Carrier-free chemo-phototherapeutic nanomedicines with endo/lysosomal escape function enhance the therapeutic effect of drug molecules in tumors.

Carrier-free nanomedicines offer advantages of extremely high drug loading capacity (>80%), minimal non-drug constituent burden, and facile preparation processes. Numerous studies have proved that multimodal cancer therapy can enhance chemotherapy efficiency and mitigate multi-drug resistance (MDR) through synergistic therapeutic effects. Upon penetration into the tumor matrix, nanoparticles (NPs) are anticipated to be uptaken by cancer cells, primarily through clathrin-meditated endocytosis pathways, leading to their accumulation in endosomes/lysosomes within cells. However, endo/lysosomes exhibit a highly degradative environment for organic NPs and drug molecules, often resulting in treatment failure. Hence, this study designed a lysosomal escape mechanism with carrier-free nanomedicine, combining the chemotherapeutic drug, curcumin (Cur), and the photothermal/photodynamic therapeutic drug, indocyanine green (ICG), for synergistic cancer treatment (ICG-Cur NPs) via a facile preparation process. To facilitate endo/lysosomal escape, ICG-Cur NPs were modified with metal-phenolic networks (MPNs) of different thickness. The results indicate that a thick MPN coating promotes rapid endo/lysosomal escape of ICG-Cur NPs within 4 h and enhances the photothermal conversion efficiency of ICG-Cur NPs by 55.8%, significantly improving anticancer efficacy in both chemo- and photo-therapies within 3D solid tumor models. This finding underscores the critical role of endo/lysosomal escape capacity in carrier-free drug NPs for therapeutic outcomes and offers a facile solution to achieve it.

Experimental study of the effect of pump pulse duration on liquid crystal laser performance.

Much work has been done to understand the factors that impact photonic band-edge liquid crystal (LC) laser threshold and slope efficiency, two parameters often stated to quantify performance. Conventionally, LC lasers are optically pumped using Q-switched lasers with a fixed pulse duration, and thus the effect of pump pulse duration on LC laser performance has received little attention. While some studies have been published at different pump pulse durations, these use different laser sources and experimental conditions, making the data incomparable. By exploiting a recent breakthrough in laser diode pumping, our experimental results prove and quantify the detrimental effect of an increase in pump pulse duration on LC laser performance. We also show that the dependency of threshold on pump pulse duration depends on how threshold is defined, owing to an ambiguity in the definition of pulse energy in systems where peak power and pulse duration can be independently controlled. For improved comparison within the literature on LC laser device performance, we thus propose an alternative convention, whereby threshold is stated in units of peak power density.

Characterisation of Antennal Sensilla and Electroantennography Responses of the Dung Beetles Bubas bison, Onitis aygulus and Geotrupes spiniger (Coleoptera: Scarabaeoidea) to Dung Volatile Organic Compounds.

Locating sporadically distributed food resources and mate finding are strongly aided by volatile cues for most insects, including dung beetles. However, there is limited information on the olfactory ecology of dung beetles. We conducted a scanning electron microscopy study on the morphology and distribution of the antennal sensilla of three introduced dung beetle species in Australia: Geotrupes spiniger (Coleoptera: Geotrupidae), Bubas bison and Onitis aygulus (Coleoptera: Scarabaeidae). Three main morphological types of antennal sensilla were identified: sensilla trichodea (ST), sensilla basiconica (SB) and sensilla chaetica (SCh). Distinct variations of SB distribution were observed in B. bison and G. spiniger and on different lamellar surfaces in both sexes of all three species. Sexual dimorphism in antennal sensilla distribution or their abundance was not evident. To complement the morphological characterisation of sensilla, electroantennography (EAG) was carried out to construct EAG response profiles of the three species to selected dung volatiles. An initial study revealed that antennae of all species were sensitive to a mix of phenol, skatole, indole, p-cresol, butanone and butyric acid, common components of livestock dung headspace. In addition to these six compounds, dimethyl sulfide, dimethyl disulfide, eucalyptol and toluene were tested for antennal activity. All compounds evoked measurable EAG responses, confirming antennal sensitivity. Geotrupes spiniger exhibited significant responses to all the compounds compared to the control, whereas B. bison and O. aygulus only responded to a subset of compounds. A comparison of relative EAG amplitudes revealed highly significant responses to p-cresol in G. spiniger and to skatole in B. bison. Geotrupes spiniger displayed differential responses to all the compounds. Pooled EAG data suggest highly significant differences in responses among the three species and among compounds. Our findings suggest that a blend of volatiles may offer potential for the trapping of dung beetles, thereby avoiding the use of dung baits that are inconvenient, inconsistent and may pose a threat to farm biosecurity.

Non-covalent binding tags for batch and flow biocatalysis.

Enzyme immobilization offers considerable advantage for biocatalysis in batch and continuous flow reactions. However, many currently available immobilization methods require that the surface of the carrier is chemically modified to allow site specific interactions with their cognate enzymes, which requires specific processing steps and incurs associated costs. Two carriers (cellulose and silica) were investigated here, initially using fluorescent proteins as models to study binding, followed by assessment of industrially relevant enzyme performance (transaminases and an imine reductase/glucose oxidoreductase fusion). Two previously described binding tags, the 17 amino acid long silica-binding peptide from the Bacillus cereus CotB protein and the cellulose binding domain from the Clostridium thermocellum, were fused to a range of proteins without impairing their heterologous expression. When fused to a fluorescent protein both tags conferred high avidity specific binding with their respective carriers (low nanomolar Kd values). The CotB peptide (CotB1p) induced protein aggregation in the transaminase and imine reductase/glucose oxidoreductase fusions when incubated with the silica carrier. The Clostridium thermocellum cellulose binding domain (CBDclos) allowed immobilization of all the proteins tested, but immobilization led to loss of enzymatic activity in the transaminases (< 2-fold) and imine reductase/glucose oxidoreductase fusion (> 80%). A transaminase-CBDclos fusion was then successfully used to demonstrate the application of the binding tag in repetitive batch and a continuous-flow reactor.

Lagovirus Non-structural Protein p23: A Putative Viroporin That Interacts With Heat Shock Proteins and Uses a Disulfide Bond for Dimerization.

The exact function(s) of the lagovirus non-structural protein p23 is unknown as robust cell culture systems for the Rabbit haemorrhagic disease virus (RHDV) and other lagoviruses have not been established. Instead, a range of in vitro and in silico models have been used to study p23, revealing that p23 oligomerizes, accumulates in the cytoplasm, and possesses a conserved C-terminal region with two amphipathic helices. Furthermore, the positional homologs of p23 in other caliciviruses have been shown to possess viroporin activity. Here, we report on the mechanistic details of p23 oligomerization. Site-directed mutagenesis revealed the importance of an N-terminal cysteine for dimerization. Furthermore, we identified cellular interactors of p23 using stable isotope labeling with amino acids in cell culture (SILAC)-based proteomics; heat shock proteins Hsp70 and 110 interact with p23 in transfected cells, suggesting that they 'chaperone' p23 proteins before their integration into cellular membranes. We investigated changes to the global transcriptome and proteome that occurred in infected rabbit liver tissue and observed changes to the misfolded protein response, calcium signaling, and the regulation of the endoplasmic reticulum (ER) network. Finally, flow cytometry studies indicate slightly elevated calcium concentrations in the cytoplasm of p23-transfected cells. Taken together, accumulating evidence suggests that p23 is a viroporin that might form calcium-conducting channels in the ER membranes.

Selection for early shoot vigour in wheat increases root hair length but reduces epidermal cell size of roots and leaves.

Six cycles of recurrent selection for early shoot vigour in wheat resulted in significant increases in leaf width and shoot biomass. Here, in replicated controlled-environment studies, the effect of early shoot vigour on root biomass, rhizosheath size, root hair length, and cell size in the roots and leaves was examined across different cycles of selection. Increased shoot vigour was associated with greater root biomass, larger rhizosheath size, and longer root hairs. Our findings demonstrate that rhizosheath size was a reliable surrogate for root hair length in this germplasm. Examination of the root epidermis revealed that the 'cell body' of the trichoblasts (hair-forming cells) and the atrichoblasts (non-hair-forming cells) decreased in size as shoot vigour increased. Therefore, in higher vigour germplasm, longer root hairs emerged from smaller trichoblasts so that total trichoblast volume (root hair plus cell body) was generally similar regardless of shoot vigour. Similarly, the sizes of the four main cell types on the leaf epidermis became progressively smaller as shoot vigour increased, which also increased stomatal density. The relationship between shoot vigour and root traits is considered, and the potential contribution of below-ground root traits to performance and competitiveness of high vigour germplasm is discussed.

Morpholino Oligonucleotide Cross-Linked Hydrogels as Portable Optical Oligonucleotide Biosensors.

Morpholino Oligonucleotides (MOs), an uncharged DNA analogue, are functionalized with an acrylamide moiety and incorporated into polymer hydrogels as responsive cross-links for microRNA sequence detection. The MO cross-links can be selectively cleaved by a short target analyte single-stranded DNA (ssDNA) sequence based on microRNA, inducing a distinct swelling response measured optically. The MO cross-links offer significant improvement over DNA based systems through improved thermal stability, no salt requirement and 1000-fold improved sensitivity over a comparative biosensor, facilitating a wider range of sensing conditions. Analysis was also achieved using a mobile phone camera, demonstrating portability.

Overcoming repetition rate limitations in liquid crystal laser systems.

Liquid crystal lasers have advantageous features, including continuous wavelength tuning at low cost. Although many potential applications have been highlighted, use of these lasers is not widespread, partially due to performance limitations. This paper presents a method of overcoming repetition rate limitations. A rapidly spinning stage is used to allow operation of a LC laser at 10 kHz: two orders of magnitude greater than possible with a static cell. Average power outputs of up to 3.5 mW are achieved along with an improvement in emission stability. Lastly, a mechanical wavelength-switching method is demonstrated. The spinning cell approach will enable research into the use of liquid crystal lasers in fluorescence imaging and display applications.

Nanocomposite-Based Microstructured Piezoresistive Pressure Sensors for Low-Pressure Measurement Range.

Piezoresistive pressure sensors capable of detecting ranges of low compressive stresses have been successfully fabricated and characterised. The 5.5 × 5 × 1.6 mm³ sensors consist of a planar aluminium top electrode and a microstructured bottom electrode containing a two-by-two array of truncated pyramids with a piezoresistive composite layer sandwiched in-between. The responses of two different piezocomposite materials, a Multiwalled Carbon Nanotube (MWCNT)-elastomer composite and a Quantum Tunneling Composite (QTC), have been characterised as a function of applied pressure and effective contact area. The MWCNT piezoresistive composite-based sensor was able to detect pressures as low as 200 kPa. The QTC-based sensor was capable of detecting pressures as low as 50 kPa depending on the contact area of the bottom electrode. Such sensors could find useful applications requiring the detection of small compressive loads such as those encountered in haptic sensing or robotics.

Mechanisms of endosperm initiation.

KEY MESSAGE: Overview of developmental events and signalling during central cell maturation and early endosperm development with a focus on mechanisms of sexual and autonomous endosperm initiation. Endosperm is important for seed viability and global food supply. The mechanisms regulating the developmental transition between Female Gametophyte (FG) maturation and early endosperm development in angiosperms are difficult to study as they occur buried deep within the ovule. Knowledge of the molecular events underlying this developmental window of events has significantly increased with the combined use of mutants, cell specific markers, and plant hormone sensing reporters. Here, we review recent discoveries concerning the developmental events and signalling of FG maturation, fertilization, and endosperm development. We focus on the regulation of the initiation of endosperm development with and without fertilization in Arabidopsis and the apomict Hieracium, comparing this to what is known in monocots where distinct differences in developmental patterning may underlie alternative mechanisms of suppression and initiation. The Polycomb Repressive Complex 2 (PRC2), plant hormones, and transcription factors are iteratively involved in early fertilization-induced endosperm formation in Arabidopsis. Auxin increases and PRC2 complex inactivation can also induce fertilization-independent endosperm proliferation in Arabidopsis. Function of the PRC2 complex member FERTILIZATION-INDEPENDENT ENDOSPERM and two loci AutE and LOP are required for autonomous endosperm development in apomictic Hieracium. A comparative understanding of cues required for early endosperm development will facilitate genetic engineering approaches for the development of resilient seed crops, especially if an option for fertilization-independent endosperm formation was possible to combat stress-induced crop failure.

Micro- and nano-structure based oligonucleotide sensors.

This paper presents a review of micro- and nano-structure based oligonucleotide detection and quantification techniques. The characteristics of such devices make them very attractive for Point-of-Care or On-Site-Testing biosensing applications. Their small scale means that they can be robust and portable, their compatibility with modern CMOS electronics means that they can easily be incorporated into hand-held devices and their suitability for mass production means that, out of the different approaches to oligonucleotide detection, they are the most suitable for commercialisation. This review discusses the advantages of micro- and nano-structure based sensors and covers the various oligonucleotide detection techniques that have been developed to date. These include: Bulk Acoustic Wave and Surface Acoustic Wave devices, micro- and nano-cantilever sensors, gene Field Effect Transistors, and nanowire and nanopore based sensors. Oligonucleotide immobilisation techniques are also discussed.

Adaptive holographic pumping of thin-film organic lasers.

In this Letter, we use a reconfigurable hologram to dynamically control the position of incidence of the pump beam onto a liquid-crystal dye-based laser. The results show that there is an increase in the stability of the laser output with time and the average power when compared with the output of the same laser when it is optically excited using a static pump beam. This technique also provides additional functionality, such as wavelength tuning and spatial shaping of the pump beam, both of which are demonstrated here.

Increasing the flexoelastic ratio of liquid crystals using highly fluorinated ester-linked bimesogens.

We present experimental results on the bulk flexoelectric coefficients e and effective elastic coefficients K of non-symmetric bimesogenic liquid crystals when the number of terminal and lateral fluoro substituents is increased. These coefficients are of importance because the flexoelastic ratio e/K governs the magnitude of flexoelectro-optic switching in chiral nematic liquid crystals. The study is carried out for two different types of linkage in the flexible spacer chain that connects the separate mesogenic units: these are either an ether or an ester unit. It is found that increasing the number of fluorine atoms on the mesogenic units typically leads to a small increase in e and a decrease in K, resulting in an enhancement of e/K. The most dramatic increase in e/K, however, is observed when the linking group is changed from ether to ester units, which can largely be attributed to an increase in e. Increasing the number of fluorine atoms does, however, increase the viscoelastic ratio and therefore leads to a concomitant increase in the response time. This is observed for both types of linkage, although the ester-linked compounds exhibit smaller viscoelastic ratios compared with their ether-linked counterparts. Highly fluorinated ester-linked compounds are also found to exhibit lower transition temperatures and dielectric anisotropies. As a result, these compounds are promising materials for use in electro-optic devices.

Analysis of grain characters in temperate grasses reveals distinctive patterns of endosperm organization associated with grain shape.

Members of the core pooids represent the most important crops in temperate zones including wheat, barley, and oats. Their importance as crops is largely due to the grain, particularly the storage capabilities of the endosperm. In this study, a comprehensive survey of grain morphology and endosperm organization in representatives of wild and cultivated species throughout the core pooids was performed. As sister to the core pooid tribes Poeae, Aveneae, Triticeae, and Bromeae within the Pooideae subfamily, Brachypodium provides a taxonomically relevant reference point. Using macroscopic, histological, and molecular analyses distinct patterns of grain tissue organization in these species, focusing on the peripheral and modified aleurone, are described. The results indicate that aleurone organization is correlated with conventional grain quality characters such as grain shape and starch content. In addition to morphological and organizational variation, expression patterns of candidate gene markers underpinning this variation were examined. Features commonly associated with grains are largely defined by analyses on lineages within the Triticeae and knowledge of grain structure may be skewed as a result of the focus on wheat and barley. Specifically, the data suggest that the modified aleurone is largely restricted to species in the Triticeae tribe.

A FILAMENTOUS FLOWER orthologue plays a key role in leaf patterning in opium poppy.

The plant-specific YABBY genes were initially defined by their roles in determining abaxial/adaxial cell fate in lateral organs of eudicots, and repressing meristematic genes in differentiating tissues such as leaves. In Arabidopsis thaliana FILAMENTOUS FLOWER (FIL) is also required for inflorescence and floral meristem establishment and flower development in a pathway involving the floral transition and identity genes. Here we describe the characterization of a FIL orthologue from the basal eudicot, Papaver somniferum (the opium poppy), and demonstrate a role for the gene in patterning the highly lobed leaf of the poppy. Silencing of PapsFIL using viral-induced gene silencing resulted in leaves of reduced laminar area, more pronounced margin serration and, in some cases, leaf bifurcation. In contrast, the gene does not appear to affect the development of the flower, and these variations in function are discussed in relation to its taxonomic position as a basal eudicot and its determinate growth habit.

Nanophotonic three-dimensional microscope.

Three-dimensional (3D) optical microscopy based on integral imaging techniques is limited mainly by diffraction effects and the pitch of the microlens array used to sample the specimen. We integrate nanotechnology to the integral imaging technique and demonstrate a nanophotonic 3D microscope, where a nanophotonic lens array is used to finely sample the specimen. The resolution limitation due to diffraction is reduced by capturing images before the diffraction effects predominate and hence overcomes the bottleneck of achieving high resolution in an integral imaging 3D microscope.

Alternate transcripts of a floral developmental regulator have both distinct and redundant functions in opium poppy.

BACKGROUND AND AIMS: The MADS-box transcription factor AGAMOUS (AG) is an important regulator of stamen and fruit identity as well as floral meristem determinacy in a number of core eudicots and monocots. However, its role outside of these groups has not been assessed explicitly. Examining its role in opium poppy, a basal eudicot, could uncover much about the evolution and development of flower and fruit development in the angiosperms. METHODS: AG orthologues were isolated by degenerate RT-PCR and the gene sequence and structure examined; gene expression was characterized using in situ hybridization and the function assessed using virus-induced gene silencing. KEY RESULTS: In opium poppy, a basal eudicot, the AGAMOUS orthologue is alternatively spliced to produce encoded products that vary at the C-terminus, termed PapsAG-1 and PapsAG-2. Both transcripts are expressed at high levels in stamens and carpels. The functional implications of this alternative transcription were examined using virus-induced gene silencing and the results show that PapsAG-1 has roles in stamen and carpel identity, reflecting those found for Arabidopsis AG. In contrast, PapsAG-2, while displaying redundancy in these functions, has a distinctive role in aspects of carpel development reflected in septae, ovule and stigma defects seen in the loss-of-function line generated. CONCLUSIONS: These results describe the first explicit functional analysis of an AG-clade gene in a basal eudicot; illustrate one of the few examples of the functional consequences of alternative splicing in transcription factors and reveal the importance of alternative transcription, as well as gene duplication, as a driving force in evolution.

Paintable band-edge liquid crystal lasers.

In this paper we demonstrate photonic band-edge laser emission from emulsion-based polymer dispersed liquid crystals. The lasing medium consists of dye-doped chiral nematic droplets dispersed within a polymer matrix that spontaneously align as the film dries. Such lasers can be easily formed on single substrates with no alignment layers. The system combines the self-organizing periodic structure of chiral nematic liquid crystals with the simplicity of the emulsion procedure so as to produce a material that retains the emission characteristics of band-edge lasers yet can be readily coated. Sequential and stacked layers demonstrate the possibility of achieving simultaneous multi-wavelength laser output from glass, metallic, and flexible substrates.

Endosperm development in Brachypodium distachyon.

Grain development and its evolution in grasses remains poorly understood, despite cereals being our most important source of food. The grain, for which many grass species have been domesticated, is a single-seeded fruit with prominent and persistent endosperm. Brachypodium distachyon, a small wild grass, is being posited as a new model system for the temperate small grain cereals, but little is known about its endosperm development and how this compares with that of the domesticated cereals. A cellular and molecular map of domains within the developing Brachypodium endosperm is constructed. This provides the first detailed description of grain development in Brachypodium for the reference strain, Bd21, that will be useful for future genetic and comparative studies. Development of Brachypodium grains is compared with that of wheat. Notably, the aleurone is not regionally differentiated as in wheat, suggesting that the modified aleurone region may be a feature of only a subset of cereals. Also, the central endosperm and the nucellar epidermis contain unusually prominent cell walls that may act as a storage material. The composition of these cell walls is more closely related to those of barley and oats than to those of wheat. Therefore, although endosperm development is broadly similar to that of temperate small grain cereals, there are significant differences that may reflect its phylogenetic position between the Triticeae and rice.

High-speed switchable lens enables the development of a volumetric stereoscopic display.

Stereoscopic displays present different images to the two eyes and thereby create a compelling three-dimensional (3D) sensation. They are being developed for numerous applications including cinema, television, virtual prototyping, and medical imaging. However, stereoscopic displays cause perceptual distortions, performance decrements, and visual fatigue. These problems occur because some of the presented depth cues (i.e., perspective and binocular disparity) specify the intended 3D scene while focus cues (blur and accommodation) specify the fixed distance of the display itself. We have developed a stereoscopic display that circumvents these problems. It consists of a fast switchable lens synchronized to the display such that focus cues are nearly correct. The system has great potential for both basic vision research and display applications.