Localization of apolipoprotein A-I epitopes involved in the activation of lecithin: Cholesterol acyltransferase

Eight murine monoclonal antibodies (Mab) to apolipoprotein A-I were characterized for their epitopes and for their ability to interfere with lecithin:cholesterol acyltransferase (LCAT) activation mediated by apo apoA-I using a synthetic substrate. Using overlapping synthetic peptides we have identified six continuous epitopes that span amino acids 1-10 (Mab A-I-19), 96-101 (Mab A-I-15), 133-141 (Mab A-I-5), 140-145 (Mab A-I-9), 144-148 (Mab A-I-8), and 167-174 (Mab A-I-57). Furthermore, antibodies A-I-11 and A-I-16 recognized discontinuous epitopes, namely amino acids 124-128 and 144-148. When antibodies were tested for their ability to inhibit LCAT activation, an inhibitory effect was observed with those whose epitopes covered the area of apoA-I encompassing amino acids 96-174. From these data we conclude that several areas of apoA-I spanning the middle region of the apolipoprotein act in concert to stimulate LCAT activity, possibly by cooperative interaction with the enzyme

Restoring soil functionality in degraded areas within vineyards by organic treatments: the experimental layout of the RESOLVE Core-organic+ project

Degraded areas are frequent in vineyards, even if managed by organic farming, mainly caused by soil truncation, soil erosion, or salts enrichment. Recovering strategies implemented in 19 degraded vineyards in 5 countries within the RESOLVE project concerned: (i) composted organic amendments, or seeding of cover crops (including legumes) for (ii) green manure or (iii) dry mulch. The relationships between vineyard management and pedo-climatic conditions are useful to better understand other contributions dealing with RESOLVE project reported in the present special issue. The results suggested that the nature of degradation requires optimum seedbed preparation to grow green manure crops. Whereas dry mulching plants needs less tillage operations (neither incorporation nor sowing, if self-reseeding plants), helping the recovery of soil functionality. The potential soil erosion by water was esteemed for the 38 plots, confirming that is a common agent of land degradation in vineyards

Monoclonal antibodies to human low density lipoprotein identify distinct areas on apolipoprotein B-100 relevant to the low density lipoprotein-receptor interaction.

We have characterized the epitopes for ten murine monoclonal antibodies (Mabs) to human low density lipoprotein (LDL) and studied their ability to interfere with the LDL-receptor interaction. The epitopes for the antibodies were defined by using the following approaches: 1) interaction with apoB-48; 2) interaction with apoB-100 thrombolytic fragments; and 3) interaction with beta-galactosidase-apoB fusion proteins spanning different areas of the apoB-100 sequence. The results obtained are consistent with the following map of epitopes: Mab 6E, amino acids (aa) 1-1297, Mabs 5A and 6B, aa 1480-1693, Mabs 2A, 7A, 3B, and 4B, aa 2152-2377, Mabs 8A and 9A, aa 2657-3248 and 3H, aa 4082-4306. Four Mabs (2A, 5A, 7A, and 9A) whose epitopes are located in three different areas of apoB, dramatically reduced (up to 95%) the LDL-receptor interaction on cultured human fibroblasts; Fab fragments were as effective as the whole antibodies. Mab 3H, on the other hand, increased LDL binding up to threefold. These findings are consistent with the hypothesis that several areas of apoB-100 are involved independently or in concert in modulating the apoprotein B conformation required for interaction with the LDL receptor

The meningococcal vaccine antigen GNA2091 is an analogue of YraP and plays key roles in outer membrane stability and virulence

K.L.S. was supported by the Australian National Health and Medical Research Council (NHMRC) C. J. Martin Fellowship and Career Development Fellowship. A.F.H. was supported by a Marie Curie Fellowship (PIEF-GA-2012-328377). F.O., L.F., and S.B. were recipients of Novartis fellowships from the Ph.D. program of the University of Siena (Siena, Italy) and University of Bologna (Bologna, Italy), respectively.GNA2091 is one of the components of the 4-component meningococcal serogroup B vaccine (4CMenB) vaccine and is highly conserved in all meningococcal strains. However, its functional role has not been fully characterized. Here we show that nmb2091 is part of an operon and is cotranscribed with the nmb2089, nmb2090, and nmb2092 adjacent genes, and a similar but reduced operon arrangement is conserved in many other gram-negative bacteria. Deletion of the nmb2091 gene causes an aggregative phenotype with a mild defect in cell separation; differences in the outer membrane composition and phospholipid profile, in particular in the phosphoethanolamine levels; an increased level of outer membrane vesicles; and deregulation of the zinc-responsive genes such as znuD. Finally, the Δ2091 strain is attenuated with respect to the wild-type strain in competitive index experiments in the infant rat model of meningococcal infection. Altogether these data suggest that GNA2091 plays important roles in outer membrane architecture, biogenesis, homeostasis, and in meningococcal survival in vivo, and amodel for its role is discussed. These findings highlight the importance of GNA2091 as a vaccine component.PostprintPeer reviewe

Assessment of soil ecosystem in degraded areas of vineyards after organic treatments

In Italian vineyards, it is quite common to have areas characterized by problems in vine health, grape production and quality, often caused by improper land preparation before vine plantation and/or management. Causes for soil malfunctioning can include reduced contribution of the soil fauna to the ecosystem services such as nutrient cycles and organic matter turnover. ReSolVe is a transnational and interdisciplinary project, supported by Core-Organic+ program, aimed at testing the effects of selective agronomic strategies for restoring optimal soil functionality in degraded areas within organic vineyard. For this purpose, the evaluation and biomonitoring of the abundance of soil mesofauna, nematodes and microarthropods, represents an efficient tool to characterize the effects of crop management on soil quality. Assessing enzyme activities involved in the main biogeochemical cycling of C, N, P and S can also provide indication of soil functions and health status. Italian experimental plots are situated in two commercial farms in Tuscany: i) Fontodi, Panzano in Chianti (FI), which has been managed organically for more than 20 years and ii) San Disdagio, Roccastrada (GR), under organic farming since 2014. In each farm, three plots (250 m2 each) in the degraded areas and three relative control plots in the non-degraded areas were selected. The different restoring strategies implemented in each area were: i) compost, produced on farm by manure + pruning residue + grass, ii) faba bean and winter barley green manure, iii) dry mulching after sowing with Trifolium squarrosum L. Each treated and control plot has been studied for soil nematodes, microarthropods, enzymatic activity, and organic matter turnover using tea-bag index, as well as total organic carbon (TOC) and total nitrogen (TN). Soil sampling was carried out to 0-30 cm depth for TOC, TN, enzymes and nematodes and to 10 cm for microarthropods. Tea-bag index was determined following the Keuskamp et al. method (2013), in order to gather data on decomposition rate and litter stabilisation by using commercially available tea bags as standardised test kits. The extraction of nematodes and microarthropods were performed by the Bermann method and the Berlese- Tullgren selector, respectively. The biological soil quality was evaluated by the Maturity Index of nematodes (MI) and Biological Soil Quality index of microarthropods (QBSar). The results from soil sampling before restoring showed significantly lower values of SOC and TN in degraded areas, but no significant differences between degraded and non-degraded areas for enzymes, QBSar, nematode abundance and MI. Fontodi farm, under organic management since many years, showed significantly higher abundance of microarthropods, nematodes and enzymes than San Disdagio farm. The application of restoration techniques in 2016 showed a significant increase of TOC and TN only under compost addition treatment. As regards microarthropod communities, all the treatments showed a sensible increase in abundance and the conservation of high QBSar values. All the treatments increased the fungal feeder activity of nematodes and decreased the number of plant parasitic nematodes taxa. The major pest of grapes, the virus-vector Xiphinema index (Longidoridae), disappeared in the treated plots, whereas it remained in the control plots

Reading tea leaves worldwide: Decoupled drivers of initial litter decomposition mass‐loss rate and stabilization

The breakdown of plant material fuels soil functioning and biodiversity. Currently, process understanding of global decomposition patterns and the drivers of such patterns are hampered by the lack of coherent large-scale datasets. We buried 36,000 individual litterbags (tea bags) worldwide and found an overall negative correlation between initial mass-loss rates and stabilization factors of plant-derived carbon, using the Tea Bag Index (TBI). The stabilization factor quantifies the degree to which easy-to-degrade components accumulate during early-stage decomposition (e.g. by environmental limitations). However, agriculture and an interaction between moisture and temperature led to a decoupling between initial mass-loss rates and stabilization, notably in colder locations. Using TBI improved mass-loss estimates of natural litter compared to models that ignored stabilization. Ignoring the transformation of dead plant material to more recalcitrant substances during early-stage decomposition, and the environmental control of this transformation, could overestimate carbon losses during early decomposition in carbon cycle models

Reading tea leaves worldwide: decoupled drivers of initial litter decomposition mass-loss rate and stabilisation

The breakdown of plant material fuels soil functioning and biodiversity. Currently, process understanding of global decomposition patterns and the drivers of such patterns are hampered by the lack of coherent large-scale datasets. We buried 36,000 individual litterbags (tea bags) worldwide and found an overall negative correlation between initial mass-loss rates and stabilization factors of plant-derived carbon, using the Tea Bag Index (TBI). The stabilization factor quantifies the degree to which easy-to-degrade components accumulate during early-stage decomposition (e.g. by environmental limitations). However, agriculture and an interaction between moisture and temperature led to a decoupling between initial mass-loss rates and stabilization, notably in colder locations. Using TBI improved mass-loss estimates of natural litter compared to models that ignored stabilization. Ignoring the transformation of dead plant material to more recalcitrant substances during early-stage decomposition, and the environmental control of this transformation, could overestimate carbon losses during early decomposition in carbon cycle models

Reading tea leaves worldwide: decoupled drivers of initial litter decomposition mass‐loss rate and stabilization

The breakdown of plant material fuels soil functioning and biodiversity. Currently, process understanding of global decomposition patterns and the drivers of such patterns are hampered by the lack of coherent large‐scale datasets. We buried 36,000 individual litterbags (tea bags) worldwide and found an overall negative correlation between initial mass‐loss rates and stabilization factors of plant‐derived carbon, using the Tea Bag Index (TBI). The stabilization factor quantifies the degree to which easy‐to‐degrade components accumulate during early‐stage decomposition (e.g. by environmental limitations). However, agriculture and an interaction between moisture and temperature led to a decoupling between initial mass‐loss rates and stabilization, notably in colder locations. Using TBI improved mass‐loss estimates of natural litter compared to models that ignored stabilization. Ignoring the transformation of dead plant material to more recalcitrant substances during early‐stage decomposition, and the environmental control of this transformation, could overestimate carbon losses during early decomposition in carbon cycle models