Detection of the Progression of Anthesis in Field-Grown Maize Tassels: A Case Study

The tassel of the maize plant is responsible for the production and dispersal of pollen for subsequent capture by the silk (stigma) and fertilization of the ovules. Both the amount and timing of pollen shed are physiological traits that impact the production of a hybrid seed. This study describes an automated end-to-end pipeline that combines deep learning and image processing approaches to extract tassel flowering patterns from time-lapse camera images of plants grown under field conditions. Inbred lines from the SAM and NAM diversity panels were grown at the Curtiss farm at Iowa State University, Ames, IA, during the summer of 2016. Using a set of around 500 pole-mounted cameras installed in the field, images of plants were captured every 10 minutes of daylight hours over a three-week period. Extracting data from imaging performed under field conditions is challenging due to variabilities in weather, illumination, and the morphological diversity of tassels. To address these issues, deep learning algorithms were used for tassel detection, classification, and segmentation. Image processing approaches were then used to crop the main spike of the tassel to track reproductive development. The results demonstrated that deep learning with well-labeled data is a powerful tool for detecting, classifying, and segmenting tassels. Our sequential workflow exhibited the following metrics: mAP for tassel detection was 0.91, F1 score obtained for tassel classification was 0.93, and accuracy of semantic segmentation in creating a binary image from the RGB tassel images was 0.95. This workflow was used to determine spatiotemporal variations in the thickness of the main spike—which serves as a proxy for anthesis progression.

Fabrication and Characterization of Simple Structure Fluidic-Based Memristor for Immunosensing of NS1 Protein Application

Non-structural protein 1 (NS1 protein) is becoming a commonplace biomarker for the diagnostic of early detection of dengue. In this study, we sought to use a label-free approach of detecting NS1 protein by harnessing fluidic-based memristor sensor. The sensor was fabricated using sol-gel spin coating technique, by which TiO2 thin film is coated on the surface of Indium tin oxide (ITO) and a glass substrate. The sensor was then functionalized with glycidoxypropyl-trimethoxysilane (GPTS), acting as antibody for NS1. The addition of the target NS1 formed an antibody-antigen complex which altered the physical and electrical properties in sensing region. Sensing of the sensor is incumbent upon the measurement of Off-On resistance ratio. Imaging with Field Emission Scanning Electron Microscope (FESEM) evinced the successful immobilization of the antibody and the subsequent capture of the NS1 protein by the immobilized antibody. The detection limit actualized by the developed sensor was 52 nM and the diameter of 2 mm gives the most optimal measurement. The developed sensor demonstrated an immense potential towards the development of label-free diagnostic of early dengue infection.

Establishment of Proximity-dependent Biotinylation Approaches in Different Plant Model Systems

Proximity-dependent biotin labelling (PDL) uses a promiscuous biotin ligase (PBL) or a peroxidase fused to a protein of interest. This enables covalent biotin labelling of proteins and allows subsequent capture and identification of interacting and neighbouring proteins without the need for the protein complex to remain intact. To date, only few papers report on the use of PDL in plants. Here we present the results of a systematic study applying a variety of PDL approaches in several plant systems using various conditions and bait proteins. We show that TurboID is the most promiscuous variant in several plant model systems and establish protocols which combine Mass Spectrometry-based analysis with harsh extraction and washing conditions. We demonstrate the applicability of TurboID in capturing membrane-associated protein interactomes using Lotus japonicus symbiotically active receptor kinases as test-case. We further benchmark the efficiency of various PBLs in comparison with one-step affinity purification approaches. We identified both known as well as novel interactors of the endocytic TPLATE complex. We furthermore present a straightforward strategy to identify both non-biotinylated as well as biotinylated peptides in a single experimental setup. Finally, we provide initial evidence that our approach has the potential to infer structural information of protein complexes.

Mechanisms of exciton photoluminescence quenching in the electric field of a standing surface acoustic wave

Mechanisms of exciton photoluminescence (PL) quenching in the longitudinal electric field of a standing surface acoustic wave (SAW) have been studied by the example of type II GaAs/AlAs superlattices (SLs). Such SLs with a long lifetime of nonequilibrium carriers have allowed examining the influence of the SAW electric field on the excitonic PL both under the continuous and impulse laser excitations. It has been found that the mechanisms of the interaction of excitons and a SAW electric field depend upon the kinetic energy of excitons and carriers. As for hot excitons and carriers, the standing SAW electric field causes the impact ionization of excitons with a subsequent capture of free carriers at the nonradiative recombination centers, which results in a decrease in the steady-state exciton PL. As for cold excitons and carriers, the impact of excitons with the carriers accelerated by the SAW electric field results mainly in exciton delocalization from the levels of quantum wells formed due to interface roughness with a subsequent capture of excitons at the nonradiative recombination centers, which leads to the acceleration of the PL kinetics.

The Road to Madness

The NCAA basketball tournament is one of the iconic events in American sports. Now known for its size (sixty-eight teams) and myriad upsets, before the tournament expanded in 1974, it consisted of twenty-five teams (one per college conference) and had been dominated by one team, the UCLA Bruins, for over a decade. By the time the 1973-74 season started, coach John Wooden and UCLA had become symbols of success and stability during a time of cultural, political, and economic turmoil in the United States. But North Carolina State’s defeat of UCLA in the 1974 tournament--and subsequent capture of the title--marked the end of a basketball dynasty, and gave birth to the cultural touchstone now known as March Madness.In WHEN MARCH BECAME MADNESS, J. Samuel Walker and Randy Roberts offer the first in-depth, historical account of the pivotal 1973-74 college basketball season and the controversial decisions that led to the expansion of the collegiate tournament. Blending oral history and extensive research in primary sources, Walker and Roberts provide a richly detailed chronicle of the players, coaches, universities, and public figures instrumental in creating one of the nation’s major sporting events.

Synthesis and Characterization of Functionalized Magnetic Nanoparticles for the Detection of Pesticide

We synthesized magnetic nanoparticles (MNPs) by mixing aqueous solutions of 3d transition metal chlorides (MCl2·nH2O) and a sodium metasilicate nonahydrate (Na2SiO3·9H2O) in order to produce monodispersed MNPs in a single step. The particle size can be controlled by adjusting the annealing temperature. We characterized the MNPs by X-ray diffraction (XRD), superconducting quantum interference device (SQUID), transmission electron microscopy (TEM), Fourier transform infrared (FT-IR), and zeta-potential measurement. Paramagnetic and superparamagnetic behaviors were found for the obtained samples depending on the particle size (d=3.0–4.6 nm). The synthesized MNPs could be modified with the amino-, phenyl-, and carboxy- groups on MNPs' surface by silanization procedure, respectively. The purpose of functionalizing the surface of the nanoscale magnetic particles was to realize subsequent capture and detection with desired other molecules by nanoparticle assisted laser ionization/desorption mass spectrometry.

Behavioural responses of brushtail possums to live trapping and implications for trap-catch correction

Context The behavioural response of animals to repeated trapping has implications for correction of population and monitoring indices that use catch per unit effort. Failure to account for sprung traps introduces biases into estimates of relative abundance. The time when animals get caught in live traps is often ignored, but it can provide important information about temporal movement patterns relevant to this issue. Aims We assessed changes in the behaviour of brushtail possums (Trichosurus vulpecula), a nocturnal marsupial, in response to repeated trapping and evaluated the potential benefit of correcting a commonly used index of abundance by using time-of-capture information. Methods Possums were live-trapped for three nights each month over a 20-month period in baited cage traps in a 6-ha area of native lowland forest in the southern North Island, New Zealand. Trapped possums were individually identified on first capture. Timing devices were attached to the traps to measure how long after sunset traps were sprung and how that time related to the duration of the trap-night (sunset to sunrise). Key results Possums were trapped, on average, ~1.25 h after sunset. Traps triggered other than by possums were sprung on average 1–2 h later. Possums caught on the first night of a trapping session were caught significantly earlier than those caught on subsequent nights. Previous capture influenced the time of subsequent capture in a trapping session in complex ways, and recapture times were generally earlier than times of first capture. Possums were captured, on average, after 11% of the duration of a trap-night and traps were triggered by animals other than possums, on average, after 22% of the duration of a trap-night. Conclusions The data on time of capture of possums and triggering of sprung traps suggested a need to alter the commonly used correction factor for population indices for possums, because, on average, traps were sprung for significantly more of each trapping interval (i.e. trap-night) than the half a trap-night assumed in the correction factor. Implications Better understanding of possum foraging behaviour is a key to more effective control using traps. In that context, more research is needed to understand the reasons for individual differences in trappability. Although it is theoretically desirable to account for sprung traps when trapping is used to index populations, to reduce biases in estimates of relative abundance, correction of the standardised residual trap-catch index for possums is probably unimportant in practical terms, because most possum control reduces numbers to levels (2–5% trap catch) at which correction of the index is unimportant. The principal exception to this is likely to be when there is a high level of non-target interference from rodents.

The Trans-Golgi Network Accessory Protein p56 Promotes Long-Range Movement of GGA/Clathrin-containing Transport Carriers and Lysosomal Enzyme Sorting

The sorting of acid hydrolase precursors at the trans-Golgi network (TGN) is mediated by binding to mannose 6-phosphate receptors (MPRs) and subsequent capture of the hydrolase-MPR complexes into clathrin-coated vesicles or transport carriers (TCs) destined for delivery to endosomes. This capture depends on the function of three monomeric clathrin adaptors named GGAs. The GGAs comprise a C-terminal “ear” domain that binds a specific set of accessory proteins. Herein we show that one of these accessory proteins, p56, colocalizes and physically interacts with the three GGAs at the TGN. Moreover, overexpression of the GGAs enhances the association of p56 with the TGN, and RNA interference (RNAi)-mediated depletion of the GGAs decreases the TGN association and total levels of p56. RNAi-mediated depletion of p56 or the GGAs causes various degrees of missorting of the precursor of the acid hydrolase, cathepsin D. In the case of p56 depletion, this missorting correlates with decreased mobility of GGA-containing TCs. Transfection with an RNAi-resistant p56 construct, but not with a p56 construct lacking the GGA-ear–interacting motif, restores the mobility of the TCs. We conclude that p56 tightly cooperates with the GGAs in the sorting of cathepsin D to lysosomes, probably by enabling the movement of GGA-containing TCs.

Photochromic Ag-TiO2 Thin Films Fabricated by Dual-Target Helicon Magnetron Sputtering

Photochromic material Ag-TiO2 thin films are fabricated on quartz substrate by dual -target helicon magnetron sputtering. The phototchromic behavior is investigated for the sample loaded with 90% Ag. Spheres, ellipsoids and polyhedra shape of Ag particles with wide range size (5∼100 nm) are dispersed in the TiO2 amorphous matrix observed by transmission electron microscopy. The spectral hole burned by the irradiation of laser at the wavelength 532 nm can be explained by a particle-plasmon-assisted electron transfer from Ag nanoparticles to TiO2 and subsequent trapping by adsorbed molecular oxygen. Moreover, the mechanism of the slow recovery after photochromism is suggested as a slow thermal release of electrons from oxygen trapping centers and subsequent capture into the Ag nanoparticles.

Development of a High-Throughput Scintillation Proximity Assay for the Identification of C-Domain Translational Initiation Factor 2 Inhibitors

Translational initiation factor 2 (IF2) is the largest of the 3 factors required for translation initiation in prokaryotes and has been shown to be essential in Escherichia coli. It stimulates the binding of fMet-tRNAfMet to the 30S ribosomal subunit in the presence of GTP. The selectivity is achieved through specific recognition of the tRNAfMet blocked α-amino group. IF2 is composed of 3 structural domains: N-domain, whose function is not known; G-domain, which contains the GTP/GDP binding site and the GTPase catalytic center; and C-domain, which recognizes and binds fMet-tRNAfMet. Its activity is strictly bacteria specific and highly conserved among prokaryotes. So far, antibiotics targeting IF2 function are not known, and this makes it an ideal target for new drugs with mechanisms of resistance not yet developed. A few assays have been developed in the past, which allow the detection of IF2 activity either directly or indirectly. In both instances, the assays are based on radioactive detection and do not allow for high throughput because of the need for separation or solvent extraction steps. The authors describe a novel biochemical assay for IF2 that exploits the molecular recognition of fMet-tRNAfMet by the C-domain. The assay is based on the incubation of biotinyl-IF2 with fMet-tRNAfMet and the subsequent capture of the radiolabeled complex by streptavidin-coated beads, exploiting the scintillation proximity assay (SPA) technology. The assay has been designed in an automatable, homogeneous, miniaturized fashion suitable for high-throughput screening and is rapid, sensitive, and robust to dimethyl sulfoxide (DMSO) up to 10% v/v. The assay, used to screen a limited chemical collection of about 5000 compounds and a subset of compounds originated by a 2-D substructural search, has shown to be able to detect potential IF2 inhibitors.