Are gardens effective in butterfly conservation? A case study with the pipevine swallowtail, <Emphasis Type="Italic">Battus philenor</Emphasis>

Gardens with nectar sources and larval host plants have been proposed to stem the decline in butterfly abundance caused by habitat loss. However, no study has provided evidence that gardens benefit butterflies. We examined the use of natural sites and gardens in the San Francisco bay area by the butterfly, Battus philenor. We found that natural sites were more likely to attract adult B. philenor, received more oviposition, and had higher juvenile survival than gardens sites. Butterflies were more likely to be present in gardens with established populations of the host plant, Aristolochia californica, growing in the sun. Battus philenor are unlikely to visit gardens with host plants planted within the past 7 years. Gardens between the ages of 8–40 years received oviposition, but did not always support completion of larval development of B. philenor. In gardens with host plants over 40 years of age, B. philenor consistently survived from egg to the adult stage. Natural enemy induced mortality of eggs did not differ between garden and natural sites, but overall egg survival was lower in gardens than at natural sites. It is unlikely that gardens serve as 'refugia' for B. philenor in years when populations in natural sites experience low survival or low fecundity. Even in gardens capable of supporting larvae to maturity, the density of eggs and survival rates were lower than in natural populations of the host plant suggesting that gardens were not optimal habitats. Therefore, without evidence that juvenile abundance and survival rates in gardens matches or exceeds that in natural sites, it is most likely that gardens act as population sinks for B. philenor.     

蓝藻病毒(噬藻体)的研究进展

蓝藻(Bule-green algae)是一类原核生物,具有细菌的一些特征,因此又常称为蓝细菌(Cyanobacterium),相应地,把感染蓝细菌的病毒称为噬藻体(Cyanophage)[1～2],这是由于噬藻体与噬菌体非常相似的缘故.除蓝藻外,所有其它的藻类均是真核生物,通常将感染真核藻类的病毒称作"藻病毒"(Phycovirus)[3],它们的绝大多数是多角体的粒子(Polyhedral particles),只有个别如珊瑚轮藻(Chara corallina)病毒是杆状的[4].真核藻类病毒和病毒类粒子(VLPs)前文有过综述[4],蓝藻病毒或噬藻体则完全不同于真核藻类的病毒,二者是藻类病毒的重要组成部分,蓝藻病毒的研究情况有必要专门介绍.     

<Emphasis Type="Italic">Ochrobactrum</Emphasis> sp. MPV1 from a dump of roasted pyrites can be exploited as bacterial catalyst for the biogenesis of selenium and tellurium nanoparticles

Background

Bacteria have developed different mechanisms for the transformation of metalloid oxyanions to non-toxic chemical forms. A number of bacterial isolates so far obtained in axenic culture has shown the ability to bioreduce selenite and tellurite to the elemental state in different conditions along with the formation of nanoparticles—both inside and outside the cells—characterized by a variety of morphological features. This reductive process can be considered of major importance for two reasons: firstly, toxic and soluble (i.e. bioavailable) compounds such as selenite and tellurite are converted to a less toxic chemical forms (i.e. zero valent state); secondly, chalcogen nanoparticles have attracted great interest due to their photoelectric and semiconducting properties. In addition, their exploitation as antimicrobial agents is currently becoming an area of intensive research in medical sciences.

Results

In the present study, the bacterial strain Ochrobactrum sp. MPV1, isolated from a dump of roasted arsenopyrites as residues of a formerly sulfuric acid production near Scarlino (Tuscany, Italy) was analyzed for its capability of efficaciously bioreducing the chalcogen oxyanions selenite (SeO₃ ^2?) and tellurite (TeO₃ ^2?) to their respective elemental forms (Se⁰ and Te⁰) in aerobic conditions, with generation of Se- and Te-nanoparticles (Se- and TeNPs). The isolate could bioconvert 2 mM SeO₃ ^2? and 0.5 mM TeO₃ ^2? to the corresponding Se⁰ and Te⁰ in 48 and 120 h, respectively. The intracellular accumulation of nanomaterials was demonstrated through electron microscopy. Moreover, several analyses were performed to shed light on the mechanisms involved in SeO₃ ^2? and TeO₃ ^2? bioreduction to their elemental states. Results obtained suggested that these oxyanions are bioconverted through two different mechanisms in Ochrobactrum sp. MPV1. Glutathione (GSH) seemed to play a key role in SeO₃ ^2? bioreduction, while TeO₃ ^2? bioconversion could be ascribed to the catalytic activity of intracellular NADH-dependent oxidoreductases. The organic coating surrounding biogenic Se- and TeNPs was also characterized through Fourier-transform infrared spectroscopy. This analysis revealed interesting differences among the NPs produced by Ochrobactrum sp. MPV1 and suggested a possible different role of phospholipids and proteins in both biosynthesis and stabilization of such chalcogen-NPs.

Conclusions

In conclusion, Ochrobactrum sp. MPV1 has demonstrated to be an ideal candidate for the bioconversion of toxic oxyanions such as selenite and tellurite to their respective elemental forms, producing intracellular Se- and TeNPs possibly exploitable in biomedical and industrial applications.

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Lactobacillli expressing llama VHH fragments neutralise <Emphasis Type="Italic">Lactococcus</Emphasis>phages

Background  

Bacteriophages infecting lactic acid bacteria (LAB) are widely acknowledged as the main cause of milk fermentation failures. In this study, we describe the surface-expression as well as the secretion of two functional llama heavy-chain antibody fragments, one binding to the major capsid protein (MCP) and the other to the receptor-binding proteins (RBP) of the lactococcal bacteriophage p2, by lactobacilli in order to neutralise lactococcal phages.     

Inhibition of fatty acid synthase suppresses U-2 OS cell invasion and migration via downregulating the activity of HER2/PI3K/AKT signaling pathway in vitro

FASN plays an important role in the malignant phenotype of various tumors. Our previous studies show that inhibition FASN could induce apoptosis and inhibit proliferation in human osteosarcoma (OS) cell in vivo and vitro. The aim in this study was to investigate the effect of inhibition FASN on the activity of HER2/PI3K/AKT axis and invasion and migration of OS cell. The expression of FASN, HER2 and p-HER2(Y1248) proteins was detected by immunohistochemistry in OS tissues from 24 patients with pulmonary metastatic disease, and the relationship between FASN and p-HER2 as well as HER2 was investigated. The results showed that there was a positive correlation between FASN and HER2 as well as p-HER2 protein expression. The U-2 OS cells were transfected with either the FASN specific RNAi plasmid or the negative control RNAi plasmid. FASN mRNA was measured by RT-PCR. Western blot assays was performed to examine the protein expression of FASN, HER2, p-HER2(Y1248), PI3K, Akt and p-Akt (Ser473). Migration and invasion of cells were investigated by wound healing and transwell invasion assays. The results showed that the activity of HER2/PI3K/AKT signaling pathway was suppressed by inhibiting FASN. Meanwhile, the U-2OS cells migration and invasion were also impaired by inhibiting the activity of FASN/HER2/PI3K/AKT. Our results indicated that inhibition of FASN suppresses OS cell invasion and migration via down-regulation of the “HER2/PI3K/AKT” axis in vitro. FASN blocker may be a new therapeutic strategy in OS management.     

Enhanced cellulase production by <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> by cultivation on glycerol followed by induction on cellulosic substrates

The use of glycerol obtained as an intermediate of the biodiesel manufacturing process as carbon source for microbial growth is a potential alternative strategy for the production of enzymes and other high-value bioproducts. This work evaluates the production of cellulase enzymes using glycerol for high cell density growth of Trichoderma harzianum followed by induction with a cellulosic material. Firstly, the influence of the carbon source used in the pre-culture step was investigated in terms of total protein secretion and fungal morphology. Enzymatic productivity was then determined for cultivation strategies using different types and concentrations of carbon source, as well as different feeding procedures (batch and fed-batch). The best strategy for cellulase production was then further studied on a larger scale using a stirred tank bioreactor. The proposed strategy for cellulase production, using glycerol to achieve high cell density growth followed by induction with pretreated sugarcane bagasse, achieved enzymatic activities up to 2.27 ± 0.37 FPU/mL, 106.40 ± 8.87 IU/mL, and 9.04 ± 0.39 IU/mL of cellulase, xylanase, and β-glucosidase, respectively. These values were 2 times higher when compared to the control experiments using glucose instead of glycerol. This novel strategy proved to be a promising approach for improving cellulolytic enzymes production, and could potentially contribute to adding value to biomass within the biofuels sector.     

<Emphasis Type="Italic">Trichomonas vaginalis</Emphasis> metalloproteinase TvMP50 is a monomeric Aminopeptidase P-like enzyme

Previously, metalloproteinase was isolated and identified from Trichomonas vaginalis, belonging to the aminopeptidase P-like metalloproteinase subfamily A/B, family M24 of clan MG, named TvMP50. The native and recombinant TvMP50 showed proteolytic activity, determined by gelatin zymogram, and a 50 kDa band, suggesting that TvMP50 is a monomeric active enzyme. This was an unexpected finding since other Xaa-Pro aminopeptidases/prolidases are active as a biological unit formed by dimers/tetramers. In this study, the evolutionary history of TvMP50 and the preliminary crystal structure of the recombinant enzyme determined at 3.4 Å resolution is reported. TvMP50 was shown to be a type of putative, eukaryotic, monomeric aminopeptidase P, and the crystallographic coordinates showed a monomer on a “pseudo-homodimer” array on the asymmetric unit that resembles the quaternary structure of the M24B dimeric family and suggests a homodimeric aminopeptidase P-like enzyme as a likely ancestor. Interestingly, TvMP50 had a modified N-terminal region compared with other Xaa-Pro aminopeptidases/prolidases with three-dimensional structures; however, the formation of the standard dimer is structurally unstable in aqueous solution, and a comparably reduced number of hydrogen bridges and lack of saline bridges were found between subunits A/B, which could explain why TvMP50 portrays monomeric functionality. Additionally, we found that the Parabasalia group contains two protein lineages with a “pita bread” fold; the ancestral monomeric group 1 was probably derived from an ancestral dimeric aminopeptidase P-type enzyme, and group 2 has a probable dimeric kind of ancestral eukaryotic prolidase lineage. The implications of such hypotheses are also presented.     

Znf179 E3 ligase-mediated TDP-43 polyubiquitination is involved in TDP-43- ubiquitinated inclusions (UBI) (+)-related neurodegenerative pathology

Background

The brain predominantly expressed RING finger protein, Znf179, is known to be important for embryonic neuronal differentiation during brain development. Downregulation of Znf179 has been observed in motor neurons of adult mouse models for amyotrophic lateral sclerosis (ALS), yet the molecular function of Znf179 in neurodegeneration has never been previously described. Znf179 contains the classical C3HC4 RING finger domain, and numerous proteins containing C3HC4 RING finger domain act as E3 ubiquitin ligases. Hence, we are interested to identify whether Znf179 possesses E3 ligase activity and its role in ALS neuropathy.

Methods

We used in vivo and in vitro ubiquitination assay to examine the E3 ligase autoubiquitination activity of Znf179 and its effect on 26S proteasome activity. To search for the candidate substrates of Znf179, we immunoprecipitated Znf179 and subjected to mass spectrometry (MS) analysis to identify its interacting proteins. We found that ALS/ FTLD-U (frontotemporal lobar degeneration (FTLD) with ubiquitin inclusions)-related neurodegenerative TDP-43 protein is the E3 ligase substrate of Znf179. To further clarify the role of E3 ubiquitin ligase Znf179 in neurodegenerative TDP-43-UBI (ubiquitinated inclusions) (+) proteinopathy, the effect of Znf179-mediated TDP-43 polyubiquitination on TDP-43 protein stability, aggregate formation and nucleus/cytoplasm mislocalization were evaluated in vitro cell culture system and in vivo animal model.

Results

Here we report that Znf179 is a RING E3 ubiquitin ligase which possesses autoubiquitination feature and regulates 26S proteasome activity through modulating the protein expression levels of 19S/20S proteasome subunits. Our immunoprecipitation assay and MS analysis results revealed that the neuropathological TDP-43 protein is one of its E3 ligase substrate. Znf179 interactes with TDP-43 protein and mediates polyubiquitination of TDP-43 in vitro and in vivo. In neurodegenerative TDP-43 proteinopathy, we found that Znf179-mediated polyubiquitination of TDP-43 accelerates its protein turnover rate and attenuates insoluble pathologic TDP-43 aggregates, while knockout of Znf179 in mouse brain results in accumulation of insoluble TDP-43 and cytosolic TDP-43 inclusions in cortex, hippocampus and midbrain regions.

Conclusions

Here we unveil the important role for the novel E3 ligase Znf179 in TDP-43-mediated neuropathy, and provide a potential therapeutic strategy for combating ALS/ FTLD-U neurodegenerative pathologies.

     

Opening myrmecochory’s black box: what happens inside the ant nest?

In the process of seed dispersal by ants (myrmecochory), foragers bring diaspores back to their nest, then eat the elaiosome and usually reject viable seeds outside the nest. Here, we investigate what happens inside the nest, a barely known stage of the myrmecochory process, for two seed species (Viola odorata, Chelidonium majus) dispersed either by the insectivorous ant Myrmica rubra or by the aphid-tending ant Lasius niger. Globally, elaiosome detachment decreased ants’ interest towards seeds and increased their probability of rejecting them. However, we found marked differences in seed management by ants inside the nest. The dynamics of elaiosome detachment were ant- and plant-specific whereas the dynamic of seed rejection were mainly ant-specific. Seeds remained for a shorter period of time inside the nest of the carnivorous ant Myrmica rubra than in Lasius niger nest. Thus, elaiosome detachment and seed rejection were two competing dynamics whose relative efficiency leads to variable outcomes in terms of types of dispersed items and of nutrient benefit to the ants. This is why some seeds remained inside the nest even without an elaiosome, and conversely, some seeds were rejected with an elaiosome still attached. Fresh seeds may be deposited directly in contact with the larvae. However, the dynamics of larvae-seeds contacts were also highly variable among species. This study illustrates the complexity and variability of the ecological network of ant–seed interactions.