A large-scale evaluation of peptide vaccines against foot-and-mouth disease: lack of solid protection in cattle and isolation of escape mutants.

A large-scale vaccination experiment involving a total of 138 cattle was carried out to evaluate the potential of synthetic peptides as vaccines against foot-and-mouth disease. Four types of peptides representing sequences of foot-and-mouth disease virus (FMDV) C3 Argentina 85 were tested: A, which includes the G-H loop of capsid protein VP1 (site A); AT, in which a T-cell epitope has been added to site A; AC, composed of site A and the carboxy-terminal region of VP1 (site C); and ACT, in which the three previous capsid motifs are colinearly represented. Induction of neutralizing antibodies, lymphoproliferation in response to viral antigens, and protection against challenge with homologous infectious virus were examined. None of the tested peptides, at several doses and vaccination schedules, afforded protection above 40%. Protection showed limited correlation with serum neutralization activity and lymphoproliferation in response to whole virus. In 12 of 29 lesions from vaccinated cattle that were challenged with homologous virus, mutant FMDVs with amino acid substitutions at antigenic site A were identified. This finding suggests the rapid generation and selection of FMDV antigenic variants in vivo. In contrast with previous studies, this large-scale vaccination experiment with an important FMDV host reveals considerable difficulties for vaccines based on synthetic peptides to achieve the required levels of efficacy. Possible modifications of the vaccine formulations to increase protective activity are discussed.     

Effects of a severe drought on Quercus ilex radial growth and xylem anatomy

We assessed the response of Quercus ilex subsp. ballota to the severe summer drought recorded in 1994 in NE Spain through the study of changes in radial growth and wood anatomy. We selected a coppice stand in the Iberian Peninsula, which is characterized by a Mediterranean climate under continental influence. We measured internode length, tree-ring width, mean and maximum vessel diameter, and vessel density for 1981–1997. The annual predicted hydraulic conductance (K_h) was calculated following Hagen-Poisseuille's law. We compared the tree-ring width, vessel diameter and K_h of Q. ilex subsp. ballota and co-existing ring-porous oaks (Q. faginea, Q. pyrenaica) for a dry summer (1994) and a wet summer (1997). To evaluate the drought-resistance of xylem for Q. ilex subsp. ballota (dominant under continental conditions) and Q. ilex subsp. ilex (dominant in mild areas) we determined vulnerability curves. Dimensionless indices of internode length, tree-ring width, and vessel density were compared with climatic data (monthly total precipitation and mean temperature) using correlation analyses. Internode length, tree-ring width, K_h, and mean and maximum vessel diameter declined in 1994. According to vulnerability curves, Q. ilex subsp. ballota showed a greater drought resistance than Q. ilex subsp. ilex. During the year of growth, we found a positive influence of January and June–August precipitation on the internode length, tree-ring width, and vessel density. The response of Q. ilex subsp. ballota radial-growth to summer drought was comparable to that of Q. faginea latewood. Overall, growth and wood anatomy of Q. ilex subsp. ballota showed a plastic response to drought.     

Summer-drought constrains the phenology and growth of two coexisting Mediterranean oaks with contrasting leaf habit: implications for their persistence and reproduction

This study analyses how coexisting evergreen and deciduous oaks adjust their phenology to cope with the stressful Mediterranean summer conditions. We test the hypothesis that the vegetative and reproductive growth of the winter deciduous (Quercus faginea Lam.) is more affected by summer drought than that of the evergreen [Quercus ilex L. subsp. ballota (Desf.) Samp.]. First, we assessed the complete aboveground phenology of both species during two consecutive years. Shoot and litter production and bud, acorn and secondary growth were monitored monthly. Second, we identified several parameters affected by summer conditions: apical bud size, individual leaf area (LA), leaf mass per area (LMA) and acorn yield in both species, and leaf-fall in Q. faginea; and analysed their variation over 10 years. Q. ilex performed up to 25% of shoot growth and most leaf development during summer, whereas Q. faginea completed most of both phenophases during spring. Secondary growth was arrested in summer under drought conditions. Approximately, 30–40% of bud and 40–50% of acorn growth was undertaken during summer in both species. Summer drought related to differences in LA, LMA and leaf senescence, but not to acorn yield. Both species had similar year-to-year patterns of acorn production, though yields were always lower in Q. faginea. Bud size decreased severely in both species during extremely dry years. In Q. ilex, bud size tended to alternate between years of large and small buds, and these patterns were followed by opposite trends in stem length. In Q. faginea, bud size was more stable through time. Q. ilex was more phenologically active during summer than Q. faginea, indicating a higher tolerance to drought. Furthermore, bud and fruit growth (the only two phenophases that both species performed during summer) were more severely affected by summer drought in Q. faginea than in the evergreen. The differential effects of summer drought on key phenophases for the persistence (bud growth) and colonization ability (fruit production) of both species may have consequences for their coexistence.     

Peptide chemical ligation inside living cells: in vivo generation of a circular protein domain.

Here we describe the first example of a peptide chemical ligation reaction performed inside a living cell. A cell-based native chemical ligation approach was developed and used to generate a circular version of the N-terminal Src homology 3 (SH3) domain from the murine c-Crk adapter protein inside Escherichia coli cells. The in vivo cyclization reaction was extremely efficient and the resulting circular protein domain was fully biologically active and able to adopt the native SH3 folded structure. This work represents an important step towards the in vivo generation of small backbone cyclic peptides for use in basic biological research.     

3,4-Methylenedioxymethamphetamine increases interleukin-1beta levels and activates microglia in rat brain: studies on the relationship with acute hyperthermia and 5-HT depletion

3,4-Methylenedioxymethamphetamine (MDMA) administration to rats produces acute hyperthermia and 5-HT release. Interleukin-1beta (IL-1beta) is a pro-inflammatory pyrogen produced by activated microglia in the brain. We examined the effect of a neurotoxic dose of MDMA on IL-1beta concentration and glial activation and their relationship with acute hyperthermia and 5-HT depletion. MDMA, given to rats housed at 22 degrees C, increased IL-1beta levels in hypothalamus and cortex from 1 to 6 h and [(3)H]-(1-(2-chlorophenyl)-N-methyl-N-(1-methylpropyl)3-isoquinolinecarboxamide) binding between 3 and 48 h. Increased immunoreactivity to OX-42 was also detected. Rats became hyperthermic immediately after MDMA and up to at least 12 h later. The IL-1 receptor antagonist did not modify MDMA-induced hyperthermia indicating that IL-1beta release is a consequence, not the cause, of the rise in body temperature. When MDMA was given to rats housed at 4 degrees C, hyperthermia was abolished and the IL-1beta increase significantly reduced. The MDMA-induced acute 5-HT depletion was prevented by fluoxetine coadministration but the IL-1beta increase and hyperthermia were unaffected. Therefore, the rise in IL-1beta is not related to the acute 5-HT release but is linked to the hyperthermia. Contrary to IL-1beta levels, microglial activation is not significantly modified when hyperthermia is prevented, suggesting that it might be a process not dependent on the hyperthermic response induced by MDMA.     

Lignin-Derived Compounds as Efficient Laccase Mediators for Decolorization of Different Types of Recalcitrant Dyes

Ten phenols were selected as natural laccase mediators after screening 44 different compounds with a recalcitrant dye (Reactive Black 5) as a substrate. Their performances were evaluated at different mediator/dye ratios and incubation times (up to 6 h) by the use of Pycnoporus cinnabarinus and Trametes villosa laccases and were compared with those of eight known synthetic mediators (including -NOH- compounds). Among the six types of dyes assayed, only Reactive Blue 38 (phthalocyanine) was resistant to laccase-mediator treatment under the conditions used. Acid Blue 74 (indigoid dye), Reactive Blue 19 (anthraquinoid dye), and Aniline Blue (triarylmethane-type dye) were partially decolorized by the laccases alone, although decolorization was much more efficient and rapid with mediators, whereas Reactive Black 5 (diazo dye) and Azure B (heterocyclic dye) could be decolorized only in the presence of mediators. The efficiency of each natural mediator depended on the type of dye to be treated but, with the only exception being Azure B (<50% decolorization), nearly complete decolorization (80 to 100%) was attained in all cases. Similar rates were attained with the best synthetic mediators, but the reactions were significantly slower. Phenolic aldehydes, ketones, acids, and esters related to the three lignin units were among the best mediators, including p-coumaric acid, vanillin, acetovanillone, methyl vanillate, and above all, syringaldehyde and acetosyringone. The last two compounds are especially promising as ecofriendly (and potentially cheap) mediators for industrial applications since they provided the highest decolorization rates in only 5 to 30 min, depending on the type of dye to be treated.     

Competition and drought limit the response of water-use efficiency to rising atmospheric carbon dioxide in the Mediterranean fir <Emphasis Type="Italic">Abies pinsapo</Emphasis>

The gas-exchange and radial growth responses of conifer forests to climatic warming and increasing atmospheric CO₂ have been widely studied. However, the modulating effects of variables related to stand structure (e.g., tree-to-tree competition) on those responses are poorly explored. The basal-area increment (BAI) and C isotope discrimination (C stable isotope ratio; δ¹³C) in the Mediterranean fir Abies pinsapo were investigated to elucidate the influences of stand competition, atmospheric CO₂ concentrations and climate on intrinsic water-use efficiency (WUEi). We assessed the variation in δ¹³C of tree-rings from dominant or co-dominant trees subjected to different degrees of competition. A high- (H) and a low-elevation (L) population with contrasting climatic constraints were studied in southern Spain. Both populations showed an increase in long-term WUEi. However, this increase occurred more slowly at the L site, where a decline of BAI was also observed. Local warming and severe droughts have occurred in the study area over the past 30 years, which have reduced water availability more at lower elevations. Contrastingly, trees from the H site were able to maintain high BAI values at a lower cost in terms of water consumption. In each population, trees subjected to a higher degree of competition by neighboring trees showed lower BAI and WUEi than those subjected to less competition, although the slopes of the temporal trends in WUEi were independent of the competitive micro-environment experienced by the trees. The results are consistent with an increasing drought-induced limitation of BAI and a decreasing rate of WUEi improvement in low-elevation A. pinsapo forests. This relict species might not be able to mitigate the negative effects of a decrease in water availability through a reduction in stomatal conductance, thus leading to a growth decline in the more xeric sites. An intense and poorly asymmetric competitive environment at the stand level may also act as an important constraint on the adaptive capacity of these drought-sensitive forests to climatic warming. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Interacting effects of changes in climate and forest cover on mortality and growth of the southernmost European fir forests

Aims The combined effects of changes in climate and land use on tree mortality and growth patterns have rarely been addressed. Relict tree species from the Mediterranean Basin serve as appropriate models to investigate these effects, since they grow in climatically stressed areas which have undergone intense cover changes. The aim is to use climate, aerial photographs, stand structure and radial‐growth data to explain the mortality and historical patterns of growth of Abies pinsapo in the area where this relict species was first protected. Location Sierra de las Nieves, West Baetic Range, southern Spain. Methods We assessed variations of tree cover in A. pinsapo forests through image analyses of aerial photographs spanning the last 50 years. We sampled 31 stands to assess current altitudinal patterns of forest structure and mortality. We evaluated the relationships between radial growth and regional climate using linear models in three sites at different elevations. Results Regional warming and a decrease in precipitation were detected. Forest tree cover increased at all elevations from 1957 until 1991, but it afterwards decreased below 1100 m. Currently, the likelihood of tree mortality increases downwards and is associated with dense, closed stands with a low living basal area. In contrast to previous droughts, a sharp synchronized reduction in tree growth, not fully accounted for in linear climate–growth models, occurred at low elevations in 1994–95, but not upwards. It was preceded by a weakening of the negative association between low‐elevation growth and water deficit since the late 1970s. Conclusions The intense densification of A. pinsapo forests following strict protection measures in the late 1950s enhanced the vulnerability of climate‐sensitive A. pinsapo forests to recent drier conditions. Such abrupt land‐use changes help to explain recent patterns of mortality and growth decline in low‐elevation A. pinsapo forests.