<Emphasis Type="Italic">Salicornia ramosissima</Emphasis> population dynamics and tolerance of salinity

Field and greenhouse studies have been conducted to clarify aspects of population dynamics and NaCl tolerance of Salicornia ramosissima J. Woods. Two populations, Varela and Verdemilho, were monitored in the field during two consecutive life cycles and aspects of their morphology and density were recorded monthly. In the laboratory seedlings were exposed to different salinity for 10 weeks and growth and mortality rate were recorded weekly. The growth of the populations differed significantly, possibly because of the different salinities of the two sampling sites and/or genetic adaptations of the two populations to the environmental conditions. The absence of a significant correlation between sediment salinity and stem elongation suggested, however, that salinity, alone was not responsible for the differences observed and was possibly associated with other factors, because of nutritional, edaphic, and microclimatic conditions. S. ramosissima did not develop well in conditions of elevated or moderate salinity; its growth was optimum at low salinity. Optimum development of S. ramosissima may, nevertheless, depend on the total number of large seeds in a population seed bank, because of their greater success in germination and germinability under stress conditions than small seeds.     

High frequency one-step gene replacement in Trichoderma reesei. II. Effects of deletions of individual cellulase genes

Four cellulase genes of Trichoderma reesei, cbh1, cbh2, egl1 and egl2, have been replaced by the amdS marker gene. When linear DNA fragments and flanking regions of the corresponding cellulase locus of more than 1 kb were used, the replacement frequencies were high, ranging from 32 to 52%. Deletion of the major cellobiohydrolase 1 gene led to a 2-fold increase in the production of cellobiohydrolase II; however, replacement of the cbh2 gene did not affect the final cellulase levels and deletion of egl1 or egl2, slightly increased production of both cellobiohydrolases. Based on our results, endoglucanase II accounts for most of the endoglucanase activity produced by the hypercellulolytic host strain. Furthermore, loss of the egl2, gene causes a significant drop in the filter paper-hydrolysing activity, indicating that endoglucanase II has an important role in the total hydrolysis of cellulose.     

Bacterial Exposures and Associations with Atopy and Asthma in Children

Background

The increase in prevalence of asthma and atopic diseases in Western countries has been linked to aspects of microbial exposure patterns of people. It remains unclear which microbial aspects contribute to the protective farm effect.

Objective

The objective of this study was to identify bacterial groups associated with prevalence of asthma and atopy, and to quantify indoor exposure to some of these bacterial groups.

Methods

A DNA fingerprinting technique, denaturing gradient gel electrophoresis (DGGE), was applied to mattress dust samples of farm children and control children in the context of the GABRIEL Advanced study. Associations between signals in DGGE and atopy, asthma and other allergic health outcomes were analyzed. Quantitative DNA based assays (qPCR) for four bacterial groups were applied on the dust samples to seek quantitative confirmation of associations indicated in DNA fingerprinting.

Results

Several statistically significant associations between individual bacterial signals and also bacterial diversity in DGGE and health outcomes in children were observed. The majority of these associations showed inverse relationships with atopy, less so with asthma. Also, in a subsequent confirmation study using a quantitative method (qPCR), higher mattress levels of specifically targeted bacterial groups - Mycobacterium spp., Bifidobacteriaceae spp. and two different clusters of Clostridium spp. - were associated with a lower prevalence of atopy.

Conclusion

DNA fingerprinting proved useful in identifying bacterial signals that were associated with atopy in particular. These findings were quantitatively confirmed for selected bacterial groups with a second method. High correlations between the different bacterial exposures impede a clear attribution of protective effects to one specific bacterial group. More diverse bacterial flora in mattress dust may link to microbial exposure patterns that protect against development of atopic diseases.     

Mechanistic studies of electrophilic protease inhibitors of full length hepatic C virus (HCV) NS3

The inhibition mechanism of electrophilic peptide-based protease inhibitors of full-length hepatitis C virus (HCV) NS3 has been investigated by determining the K_i-values for a series of compounds differing in the electrophilicity and acidity of the C-terminal residue at pH-values above and below the pK_a of the catalytic histidine (6.85) and at two different ionic strengths. Electrophilic compounds with a pentafluoroethyl ketone group showed stronger inhibition at pH 8 than pH 6, as expected for a mechanism requiring an unprotonated catalytic histidine. However, the difference was only significant at high ionic strength. In contrast, electrophilic compounds with an acidic C-terminal group or a cyclic P1 residue showed a lower inhibitory effect at pH 8 than at pH 6, inconsistent with a mechanism-based inhibition. Moreover, all electrophilic compounds had an unexpectedly strong inhibition at pH 6, when mechanism-based inhibition is unlikely. The results suggest that for some of the electrophilic compounds the reactive group may not be properly positioned in the active site and that binding of these inhibitors is a result of non-covalent interactions. The nature of these interactions is discussed.     

High mobility group box protein 1 in complex with lipopolysaccharide or IL-1 promotes an increased inflammatory phenotype in synovial fibroblasts

Introduction  

In addition to its direct proinflammatory activity, extracellular high mobility group box protein 1 (HMGB1) can strongly enhance the cytokine response evoked by other proinflammatory molecules, such as lipopolysaccharide (LPS), CpG-DNA and IL-1β, through the formation of complexes. Extracellular HMGB1 is abundant in arthritic joint tissue where it is suggested to promote inflammation as intra-articular injections of HMGB1 induce synovitis in mice and HMGB1 neutralizing therapy suppresses development of experimental arthritis. The aim of this study was to determine whether HMGB1 in complex with LPS, interleukin (IL)-1α or IL-1β has enhancing effects on the production of proinflammatory mediators by rheumatoid arthritis synovial fibroblasts (RASF) and osteoarthritis synovial fibroblasts (OASF). Furthermore, we examined the toll-like receptor (TLR) 4 and IL-1RI requirement for the cytokine-enhancing effects of the investigated HMGB1-ligand complexes.     

Egg quality of Japanese quail depends on layer age and storage time

The aim of the study was to analyse egg quality changes of Japanese quail during the reproductive season and after several days of storage. A significantly higher egg weight was recorded in week 9 than in week 25 and week 31 of age ofbirds. The egg specific gravity value of eggs also decreased with the age of quails. The smallest yolk weight and its percentage proportion in egg mass were observed in week 25. Eggs laid in weeks 9 and 25 were characterised by similar and lower (P < or = 0.05) white weight in comparison to those examined on the last date. The highest white proportion in egg weight was recorded during the 25th week. The value of this trait was similar or smaller on the remaining dates. Similar and lower values of the white index and Haugh units, in comparison with the first date, were determined in eggs laid on weeks 25 and 31, whereas similar and greater eggshell thickness than in the 9th week of age was recorded on the last two dates of examination. Lower (P < or = 0.05) egg weight was observed after 5 and 8 than after 0 (measured on the day of laying) days of storage. Likewise, smaller egg specific gravity was recorded after three consecutive periods of storage in comparison with the eggs examined on the day of laying. Eggs examined after 3, 5 and 8 days of storage, in comparison with those stored for 0 days, were characterised by lower yolk index. Significantly smaller weight and white index than after 0 days were determined in eggs stored, respectively, for 5 and 8 as well as for 3, 5 and 8 days. Similarly, smaller numbers of Haugh units were determined after 3 consecutive storage periods in comparison with the eggs examined on the day of laying. Eggs stored for 0 and 3 days were characterised by a greater eggshell weight than those examined after 5 and 8 days. The weight of Japanese quail eggs decreased significantly from day 5 onwards, whereas the egg specific gravity-- from the 3rd day of storage. Yolk and white quality began to deteriorate already after 3 days of egg storage. It appears, therefore, that egg storage of Japanese quails before hatching lasting more than 3 days in the analysed conditions deteriorated the eggshell quality and internal egg traits.     

Structural analogues of ZAPA as GABAA agonists

Structural analogues of ZAPA, Z-3-[(aminoiminomethyl)thio]prop-2-enoic acid, an isothiouronium analogue of GABA, are potent GABA_A agonists as seen in the isolated guinea-pig ileum and in the facilitation of [³H]diazepam binding to rat brain membranes. Compounds with guanidino or amidine groups replacing the amino functionality of GABA were also found to be active. The highest activity was displayed by the isothiouronium salts in which the conformational flexibility of the molecule is restricted by a Z-substituted carbon–carbon double bond. A series of bis-isothiouronium compounds was prepared from aliphatic ,ω-bis-thioureas as mixtures of E and Z adducts. Maximum GABA_A agonist activity for this series was found with a C₆–C₈ carbon chain, and the results were consistent with an interaction at the GABA_A receptor with only one end of the molecule, rather than the more potent effect expected of a molecule bridging two active sites. GABA_A antagonist/partial agonist activity was observed on the guinea-pig isolated ileum for a number of different analogue types, with the most potent being bis-isothiouronium derivatives. None of the substituted derivatives of ZAPA was as active as ZAPA itself, and maximum GABA_A activity was found in the n-pentyl and n-hexyl analogues.     

Recovery of species richness and conservation of native Atlantic forest trees in the cacao plantations of southern Bahia in Brazil

The Atlantic forests of southern Bahia in Brazil present great species richness and a high degree of endemism. A large part of these native forests were transformed into cacao plantations in an agroforestry system known locally as cabrucas, where native trees were culled and cacao was planted under the shade of remaining trees. The present study analyzed the influence of time of implantation (age) and time of abandonment of management practices on tree species diversity of cabruca plantations to evaluate the capacity for conservation and recovery of species richness of native Atlantic Forest trees in cabrucas. Phytosociological surveys were conducted in five cabrucas with different conditions of age and state of abandonment. All trees, including hemiepiphytes and excluding the cacao plants, with a minimum stem diameter of 10 cm at breast height, were surveyed within a 3-ha sampling area in each plantation. A total of 2514 individual trees belonging to 293 species and 52 families were recorded in the five cabrucas. The Shannon diversity index varied from 3.31 to 4.22 among the cabrucas and was positively correlated with the time of abandonment (r = 0.97). The new cabrucas showed the highest values of estimated total richness (Chao) and the highest proportion of late successional species than the old ones. All areas preserved a very high proportion of native forest species while the three old cabrucas showed a higher proportion of exotic species than the two new ones. Thus the exotic species seem to replace more of the native species in the long run because of management practices and local preferences. The cabrucas presented also a high capacity for the regeneration of tree species richness after abandonment. Simple alterations in management practices could improve the recruitment of late successional species in these areas. Economic incentives may be necessary for the farmers to adopt management practices to retain native species which bring no economic returns.