UV-B light contributes directly to the synthesis of chiloglottone floral volatiles

Background and Aims Australian sexually deceptive Chiloglottis orchids attract their specific male wasp pollinators by means of 2,5-dialkylcyclohexane-1,3-diones or ‘chiloglottones’, representing a newly discovered class of volatiles with unique structures. This study investigated the hypothesis that UV-B light at low intensities is directly required for chiloglottone biosynthesis in Chiloglottis trapeziformis.Methods Chiloglottone production occurs only in specific tissue (the callus) of the labellum. Cut buds and flowers, and whole plants with buds and flowers, sourced from the field, were kept in a growth chamber and interactions between growth stage of the flowers and duration and intensity of UV-B exposure on chiloglottone production were studied. The effects of the protein synthesis inhibitor cycloheximide were also examined.Key Results Chiloglottone was not present in buds, but was detected in buds that were manually opened and then exposed to sunlight, or artificial UV-B light for ≥5 min. Spectrophotometry revealed that the sepals and petals blocked UV-B light from reaching the labellum inside the bud. Rates of chiloglottone production increased with developmental stage, increasing exposure time and increasing UV-B irradiance intensity. Cycloheximide did not inhibit the initial production of chiloglottone within 5 min of UV-B exposure. However, inhibition of chiloglottone production by cycloheximide occurred over 2 h of UV-B exposure, indicating a requirement for de novo protein synthesis to sustain chiloglottone production under UV-B.Conclusions The sepals and petals of Chiloglottis orchids strongly block UV-B wavelengths of light, preventing chiloglottone production inside the bud. While initiation of chiloglottone biosynthesis requires only UV-B light, sustained chiloglottone biosynthesis requires both UV-B and de novo protein biosynthesis. The internal amounts of chiloglottone in a flower reflect the interplay between developmental stage, duration and intensity of UV-B exposure, de novo protein synthesis, and feedback loops linked to the starting amount of chiloglottone. It is concluded that UV-B light contributes directly to chiloglottone biosynthesis. These findings suggest an entirely new and unexpected biochemical reaction that might also occur in taxa other than these orchids.     

Genomic analysis identifies candidate pathogenic variants in 9 of 18 patients with unexplained West syndrome

West syndrome, which is narrowly defined as infantile spasms that occur in clusters and hypsarrhythmia on EEG, is the most common early-onset epileptic encephalopathy (EOEE). Patients with West syndrome may have clear etiologies, including perinatal events, infections, gross chromosomal abnormalities, or cases followed by other EOEEs. However, the genetic etiology of most cases of West syndrome remains unexplained. DNA from 18 patients with unexplained West syndrome was subjected to microarray-based comparative genomic hybridization (array CGH), followed by trio-based whole-exome sequencing in 14 unsolved families. We identified candidate pathogenic variants in 50 % of the patients (n = 9/18). The array CGH revealed candidate pathogenic copy number variations in four cases (22 %, 4/18), including an Xq28 duplication, a 16p11.2 deletion, a 16p13.1 deletion and a 19p13.2 deletion disrupting CACNA1A. Whole-exome sequencing identified candidate mutations in known epilepsy genes in five cases (36 %, 5/14). Three candidate de novo mutations were identified in three cases, with two mutations occurring in two new candidate genes (NR2F1 and CACNA2D1) (21 %, 3/14). Hemizygous candidate mutations in ALG13 and BRWD3 were identified in the other two cases (14 %, 2/14). Evaluating a panel of 67 known EOEE genes failed to identify significant mutations. Despite the heterogeneity of unexplained West syndrome, the combination of array CGH and whole-exome sequencing is an effective means of evaluating the genetic background in unexplained West syndrome. We provide additional evidence for NR2F1 as a causative gene and for CACNA2D1 and BRWD3 as candidate genes for West syndrome.     

Molecular dynamics simulation of the melting processes of core–shell and pure nanoparticles

Core–shell nanoparticles are nanosized particles that consist of a core and a shell, constructed from different metallic elements. Core–shell nanoparticles have received extensive attention, owing to their various potential applications such as paints, optical films and catalysts. Herein, we investigate the melting behaviours of different core–shell nanoparticles under continuous heating using molecular dynamics simulation. Different metallic elements were examined as core–shell and pure nanoparticles. Five different processes were observed during the melting of core–shell nanoparticles. In contrast, only one process was identified during the melting of pure nanoparticles. These processes were influenced by the nanoparticle size, shell thickness and differences between the lattice constants and melting point temperatures of the metallic elements. Our simulation provides microscopic insights into the melting behaviours of existing and proposed core–shell nanoparticles that would be highly beneficial towards the fabrication of materials with different chemical coatings.     

Evaluation of glyphosate application in regulating the reproduction of riparian black locust (Robinia pseudoacacia L.) after clear-cutting,and the possibility of leaching into soil

Black locust (Robinia pseudoacacia L.)—an invasive alien species in riparian forests—is becoming more prevalent in many rivers of eastern Japan. Riparian black locust forests are typically cut down to maintain river-flow capacity. However, such forests often reproduce rapidly by stump sprouting and root suckering and regenerate by germination. Thus, more effective riparian forest management approaches are required. To regulate the reproduction of black locust forests after clear-cutting, we examined the regrowth-inhibiting effects of glyphosate herbicide application to stumps, in accordance with current river management protocol (i.e., winter logging operation). Further, we investigated the concentrations of glyphosate leaching into the soil at a depth of 30 cm in a riparian area of the Tenryu River. Our results showed that glyphosate application to stumps completely inhibited stump sprouting but not root suckering or seedling germination. The glyphosate concentration leaching into the soil reached a maximum (2.6 ± 0.7 mg kg^?1, mean ± standard error) on day 1 after the application, and subsequently declined to below the detection limit on day 2. Thus, the rapid degradation of glyphosate was confirmed, despite the fact that the herbicide leached into the soil after application to the stumps. The glyphosate application has limited effectiveness against root suckering and germination of riparian black locust forests after clear-cutting in winter, in accordance with the current river management protocol.     

Backbone 1H, 13C,and 15N assignments of yeast Ump1, an intrinsically disordered protein that functions as a proteasome assembly chaperone

Eukaryotic proteasome assembly is a highly organized process mediated by several proteasome-specific chaperones, which interact with proteasome assembly intermediates. In yeast, Ump1 and Pba1-4 have been identified as assembly chaperones that are dedicated to the formation of the proteasome 20S catalytic core complex. The crystal structures of Pba chaperones have been reported previously, but no detailed information has been provided for the structure of Ump1. Thus, to better understand the mechanisms underlying Ump1-mediated proteasome assembly, we characterized the conformation of Ump1 in solution using NMR. Backbone chemical shift data indicated that Ump1 is an intrinsically unstructured protein and largely devoid of secondary structural elements.     

Environmental radioactivity damages the DNA of earthworms of Fukushima Prefecture,Japan

Radioactive materials released in 2011 during the accident at Japan's Fukushima nuclear power plant are expected to become concentrated in wildlife via the food web, triggering DNA damage. However, the extent of this wildlife DNA damage remains unknown. We explored the effects of exposure to environmental radiation on the DNA of wild boars (Sus scrofa leucomystax) and earthworms (Megascolecidae) and examined the modes of transfer of radioactivity to these animals. We used a comet assay to analyze the DNA damage in animals collected from regions with low (0.28 μSv/h) and high (2.85 μSv/h) levels of atmospheric radiation. We constructed a model food web featuring the wild boar as the top predator, and we measured the radioactivity levels in soil, plant material, earthworms, and wild boar. The extent of DNA damage in wild boars did not differ significantly between animals captured in the two regions, but earthworms from the “high-dose” region had a significantly greater extent of DNA damage than did those from the “low-dose” region. Radioactive material can be accumulated either directly from the soil or indirectly via plant ingestion. Wildlife inhabiting radioactively contaminated areas is at high risk of DNA damage although the risk varies among species because of the differences in foraging habits. The long-term results of the observed DNA damage must be studied.     

Stereoselectivity of Isoflurane in Adhesion Molecule Leukocyte Function-Associated Antigen-1

Background

Isoflurane in clinical use is a racemate of S- and R-isoflurane. Previous studies have demonstrated that the effects of S-isoflurane on relevant anesthetic targets might be modestly stronger (less than 2-fold) than R-isoflurane. The X-ray crystallographic structure of the immunological target, leukocyte function-associated antigen-1 (LFA-1) with racemic isoflurane suggested that only S-isoflurane bound specifically to this protein. If so, the use of specific isoflurane enantiomers may have advantage in the surgical settings where a wide range of inflammatory responses is expected to occur. Here, we have further tested the hypothesis that isoflurane enantioselectivity is apparent in solution binding and functional studies.

Methods

First, binding of isoflurane enantiomers to LFA-1 was studied using 1-aminoanthracene (1-AMA) displacement assays. The binding site of each enantiomer on LFA-1 was studied using the docking program GLIDE. Functional studies employed the flow-cytometry based ICAM binding assay.

Results

Both enantiomers decreased 1-AMA fluorescence signal (at 520 nm), indicating that both competed with 1-AMA and bound to the αL I domain. The docking simulation demonstrated that both enantiomers bound to the LFA-1 “lovastatin site.” ICAM binding assays showed that S-isoflurane inhibited more potently than R-isoflurane, consistent with the result of 1-AMA competition assay.

Conclusions

In contrast with the x-ray crystallography, both enantiomers bound to and inhibited LFA-1. S-isoflurane showed slight preference over R-isoflurane.