bluuebunny/crossref_metadata_2025 huggingface dataset:
Intracranial Arterial Calcification and Intracranial Atherosclerosis: Close but Different
Authors: Du Heng , Li Jia , Yang Wenjie , Bos Daniel , Zheng Lu , Wong Lawrence Ka Sing , Leung Thomas W. , Chen Xiangyan
Published: February 2022

Background and Purpose

Intracranial arterial calcification (IAC) may be present in the intimal or medial arterial layer. This study aimed to elucidate the link between the calcification and atherosclerotic disease in the intracranial vasculature.

Methods

Consecutive patients with acute ischemic stroke were included. Bilateral intracranial segment of the internal carotid artery, M1 segment of the middle cerebral artery, intracranial segment of the vertebral artery, and the basilar artery were visualized by the multi-detector computed tomography (CT) and vessel-wall magnetic resonance imaging (vwMRI) within 14 days after stroke onset. IAC was into the intimal or medial pattern. Subsequently, on the vwMRI, we assessed the luminal stenosis, eccentricity, plaque burden, and intraplaque hemorrhage (IPH) as markers of atherosclerosis at each IAC site.

Results

Among 69 patients with stroke, IAC was identified in 35% of (161/483) artery segments, of which 61.5% were predominantly intimal calcification and 38.5% were predominantly medial calcification. About 79.8% of intimal calcifications and 64.5% of medial calcifications co-existed with atherosclerotic plaques. Intimal calcification was associated with luminal stenosis (p = 0.003) caused by atherosclerotic lesions. Compared with the medial IAC, intimal IAC was more often accompanied by eccentric plaques (p = 0.02), larger plaque burden (p = 0.001), and IPH (p = 0.001).

Conclusion

Our multimodal imaging-based comparison study on intracranial arteriosclerosis demonstrated that intimal IAC, compared with medial IAC, was more often accompanied by the luminal stenosis, larger plaque burden, eccentricity, and IPH, providing strong evidence for clinical evaluation on the mechanism, risk, and prognosis of ischemic stroke.
Ankk1 Loss of Function Disrupts Dopaminergic Pathways in Zebrafish
Authors: Leggieri Adele , García-González Judit , Torres-Perez Jose V. , Havelange William , Hosseinian Saeedeh , Mech Aleksandra M. , Keatinge Marcus , Busch-Nentwich Elisabeth M. , Brennan Caroline H.
Published: February 2022

Ankyrin repeat and kinase domain containing 1 (ANKK1) is a member of the receptor-interacting protein serine/threonine kinase family, known to be involved in cell proliferation, differentiation and activation of transcription factors. Genetic variation within the ANKK1 locus is suggested to play a role in vulnerability to addictions. However, ANKK1 mechanism of action is still poorly understood. It has been suggested that ANKK1 may affect the development and/or functioning of dopaminergic pathways. To test this hypothesis, we generated a CRISPR-Cas9 loss of function ankk1 zebrafish line causing a 27 bp insertion that disrupts the ankk1 sequence introducing an early stop codon. We found that ankk1 transcript levels were significantly lower in ankk1 mutant (ankk127ins) fish compared to their wild type (ankk1+/+) siblings. In ankk1+/+ adult zebrafish brain, ankk1 protein was detected in isocortex, hippocampus, basolateral amygdala, mesencephalon, and cerebellum, resembling the mammalian distribution pattern. In contrast, ankk1 protein was reduced in the brain of ankk127ins/27ins fish. Quantitative polymerase chain reaction analysis revealed an increase in expression of drd2b mRNA in ankk127ins at both larval and adult stages. In ankk1+/+ adult zebrafish brain, drd2 protein was detected in cerebral cortex, cerebellum, hippocampus, and caudate homolog regions, resembling the pattern in humans. In contrast, drd2 expression was reduced in cortical regions of ankk127ins/27ins being predominantly found in the hindbrain. No differences in the number of cell bodies or axonal projections detected by anti-tyrosine hydroxylase immunostaining on 3 days post fertilization (dpf) larvae were found. Behavioral analysis revealed altered sensitivity to effects of both amisulpride and apomorphine on locomotion and startle habituation, consistent with a broad loss of both pre and post synaptic receptors. Ankk127ins mutants showed reduced sensitivity to the effect of the selective dopamine receptor antagonist amisulpride on locomotor responses to acoustic startle and were differentially sensitive to the effects of the non-selective dopamine agonist apomorphine on both locomotion and habituation. Taken together, our findings strengthen the hypothesis of a functional relationship between ANKK1 and DRD2, supporting a role for ANKK1 in the maintenance and/or functioning of dopaminergic pathways. Further work is needed to disentangle ANKK1’s role at different developmental stages.
Linking Molecular Mechanisms and Evolutionary Consequences of Resource Polyphenism
Authors: Levis Nicholas A. , Ragsdale Erik J.
Published: February 2022

Resource polyphenism—the occurrence of environmentally induced, discrete, and intraspecific morphs showing differential niche use—is taxonomically widespread and fundamental to the evolution of ecological function where it has arisen. Despite longstanding appreciation for the ecological and evolutionary significance of resource polyphenism, only recently have its proximate mechanisms begun to be uncovered. Polyphenism switches, especially those influencing and influenced by trophic interactions, offer a route to integrating proximate and ultimate causation in studies of plasticity, and its potential influence on evolution more generally. Here, we use the major events in generalized polyphenic development as a scaffold for linking the molecular mechanisms of polyphenic switching with potential evolutionary outcomes of polyphenism and for discussing challenges and opportunities at each step in this process. Not only does the study of resource polyphenism uncover interesting details of discrete plasticity, it also illuminates and informs general principles at the intersection of development, ecology, and evolution.
An Increased Anticholinergic Drug Burden Index Score Negatively Affect Nutritional Status in Older Patients Without Dementia
Authors: Ates Bulut Esra , Erken Neziha , Kaya Derya , Dost Fatma Sena , Isik Ahmet Turan
Published: February 2022

Introduction/Aim

Anticholinergic drugs, which have severe central and peripheric side effects, are frequently prescribed to older adults. Increased anticholinergic drug burden is associated with poor physical and cognitive functions. On the other side, the impact of anticholinergics on nutritional status is not elaborated in the literature. Therefore, this study was aimed to investigate the effect of the anticholinergic burden on nutrition.

Materials and Methods

Patients who underwent comprehensive geriatric assessment (CGA) 6 months apart were included in the study. Patients diagnosed with dementia were excluded because of the difference in the course of cognition, physical performance and nutrition. Nutritional status and global cognition were evaluated using Mini Nutritional Assessment-short form (MNA-SF), Mini-Mental State Examination (MMSE). Anticholinergic drug burden was assessed with the Drug Burden Index (DBI), enabling a precise dose-related cumulative exposure. Patients were divided into three groups according to DBI score: 0, no DBI exposure; 0–1, low risk; and ≥1, high risk. Regression analysis was performed to show the relationship between the difference in CGA parameters and the change in DBI score at the sixth month.

Results

A total of 423 patients were included in the study. Participants' mean age was 79.40 ± 7.50, and 68.6% were female. The DBI 0 score group has better MMSE and MNA-SF scores and a lower rate of falls, polypharmacy, malnutrition, and risk of malnutrition in the baseline. Having malnutrition or risk of malnutrition is 2.21 times higher for every one-unit increase in DBI score. Additionally, during the 6-month follow-up, increased DBI score was associated with decreased MNA-SF and MMSE score, albumin.

Conclusions

The harmful effects of anticholinergics may be prevented because anticholinergic activity is a potentially reversible factor. Therefore, reducing exposure to drugs with anticholinergic activity has particular importance in geriatric practice.
The Solute Carrier MFSD1 Decreases the Activation Status of β1 Integrin and Thus Tumor Metastasis
Authors: Roblek Marko , Bicher Julia , van Gogh Merel , György Attila , Seeböck Rita , Szulc Bozena , Damme Markus , Olczak Mariusz , Borsig Lubor , Siekhaus Daria E.
Published: February 2022

Solute carriers are increasingly recognized as participating in a plethora of pathologies, including cancer. We describe here the involvement of the orphan solute carrier Major Facilitator Superfamily Domain-containing protein 1 (MFSD1) in the regulation of tumor cell migration. Loss of MFSD1 enabled higher levels of metastasis in experimental and spontaneous metastasis mouse models. We identified an increased migratory potential in MFSD1−/− tumor cells which was mediated by increased focal adhesion turnover, reduced stability of mature inactive β1 integrin, and the resulting increased integrin activation index. We show that MFSD1 promoted recycling to the cell surface of endocytosed inactive β1 integrin and thereby protected β1 integrin from proteolytic degradation; this led to dampening of the integrin activation index. Furthermore, downregulation of MFSD1 expression was observed during the early steps of tumorigenesis, and higher MFSD1 expression levels correlate with a better cancer patient prognosis. In sum, we describe a requirement for endolysosomal MFSD1 in efficient β1 integrin recycling to suppress tumor cell dissemination.
Blockade of Uttroside B-Induced Autophagic Pro-Survival Signals Augments Its Chemotherapeutic Efficacy Against Hepatocellular Carcinoma
Authors: Nath Lekshmi R. , Swetha Mundanattu , Vijayakurup Vinod , Thangarasu Arun Kumar , Haritha Nair Hariprasad , Shabna Anwar , Aiswarya Sreekumar U. , Rayginia Tennyson P. , Keerthana C. K. , Kalimuthu Kalishwaralal , Sundaram Sankar , Lankalapalli Ravi Shankar , Pillai Sreekumar , Towner Rheal , Isakov Noah , Anto Ruby John
Published: February 2022

Our previous study has demonstrated that Uttroside B (Utt-B), a saponin isolated from the leaves of Solanum nigrum Linn induces apoptosis in hepatic cancer cells and exhibits a remarkable growth inhibition of Hepatocellular Carcinoma (HCC). Our innovation has been granted a patent from the US (US 2019/0160088A1), Canada (3,026,426.), Japan (JP2019520425) and South Korea (KR1020190008323) and the technology have been transferred commercially to Q Biomed, a leading US-based Biotech company. Recently, the compound received approval as ‘Orphan Drug’ against HCC from US FDA, which reveals the clinical relevance of evaluating its antitumor efficacy against HCC. In the present study, we report that Utt-B promotes pro-survival autophagy in hepatic cancer cells as evidenced by the increased expression of autophagy-related proteins, including LC3-II, Beclin1, ATG 5, and ATG 7, as well as a rise in the autophagic flux. Hence, we investigated whether Utt-B-induced autophagic response is complementing or contradicting its apoptotic program in HCC. Inhibition of autophagy using the pharmacological inhibitors, Bafilomycin A1(Baf A1), and 3-methyl adenine (3-MA), and the biological inhibitor, Beclin1 siRNA, significantly enhances the apoptosis of hepatic cancer cells and hence the cytotoxicity induced by Utt-B. We also found increased expression of autophagy markers in Utt-B-treated xenografts derived from HCC. We further analyzed whether the antimalarial drug, Chloroquine (Cqn), a well-known autophagy inhibitor, can enhance the anticancer effect of Utt-B against HCC. We found that inhibition of autophagy using Cqn significantly enhances the antitumor efficacy of Utt-B in vitro and in vivo, in NOD SCID mice bearing HCC xenografts. Taken together, our results suggest that the antitumor effect of Utt-B against HCC can be further enhanced by blocking autophagy. Furthermore, Utt-B in combination with Cqn, a clinically approved drug, if repurposed and used in a combinatorial regimen with Utt-B, can further improve the therapeutic efficacy of Utt-B against HCC.
The Roles of IRF-8 in Regulating IL-9-Mediated Immunologic Mechanisms in the Development of DLBCL: A State-of-the-Art Literature Review
Authors: Cai Mingyue , Chen Na
Published: February 2022

Interferon regulatory factor 8 (IRF-8) is a transcription suppressor that functions through associations with other transcription factors, contributing to the growth and differentiation of bone marrow cells and the activation of macrophages. IRF-8 expression profoundly affects pathogenic processes ranging from infections to blood diseases. Interleukin-9 (IL-9) is a multipotent cytokine that acts on a variety of immune cells by binding to the IL-9 receptor (IL-9R) and is involved in a variety of diseases such as cancer, autoimmune diseases, and other pathogen-mediated immune regulatory diseases. Studies have shown that IL-9 levels are significantly increased in the serum of patients with diffuse large B-cell lymphoma (DLBCL), and IL-9 levels are correlated with the DLBCL prognostic index. The activator protein-1 (AP-1) complex is a dimeric transcription factor that plays a critical role in cellular proliferation, apoptosis, angiogenesis, oncogene-induced transformation, and invasion by controlling basic and induced transcription of several genes containing the AP-1 locus. The AP-1 complex is involved in many cancers, including hematological tumors. In this report, we systematically review the precise roles of IL-9, IRF-8, and AP-1 in tumor development, particularly with regard to DLBCL. Finally, the recent progress in IRF-8 and IL-9 research is presented; the possible relationship among IRF-8, IL-9, and AP-1 family members is analyzed; and future research prospects are discussed.
Selection Strategy of Jaw Tracking in VMAT Planning for Lung SBRT
Authors: Sun Wuji , Shi Yinghua , Li Yu , Ge Chao , Yang Xu , Xia Wenming , Chen Kunzhi , Wang Libo , Dong Lihua , Wang Huidong
Published: February 2022

Purpose

This study aimed to investigate the dosimetric effect and delivery reliability of jaw tracking (JT) with increasing planning target volume (PTV) for lung stereotactic body radiation therapy (SBRT) plans. A threshold of PTV was proposed as a selection criterion between JT and fixed-jaw (FJ) techniques.

Methods

A total of 28 patients with early-stage non-small-cell lung cancer were retrospectively included. The PTVs ranged from 4.88 cc to 68.74 cc, prescribed with 48 Gy in four fractions. Three-partial-arc volumetric modulated arc therapy (VMAT) plans with FJ and with JT were created for each patient with the same optimization objectives. These two sets of plans were compared using metrics, including conformity index (CI), V50%, R50%, D2cm, dose–volume parameters of organs at risk, and monitor units (MUs). The ratio of small subfields (<3 cm in either dimension), %SS, was acquired as a surrogate for the small-field uncertainty. Statistical analyses were performed to evaluate the correlation between the differences in these parameters and the PTV.

Results

The V50%, R50%, D2cm, and V20Gy, D1,500cc, and D1,000cc of the lung showed a statistically significant improvement in JT plans as opposed to FJ plans, while the number of MU in JT plans was higher by an average of 1.9%. Between FJ and JT plans, the PTV was strongly correlated with the differences in V50%, moderately correlated with those in V20Gy of the lung, and weakly correlated with those in D2cm and D1,500cc of the lung. By using JT, %SS was found to be negatively correlated with the PTV, and the PTV should be at least approximately 12.5 cc for an expected %SS <50%, which was 15 cc for a %SS <20% and 20 cc for a %SS <5%.

Conclusions

Considering the dosimetric differences and small-field uncertainties, JT could be selected using a PTV threshold, such as 12.5, 15, or 20 cc, on the basis of the demand of delivery reliability for lung SBRT.
Gene Expression Profiling in Multiple Myeloma: Redefining the Paradigm of Risk-Adapted Treatment
Authors: Cerchione Claudio , Usmani Saad Z. , Stewart A. Keith , Kaiser Martin , Rasche Leo , Kortüm Martin , Mateos María-Victoria , Spencer Andrew , Sonneveld Pieter , Anderson Kenneth C.
Published: February 2022

Multiple myeloma is a blood cancer characterized by clonal proliferation of plasma cells in the bone marrow. In recent years, several new drugs have been added to the therapeutic landscape of multiple myeloma, which have contributed to increased survival rates. However, while the use of therapeutics has evolved, there is still a group of high-risk patients who do not benefit from current treatment strategies. Risk stratification and risk-adapted treatment are crucial to identify the group of patients with urgent need for novel therapies. Gene expression profiling has been introduced as a tool for risk stratification in multiple myeloma based on the genetic make-up of myeloma cells. In this review we discuss the challenge of defining the high-risk multiple myeloma patient. We focus on the standardized analysis of myeloma cancer cells by gene expression profiling and describe how gene expression profiling provides additional insights for optimal risk-adapted treatment of patients suffering from multiple myeloma.
Mining GEO and TCGA Database for Immune Microenvironment of Lung Squamous Cell Carcinoma Patients With or Without Chemotherapy
Authors: Qiang Huiping , Li Jiaqi , Chang Qing , Shen Yinchen , Qian Jialin , Chu Tianqing
Published: February 2022

Background

Chemotherapy is the main treatment for patients with lung squamous cell carcinoma (LUSC). However, how chemotherapy affects their immune system is rarely reported. This study was aimed to compare the differences in the immune microenvironment of LUSC patients with or without chemotherapy.

Methods

A total of 494 LUSC samples were obtained from The Cancer Genome Atlas (TCGA) database. The immune cell infiltration was evaluated by the ssGSEA algorithm, and the tumor subtype was assayed by ConsensusClusterPlus. The differences in tumor mutation burden (TMB) and clinical information between the two types were then compared. Additionally, the differentially expressed genes (DEGs) between two types were analyzed and hub genes were validated in the GEO database.

Results

LSCC samples in TCGA were divided into three subtypes. Then, combining the tumor subtype and immune scores, the samples were divided into hot and cold tumors. Regardless of whether LUSC patients received chemotherapy, the survival of the hot tumor group was not significantly prolonged compared with that of the cold tumor group. For LUSC patients who received chemotherapy, the TMB value in hot tumor group was significantly higher. Total 501 DEGs were identified between two groups. The high expressions of hub genes CD19, CTLA4, FCGR3B, CD80, IL-10, etc. were also validated in the GSE37745 dataset.

Conclusion

Chemotherapy does not affect the survival and prognosis of LUSC patients, but it significantly increases the TMB value of patients with hot tumor. The DEGs, especially hub genes, such as CD19, CTLA4, and FCGR3B, may serve as biomarkers to distinguish cold and hot tumors in LUSC.
Lipidomics Indicates the Hepatotoxicity Effects of EtOAc Extract of Rhizoma Paridis
Authors: Li Chaofeng , Wang Mingshuang , Fu Tingting , Li Zhiqi , Chen Yang , He Tao , Feng Dan , Wang Zhaoyi , Fan Qiqi , Chen Meilin , Zhang Honggui , Lin Ruichao , Zhao Chongjun
Published: February 2022

Rhizoma Paridis is a traditional Chinese medicine commonly used in the clinical treatment of gynecological diseases. Previous studies have shown that aqueous extracts of Rhizoma Paridis exhibit some hepatotoxicity to hepatocytes. Here, using lipidomics analysis, we investigated the potential hepatotoxicity of Rhizoma Paridis and its possible mechanism. The hepatic damaging of different solvent extracts of Rhizoma Paridis on zebrafish larvae were determined by a combination of mortality dose, biochemical, morphological, and functional tests. We found that ethyl acetate extracts (AcOEtE) were the most toxic fraction. Notably, lipidomic responsible for the pharmacological effects of AcOEtE were investigated by Q-Exactive HF-X mass spectrometer (Thermo Scientific high-resolution) coupled in tandem with a UHPLC system. Approximately 1958 unique spectral features were detected, of which 325 were identified as unique lipid species. Among these lipid species, phosphatidylethanolamine cardiolipin Ceramide (Cer), lysophosphatidylinositol sphingosine (Sph), etc., were significantly upregulated in the treated group. Pathway analysis indicates that Rhizoma Paridis may cause liver damage via interfering with the glycerophospholipid metabolism. Collectively, this study has revealed previously uncharacterized lipid metabolic disorder involving lipid synthesis, metabolism, and transport that functionally determines hepatic fibrosis procession.
The Efficacy and Mid-term Durability of Urethral Sphincter Injections of Platelet-Rich Plasma in Treatment of Female Stress Urinary Incontinence
Authors: Chiang Ching-Hsiang , Kuo Hann-Chorng
Published: February 2022

Aims: This study investigated the therapeutic effect of repeated urethral sphincter injections of autologous platelet-rich plasma (PRP) in treatment of stress urinary incontinence (SUI) in women due to intrinsic sphincter deficiency (ISD) refractory to medical treatment or after the first anti-incontinence surgery.

Methods: Twenty-six women with SUI due to urodynamically proven ISD were prospectively enrolled. Five milliliters of PRP (2.5–5 times of the platelet concentrations in peripheral blood) were injected into the external sphincter at 5 sites, with 4 treatments at monthly interval. The primary end-point was post-treatment Global Response Assessment (GRA, scored 0–3) score after four PRP treatments. A GRA ≥ 2 was considered as a successful result. The secondary endpoints included changes in visual analog scale (VAS) of SUI and urodynamic parameters. The follow-up date was 12 months after the fourth PRP treatment.

Results: The mean age was 61.7 ± 15.3 years. The overall success rate was 50% with the post-treatment mean GRA of 1.5 ± 1.1. Complete dryness was achieved in 12 patients (46.2%) after the PRP treatment, and 7 (26.9%) kept total continence at 12 months. The mean VAS of SUI score decreased significantly from 6.4 ± 2.3 to 3.9 ± 2.3 after treatment (p < 0.001). The abdominal leak point pressure increased significantly from 117.5 ± 63.8 to 133.6 ± 61.7 cmH2O (p = 0.045). No perioperative adverse events or severe complications occurred, except 1 (3.8%) patient reported straining to void which was self-limited.

Conclusion: Repeated urethral sphincter injections of autologous PRP are a safe procedure that provides significant reduction in the severity of female SUI and a mid-term durability, suggesting PRP treatment is effective to increase urethral sphincter resistance for female SUI.
Operation of the Atypical Canonical Bone Morphogenetic Protein Signaling Pathway During Early Human Odontogenesis
Authors: Hu Xiaoxiao , Lin Chensheng , Ruan Ningsheng , Huang Zhen , Zhang Yanding , Hu Xuefeng
Published: February 2022

Bone morphogenetic protein (BMP) signaling plays essential roles in the regulation of early tooth development. It is well acknowledged that extracellular BMP ligands bind to the type I and type II transmembrane serine/threonine kinase receptor complexes to trigger the BMP signaling pathway. Then, the receptor-activated Smad1/5/8 in cytoplasm binds to Smad4, the central mediator of the canonical BMP signaling pathway, to form transfer complexes for entering the nucleus and regulating target gene expression. However, a recent study revealed the functional operation of a novel BMP-mediated signaling pathway named the atypical BMP canonical signaling pathway in mouse developing tooth, which is Smad1/5/8 dependent but Smad4 independent. In this study, we investigated whether this atypical BMP canonical signaling is conserved in human odontogenesis. We showed that pSMAD1/5/8 is required for the expression of Msh homeobox 1 (MSX1), a well-defined BMP signaling target gene, in human dental mesenchyme, but the typical BMP canonical signaling is in fact not operating in the early human developing tooth, as evidenced by the absence of pSMAD1/5/8-SMAD4 complexes in the dental mesenchyme and translocation of pSMAD1/5/8, and the expression of MSX1 induced by BMP4 is mothers against decapentaplegic homolog 4 (SMAD4)-independent in human dental mesenchymal cells. Moreover, integrative analysis of RNA-Seq data sets comparing the transcriptome profiles of human dental mesenchymal cells with and without SMAD4 knockdown by siRNA displays unchanged expression profiles of pSMAD1/5/8 downstream target genes, further affirming the functional operation of the atypical canonical BMP signaling pathway in a SMAD1/5/8-dependent but SMAD4-independent manner in the dental mesenchyme during early odontogenesis in humans.
Changes in Gene Expression in Leaves of Cacao Genotypes Resistant and Susceptible to Phytophthora palmivora Infection
Authors: Baruah Indrani K. , Ali Shahin S. , Shao Jonathan , Lary David , Bailey Bryan A.
Published: February 2022

Black pod rot, caused by Phytophthora palmivora, is a devastating disease of Theobroma cacao L. (cacao) leading to huge losses for farmers and limiting chocolate industry supplies. To understand resistance responses of cacao leaves to P. palmivora, Stage 2 leaves of genotypes Imperial College Selection 1 (ICS1), Colección Castro Naranjal 51 (CCN51), and Pound7 were inoculated with zoospores and monitored for symptoms up to 48 h. Pound7 consistently showed less necrosis than ICS1 and CCN51 48 h after inoculation. RNA-Seq was carried out on samples 24 h post inoculation. A total of 24,672 expressed cacao genes were identified, and 2,521 transcripts showed induction in at least one P. palmivora-treated genotype compared to controls. There were 115 genes induced in the P. palmivora-treated samples in all three genotypes. Many of the differentially expressed genes were components of KEGG pathways important in plant defense signal perception (the plant MAPK signaling pathway, plant hormone signal transduction, and plant pathogen interactions), and plant defense metabolite biosynthesis (phenylpropanoid biosynthesis, α-linolenic acid metabolism, ethylene biosynthesis, and terpenoid backbone biosynthesis). A search of putative cacao resistance genes within the cacao transcriptome identified 89 genes with prominent leucine-rich repeat (LRR) domains, 170 protein kinases encoding genes, 210 genes with prominent NB-ARC domains, 305 lectin-related genes, and 97 cysteine-rich RK genes. We further analyzed the cacao leaf transcriptome in detail focusing on gene families-encoding proteins important in signal transduction (MAP kinases and transcription factors) and direct plant defense (Germin-like, ubiquitin-associated, lectin-related, pathogenesis-related, glutathione-S-transferases, and proteases). There was a massive reprogramming of defense gene processes in susceptible cacao leaf tissue after infection, which was restricted in the resistant genotype Pound7. Most genes induced in Pound7 were induced in ICS1/CCN51. The level of induction was not always proportional to the infection level, raising the possibility that genes are responding to infection more strongly in Pound7. There were also defense-associated genes constitutively differentially expressed at higher levels in specific genotypes, possibly providing a prepositioned defense. Many of the defense genes occur in blocks where members are constitutively expressed at different levels, and some members are induced by Ppal infection. With further study, the identified candidate genes and gene blocks may be useful as markers for breeding disease-resistant cacao genotypes against P. palmivora.
Genetic Transformation of Recalcitrant Upland Switchgrass Using Morphogenic Genes
Authors: Xu Nuoya , Kang Minjeong , Zobrist Jacob D. , Wang Kan , Fei Shui-zhang
Published: February 2022

Switchgrass (Panicum virgatum) is an excellent feedstock for biofuel production. While genetic transformation is routinely done in lowland switchgrass, upland cultivars remain recalcitrant to genetic transformation. Here we report the establishment of an efficient and reproducible transformation protocol for two upland cultivars, ‘Summer’ and ‘Blackwell’, by ectopic overexpression of morphogenic genes, Baby boom (Bbm) and Wuschel2 (Wus2). Two auxotrophic Agrobacterium strains, LBA4404Thy- and EHA105Thy-, each harboring the same construct containing ZmBbm, ZmWus2, and a green fluorescence protein (GFP) gene, ZsGreen1, were used to infect immature leaf segments derived from in vitro grown seedlings. The Agrobacterium strains also contain a transformation helper plasmid that carry additional copies of Agrobacterium virulence genes. GFP-expressing calli were identified and selected for regeneration. The highest transformation efficiency of 6% was obtained for the tetraploid cultivar Summer when LBA4404Thy- was used for infection, which is twice of that for the octoploid cultivar Blackwell. LBA4404Thy- consistently outperformed EHA105Thy- on transformation frequency across the two cultivars. Fifteen randomly selected putative transgenic plants of Summer and Blackwell, representing independent callus events, were confirmed as transgenic by the presence of the transgene, ZmAls, and the absence of AtuFtsZ, a chromosomal gene specific to the Agrobacterium strain LBA4404 using polymerase chain reaction. Transgene integration and expression was further confirmed by the detection of GFP in roots, and the resistance to herbicide injury to leaves of selected putative transgenic plants. The ZmBbm and ZmWus2 genes were successfully removed from 40 to 33.3% of the transgenic plants of Summer and Blackwell, respectively, via the Cre-Lox recombination system upon heat treatment of GFP-expressing embryogenic calli. Our successful transformation of recalcitrant upland switchgrass provides a method for gene function analysis and germplasm enhancement via biotechnology.
Integrated Metabolomic and Transcriptomic Analysis of the Flavonoid Accumulation in the Leaves of Cyclocarya paliurus at Different Altitudes
Authors: Du Zhaokui , Lin Weida , Yu Binbin , Zhu Jinxing , Li Junmin
Published: February 2022

Cyclocarya paliurus is a medicinal plant containing flavonoids, triterpenoids, polyphenolics, polysaccharides, and other compounds with diverse biological functions. C. paliurus is distributed across altitudes ranging from 400 to 1,000 m. However, little is known about the effect of altitude on metabolite accumulation in C. paliurus. Also, the biosynthetic pathway involved in flavonoid accumulation in C. paliurus has not been fully elucidated. In this study, mature leaves of C. paliurus growing at low altitude (280 m) and high altitude (920 m) were sampled and subjected to metabolomic and transcriptomic analyses. The flavonoid content and composition were higher in the leaves of C. paliurus collected at high altitude than in those collected at low altitude. Most of the differentially accumulated metabolites (DAMs) were enriched in “flavone and flavonol biosynthesis.” The significant differentially expressed genes (DEGs) between low and high altitudes were mainly enriched in “biological process.” The most heavily enriched KEGG pathway was related to the subcategory “Oxidative phosphorylation,” indicating that complicated biological processes are involved in the response of C. paliurus to harsh environmental factors. High UV-light might be the main influencing factor among the harsh environmental factors found in high altitudes. Integrated analysis of metabolomic and transcriptomic data showed that 31 flavonoids were significantly correlated with 227 DEGs, resulting in 412 related pairs (283 positive and 129 negative) between the DEGs and flavonoids. The possible mechanisms underlying the differentially accumulation of flavonoids at different altitude might be due to variations in transport and relocation of flavonoids in C. paliurus leaves, but not different flavonoid biosynthesis pathways. The up-regulation of genes related to energy and protein synthesis might contribute to flavonoid accumulation at high altitudes. This study broadens our understanding of the effect of altitude on metabolite accumulation and biosynthesis in C. paliurus.
Virulence Biomarkers of Bursaphelenchus xylophilus: A Proteomic Approach
Authors: Cardoso Joana M. S. , Anjo Sandra I. , Manadas Bruno , Silva Hugo , Abrantes Isabel , Nakamura Katsunori , Fonseca Luís
Published: February 2022

The pinewood nematode (PWN), Bursaphelenchus xylophilus, one of the most serious forest pests worldwide, is considered the causal agent of the pine wilt disease (PWD). The main host species belong to the genus Pinus, and a variation in the susceptibility of several pine species to PWN infection is well-known. It is also recognized that there is variation in the virulence among B. xylophilus isolates. In the present study, we applied a quantitative mass spectrometry-based proteomics approach to perform a deep characterization of proteomic changes across two B. xylophilus isolates with different virulence from different hosts and geographical origins. A total of 1,456 proteins were quantified and compared in the two isolates secretomes, and a total of 2,741 proteins were quantified and compared in the nematode proteomes in pine tree extract and fungus stimuli conditions. From the proteomic analyses, a group of proteins was selected and identified as potential virulence biomarkers and shed light on putative most pathogenic proteins of this plant-parasitic nematode. Proteomic data are available via ProteomeXchange with identifier PXD029377.
An Improved and Simplified Propagation System for Pollen-Free Sugi (Cryptomeria japonica) via Somatic Embryogenesis
Authors: Maruyama Tsuyoshi E. , Tsuruta Momi , Ueno Saneyoshi , Kawakami Kiyohisa , Bamba Yukiko , Moriguchi Yoshinari
Published: February 2022

Sugi (Japanese cedar, Cryptomeria japonica) is the most important forestry tree species in Japan, covering 44% of the total artificial forest area. Large amounts of pollen released from these forests each spring cause allergic reactions in approximately 40% of the population, which are a serious social and public health problem in Japan. As a countermeasure, there is an urgent need to reforest using male-sterile plants (MSPs; pollen-free plants); however, the production of MSPs via conventional methods is inefficient, time consuming, and requires considerable resources in terms of labor and space. In the present paper, we described an improved and simplified methodology for the efficient propagation of pollen-free Japanese cedar, combining the use of genetic markers (marker-assisted selection or marker-aided selection) for the early selection of male-sterile genotypes and the use of somatic embryogenesis (SE) for the clonal mass propagation of seedlings. We describe all the stages involved in the production process of somatic seedlings. Our results demonstrated that this methodology easily and efficiently produces MSPs with a discrimination rate of 100% in a short period of time. Production of 243.6 ± 163.6 cotyledonary embryos per plate, somatic embryo germination, and plantlet conversion frequencies of 87.1 ± 11.9% and 84.8 ± 12.6%, respectively, and a 77.6 ± 12.1% survival rate after ex vitro acclimatization was achieved. Moreover, we also describe an easy method for the collection of somatic embryos prior to germination, as well as an efficient and practical method for their storage at 5°C. Finally, a representative schedule for the propagation of pollen-free sugi somatic seedlings is presented as a reference for practical uses. This methodology will definitively help to accelerate the production of C. japonica MSPs across Japan.
Mapping Floral Genetic Architecture in Prunus mume, an Ornamental Woody Plant
Authors: Li Mingyu , Sang Mengmeng , Wen Zhenying , Meng Juan , Cheng Tangren , Zhang Qixiang , Sun Lidan
Published: February 2022

Floral traits are both evolutionarily and economically relevant for ornamental plants. However, their underlying genetic architecture, especially in woody ornamental plants, is still poorly understood. We perform mapping experiments aimed at identifying specific quantitative trait loci (QTLs) that control the size, shape, architecture, color, and timing of flowers in mei (Prunus mume). We find that the narrow region of chromosome 1 (5–15 Mb) contains a number of floral QTLs. Most QTLs detected from this mapping study are annotated to candidate genes that regulate various biological functions toward the floral formation. We identify strong pleiotropic control on different aspects of flower morphology (including shape, petal number, pistil number, petal color, and calyx color) and flower timing, but find different genetic systems that mediate whether a flower produces pistils and how many pistils a flower produces. We find that many floral QTLs display pleiotropic effects on shoot length growth but shoot radial growth, implicating a possible association of floral display with light capture. We conduct a transcriptomic study to characterize the genomic signature of floral QTLs expressed in mei. Our mapping results about the genetic control of floral features make it promising to select superior varieties for mei carrying flowers of ornamental value.
Cytosolic Glutamine Synthetase GS1;3 Is Involved in Rice Grain Ripening and Germination
Authors: Fujita Takayuki , Beier Marcel Pascal , Tabuchi-Kobayashi Mayumi , Hayatsu Yoshitaka , Nakamura Haruka , Umetsu-Ohashi Toshiko , Sasaki Kazuhiro , Ishiyama Keiki , Murozuka Emiko , Kojima Mikiko , Sakakibara Hitoshi , Sawa Yuki , Miyao Akio , Hayakawa Toshihiko , Yamaya Tomoyuki , Kojima Soichi
Published: February 2022

Ammonium is combined with glutamate to form glutamine. This reaction is catalyzed by glutamine synthetase (GS or GLN). Plants harbor several isoforms of cytosolic GS (GS1). Rice GS1;3 is highly expressed in seeds during grain filling and germination, suggesting a unique role in these processes. This study aimed to investigate the role of GS1;3 for rice growth and yield. Tos17 insertion lines for GS1;3 were isolated, and the nitrogen (N), amino acid, and ammonium contents of GS1;3 mutant grains were compared to wild-type grains. The spatiotemporal expression of GS1;3 and the growth and yield of rice plants were evaluated in hydroponic culture and the paddy field. Additionally, the stable isotope of N was used to trace the foliar N flux during grain filling. Results showed that the loss of GS1;3 retarded seed germination. Seeds of GS1;3 mutants accumulated glutamate but did not show a marked change in the level of phytohormones. The expression of GS1;3 was detected at the beginning of germination, with limited promoter activity in seeds. GS1;3 mutants showed a considerably decreased ripening ratio and decreased N efflux in the 12th leaf blade under N deficient conditions. The β-glucuronidase gene expression under control of the GS1;3 promoter was detected in the vascular tissue and aleurone cell layer of developing grains. These data suggest unique physiological roles of GS1;3 in the early stage of seed germination and grain filling under N deficient conditions in rice.
The Students’ Flow Experience With the Continuous Intention of Using Online English Platforms
Authors: Zhao Hong , Khan Asif
Published: February 2022

Built on the integrated theoretical framework of antecedents of flow and expectation confirmation model (ECM), this research investigates the way flow experience drives the online students’ intention to engage in online English teaching platforms. This study focused on the online students engaged in online English learning platforms in Taiwan. A total of 500 online students were selected. An online survey was conducted with the help of a marketing research agency located in Taiwan. According to the results, the online students’ flow experience was found to be in a significant relationship with continuous intention. The antecedents, including perceived enjoyment, challenge, and situational involvement, were found to be in a positive relationship with flow experience; however, confirmation and perceived vividness did not have significant effects on the flow. Furthermore, flow and confirmation were found to be in a significant relationship with perceived usefulness and satisfaction. Moreover, perceived usefulness was found to be in a significant relationship with satisfaction and continuous intention. Lastly, satisfaction was found to be in a significant relationship with continuous intention. Finally, the questions proposed in this research with their empirical findings offer profound understanding for establishing a well-devised online English learning platform that can motivate online learning. These results and managerial implications for online English language platforms are innovative and significant in practice.
Analysis of the Emotional Exhaustion Derived From Techno-Stress in the Next Generation of Qualified Employees
Authors: Buenadicha-Mateos María , Sánchez-Hernández María Isabel , González-López Óscar Rodrigo
Published: February 2022

This study analyses the emotional exhaustion of students inhigher education, derived from the extremely technology-relatedstrain associated to the current COVID-19 pandemic in a conservation of resources’ approach. Technostress, as source of emotional exhaustion, was investigated in a sample of 333 students in a medium size public university in Spain. Data was collected in May 2020, during the COVID lockdown. After literature review, a structural model was developed, linking technostress with emotional exhaustion. Results confirm the expected cause-effect relationships. In addition, the study reveals two mediator variables that must be considered when managing students′ suffering, perceived stress and intrapersonal conflicts. This study contributes to the academic literature in the field of managing and mitigating suffering. They do so by providing both new knowledge and empirical evidence on the effects of technostress in the new generations that will soon join the working life.
Symptom Patterns of the Occurrence of Depression and Anxiety in a Japanese General Adult Population Sample: A Latent Class Analysis
Authors: Lei Huijie , Chen Chong , Hagiwara Kosuke , Kusumi Ichiro , Tanabe Hajime , Inoue Takeshi , Nakagawa Shin
Published: February 2022

Background

Given the high comorbidity and shared risk factors between depression and anxiety, whether they represent theoretically distinct disease entities or are just characteristics of a common negative affect dimension remains debated. Employing a data-driven and person-centered approach, the present study aims to identify meaningful and discrete symptom patterns of the occurrence of depression and anxiety.

Methods

Using data from an adult sample from the Japanese general population (n = 403, including 184 females, age = 42.28 ± 11.87 years), we applied latent class analysis to identify distinct symptom patterns of depression (PHQ-9) and anxiety (STAI Y1). To empirically validate the derived class memberships, we tested the association between the derived classes and personal profiles including childhood experiences, life events, and personality traits.

Results

The best-fitting solution had four distinct symptom patterns or classes. Whereas both Class 1 and 2 had high depression, Class 1 showed high anxiety due to high anxiety-present symptoms (e.g., “I feel nervous”) while Class 2 showed moderate anxiety due to few anxiety-absent symptoms (e.g., “I feel calm”). Class 3 manifested mild anxiety symptoms due to lacking responses on anxiety-absent items. Class 4 manifested the least depressive and anxiety-present symptoms as well as the most anxiety-absent symptoms. Importantly, whereas both Class 1 and 2 had higher childhood neglect and reduced reward responsiveness, etc. compared to Class 4 (i.e., the most healthy class), only Class 1 had greater negative affect and reported more negative life events.

Conclusions

To our knowledge, this is the first latent class analysis that examined the symptom patterns of depression and anxiety in Asian subjects. The classes we identified have distinct features that confirm their unique patterns of symptom endorsement. Our findings may provide insights into the etiology of depression, anxiety, and their comorbidity.
Measurement Invariance of the Questionnaire on the Internal Stigma of Internet Surfing Among Sino-Australian Undergraduates
Authors: Luo Jie , Ge Ying , Hao Ji-chun , Wilkinson Ross B. , Wenger Jay L.
Published: February 2022

Background

The stigma of internet surfing is a relatively new area of study arising from the popularity of the internet. The Questionnaire on the Internal Stigma of Internet Surfing-9 (QISIS-9) was developed for the Chinese culture, so its suitability for use in other cultural contexts is uncertain. This paper examines the measurement invariance of the QISIS-9 among Sino-Australian undergraduates to verify the cross-cultural measurement invariance of QISIS-9 and promote cross-cultural (nationality) research regarding the internal stigma of internet surfing.

Methods

The Internal Stigma of Internet Surfing-9 (QISIS-9) was used to assess 200 Chinese undergraduates (50% female, Mage = 19.78) and 204 Australian undergraduates (76% female, Mage = 21.10), respectively.

Results

A confirmatory factor analysis (CFA) indicated that the single-factor model of QISIS-9 is acceptable with both Chinese and Australian undergraduates. However, the factor loading of Item 9, to which a reverse score is assigned, is not ideal for both samples. Thus, the item should be deleted. According to a multigroup confirmatory factor analysis (MCFA), QISIS-8, the revised version of QISIS-9, meets the strict measurement invariance among the Chinese and Australian participants. The QISIS-8 demonstrated appropriate internal consistency in the scores for both the Chinese and Australian undergraduates.

Conclusion

The new QISIS-8 can effectively assess the internal stigma of internet surfing among Chinese and Australian undergraduates, and it provides a frame of reference for further cross-cultural (border) comparisons.