Post-Seizure α-Tocopherol Treatment Decreases Neuroinflammation and Neuronal Degeneration Induced by Status Epilepticus in Rat Hippocampus

Vitamin E (as α-tocopherol, α-T) was shown to have beneficial effects in epilepsy, mainly ascribed to its antioxidant properties. Besides radical-induced neurotoxicity, neuroinflammation is also involved in the pathophysiology of epilepsy, since neuroglial activation and cytokine production exacerbate seizure-induced neurotoxicity and contribute to epileptogenesis. We previously showed that α-T oral supplementation before inducing status epilepticus, markedly reduces astrocytic and microglial activation, neuronal cell death and oxidative stress in the hippocampus, as observed 4 days after seizure. In order to evaluate the possibility that such a neuroprotective and anti-inflammatory effect may also provide a strategy for an acute intervention in epilepsy, in this study, seizures were induced by single intaperitoneal injection of kainic acid and, starting from 3 h after status epilepticus, rats were treated with an intraperitoneal bolus of α-T (250 mg/kg b.w.; once a day) for 4 days, that was the time after which morphological and biochemical analyses were performed on hippocampus. Post-seizure α-T administration significantly reduced astrocytosis and microglia activation, and decreased neuron degeneration and spine loss; these effects were associated with the presence of a lowered lipid peroxidation in hippocampus. These results confirm and further emphasize the anti-inflammatory and neuroprotective role of α-T in kainic acid-induced epilepsy. Moreover, the findings show that post-seizure treatment with α-T provides an effective secondary prevention against post-seizure inflammation-induced brain damages and possibly against their epileptogenic effects.     

Dramatic Potentiation of the Antiviral Activity of HIV Antibodies by Cholesterol Conjugation

The broadly neutralizing antibodies HIV 2F5 and 4E10, which bind to overlapping epitopes in the membrane-proximal external region of the fusion protein gp41, have been proposed to use a two-step mechanism for neutralization; first, they bind and preconcentrate at the viral membrane through their long, hydrophobic CDRH3 loops, and second, they form a high affinity complex with the protein epitope. Accordingly, mutagenesis of the CDRH3 can abolish their neutralizing activity, with no change in the affinity for the peptide epitope. We show here that we can mimic this mechanism by conjugating a cholesterol group outside of the paratope of an antibody. Cholesterol-conjugated antibodies bind to lipid raft domains on the membrane, and because of this enrichment, they show increased antiviral potency. In particular, we find that cholesterol conjugation (i) rescues the antiviral activity of CDRH3-mutated 2F5, (ii) increases the antiviral activity of WT 2F5, (iii) potentiates the non-membrane-binding HIV antibody D5 10–100-fold (depending on the virus strain), and (iv) increases synergy between 2F5 and D5. Conjugation can be made at several positions, including variable and constant domains. Cholesterol conjugation therefore appears to be a general strategy to boost the potency of antiviral antibodies, and, because membrane affinity is engineered outside of the antibody paratope, it can complement affinity maturation strategies.     

PI3K‐p110‐alpha‐subtype signalling mediates survival,proliferation and neurogenesis of cortical progenitor cells via activation of mTORC2

Development of the cerebral cortex is controlled by growth factors among which transforming growth factor beta (TGFβ) and insulin‐like growth factor 1 (IGF1) have a central role. The TGFβ‐ and IGF1‐pathways cross‐talk and share signalling molecules, but in the central nervous system putative points of intersection remain unknown. We studied the biological effects and down‐stream molecules of TGFβ and IGF1 in cells derived from the mouse cerebral cortex at two developmental time points, E13.5 and E16.5. IGF1 induces PI3K, AKT and the mammalian target of rapamycin complexes (mTORC1/mTORC2) primarily in E13.5‐derived cells, resulting in proliferation, survival and neuronal differentiation, but has small impact on E16.5‐derived cells. TGFβ has little effect at E13.5. It does not activate the PI3K‐ and mTOR‐signalling network directly, but requires its activity to mediate neuronal differentiation specifically at E16.5. Our data indicate a central role of mTORC2 in survival, proliferation as well as neuronal differentiation of E16.5‐derived cortical cells. mTORC2 promotes these cellular processes and is under control of PI3K‐p110‐alpha signalling. PI3K‐p110‐beta signalling activates mTORC2 in E16.5‐derived cells but it does not influence cell survival, proliferation and differentiation. This finding indicates that different mTORC2 subtypes may be implicated in cortical development and that these subtypes are under control of different PI3K isoforms.

     

Charge Transport Anisotropy in Highly Oriented Thin Films of the Acceptor Polymer P(NDI2OD‐T2)

The nanomorphology of the high mobility polymer poly{[N,N′‐bis(2‐octyldodecyl)‐1,4,5,8‐naphthalenedicarboximide‐2,6‐diyl]‐alt‐5,5′‐(2,2′‐bithiophene)} P(NDI2OD‐T2) in thin films is explored as a function of different annealing conditions and correlated to optical and electrical properties. While nanofibrils with face‐on orientation in form I are obtained directly after spin‐coating and annealing below the melt transition temperature, clear evidence of lamellar structures is found after melt‐annealing followed by slow cooling to room temperature. Interestingly these structural changes are accompanied by distinct changes in the absorption patterns. Electron diffraction measurements further show clear transitions towards predominant edge‐on oriented chains in form II upon melt‐annealing. Large‐scale alignment with dichroic ratios up to 10 and improved order is achieved by high temperature rubbing and subsequent post‐rubbing annealing. These highly oriented morphologies allow anisotropic in‐plane charge transport to be probed with top‐gate transistors parallel and perpendicular to the polymer chain direction. Mobilities up to 0.1 cm² V^‐1 s^‐1 are observed parallel to the polymer chain, which is up to 10 times higher than those perpendicular to the polymer chain.     

Species’ traits influence ground beetle responses to farm and landscape level agricultural intensification in Europe

Agricultural intensification may result in important shifts in insect community composition and function, but this remains poorly explored. Studying how groups of species with shared traits respond to local and landscape scale land-use management can reveal mechanisms behind such observed impacts. We tested if ground beetles (Coleoptera: Carabidae) divided into trait groups based on body sizes, wing morphologies and dietary preferences respond differently to farming practise (organic and conventional), farming intensity (measured as yield) and landscape complexity (measured as the proportion of arable land within a 1,000 m radius) across Europe. We used data from 143 farms in five regions in northern and central Europe. Organic farms did not differ in abundance or richness of any trait group compared to conventional farms. As farm scale intensity (yield) increased, overall abundance of beetles decreased, but abundances of small and medium sized beetles, as well as that of wingless beetles, were unaffected. Overall species richness was not affected by yield, whereas consideration of traits revealed that phytophagous and omnivorous beetles were less species rich on farms with high yields. Increasing the proportion of arable land in the landscape increased overall beetle abundance. This was driven by an increase in omnivorous beetles. The total species richness was not affected by an increase in the proportion arable land, although the richness of wingless beetles was found to increase. Potential effects on ecosystem functioning need to be taken into account when designing schemes to maintain agricultural biodiversity, because species with different ecological traits respond differently to local management and landscape changes.     

The C2 fragment from Neisseria meningitidis antigen NHBA increases endothelial permeability by destabilizing adherens junctions

Neisseria meningitidis is a human pathogen that can cause fatal sepsis and meningitis once it reaches the blood stream and the nervous system. Here we demonstrate that a fragment, released upon proteolysis of the surface‐exposed protein Neisserial Heparin Binding Antigen (NHBA), by the bacterial protease NalP, alters the endothelial permeability by inducing the internalization of the adherens junction protein VE‐cadherin. We found that C2 rapidly accumulates in mitochondria where it induces the production of reactive oxygen species: the latter are required for the phosphorylation of the junctional protein and for its internalization that, in turn, is responsible for the endothelial leakage. Our data support the notion that the NHBA‐derived fragment C2 might contribute to the extensive vascular leakage typically associated with meningococcal sepsis.     

Nailfold videocapillaroscopy micro-haemorrhage and giant capillary counting as an accurate approach for a steady state definition of disease activity in systemic sclerosis

Introduction

Nailfold videocapillaroscopy (NVC) in systemic sclerosis (SSc) is a procedure commonly used for patient classification and subsetting, but not to define disease activity (DA). This study aimed to evaluate whether the number of micro-haemorrhages (MHE), micro-thrombosis (MT), giant capillaries (GC), and normal/dilated capillaries (Cs) in NVC could predict DA in SSc.

Methods

Eight-finger NVC was performed in 107 patients with SSc, and the total number of MHE/MT, GC, and the mean number of Cs were counted and defined as number of micro-haemorrhages (NEMO), GC and Cs scores, respectively. The European Scleroderma Study Group (ESSG) index constituted the gold standard for DA assessment, and scores ≥3.5 and =3 were considered indicative of high and moderate activity, respectively.

Results

NEMO and GC scores were positively correlated with ESSG index (R = 0.65, P <0.0001, and R = 0.47, P <0.0001, respectively), whilst Cs score showed a negative correlation with that DA index (R = −0.30, P <0.001). The area under the curve (AUC) of receiver operating characteristic plots, obtained by NEMO score sensitivity and specificity values in classifying patients with ESSG index ≥3.5, was significantly higher than the corresponding AUC derived from either GC or Cs scores (P <0.03 and P <0.0006, respectively). A modified score, defined by the presence of a given number of MHE/MT and GC, had a good performance in classifying active patients (ESSG index ≥3, sensitivity 95.1%, specificity 84.8%, accuracy 88.7%).

Conclusions

MHE/MT and GC appear to be good indicators of DA in SSc, and enhances the role of NVC as an easy technique to identify active patients.     

The identification and localization of two intermediate filament proteins in the tunic of Styela plicata (Tunicata, Styelidae)

The intermediate filament (IF) proteins Styela C and Styela D from the tunicate Styela (Urochordata) are co-expressed in all epidermal cells and they are thought to behave as type I and type II keratins. These two IF proteins, Styela C and Styela D, were identified in immunoblots of proteins isolated from the tunic of Styela plicata. The occurrence and distribution of these proteins within the tunic of this ascidian was examined by means of immunofluorescence and immunoperoxidase techniques, using anti-Styela C and anti-Styela D antibodies. In addition, immuno-electron microscopy of the tunic showed that the two proteins are located in the cuticle layer and in the tunic matrix. These results represent the first data about the presence of IF proteins in the tunic of adult ascidian S. plicata. The possible involvement of these IF proteins in reinforcing the integrity of the tunic, that represents the interface between the animal body and the external environment, is discussed.     

Field assessment of host plant specificity and potential effectiveness of a prospective biological control agent, Aceria salsolae, of Russian thistle, Salsola tragus

The eriophyid mite, Aceria salsolae de Lillo and Sobhian, is being evaluated as a prospective classical biological control agent of invasive alien tumbleweeds, including Salsola tragus, S. collina, S. paulsenii and S. australis, in North America. Previous laboratory experiments to determine the host specificity of the mite indicated that it could sometimes persist and multiply on some nontarget plants, including Bassia hyssopifolia and B. scoparia. These are both European plants whose geographic range overlaps that of the mite, but the mite has never been observed on them in the field. A field experiment was conducted in Italy to determine if the mite would infest and damage these plants under natural outdoor conditions. The results indicate that this mite does not attain significant populations on these nontarget plants nor does it significantly damage them. Salsola tragus was heavily infested by A. salsolae, and plant size was negatively correlated to the level of infestation. Although S. kali plants were also infested, their size did not appear to be affected by the mites. The other nontarget plants were not as suitable for the mite in the field as in previous laboratory experiments. We conclude that there would be no significant risk to nontarget plants as a result of using A. salsolae as a biological agent to control Salsola species in North America.