Tree canopy cover and natural regeneration into strictly-protected forest areas: the MaB reserve of Montedimezzo (Isernia,Italy).

The ecological analysis and the analysis of mechanisms underlying the natural regeneration process into strictly protected forest areas, is basic to the understanding of ecosystem functioning and to estimate the recovery ability as a function of time and environmental changes. Aim of the paper is to analyze the spatial-temporal dynamics of tree species regeneration into mixed Turkey oak-common beech stands growing undisturbed since more than one half- century. The ability of first establishment and following growth of natural regeneration related to the ecological parameters, the structural features of standing crop and its development stages, are analyzed in detail. The study was carried out into two permanent monitoring plots (A) and (B) where two different tree species play the main functional role. In (A) common beech is dominant (Fagus sylvatica L.-elevation 1040 m a.s.l.); in (B) Turkey oak is prevailing (Quercus cerris L.-elevation 940 m a.s.l.). In each plot, the analysis of vertical stand structure, of natural regeneration pattern and ecological surveys (LAI, trasmittance, soil moisture content, canopy interception of rainfall) were undertaken by monthly surveys from May to October into 57 sub-plots each 1m2 wide over the period 2005-2007. Results highlighted that (B), Turkey oak prevailing, shows a more complex structure, a higher tree canopy thickness and the lack of gaps. The area shows therefore a lower trasmittance and a higher LAI value; throughfall and soil moisture content are also reduced as compared to (A). As for the natural regeneration pattern, further to a first stage when seedlings mortality is high, their survival rate is being kept high and steady over time in (A). A further mortality peak has been detected vice versa over the following summer season in (B). The different main tree species composition and radiation regime seem to be the basic reasons of the dynamics observed as for mortality and survival rates. The way of seedlings establishment points out that optimal ecological conditions are present for the production and germination of seeds as well, but that the limiting factor is the reduced radiation amount that reaches the forest floor.

Erratum: Lipase-catalyzed regioselective acylation of resorcin[4]arenes (Journal of Molecular Catalysis B: Enzymatic (2002) 16 (24-247) PII: S1381117701000686)

10Immobilized lipase from Mucor miehei (RML) catalyzed the regioselective acylation of the C-2 side-chain of the C-alkyl resorcin[4]arene tetra-alcohol 1 in the 1,2-alternate form in organic solvents using vinyl acetate as acylating reagent. The influence of reaction parameters and solvent choice were also studied. Docking simulations allowed the determination of the binding geometry of 1, revealing the importance of Trp88 residue in stabilizing the Michaelis–Menten complex between enzyme and substrate.reservedmixedBotta, Bruno; Zappia, Giovanni; Tafi, Andrea; Botta, Maurizio; Manetti, Fabrizio; Cernia, Enrico; Milana, Giuliana; Palocci, Cleofe; Soro, Simonetta; Delle Monache, GiulianoBotta, Bruno; Zappia, Giovanni; Tafi, Andrea; Botta, Maurizio; Manetti, Fabrizio; Cernia, Enrico; Milana, Giuliana; Palocci, Cleofe; Soro, Simonetta; Delle Monache, Giulian

Absence of protein A expression is associated with higher capsule production in staphylococcal isolates

Staphylococcus aureus is a major human pathogen, and a leading cause of soft tissue and blood stream infections. One of the causes of its success as a pathogen is the peculiar array of immune evasion factors through which the bacterium avoids host defenses, where the staphylococcal protein A (SpA) plays a major role thanks to its IgG binding activities. Moreover, SpA has recently been proposed as a promising vaccine antigen. In this study, we evaluated the expression of SpA in a collection of staphylococcal strains, about 7% of which did not express SpA (SpA- strains), despite the presence of the gene. By a comparative genomic analysis, we identified that a mutation in the spa 5′ UTR sequence affecting the RBS is responsible for the loss of SpA in a subset of SpA- strains. Using a high-throughput qRT-PCR approach on a selected panel of virulence-related genes, we identified that the SpA- phenotype is associated with lower spa transcript levels and increased expression and production of capsule as well as other changes in the transcription of several key virulence factors. Our data suggest that the SpA- phenotype has occurred in geographically distinct strains through different molecular mechanisms including both mutation, leading likely to translation alterations, and transcriptional deregulation. Furthermore, we provide evidence that SpA- strains are highly susceptible to phagocytic uptake mediated by anti-capsule antibodies. These data suggest that S. aureus may alter its virulence factor expression pattern as an adaptation to the host or environment. Vaccination strategies targeting both SpA and capsule could therefore result in broader coverage against staphylococcal isolates than SpA alone

Virulence Gene Expression of Staphylococcus aureus in Human Skin

Staphylococcus aureus is the main cause of human skin and soft tissue infections. However, S. aureus pathogenicity within the skin is not fully characterized. Here, we implemented an S. aureus cutaneous infection model using human skin explants and performed a time-course infection to study the gene expression profile of a large panel of virulence-related factors of S. aureus USA300 LAC strain, by high-throughput RT-PCR. We pinpointed the genes that were differentially regulated by the bacteria in the skin tissues and identified 12 virulence factors that were upregulated at all time points assessed. Finally, using confocal microscopy, we show that the expression of alpha-hemolysin by S. aureus varies dependent on the skin niche and that the bacteria preferentially accumulates inside sweat glands and ducts. Taken together, our study gives insights about the pathogenic lifestyle of S. aureus within human skin tissues, which may contribute for the development of anti-S. aureus therapeutic strategies

Prevalence of M75 Streptococcus Pyogenes strains harboring slaa gene in patients affected by pediatric obstructive sleep apnea syndrome in Central Italy

Recently we reported an association between pediatric obstructive sleep apnea syndrome (OSAS) and Group A streptococcus (GAS) sub-acute chronic tonsil colonization. We showed that GAS may contribute to tonsil hyperplasia via a streptolysin O (SLO)-dependent cysteinyl leukotrienes (CysLTs) production, which can trigger T and B cell proliferation. In the present study, we characterized the GAS strains isolated from pediatric OSAS patients in comparison with a panel of age and sex matched GAS strains unrelated to OSAS, but isolated in the same area and during the same period ranging from 2009 to 2013. We found that slaA gene, previously reported to be associated to CysLTs production pathway, was significantly associated to GAS OSAS strains. Moreover, the most numerous group (32%) of the GAS OSAS strains belonged to M75 type, and 6 out of 7 of these strains harbored the slaA gene. Multilocus Sequence Typing (MLST) experiments demonstrated that the clone emm75/ST49/ smeZ, slaA was associated to OSAS cases. In conclusion, we found an association between slaA gene and the GAS OSAS strains, and we showed that the clone emm75/ST49 harboring genes smeZ and slaA was exclusively isolated from patients affected by OSAS, thus suggesting that this genotype might be associated to the pathogenesis of OSAS, although further studies are needed to elucidate the possible role of SlaA in tonsil hypertrophy development

Functionalized carbon nanotubes as immunomodulator systems

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Radiation of the Tnt1 retrotransposon superfamily in three Solanaceae genera

Abstract\ud \ud \ud \ud Background\ud \ud Tnt1 was the first active plant retrotransposon identified in tobacco after nitrate reductase gene disruption. The Tnt1 superfamily comprises elements from Nicotiana (Tnt1 and Tto1) and Lycopersicon (Retrolyc1 and Tlc1) species. The study presented here was conducted to characterise Tnt1-related sequences in 20 wild species of Solanum and five cultivars of Solanum tuberosum.\ud \ud \ud \ud Results\ud \ud Tnt1-related sequences were amplified from total genomic DNA using a PCR-based approach. Purified fragments were cloned and sequenced, and clustering analysis revealed three groups that differ in their U3 region. Using a network approach with a total of 453 non-redundant sequences isolated from Solanum (197), Nicotiana (140) and Lycopersicon (116) species, it is demonstrated that the Tnt1 superfamily can be treated as a population to resolve previous phylogenetic multifurcations. The resulting RNAseH network revealed that sequences group according to the Solanaceae genus, supporting a strong association with the host genome, whereas tracing the U3 region sequence association characterises the modular evolutionary pattern within the Tnt1 superfamily. Within each genus, and irrespective of species, nearly 20% of Tnt1 sequences analysed are identical, indicative of being part of an active copy. The network approach enabled the identification of putative "master" sequences and provided evidence that within a genus these master sequences are associated with distinct U3 regions.\ud \ud \ud \ud Conclusion\ud \ud The results presented here support the hypothesis that the Tnt1 superfamily was present early in the evolution of Solanaceae. The evidence also suggests that the RNAseH region of Tnt1 became fixed at the host genus level whereas, within each genus, propagation was ensured by the diversification of the U3 region. Different selection pressures seemed to have acted on the U3 and RNAseH modules of ancestral Tnt1 elements, probably due to the distinct functions of these regions in the retrotransposon life cycle, resulting in both co evolution and adaptation of the element population with its host.M.E. Manetti is a recipient of a CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) Ph.D. fellowship. We thank Silvia Regina Blanco Ribeiro for sequencing and Myna Nakabashi for technical assistance. Financial support for the work presented here was provided to MAVS from FAPESP (Brazil), CNPq (Brazil) and USP (Brazil).M.E. Manetti is a recipient of a CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) Ph.D. fellowship. We thank Silvia Regina Blanco Ribeiro for sequencing and Myna Nakabashi for technical assistance. Financial support for the work presented here was provided to MAVS from FAPESP (Brazil), CNPq (Brazil) and USP (Brazil)

Protein Array Profiling of Tic Patient Sera Reveals a Broad Range and Enhanced Immune Response against Group A Streptococcus Antigens

The human pathogen Group A Streptococcus (Streptococcus pyogenes, GAS) is widely recognized as a major cause of common pharyngitis as well as of severe invasive diseases and non-suppurative sequelae associated with the existence of GAS antigens eliciting host autoantibodies. It has been proposed that a subset of paediatric disorders characterized by tics and obsessive-compulsive symptoms would exacerbate in association with relapses of GAS-associated pharyngitis. This hypothesis is however still controversial. In the attempt to shed light on the contribution of GAS infections to the onset of neuropsychiatric or behavioral disorders affecting as many as 3% of children and adolescents, we tested the antibody response of tic patient sera to a representative panel of GAS antigens. In particular, 102 recombinant proteins were spotted on nitrocellulose-coated glass slides and probed against 61 sera collected from young patients with typical tic neuropsychiatric symptoms but with no overt GAS infection. Sera from 35 children with neither tic disorder nor overt GAS infection were also analyzed. The protein recognition patterns of these two sera groups were compared with those obtained using 239 sera from children with GAS-associated pharyngitis. This comparative analysis identified 25 antigens recognized by sera of the three patient groups and 21 antigens recognized by tic and pharyngitis sera, but poorly or not recognized by sera from children without tic. Interestingly, these antigens appeared to be, in quantitative terms, more immunogenic in tic than in pharyngitis patients. Additionally, a third group of antigens appeared to be preferentially and specifically recognized by tic sera. These findings provide the first evidence that tic patient sera exhibit immunological profiles typical of individuals who elicited a broad, specific and strong immune response against GAS. This may be relevant in the context of one of the hypothesis proposing that GAS antigen-dependent induction of autoantibodies in susceptible individuals may be involved the occurrence of tic disorders