Expression of a barley cystatin gene in maize enhances resistance against phytophagous mites by altering their cysteine-proteases

Phytocystatins are inhibitors of cysteine-proteases from plants putatively involved in plant defence based on their capability of inhibit heterologous enzymes. We have previously characterised the whole cystatin gene family members from barley (HvCPI-1 to HvCPI-13). The aim of this study was to assess the effects of barley cystatins on two phytophagous spider mites, Tetranychus urticae and Brevipalpus chilensis. The determination of proteolytic activity profile in both mite species showed the presence of the cysteine-proteases, putative targets of cystatins, among other enzymatic activities. All barley cystatins, except HvCPI-1 and HvCPI-7, inhibited in vitro mite cathepsin L- and/or cathepsin B-like activities, HvCPI-6 being the strongest inhibitor for both mite species. Transgenic maize plants expressing HvCPI-6 protein were generated and the functional integrity of the cystatin transgene was confirmed by in vitro inhibitory effect observed against T. urticae and B. chilensis protein extracts. Feeding experiments impaired on transgenic lines performed with T. urticae impaired mite development and reproductive performance. Besides, a significant reduction of cathepsin L-like and/or cathepsin B-like activities was observed when the spider mite fed on maize plants expressing HvCPI-6 cystatin. These findings reveal the potential of barley cystatins as acaricide proteins to protect plants against two important mite pests

Geographical, seasonal, and depth variation in sinking particle speeds in the North Atlantic

Particle sinking velocity is considered to be a controlling factor for carbon transport to the deep sea and thus carbon sequestration in the oceans. The velocities of the material exported to depth are considered to be high in high-latitude productive systems and low in oligotrophic distributions. We use a recently developed method based on the measurement of the radioactive pair 210Po-210Pb to calculate particle sinking velocities in the temperate and oligotrophic North Atlantic during different bloom stages. Our estimates of average sinking velocities (ASVs) show that slowly sinking particles (<100?m?d?1) contribute significantly to carbon flux at all the locations except in the temperate regions during the bloom. ASVs appear to vary strongly with season, which we propose is caused by changes in the epipelagic community structure. Our results are the first field data to confirm the long-standing theory that particle sinking velocities increase with depth, with increases of up to 90% between 50 and 150?m depth

A barley cysteine-protease inhibitor reduces teh performance of two aphid species in artificial diets and transgenic arabidopsis plants

Cystatins from plants have been implicated in plant defense towards insects, based on their role as inhibitors of heterologous cysteine-proteinases. We have previously characterized thirteen genes encoding cystatins (HvCPI-1 to HvCPI-13) from barley (Hordeum vulgare), but only HvCPI-1 C68 → G, a variant generated by direct-mutagenesis, has been tested against insects. The aim of this study was to analyze the effects of the whole gene family members of barley cystatins against two aphids, Myzus persicae and Acyrthosiphon pisum. All the cystatins, except HvCPI-7, HvCPI-10 and HvCPI-12, inhibited in vitro the activity of cathepsin L- and/or B-like proteinases, with HvCPI-6 being the most effective inhibitor for both aphid species. When administered in artificial diets, HvCPI-6 was toxic to A. pisum nymphs (LC50 = 150 μg/ml), whereas no significant mortality was observed on M. persicae nymphs up to 1000 μg/ml. The effects of HvCPI-6 ingestion on A. pisum were correlated with a decrease of cathepsin B- and L-like proteinase activities. In the case of M. persicae, there was an increase of these proteolytic activities, but also of the aminopeptidase-like activity, suggesting that this species is regulating both target and insensitive enzymes to overcome the effects of the cystatin. To further analyze the potential of barley cystatins as insecticidal proteins against aphids, Arabidopsis plants expressing HvCPI-6 were tested against M. persicae. For A. pisum, which does not feed on Arabidopsis, a combined diet-Vicia faba plant bioassay was performed. A significant delay in the development time to reach the adult stage was observed in both species. The present study demonstrates the potential of barley cystatins to interfere with the performance of two aphid specie

New GOLD classification: Longitudinal data on group assignment

Rationale: Little is known about the longitudinal changes associated with using the 2013 update of the multidimensional GOLD strategy for chronic obstructive pulmonary disease (COPD). Objective: To determine the COPD patient distribution of the new GOLD proposal and evaluate how this classification changes over one year compared with the previous GOLD staging based on spirometry only. Methods: We analyzed data from the CHAIN study, a multicenter observational Spanish cohort of COPD patients who are monitored annually. Categories were defined according to the proposed GOLD: FEV1%, mMRC dyspnea, COPD Assessment Test (CAT), Clinical COPD Questionnaire (CCQ), and exacerbations-hospitalizations. One-year follow-up information was available for all variables except CCQ data. Results: At baseline, 828 stable COPD patients were evaluated. On the basis of mMRC dyspnea versus CAT, the patients were distributed as follows: 38.2% vs. 27.2% in group A, 17.6% vs. 28.3% in group B, 15.8% vs. 12.9% in group C, and 28.4% vs. 31.6% in group D. Information was available for 526 patients at one year: 64.2% of patients remained in the same group but groups C and D show different degrees of variability. The annual progression by group was mainly associated with one-year changes in CAT scores (RR, 1.138; 95%CI: 1.074-1.206) and BODE index values (RR, 2.012; 95%CI: 1.487-2.722). Conclusions: In the new GOLD grading classification, the type of tool used to determine the level of symptoms can substantially alter the group assignment. A change in category after one year was associated with longitudinal changes in the CAT and BODE index

Potential Use of a Serpin from Arabidopsis for Pest Control

Although genetically modified (GM) plants expressing toxins from Bacillus thuringiensis (Bt) protect agricultural crops against lepidopteran and coleopteran pests, field-evolved resistance to Bt toxins has been reported for populations of several lepidopteran species. Moreover, some important agricultural pests, like phloem-feeding insects, are not susceptible to Bt crops. Complementary pest control strategies are therefore necessary to assure that the benefits provided by those insect-resistant transgenic plants are not compromised and to target those pests that are not susceptible. Experimental GM plants producing plant protease inhibitors have been shown to confer resistance against a wide range of agricultural pests. In this study we assessed the potential of AtSerpin1, a serpin from Arabidopsis thaliana (L). Heynh., for pest control. In vitro assays were conducted with a wide range of pests that rely mainly on either serine or cysteine proteases for digestion and also with three non-target organisms occurring in agricultural crops. AtSerpin1 inhibited proteases from all pest and non-target species assayed. Subsequently, the cotton leafworm Spodoptera littoralis Boisduval and the pea aphid Acyrthosiphon pisum (Harris) were fed on artificial diets containing AtSerpin1, and S. littoralis was also fed on transgenic Arabidopsis plants overproducing AtSerpin1. AtSerpin1 supplied in the artificial diet or by transgenic plants reduced the growth of S. littoralis larvae by 65% and 38%, respectively, relative to controls. Nymphs of A. pisum exposed to diets containing AtSerpin1 suffered high mortality levels (LC50 = 637 µg ml−1). The results indicate that AtSerpin1 is a good candidate for exploitation in pest control

The EFSUMB Guidelines and Recommendations for Musculoskeletal Ultrasound - Part I: Extraarticular Pathologies

The first part of the guidelines and recommendations for musculoskeletal ultrasound, produced under the auspices of the European Federation of Societies for Ultrasound in Medicine and Biology (EFSUMB), provides information about the use of musculoskeletal ultrasound for assessing extraarticular structures (muscles, tendons, entheses, ligaments, bones, bursae, fasciae, nerves, skin, subcutaneous tissues, and nails) and their pathologies. Clinical applications, practical points, limitations, and artifacts are described and discussed for every structure. After an extensive literature review, the recommendations have been developed according to the Oxford Centre for Evidence-based Medicine and GRADE criteria and the consensus level was established through a Delphi process. The document is intended to guide clinical users in their daily practice

Vertical imbalance in organic carbon budgets is indicative of a missing vertical transfer during a phytoplankton bloom near South Georgia (COMICS)

The biological carbon pump, driven principally by the surface production of sinking organic matter and its subsequent remineralization to carbon dioxide (CO2) in the deep ocean, maintains atmospheric CO2 concentrations around 200 ppm lower than they would be if the ocean were abiotic. One important driver of the magnitude of this effect is the depth to which organic matter sinks before it is remineralised, a parameter we have limited confidence in measuring given the difficulty involved in balancing sources and sinks in the ocean's interior. One solution to this imbalance might be a temporal offset in which organic carbon accumulates in the mesopelagic zone (100–1000 m depth) early in the productive season before it is consumed later. Here, we develop a novel accounting method to address non-steady state conditions by estimating fluxes of particulate organic matter into, and accumulation within, distinct vertical layers in the mesopelagic zone using high-resolution spatiotemporal vertical profiles. We apply this approach to a time series of measurements made during the declining phase of a large diatom bloom in a low-circulation region of the Southern Ocean downstream of South Georgia. Our data show that the major export event led to a significant accumulation of organic matter in the upper mesopelagic zone (100–200 m depth) which declined over the following weeks, implying that temporal offsets need to be considered when compiling budgets. However, even when accounting for this accumulation, a mismatch in the vertically resolved organic carbon budget remained, implying that there are likely widespread processes that we do not yet understand that redistribute material vertically within the mesopelagic zone

Vertical imbalance in organic carbon budgets is indicative of a missing vertical transfer during a phytoplankton bloom near South Georgia (COMICS)

The biological carbon pump, driven principally by surface production and sinking of organic matter to deep water and its subsequent remineralization to CO2 maintains atmospheric CO2 around 200 ppm lower than it would be if the ocean were abiotic. One important driver of the magnitude of this effect is the depth to which organic matter sinks before it is remineralised, a parameter we have limited confidence in measuring given the difficulty involved in balancing sources and sinks in the ocean's interior. This imbalance is due, in part, to our inability to measure respiration directly and our reliance on radiotracer-based proxies. One solution to these problems might be a temporal offset in which organic carbon accumulates in the mesopelagic zone (100–1000 m depth) early in the productive season prior to it being consumed later, a situation which could lead to a net apparent sink occurring if a steady state assumption is applied as is often the approach. In this work, we develop a novel accounting method to address this issue, independent of respiration measurements, by estimating fluxes into and accumulation within distinct vertical layers in the mesopelagic. We apply this approach to a time series of measurements of particle sinking velocities and interior organic carbon concentrations made during the declining phase of a large diatom bloom in a low-circulation region of the Southern Ocean downstream of South Georgia. Our data show that the major export event led to a significant accumulation of organic matter in the upper mesopelagic (100–200 m depth) which declined over several weeks, implying that temporal offsets need to be considered when compiling budgets. However, even when accounting for this accumulation, a mismatch in the vertically resolved organic carbon budget remained, implying that there are likely widespread processes that we do not yet understand that redistribute material vertically in the mesopelagic

Distribution and outcomes of a phenotype-based approach to guide COPD management: Results from the CHAIN cohort

Rationale: The Spanish guideline for COPD (GesEPOC) recommends COPD treatment according to four clinical phenotypes: non-exacerbator phenotype with either chronic bronchitis or emphysema (NE), asthma-COPD overlap syndrome (ACOS), frequent exacerbator phenotype with emphysema (FEE) or frequent exacerbator phenotype with chronic bronchitis (FECB). However, little is known on the distribution and outcomes of the four suggested phenotypes. Objective: We aimed to determine the distribution of these COPD phenotypes, and their relation with one-year clinical outcomes. Methods: We followed a cohort of well-characterized patients with COPD up to one-year. Baseline characteristics, health status (CAT), BODE index, rate of exacerbations and mortality up to one year of follow-up were compared between the four phenotypes. Results: Overall, 831 stable COPD patients were evaluated. They were distributed as NE, 550 (66.2%); ACOS, 125 (15.0%); FEE, 38 (4.6%); and FECB, 99 (11.9%); additionally 19 (2.3%) COPD patients with frequent exacerbations did not fulfill the criteria for neither FEE nor FECB. At baseline, there were significant differences in symptoms, FEV1 and BODE index (all p<0.05). The FECB phenotype had the highest CAT score (17.1±8.2, p<0.05 compared to the other phenotypes). Frequent exacerbator groups (FEE and FECB) were receiving more pharmacological treatment at baseline, and also experienced more exacerbations the year after (all p<0.05) with no differences in one-year mortality. Most of NE (93%) and half of exacerbators were stable after one year. Conclusions: There is an uneven distribution of COPD phenotypes in stable COPD patients, with significant differences in demographics, patient-centered outcomes and health care resources use