The antimicrobial activity of lapachol and its thiosemicarbazone and semicarbazone derivatives

Lapachol was chemically modified to obtain its thiosemicarbazone and semicarbazone derivatives. These compounds were tested for antimicrobial activity against several bacteria and fungi by the broth microdilution method. The thiosemicarbazone and semicarbazone derivatives of lapachol exhibited antimicrobial activity against the bacteria Enterococcus faecalis and Staphylococcus aureus with minimal inhibitory concentrations (MICs) of 0.05 and 0.10 µmol/mL, respectively. The thiosemicarbazone and semicarbazone derivatives were also active against the pathogenic yeast Cryptococcus gattii (MICs of 0.10 and 0.20 µmol/mL, respectively). In addition, the lapachol thiosemicarbazone derivative was active against 11 clinical isolates of Paracoccidioides brasiliensis, with MICs ranging from 0.01-0.10 µmol/mL. The lapachol-derived thiosemicarbazone was not cytotoxic to normal cells at the concentrations that were active against fungi and bacteria. We synthesised, for the first time, thiosemicarbazone and semicarbazone derivatives of lapachol. The MICs for the lapachol-derived thiosemicarbazone against S. aureus, E. faecalis, C. gattii and several isolates of P. brasiliensis indicated that this compound has the potential to be developed into novel drugs to treat infections caused these microbes.     

The novel Fh8 and H fusion partners for soluble protein expression in Escherichia coli: a comparison with the traditional gene fusion technology

The Escherichia coli host system is an advantageous choice for simple and inexpensive recombinant protein production but it still presents bottlenecks at expressing soluble proteins from other organisms. Several efforts have been taken to overcome E. coli limitations, including the use of fusion partners that improve protein expression and solubility. New fusion technologies are emerging to complement the traditional solutions. This work evaluates two novel fusion partners, the Fh8 tag (8 kDa) and the H tag (1 kDa), as solubility enhancing tags in E. coli and their comparison to commonly used fusion partners. A broad range comparison was conducted in a small-scale screening and subsequently scaled-up. Six difficult-to-express target proteins (RVS167, SPO14, YPK1, YPK2, Frutalin and CP12) were fused to eight fusion tags (His, Trx, GST, MBP, NusA, SUMO, H and Fh8). The resulting protein expression and solubility levels were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis before and after protein purification and after tag removal. The Fh8 partner improved protein expression and solubility as the well-known Trx, NusA or MBP fusion partners. The H partner did not function as a solubility tag. Cleaved proteins from Fh8 fusions were soluble and obtained in similar or higher amounts than proteins from the cleavage of other partners as Trx, NusA or MBP. The Fh8 fusion tag therefore acts as an effective solubility enhancer, and its low molecular weight potentially gives it an advantage over larger solubility tags by offering a more reliable assessment of the target protein solubility when expressed as a fusion protein.     

Effect of decomposing post-fire coarse woody debris on soil fertility and nutrient availability in a Mediterranean ecosystem

Post-fire coarse woody debris can represent a valuable nutrient reservoir for a regenerating ecosystem, helping to prevent soil fertility losses after a wildfire. However, there is scarce information on its effect on soil nutrient cycling and availability. We established three study sites along an altitudinal gradient in a burnt pine forest (SE Spain). At each site we determined: (1) decomposition rates and nutrient dynamics in charred logs left on the ground, 2 and 4 years after the fire, and (2) available nutrients in the soil and in the microbial fraction below charred logs and in bare soil areas. Despite the relatively slow decay rates in this Mediterranean climate (ca. 10 % of dry weight lost after 4 years), N and P were progressively released by logs, accounting for ca. 40 and 65 % of the initial content respectively after 4 years. This implies that the total aboveground biomass of the burnt forest released around 20 kg ha^?1 of N and 2 kg ha^?1 of P during this period. The presence of post fire coarse woody debris consistently increased soil organic matter by around 18 %, total C and N by 42 and 26 %, respectively, dissolved organic C and N by 47 %, available inorganic P by 68 %, and microbial biomass and nitrogen by some 36 and 48 %, respectively. By contrast, soil bulk density decreased by ca. 18 % under logs compared to bare areas. Thus, the fire-killed wood was useful in the recovery of soil fertility and nutrient availability. Leaving the post-fire woody debris on site can enhance the biogeochemical sustainability, microbiological processes and soil ecological functioning. The detrimental effect of post-fire salvage logging on soil fertility should be therefore considered when making management decisions.     

Abscisic acid mediates hydrogen peroxide production in peanut induced by water stress

Peanut plants exposed to water stress induced by polyethylene glycol (PEG) accumulated abscisic acid (ABA) and hydrogen peroxide (H₂O₂), the increase being significant at 12 and 24 h after addition, respectively. To address the question whether the increase in H₂O₂ production was related to ABA accumulation, the peanut leaves were pretreated with ABA biosynthesis inhibitor (sodium tungstate) and then exposed to water stress. Under these conditions, a decrease of ABA and H₂O₂ content were found after 12 h. The addition of 100 μM ABA restored H₂O₂ content reaching values similar to those under water stress at 12 h. We concluded that ABA accumulation is the first signal that triggers the H₂O₂ generation in peanut during first 12 h but its subsequent production is partially ABA-independent.     

Population Composition and Ectoparasite Prevalence on Bats (Sturnira ludovici; Phyllostomidae) in Forest Fragments and Coffee Plantations of Central Veracruz,Mexico

Studies comparing the abundance of frugivorous bats in shade‐coffee plantations and forest fragments report contradictory results, and have not taken into account the landscape context in which coffee plantations are immersed. Variables of population composition such as abundance, sex proportion, and reproductive condition, together with biological tags (i.e., bat fly prevalence), can provide information about spatiotemporal dynamics of habitats used by bats. In the central part of Veracruz, Mexico, we compared population variables and ectoparasite prevalence of the highland yellow‐shouldered bat (Sturnira ludovici) in two landscapes, one dominated by shade‐coffee plantations and another by forest fragments. Comparing these attributes between these two landscapes will increase our knowledge about the role of this agro‐ecosystem in the conservation of this species, which is an important seed disperser of cloud forest vegetation. Total abundance and proportion of females was greater in forest fragments than in coffee plantations, whereas the percentage of reproductive females and bat fly prevalence was similar between landscapes. Our results show that landscapes with forest fragments harbor the greatest abundance of S. ludovici, but shade‐coffee plantations also are utilized by S. ludovici and likely adjacent forest remnants provide enough food resources for this species and other frugivores. Moreover, this study provides more evidence documenting the importance of preserving the last cloud forest fragments in the central region of Veracruz, Mexico, and suggests that using shade‐coffee plantations to connect forest fragments may be an effective way of maintaining populations of S. ludovici and likely other volant frugivores.     

Spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis (Decapoda: Dendrobranchiata)

Braga, A.L., Nakayama, C.L., Suita de Castro, L.A. and Wasielesky, W. 2011. Spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis (Decapoda: Dendrobranchiata). —Acta Zoologica (Stockholm) 00 : 1–6. The spermatozoa ultrastructure of the pink shrimp Farfantepenaeus paulensis was investigated in this morphological study. Spermatophores and spermatozoa were analyzed by electron microscopy. The pink shrimp spermatophore is divided into two regions: the appendage and the spermatophore main body, where spermatozoa are grouped in a spermatic mass. Pink shrimp spermatozoa are unistellate and are composed of main body and single spike. The spermatozoa body comprises a perinuclear cytoplasmic band, nucleus, acrosomal cap, and subacrosomal region. The spermatozoa cell mean total length was 10.71 μm, the mean body diameter was 5.56 μm, and the mean spike length and diameter were 5.15 μm and 0.85 μm, respectively.     

Identification of the rhizobial symbiont of Astragalus glombiformis in Eastern Morocco as Mesorhizobium camelthorni

Astragalus gombiformis is a desert symbiotic nitrogen-fixing legume of great nutritional value as fodder for camels and goats. However, there are no data published on the rhizobial bacteria that nodulate this wild legume in northern Africa. Thirty-four rhizobial bacteria were isolated from root nodules of A. gombifomis grown in sandy soils of the South-Eastern of Morocco. Twenty-five isolates were able to renodulate their original host and possessed a nodC gene copy. The phenotypic and genotypic characterizations carried out illustrated the diversity of the isolates. Phenotypic analysis showed that isolates used a great number of carbohydrates as sole carbon source. However, although they were isolated from arid sandy soils, the isolates do not tolerate drought stress applied in vitro. The phenotypic diversity corresponded mainly to the diversity in the use of some carbohydrates. The genetic analysis as assessed by repetitive extragenic palindromic (REP)-polymerase chain reaction (PCR) showed that the isolates clustered into 3 groups at a similarity coefficient of 81 %. The nearly-complete 16S rRNA gene sequence from a representative strain of each PCR-group showed they were closely related to members of the genus Mesorhizobium of the family Phyllobactericeae within the Alphaproteobacteria. Sequencing of the housekeeping genes atpD, glnII and recA, and their concatenated phylogenetic analysis, showed they are closely related to Mesorhizobium camelthorni. Sequencing of the symbiotic nodC gene from each strain revealed they had 83.53 % identity with the nodC sequence of the type strain M. camelthorni CCNWXJ 40-4^T.     

2-(1H-Pyrazol-4-yl)acetic acids as CRTh2 antagonists

High throughput screening identified the pyrazole-4-acetic acid substructure as CRTh2 receptor antagonists. Optimisation of the compounds uncovered a tight SAR but also identified some low nanomolar inhibitors.