Botryorhodines A-D, antifungal and cytotoxic depsidones from Botryosphaeria rhodina, an endophyte of the medicinal plant Bidens pilosa

An endophytic fungus (Botryosphaeria rhodina) was isolated from the stems of the medicinal plant Bidens pilosa (Asteraceae) that is known for its anti-inflammatory, antiseptic and antifungal effects. The ethyl acetate extract of the fungal isolate exhibits significant antifungal activity as well as potent cytotoxic and antiproliferative effects against several cancer cell lines. Activity-guided fractionation resulted in the isolation of a complex of four depsidones, botryorhodines A-D and the auxin indole carboxylic acid. Botryorhodine A and B show moderate to weak cytotoxic activities against HeLa cell lines with a CC₅₀ of 96.97 μM and 36.41 μM, respectively. In addition, they also show antifungal activity against a range of pathogenic fungi such as Aspergillus terreus (MIC 26.03 μM for botryorhodine A and 49.70 μM for B) and the plant pathogen Fusarium oxysporum (MIC 191.60 μM for botryorhodine A and 238.80 μM for B). A potential role of the endophyte in modulating fungal populations living within or attacking the host plant is discussed.     

New triterpene saponins from the roots of Acacia macrostachya (Mimosaceae)

Four new oleanane-type saponins, macrostachyaosides A, B, C, and D (1–4) were isolated from the roots of Acacia macrostachya. Their structures were elucidated on the basis of extensive 1D- and 2D-NMR data and HR-ESI-MS analyses. At concentrations of 100 μM of each compounds, none of the tested compounds caused a significant growth reduction against HL60 cells.     

Rearing performances and environmental assessment of sea cage farming in Tunisia using life cycle assessment (LCA) combined with PCA and HCPC

Purpose

The present study aims to understand the influence of rearing practices and the contributions of production phases of fish farming to their environmental impacts and determine which practices and technical characteristics can best improve the farms’ environmental performance. Another objective is to identify the influence of variability in farming practices on the environmental performances of sea cage aquaculture farms of sea bass and sea bream in Tunisia by using principal component analysis (PCA) and hierarchical clustering on principal components (HCPC) methods and then combining the classification with life cycle assessment (LCA).

Methods

The approach consisted of three major steps: (i) of the 24 aquaculture farms in Tunisia, 18 were selected which follow intensive rearing practices in sea cages of European sea bass (Dicentrarchus labrax) and gilthead sea bream (Sparus aurata) and then a typology was developed to classify the studied farms into rearing practice groups using HCPC; (ii) LCA was performed on each aquaculture farm and (iii) mean impacts and contributions of production phases were calculated for each group of farms. Impact categories included acidification, eutrophication, global warming, land occupation, total cumulative energy demand and net primary production use.

Results and discussion

Results revealed high correlation between rearing practices and impacts. The feed-conversion ratio (FCR), water column depth under the cages and cage size had the greatest influence on impact intensity. Rearing practices and fish feed were the greatest contributors to the impacts studied due to the production of fish meal and oil and the low efficiency of feed use, which generated large amounts of nitrogen and phosphorus emissions. It is necessary to optimise the diet formulation and to follow better feeding strategies to lower the FCR and improve farm performance. Water column depth greatly influenced the farms’ environmental performance due to the increase in waste dispersion at deeper depths, while shallow depths resulted in accumulation of organic matter and degradation of water quality. Cage size influences environmental performances of aquaculture farms. Thus, from an environmental viewpoint, decision makers should grant licences for farms in deeper water with larger cages and encourage them to improve their FCRs.

Conclusions

This study is the first attempt to combine the HCPC method and the LCA framework to study the environmental performance of aquacultural activity. The typology developed captures the variability among farms because it considers several farm characteristics in the classification. The LCA demonstrated that technical parameters in need of improvement are related to the technical expertise of farm managers and workers and to the location of the farm.

     

Conserved Amphipathic Helices Mediate Lipid Droplet Targeting of Perilipins 1–3

Perilipins (PLINs) play a key role in energy storage by orchestrating the activity of lipases on the surface of lipid droplets. Failure of this activity results in severe metabolic disease in humans. Unlike all other lipid droplet-associated proteins, PLINs localize almost exclusively to the phospholipid monolayer surrounding the droplet. To understand how they sense and associate with the unique topology of the droplet surface, we studied the localization of human PLINs in Saccharomyces cerevisiae, demonstrating that the targeting mechanism is highly conserved and that 11-mer repeat regions are sufficient for droplet targeting. Mutations designed to disrupt folding of this region into amphipathic helices (AHs) significantly decreased lipid droplet targeting in vivo and in vitro. Finally, we demonstrated a substantial increase in the helicity of this region in the presence of detergent micelles, which was prevented by an AH-disrupting missense mutation. We conclude that highly conserved 11-mer repeat regions of PLINs target lipid droplets by folding into AHs on the droplet surface, thus enabling PLINs to regulate the interface between the hydrophobic lipid core and its surrounding hydrophilic environment.     

Autologous transplantation of CD34+ bone marrow derived mononuclear cells in management of non-reconstructable critical lower limb ischemia

Patients with a decrease in limb perfusion with a potential threat to limb viability manifested by ischemic rest pain, ischemic ulcers, and/or gangrene are considered to have critical limb ischemia (CLI). Because of this generally poor outcome, there is a strong need for attempting any procedure to save the affected limb. The aim of this work is to evaluate the possibility to use stem cell therapy as a treatment option for patients with chronic critical lower limb ischemia with no distal run off. This study includes 20 patients with chronic critical lower limb ischemia with no distal run off who are unsuitable for vascular or endovascular option. These patients underwent stem cell therapy (SCT) by autologous transplantation of bone marrow derived mononuclear cells. 55 % of patients treated with SCT showed improvement of the rest pain after the first month, 60 % continued improvement of the rest pain after 6 months, 75 % after 1 year and 80 % after 2 years and continued without any deterioration till the third year. Limb salvage rate after STC was 80 % after the first year till the end of the second and third years. SCT can result in angiogenesis in patients with no-option CLI, providing a foundation for the application of this therapy to leg ischemia.     

Fat inclusions strongly alter membrane mechanics

Neutral lipids (NLs) are apolar oil molecules synthesized in the endoplasmic reticulum bilayer upon diverse biological stimuli. NLs synthesized are released in the hydrophobic core of the bilayer. At a critical concentration, NLs condense by phase separation and nucleate a lipid droplet (LD). After an LD forms, a fraction of NLs can be present in the bilayer but at a concentration below that of the nucleation. Here, we study whether and how the accumulation of NLs alters a lipid bilayer’s mechanical properties. In synthetic systems, we found that NLs proffer unusual bilayer stretching capacities, especially in the presence of negatively curved phospholipids. This impact becomes spectacular when an LD is contiguous with the bilayer and supplies it with NLs. The tested NLs markedly decrease the bilayer area expansion modulus and significantly increase lysis tension but had opposite effects on membrane bending rigidity. Our data unveil how NL molecules modify overall membrane mechanics, the alteration of which may be linked to pathologies or anticancer treatments targeting NLs.     

Salicylic acid alleviates mercury toxicity by preventing oxidative stress in roots of Medicago sativa

Salicylic acid (SA) as a signal molecule mediates many biotic and environmental stress-induced physiological responses in plants. In this study, we investigated the role of SA in regulating Hg-induced oxidative stress in the roots of alfalfa (Medicago sativa). Plants pretreated with 0.2 mM SA for 12 h and subsequently exposed to 10 μM Hg²⁺ for 24 h displayed attenuated toxicity to the root. The SA-promoted root growth was correlated with decreased lipid peroxidation in root cells. The ameliorating effect of SA was confirmed by the histochemical staining for the detection of loss of membrane integrity in Hg-treated roots. We show that treatment with 0.2 mM SA increased the activity of NADH oxidase, ascorbate peroxidase (APX) and peroxidase (POD) in the roots exposed Hg. However, a slightly decreased superoxide dismutase (SOD) activity was observed in SA + Hg-treated roots when compared to those of Hg treatment alone. We also measured accumulation of ascorbate (ASC), glutathione (GSH) and proline in the roots of alfalfa and found that roots treated with SA in the presence of Hg accumulated more ASC, GSH and proline than those treated with Hg only. These results suggest that exogenous SA may improve the tolerance of the plant to the Hg toxicity.     

Elimination of active trachoma after two topical mass treatments with azithromycin 1.5% eye drops

Background

Following an epidemiological study carried out in 2006 showing a high prevalence of blinding trachoma in the Far North Region of Cameroon, a trachoma elimination programme using the SAFE strategy was initiated: three yearly trachoma mass treatments were to be performed.

Methodology/Principal Findings

The entire district population (120,000 persons) was treated with azithromycin 1.5% eye drops in February 2008 and January 2009. To assess the effect of treatment on the prevalence of active trachoma, three epidemiological studies were conducted on a representative sample of children aged between 1 and 10 years. The first study was performed just prior to the first treatment, the second just prior to the 2nd treatment and the third one, one year later. The prevalence of active forms of trachoma (TF + TI) dropped from 31.5% (95%CI 26.4–37.5) before treatment to 6.3% (95%CI 4.1–9.6) one year after first treatment; a reduction of nearly 80%. One year after the second treatment, the prevalence decreased to 3.1% (95%CI 2.0–4.9), a total reduction of 90%. Furthermore, there were no more TI cases (only TF). There was no report of serious or systemic side effects. Tolerance was excellent.

Conclusions/Significance

Active trachoma mass treatment with azithromycin 1.5% eye drops is feasible, well tolerated, and effective.     

A rhamnogalacturonan lyase in the Clostridium cellulolyticum cellulosome

Clostridium cellulolyticum secretes large multienzymatic complexes with plant cell wall-degrading activities named cellulosomes. Most of the genes encoding cellulosomal components are located in a large gene cluster: cipC-cel48F-cel8C-cel9G-cel9E-orfX-cel9H-cel9J-man5K-cel9M. Downstream of the cel9M gene, a new open reading frame was discovered and named rgl11Y. Amino acid sequence analysis indicates that this gene encodes a multidomain pectinase, Rgl11Y, containing an N-terminal signal sequence, a catalytic domain belonging to family 11 of the polysaccharide lyases, and a C-terminal dockerin domain. The present report describes the biochemical characterization of a recombinant form of Rgl11Y. Rgl11Y cleaves the alpha-L-Rhap-(1-->4)-alpha-D-GalpA glycosidic bond in the backbone of rhamnogalacturonan I (RGI) via a beta-elimination mechanism. Its specific activity on potato pectic galactan and rhamnogalacturonan was found to be 28 and 3.6 IU/mg, respectively, indicating that Rgl11Y requires galactan decoration of the RGI backbone. The optimal pH of Rgl11Y is 8.5 and calcium is required for its activity. Rgl11Y was shown to be incorporated in the C. cellulolyticum cellulosome through a typical cohesin-dockerin interaction. Rgl11Y from C. cellulolyticum is the first cellulosomal rhamnogalacturonase characterized.