1342 preterm or very low birth weight and 1738 term participants from 8 countries. The mean gestational age at birth of the preterm participants was 30.2 weeks (range: 28.8-34.1 weeks), birth weight was 1280 g (range: 1098-1958 g), and age at systolic blood pressure measurement was 17.8 years (range: 6.3-22.4 years). Former preterm or very low birth weight infants had higher systolic blood pressure than term infants (pooled estimate: 2.5 mm Hg [95% CI: 1.7-3.3 mm Hg]). For the 5 highest quality studies, the systolic blood pressure difference was slightly greater, at 3.8 mm Hg (95% CI: 2.6-5.0 mm Hg). We conclude that infants who are born preterm or very low birth weight have modestly higher systolic blood pressure later in life and may be at increased risk for developing hypertension and its sequela