| 1. |
- Kumar, Ramakrishnan B., et al.
(author)
-
Arachidonic acid promotes the binding of 5-lipoxygenase on nanodiscs containing 5-lipoxygenase activating protein in the absence of calcium-ions
- 2020
-
In: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 15:7
-
Journal article (peer-reviewed)abstract
- Among the first steps in inflammation is the conversion of arachidonic acid (AA) stored in the cell membranes into leukotrienes. This occurs mainly in leukocytes and depends on the interaction of two proteins: 5-lipoxygenase (5LO), stored away from the nuclear membranes until use and 5-lipoxygenase activating protein (FLAP), a transmembrane, homotrimeric protein, constitutively present in nuclear membrane. We could earlier visualize the binding of 5LO to nanodiscs in the presence of Ca2+-ions by the use of transmission electron microscopy (TEM) on samples negatively stained by sodium phosphotungstate. In the absence of Ca2+-ions 5LO did not bind to the membrane. In the present communication, FLAP reconstituted in the nanodiscs which could be purified if the His-tag was located on the FLAP C-terminus but not the N-terminus. Our aim was to find out if 1) 5LO would bind in a Ca2+-dependent manner also when FLAP is present? 2) Would the substrate (AA) have effects on 5LO binding to FLAP-nanodiscs? TEM was used to assess the complex formation between 5LO and FLAP-nanodiscs along with, sucrose gradient purification, gel-electrophoresis and mass spectrometry. It was found that presence of AA by itself induces complex formation in the absence of added calcium. This finding corroborates that AA is necessary for the complex formation and that a Ca2+-flush is mainly needed for the recruitment of 5LO to the membrane. Our results also showed that the addition of Ca2+-ions promoted binding of 5LO on the FLAP-nanodiscs as was also the case for nanodiscs without FLAP incorporated. In the absence of added substances no 5LO-FLAP complex was formed. Another finding is that the formation of a 5LO-FLAP complex appears to induce fragmentation of 5LOin vitro.
|
|
| 2. |
- Lopez-Goldar, Xose, et al.
(author)
-
Resin acids as inducible chemical defences of pine seedlings against chewing insects
- 2020
-
In: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 15:5
-
Journal article (peer-reviewed)abstract
- Inducibility of defences in response to biotic stimuli is considered an important trait in plant resistance. In conifers, previous research has mostly focused on the inducibility of the volatile fraction of the oleoresin (mono- and sesquiterpenes), leaving the inducibility of the nonvolatile resin acids largely unexplored, particularly in response to real herbivory. Here we investigated the differences in the inducibility of resin acids in two pine species, one native from Europe (Pinus pinaster Ait.) and another from North America (Pinus radiata D. Don), in response to wounding by two European insects: a bark chewer, the pine weevil (Hylobius abietis L.), and a defoliator, the pine processionary caterpillar (Thaumetopoea pityocampa Schiff.). We quantified the constitutive (control) and induced concentrations of resin acids in the stem and needles of both pine species by gas chromatography techniques. Both pine species strongly increased the concentration of resin acids in the stem after pine weevil feeding, although the response was greater in P. pinaster than in P. radiata. However, systemic defensive responses in the needles were negligible in both pine species after pine weevil feeding in the stem. On the other hand, P. radiata locally reduced the resin acid concentration in the needles after pine caterpillar feeding, whereas in P. pinaster resin acid concentration was apparently unaffected. Nevertheless, systemic induction of resin acids was only observed in the stem of P. pinaster in response to pine caterpillar feeding. In summary, pine induced responses were found highly compartmentalized, and specific to herbivore identity. Particularly, plant defence suppression mechanisms by the pine caterpillar, and ontogenetic factors might be potentially affecting the induced response of resin acids in both pine species.
|
|
| 3. |
- Obura, Morgan, et al.
(author)
-
Post-load glucose subgroups and associated metabolic traits in individuals with type 2 diabetes : An IMI-DIRECT study
- 2020
-
In: PLoS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 15:11
-
Journal article (peer-reviewed)abstract
- AIM: Subclasses of different glycaemic disturbances could explain the variation in characteristics of individuals with type 2 diabetes (T2D). We aimed to examine the association between subgroups based on their glucose curves during a five-point mixed-meal tolerance test (MMT) and metabolic traits at baseline and glycaemic deterioration in individuals with T2D. METHODS: The study included 787 individuals with newly diagnosed T2D from the Diabetes Research on Patient Stratification (IMI-DIRECT) Study. Latent class trajectory analysis (LCTA) was used to identify distinct glucose curve subgroups during a five-point MMT. Using general linear models, these subgroups were associated with metabolic traits at baseline and after 18 months of follow up, adjusted for potential confounders. RESULTS: At baseline, we identified three glucose curve subgroups, labelled in order of increasing glucose peak levels as subgroup 1-3. Individuals in subgroup 2 and 3 were more likely to have higher levels of HbA1c, triglycerides and BMI at baseline, compared to those in subgroup 1. At 18 months (n = 651), the beta coefficients (95% CI) for change in HbA1c (mmol/mol) increased across subgroups with 0.37 (-0.18-1.92) for subgroup 2 and 1.88 (-0.08-3.85) for subgroup 3, relative to subgroup 1. The same trend was observed for change in levels of triglycerides and fasting glucose. CONCLUSIONS: Different glycaemic profiles with different metabolic traits and different degrees of subsequent glycaemic deterioration can be identified using data from a frequently sampled mixed-meal tolerance test in individuals with T2D. Subgroups with the highest peaks had greater metabolic risk.
|
|
| 4. |
- Åkerlund, Cecilia, et al.
(author)
-
Impact of duration and magnitude of raised intracranial pressure on outcome after severe traumatic brain injury : A CENTER-TBI high-resolution group study
- 2020
-
In: PLOS ONE. - : Public Library of Science. - 1932-6203. ; 15:12
-
Journal article (peer-reviewed)abstract
- Magnitude of intracranial pressure (ICP) elevations and their duration have been associated with worse outcomes in patients with traumatic brain injuries (TBI), however published thresholds for injury vary and uncertainty about these levels has received relatively little attention. In this study, we have analyzed high-resolution ICP monitoring data in 227 adult patients in the CENTER-TBI dataset. Our aim was to identify thresholds of ICP intensity and duration associated with worse outcome, and to evaluate the uncertainty in any such thresholds. We present ICP intensity and duration plots to visualize the relationship between ICP events and outcome. We also introduced a novel bootstrap technique to evaluate uncertainty of the equipoise line. We found that an intensity threshold of 18 ± 4 mmHg (2 standard deviations) was associated with worse outcomes in this cohort. In contrast, the uncertainty in what duration is associated with harm was larger, and safe durations were found to be population dependent. The pressure and time dose (PTD) was also calculated as area under the curve above thresholds of ICP. A relationship between PTD and mortality could be established, as well as for unfavourable outcome. This relationship remained valid for mortality but not unfavourable outcome after adjusting for IMPACT core variables and maximum therapy intensity level. Importantly, during periods of impaired autoregulation (defined as pressure reactivity index (PRx)>0.3) ICP events were associated with worse outcomes for nearly all durations and ICP levels in this cohort and there was a stronger relationship between outcome and PTD. Whilst caution should be exercised in ascribing causation in observational analyses, these results suggest intracranial hypertension is poorly tolerated in the presence of impaired autoregulation. ICP level guidelines may need to be revised in the future taking into account cerebrovascular autoregulation status considered jointly with ICP levels.
|
|
