A High-Throughput Screening System for Populus Wood-Associated Transcription Factors and Its Application to Lignin Regulation

Wood formation of trees is a complex and costly developmental process, whose regulatory network is involved in the protein-protein and protein-DNA interactions. To detect such interactions in wood development, we developed a high-throughput screening system with 517 Gal4-AD-wood-associated transcription factors (TFs) library from Populus alba × P. glandulosa cv “84K.” This system can be used for screening the upstream regulators and interacting proteins of targets by mating-based yeast-one hybrid (Y1H) and yeast-two-hybrid (Y2H) method, respectively. Multiple regulatory modules of lignin biosynthesis were identified based on this Populus system. Five TFs interacted with the 500-bp promoter fragment of PHENYLALANINE AMMONIA-LYASE 2 (PAL2), the first rate-limiting enzyme gene in the lignin biosynthesis pathway, and 10 TFs interacted with PaMYB4/LTF1, a key regulator of lignin biosynthesis. Some of these interactions were further validated by EMSA and BiFC assays. The TF-PaPAL2 promoter interaction and TF-PaMYB4 interaction revealed a complex mechanism governing the regulation of lignin synthesis in wood cells. Our high-throughput Y1H/Y2H screening system may be an efficient tool for studying regulatory network of wood formation in tree species.

Role of Bioactive Compounds in the Regulation of Mitochondrial Dysfunctions in Brain and Age-Related Neurodegenerative Diseases

Mitochondria are multifunctional organelles that participate in a wide range of metabolic processes, including energy production and biomolecule synthesis. The morphology and distribution of intracellular mitochondria change dynamically, reflecting a cell’s metabolic activity. Oxidative stress is defined as a mismatch between the body’s ability to neutralise and eliminate reactive oxygen and nitrogen species (ROS and RNS). A determination of mitochondria failure in increasing oxidative stress, as well as its implications in neurodegenerative illnesses and apoptosis, is a significant developmental process of focus in this review. The neuroprotective effects of bioactive compounds linked to neuronal regulation, as well as related neuronal development abnormalities, will be investigated. In conclusion, the study of secondary components and the use of mitochondrial features in the analysis of various neurodevelopmental diseases has enabled the development of a new class of mitochondrial-targeted pharmaceuticals capable of alleviating neurodegenerative disease states and enabling longevity and healthy ageing for the vast majority of people.

If an Entrepreneur Founded the DSM-5 Alternative Model of Personality Disorder: When Axis II Meets Disruptive Innovation

Personality disorder (PD) researchers proposed a highly innovative “paradigm-shifting” revamp for the Diagnostic and Statistical Manual of Mental Disorders 5th edition (DSM-5; APA, 2013). Yet, ten years later, Widiger and Hines (this issue) summarize a developmental process plagued by disagreement and stagnation, with little evidence of the field having reaped the desired benefits of this diagnostic revolution. In this commentary, we draw on principles from entrepreneurial creation, operation, and success—positioning the personality disorder scientists in the role of “disruptive innovator”—and summarize key principles from the entrepreneurial process that may be relevant in understanding the challenges and failures of the personality disorder revolution to date.

Comparative Transcriptome and WGCNA RevealKey Genes Involved in Lignocellulose Degradation in Sarcomyxa Edulis

In order to explore the molecular mechanism of Sarcomyxa edulis response to lignocelluloses degradation, the developmental transcriptomes was analyzed for six stages covering the whole developmental process, including mycelium growing to half bag (B1), mycelium in cold stimulation after full bag (B2), mycelium in primordia appearing (B3), primordia (B4), mycelium at the harvest stage (B5) and mature fruiting body (B6). A total of 6 samples were used for transcriptome sequencing, with three biological replicates. Based on the above transcriptome data, we constructed a co-expression network of weighted genes associated with extracellular enzyme physiological traits by WGCNA, and obtained 19 gene co-expression modules closely related to lignocelluloses degradation. In addition, a number of key genes involved in lignocelluloses degradation pathways were discovered from the four modules with the highest correlation with target traits. These results provide clues for further study on the molecular genetic mechanisms of Sarcomyxa edulis lignocelluloses degradation.

An ancestral genomic sequence that serves as a nucleation site for de novo gene birth

A short non-coding sequence present between the gamma-glutamyltransferase 1 (GGT1) and gamma-glutamyltransferase 5 (GGT5) genes, termed a spacer sequence has been detected in the genomes of Mus musculus, the house mouse and in Philippine tarsier, a primitive ancestral primate. It is highly conserved during primate evolution with certain sequences being totally invariant from mouse to humans. Evidence is presented to show this intergenic sequence serves as a nucleation site for the initiation of diverse genes. We also outline the birth of the human lincRNA gene BCRP3 (BCR activator of RhoGEF and GTPase 3 pseudogene) during primate evolution. The gene developmental process involves sequence initiation, addition of a complex of tandem transposable elements and addition of a segment of another gene. The sequence, initially formed in the Old World Monkeys such as the Rhesus monkey (Macaca mulatta) and the baboon (Papio anubis), develops into different primate genes before evolving into the human BCRP3 gene; it appears to also include trial and error during sequence/gene formation. The protein gene, GGT5 may have also formed by spacer sequence initiation in an ancient ancestor such as zebrafish, but spacer and GGT5 gene sequence drift during evolution produced a divergence that precludes further assessment.

Cerebral Blood Flow Monitoring in High-Risk Fetal and Neonatal Populations

Cerebrovascular pressure autoregulation promotes stable cerebral blood flow (CBF) across a range of arterial blood pressures. Cerebral autoregulation (CA) is a developmental process that reaches maturity around term gestation and can be monitored prenatally with both Doppler ultrasound and magnetic resonance imaging (MRI) techniques. Postnatally, there are key advantages and limitations to assessing CA with Doppler ultrasound, MRI, and near-infrared spectroscopy. Here we review these CBF monitoring techniques as well as their application to both fetal and neonatal populations at risk of perturbations in CBF. Specifically, we discuss CBF monitoring in fetuses with intrauterine growth restriction, anemia, congenital heart disease, neonates born preterm and those with hypoxic-ischemic encephalopathy. We conclude the review with insights into the future directions in this field with an emphasis on collaborative science and precision medicine approaches.

The ectomycorrhizal fungus Pisolithus microcarpus encodes a microRNA involved in cross-kingdom gene silencing during symbiosis

Small RNAs (sRNAs) are known to regulate pathogenic plant–microbe interactions. Emerging evidence from the study of these model systems suggests that microRNAs (miRNAs) can be translocated between microbes and plants to facilitate symbiosis. The roles of sRNAs in mutualistic mycorrhizal fungal interactions, however, are largely unknown. In this study, we characterized miRNAs encoded by the ectomycorrhizal fungus Pisolithus microcarpus and investigated their expression during mutualistic interaction with Eucalyptus grandis. Using sRNA sequencing data and in situ miRNA detection, a novel fungal miRNA, Pmic_miR-8, was found to be transported into E. grandis roots after interaction with P. microcarpus. Further characterization experiments demonstrate that inhibition of Pmic_miR-8 negatively impacts the maintenance of mycorrhizal roots in E. grandis, while supplementation of Pmic_miR-8 led to deeper integration of the fungus into plant tissues. Target prediction and experimental testing suggest that Pmic_miR-8 may target the host NB-ARC domain containing transcripts, suggesting a potential role for this miRNA in subverting host signaling to stabilize the symbiotic interaction. Altogether, we provide evidence of previously undescribed cross-kingdom sRNA transfer from ectomycorrhizal fungi to plant roots, shedding light onto the involvement of miRNAs during the developmental process of mutualistic symbioses.

IL-1β Impacts Vascular Integrity and Lymphatic Function in the Embryonic Omentum

Background: The chromatin remodeling enzyme Brahma Related Gene 1 (BRG1) regulates gene expression in a variety of rapidly differentiating cells during embryonic development. However, the critical genes that BRG1 regulates during lymphatic vascular development are unknown. Methods: We used genetic and imaging techniques to define the role of BRG1 in murine embryonic lymphatic development, although this approach inadvertently expanded our study to multiple interacting cell types. Results: We found that omental macrophages fine-tune an unexpected developmental process by which erythrocytes escaping from naturally discontinuous omental blood vessels are collected by nearby lymphatic vessels. Our data indicate that circulating fibrin(ogen) leaking from gaps in omental blood vessels can trigger inflammasome-mediated interleukin 1 beta (IL-1β) production and secretion from nearby macrophages. IL-1β destabilizes adherens junctions in omental blood and lymphatic vessels, contributing to both extravasation of erythrocytes and their uptake by lymphatics. BRG1 regulates IL-1β production in omental macrophages by transcriptionally suppressing the inflammasome trigger Receptor Interacting Protein Kinase 3 (RIPK3). Conclusions: Genetic deletion of Brg1 in embryonic macrophages leads to excessive IL-1β production, erythrocyte leakage from blood vessels, and blood-filled lymphatics in the developing omentum. Altogether, these results highlight a novel context for epigenetically-regulated crosstalk between macrophages, blood vessels, and lymphatics.

Glial Patchwork: Oligodendrocyte Progenitor Cells and Astrocytes Blanket the Central Nervous System

Tiling is a developmental process where cell populations become evenly distributed throughout a tissue. In this review, we discuss the developmental cellular tiling behaviors of the two major glial populations in the central nervous system (CNS)—oligodendrocyte progenitor cells (OPCs) and astrocytes. First, we discuss OPC tiling in the spinal cord, which is comprised of the three cellular behaviors of migration, proliferation, and contact-mediated repulsion (CMR). These cellular behaviors occur simultaneously during OPC development and converge to produce the emergent behavior of tiling which results in OPCs being evenly dispersed and occupying non-overlapping domains throughout the CNS. We next discuss astrocyte tiling in the cortex and hippocampus, where astrocytes migrate, proliferate, then ultimately determine their exclusive domains by gradual removal of overlap rather than sustained CMR. This results in domains that slightly overlap, allowing for both exclusive control of “synaptic islands” and astrocyte-astrocyte communication. We finally discuss the similarities and differences in the tiling behaviors of these glial populations and what remains unknown regarding glial tiling and how perturbations to this process may impact injury and disease.

EFL PHONOLOGY: A CASE STUDY OF ENGLISH FRICATIVE PRODUCTION BY INDONESIAN LEARNERS/FONOLOGI BAHASA INGGRIS SEBAGAI BAHASA ASING: STUDI KASUS PELAFALAN BUNYI GESER OLEH MAHASISWA INDONESIA

This study reports the difficulties 40 first-year Indonesian college students, majoring in English, had in pronouncing the English fricatives. The aim of this paper is, first, to reveal how these Indonesian EFL learners produced English fricatives, the order of difficulties, and the pronunciation constraints they experienced. The second aim is to identify the possible causes of the pronunciation difficulties. In collecting the data, two types of tasks were administered: a word-list-reading task (Task 1) and a sentence-list-reading task (Task 2). By using Wilcoxon based T-Test, it was revealed that there was a significant difference in the number of errors in Task 1 and Task 2. There was also an increase in errors in Task 2. The results show that the order of difficulties Indonesian learners had in producing fricative sounds (from the most to the least problematic) were: /v/, /ʃ/, /ð/, /θ/, /z/, /ʒ/, /f/, and /s/. It is likely that the influence of the challenging English spelling system played the most important role in the students’ errors. Other factors such as transfer of L1 and the developmental process also contributed to the errors. This research implies that teachers need to apply certain strategies to meet students’ needs. Keywords: English, fricative, Indonesian students, errors AbstrakPenelitian ini melaporkan kesulitan yang dialami mahasiswa tahun pertama Jurusan Pendidikan Bahasa Inggris dalam melafalkan bunyi geser , urutan kesulitan dan kendala dalam pelafalan bunyi tersebut. Tujuan kedua penelitian ini adalah mengidentifikasi kemungkian sebab-sebab dari kesulitan ini. Dua jenis tugas diberikan kepada siswa untuk mengumpulkan data yaitu tugas membaca daftar kata (Tugas 1) dan tugas membaca daftar kalimat (Tugas 2). Berdasarkan hasil Uji T Wilcoxon terungkap bahwa terdapat perbedaan yang signifikan dari segi jumlah kesalahan dalam mengerjakan tugas I dan tugas 2. Juga terdapat peningkatan kesalahan dalam Tugas 2. Hasil-hasil ini memperlihatkan bahwa urutan kesulitan yang dialami mahasiswa Indonesia dalam melafalkan bunyi frikatif ( dari yang paling bermasalah hingga yang kurang bermasalah ) adalah : /v/, /ʃ/, /ð/, /θ/, /z/, /ʒ/, /f/, dan /s/. Ada kecenderungan bahwa sistem ejaan Bahasa Inggris yang rumit sangat besar pengaruhnya terhadap kesalahan-kesalahan yang dibuat oleh mahasiswa. Faktor-faktor lain seperti transfer dari BI dan proses perkembangan juga berkontribusi terhadap kesalahan-kesalahan tersebut. Implikasi penelitian ini adalah dosen atau guru dapat menerapkan strategi-srategi tertentu untuk memenuhi kebutuhan mahasiswa atau siswa. Kata kunci: bahasa Inggris, bunyi geser, mahasiswa Indonesia, kesalahan