bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunogold labeling of synaptic vesicle proteins in developing hippocampal neurons
Authors: Tao-Cheng Jung-Hwa 
Published: November 2019

Abstract
Synaptic vesicles (SV) contain high concentrations of specific proteins. How these proteins are transported from soma to synapses, and how they become concentrated at SV clusters at presynaptic terminals were examined by immunogold electron microscopy in dissociated rat hippocampal neurons at 3-6 days in culture, a developmental stage when axonal transport of SV proteins is robust. In neuronal somas, labels for the SV integral membrane proteins (synaptophysin, SV2, VAMP/synaptobrevin, and synaptotagmin) were localized at Golgi complexes and other membranous structures that were dispersed in the cytoplasm as individual vesicle/vacuoles. These vesicles/vacuoles became aggregated in axons, with the size of aggregates ranging from 0.2 to 2 µm in length. Pleomorphic vesicle/vacuoles within the aggregate were typically larger (50-300 nm) than SVs, which were uniform in size at ~40 nm. These pleomorphic vesicles/vacuoles are likely transport cargos carrying SV integral membrane proteins from the soma, and sorted into axons at early developmental stages. Serial thin sections of young axons indicated that many labeled aggregates were not synaptic, and in fact, some of these axons were without dendritic contacts. In contrast, labels for two SV associated proteins, synapsin I and a-synuclein, were not localized at the Golgi complexes or associated with membranous structures in the soma, but were dispersed in the cytoplasm. However, these SV-associated proteins became highly concentrated on clusters of SV-like vesicles in axons, and such clusters were already distinctive in axons as early as 3 days in culture. These clusters consisted of ~4 - 30 vesicles in single thin sections, and the vesicles were of a uniform size (~40 nm). By serial sectioning analysis, these clusters could be part of nascent synapses or exist in axons without any dendritic contact. Importantly, the vesicles were intensely labeled for SV integral membrane proteins as well as SV associated proteins. Thus, these EM observations illustrate that the two groups of proteins, SV integral membrane and SV-associated, proceed through different routes of biosynthesis and axon transport, and are only sorted into the same final compartment, SV clusters, when they are in the axons.
Arabidopsis adaptor protein 1G2 is required for female and male gametogenesis
Authors: Zhou Yongmei , Fang Wenqin , Chen Li-Yu , Pandey Neha , Muhammad Azam Syed , Ming Ray 
Published: November 2019

Abstract
Background: The gametophyte s are essential for the productive process in angiosperms. During sexual reproduction in flowering plants, haploid spores are formed from meioses of spore mother cells. The spores then undergo mitosis and develop into female and male gametes and give rise to seeds after fertilization. Results: We identified a female sterile mutant from EMS mutagenesis, and a BC1F2 population was generated for map based cloning of the causal gene. Genome re-sequencing of mutant and non-mutant pools revealed a candidate gene, AP1G2 . Analyses of two insertions mutants, ap1g2-1 +/- in exon 7 and ap1g2-3 -/- in 3’ UTR, revealed partial female sterility. Complementation test using native promoter of AP1G2 restored the function in ap1g2-1 +/- and ap1g2-3 -/- . AP1G2 is a paralog of AP1G1 , encoding the large subunit (γ) of adaptor protein-1 (AP-1). ap1g2 mutation led to defective female and male gametophyte development was determined. In the ap1g2 mutants, the mitotic cycles and synchronic development of female gametophytes were impaired, which led to the arrest of female gametophytes at one nucleus stage FG1. Pollen development in ap1g2 was also arrested at one nucleus stage before PMI (pollen mitosis I). AP1G2 was expressed at high levels in different stages of ovule and pollens and actively dividing tissues, including shoot apical meristems, leaf primordial and root tips. Conclusions: AP1G2 was identified to have a role in the processes of female and male gametogenesis by regulating the first mitosis at one nucleus stage, and the expression pattern suggested AP1G2 is crucial for plant growth and development. Keywords: Arabidopsis, AP1G2 , megagametogenesis, microgametogenesis, development.
Different Doses of Allergen Exposure on Dendritic Cells Determine Their Genetic/Epigenetic Regulation and T Cell Differentiation
Authors: Chen Jianjun , Wang Ying , Yu Zizhong , Zhou Yue , Zhu Yun , Wang Jinhui , Fu Junmei , Yuan Yang , Chen Shan , Wang Yanjun , Cho Seong H , Yu Wenting , Gao Pei , Zhu Wanting , Kong Weijia 
Published: November 2019

Abstract
Background: It has been reported that when DCs are exposed to high-dose antigens, they can induce Th0 cells into regulatory cells (Treg) and Th1 cells. When DCs are subjected to low-dose allergen, they can drive to Th2 cells. However, the mechanisms of such dose-effect relationship are poorly understood so far. Methods: Bone marrow immature DCs (imDCs) were generated from mice and stimulated with OVA of different concentrations (0, 10, 100, 1000, 10000 μg/ml, respectively). The mDCs were then seeded and cocultured with naïve T cells for 3 days, and then the markers of different T cell subgroups were flow cytometrically tested. RNA-seq detection and DNA methylation of DCs were performed. Results: When DCs were stimulated with low-dose (10ug/ml), 77 genes were up-regulated and 87 genes down-regulated. Most activated genes were related to ribosome synthesis and ion channel inhibition but not to the immune responses and Th2 activation. At the medium-dose (100μg/ml), 339 genes were up-regulated and 168 genes down-regulated. Most activated genes involved cytokine synthesis and regulation of immune responses. At high-dose (10000μg/ml), 2794 genes were up-regulated and 1156 genes down-regulated. Tumor necrosis factor signaling pathway, MAPK signaling pathway, antigen processing and presentation signaling pathway were mostly up-regulated. The related co-stimulators, co-inhibitory molecules, inhibitory cytokines, negative regulating enzymes were highly expressed. The monocarbate, coenzyme, fatty acid, glucolipid, starch, sucrose and other metabolism-related signaling pathways were down-regulated. Conclusion: The profiles of DNA methylation and RNA synthesis of DCs varied with different doses of OVA, which serves to induce T cells to differentiate in various directions.
Genomic Variation between PRSV Resistant Transgenic SunUp and Its Progenitor Cultivar Sunset Induced by Particle Bombardment Transformation
Authors: Fang Jingping , Wood Andrew , Chen Youqiang , Yue Jingjing , Ming Ray 
Published: November 2019

Abstract
The safety of genetically transformed plants remains a subject of scrutiny. Genomic variants in PRSV resistant transgenic papaya will provide evidence to rationally address such concerns. In this study, a total of more than 74 million Illumina reads for progenitor ‘Sunset’ were mapped onto transgenic papaya ‘SunUp’ reference genome. 310,364 single nucleotide polymorphisms (SNPs), 34,071 Small Inserts/deletions (InDels) and 1,200 large structural variations (SVs) were detected between ‘Sunset’ and ‘SunUp’. Those variations have an uneven distribution across nine chromosomes in papaya. Only 0.27% of mutations were predicted to be high-impact mutations. ATP-related categories were highly enriched among these high-impact genes. The SNP mutation rate was about 8.4×10 -4 per site, comparable with the rate induced by spontaneous mutation over numerous generations. The transition-to-transversion ratio was 1.439 and the predominant mutations were C/G to T/A transitions. Spontaneous mutations were the leading cause of SNPs in transgenic papaya ‘SunUp’. A total of 3,430 nuclear plastid DNA (NUPT) and 2,764 nuclear mitochondrial DNA (NUMT) junction sites have been found in ‘SunUp’, which is proportionally higher than the predicted total NUPT and NUMT junction sites in ‘Sunset’ (3,346 and 2,745, respectively). Among all nuclear organelle DNA (norgDNA) junction sites, 96% of junction sites were shared by ‘SunUp’ and ‘Sunset’. The average identity between ‘SunUp’ specific norgDNA and corresponding organelle genomes was higher than that of norgDNA shared by ‘SunUp’ and ‘Sunset’. Six ‘SunUp’ organelle-like borders of transgenic insertions were nearly identical to corresponding sequences in organelle genomes (98.18~100%). None of the paired-end spans of mapped ‘Sunset’ reads were elongated by any ‘SunUp’ transformation plasmid derived inserts. Significant amounts of DNA were transferred from organelles to the nuclear genome during bombardment, including the six flanking sequences of the three transgenic insertions. Comparative whole-genome analyses between ‘SunUp’ and ‘Sunset’ provide a reliable estimate of genome-wide variations and evidence of organelle-to-nucleus transfer of DNA associated with biolistic transformation.
Neuropathological changes in dorsal root ganglia induced by pyridoxine in dogs
Authors: Yun Sumin , Kim Woosuk , Kang Min Soo , Kim Tae Hyeong , Kim Yoonhwan , Ahn Jin-Ok , Choi Jung Hoon , Hwang In Koo , Chung Jinyoung 
Published: November 2019

Abstract
Background: Pyridoxine (PDX), vitamin B 6 , is an essential vitamin. PDX deficiency induces various symptoms and abuse of PDX supplement also acts as a neurotoxicant that induces severe sensory neuropathy. Results: To construct a sensory neuropathy model in dogs, excess pyridoxine (PDX) was injected subcutaneously into dogs for seven days and dorsal root ganglia (DRG) were observed at 0, 1 and 4 weeks after the last PDX treatment. During the administration period, body weight loss and proprioceptive loss occurred. After the treatments were completed, electrophysiological recordings showed that the H-reflex of the treated dogs had disappeared at week 0. These phenomena persisted for four weeks exceptional body weight. CV and HE staining revealed that neurons in DRG had significant loss of large-sized neurons 0 weeks, but these neurons were recovered 4 weeks. Iba-1 and GFAP immunohistochemistry showed that reactive microgliosis and satellite glial cells were pronounced at 1 week. Conclusions: This result suggests that PDX-induced neuropathy model is reversible and can be a good experimental model for research on neuropathy in dogs.
Clinical efficacy analysis of different therapeutic methods in patients with cesarean scar pregnancy
Authors: Wu Yan , Sun Leifang , Si Yanan , Luan Xiaoli , Gao Yongmei 
Published: November 2019

Abstract
Background Cesarean scar pregnancy (CSP) is a rare form of ectopic pregnancy with a serious life-threatening situation. The purpose of this study is to analyze the clinical efficacy of five therapeutic strategies in patients with CSP. Methods A total of 135 CSP patients were included in our research and divided into five groups based on the treatment they received, including transvaginal resection (Group A), laparoscopic resection (Group B), uterine arterial embolization (UAE) combined with hysteroscopic curettage (Group C), UAE combined with uterine curettage (Group D), and hysteroscopic curettage (Group E). To investigate the clinical efficacy of these strategies, intraoperative bleeding, serum β-hCG levels and recovery time, menstruation recovery time, hormone levels at 1 month after treatment, hospital stays and costs were analyzed. Results The lowest postoperative serum β-hCG levels and the shortest serum β-hCG and menstruation recovery times were observed in group A, the next were in group B. Group C and D had small amount of blood loss. The hospital stays and costs were low in group E. In addition, no significant difference was performed at the sex hormone levels between the five groups. Conclusions Our results indicated that resection surgery and UAE have good curative effects, but high hospital costs in CSP treatment. The selection of an optimal treatment regimen for CSP should be carried out based on specific conditions of the patients.
Epidemiology of Multiple sclerosis in Saudi Arabia, a Descriptive study
Authors: Jumah Mohammed Al , Bunyan R. , Otaibi H. Al , Towaijri G. Al , Karim A. , Malik Y. Al , Kalakatawi M. , Alrajeh S. , Mejally M. Al , Gahtani H. Al , Almubarak A. , Cupler E. , Alawi S. , Qureshi S. , Naharir S. , Almalki A. , Alhazzani A. , Althubaiti I. , Alzahrani N. , Alradhi G. , Saeedi J. , Ishak S. , Almudaiheem H. , Elmetwally A. , Jedaie A. Al 
Published: November 2019

Abstract
Introduction In 2015, the first nationwide, multi-center MS registry was initiated in the Kingdom of Saudi Arabia with an objective to describe current epidemiology, disease patterns, and clinical characteristics of MS in Saudi Arabia. This article aims to report initial findings of the registry and report regional prevalence of MS. Method The registry included all MS patients with confirmed MS diagnosis according to 2010 McDonald Criteria. It aimed to identify and recruit all healthcare institutions treating MS patients in the Kingdom, and collect data such as demographics, clinical characteristics (disease onset, diagnosis, disease course, relapse rate, and disability measures), family history, and treatments. All the included sites have obtained IRB/EC approvals for participating in the registry. Results As of September 2018, the registry has included 20 hospitals from the different regions across the Kingdom and has collected comprehensive data on 2,516 patients from those hospitals, with mean age 33.42 (SD: 9.70) and 66.5% being females. The reported prevalence of MS for those hospitals is estimated to be 7.70/100,000 total population and 11.80/100,000 Saudi nationals. Assuming that the prevalence is the same across all the hospitals in the particular region, we projected the prevalence for total hospitals capable of diagnosing and treating MS and accordingly projected the total prevalence of MS as 40.40/100,000 total population and 61.95/100,000 Saudi nationals. The prevalence was reported 77.5% among people age <40 and below. The female to male ratio was 2:1. The prevalence was higher among female, young and educated individuals across all five regions of Saudi Arabia. Conclusion The prevalence of MS has significantly increased in Saudi Arabia but is still much lower than western and other neighboring countries like Kuwait, Qatar, and the UAE. However, compared to the past rates, Saudi Arabia’s projected prevalence of MS through this national study is 40.40/100,000population, putting them above the low risk zone as per Kurtzke classification. The projected prevalence is estimated to be much higher among Saudi nationals only (61.95/100,000Saudi-nationals). The prevalence was higher among female, younger, and educated individuals. Further studies are needed to asses the risk factors associated with increased prevalence in Saudi Arabia.
Duck enteritis virus UL21 is a late gene and encodes a protein that interacts with pUL16
Authors: Yang Linjiang , Wang Mingshu , Zeng Chunhui , Shi Yong , Cheng Anchun , Liu Mafeng , Zhu Dekang , Chen Shun , Jia Renyong , Yang Qiao , Wu Ying , Zhang Shaqiu , Zhao Xinxin , Huang Juan , Liu Yunya , Ou Xumin , Mao Sai , Yu Yanling , Zhang Ling , Tian Bin , Pan Leichang , Rehman Mujeeb Ur , Chen Xiaoyue 
Published: November 2019

Abstract
Background pUL21 is a conserved protein of Alphaherpesvirinae and exhibits multiple important functions. The C-terminus of pUL21 in other members of this subfamily has RNA-binding ability; this domain contributes to pseudorabies virus (PRV) retrograde axonal transport in vitro and in vivo and participates in newly replicated viral DNA packaging and intracellular virus transport. However, little is known about duck enteritis virus (DEV) pUL21. Methods In our study, recombinant pUL21 was expressed using apET-32c (+) vector in Escherichia coli BL21 cells induced with 0.4 mM isopropyl β-D-thiogalactoside for 8 h at 30°C. The antibody for indirect immunofluorescence (IFA) and western blotting (WB) analysis were then prepared. Pharmacological inhibition, WB and quantitative reverse transcription PCR (RT-qPCR) were performed. A coimmunoprecipitation (CO-IP) assay was conducted to test the interaction between pUL21 and pUL16. Results We verified that DEV UL21 is a γ2 gene and encodes a structural protein. Moreover, we observed that pUL21 localized to the nucleus and cytoplasm. DEV pUL21 interacted with pUL16 and formed a complex in transfected human embryonic kidney (HEK) 293T cells and DEV-infected duck embryo fibroblasts (DEFs). These results were further confirmed by CO-IP assays. Conclusions The DEV UL21 gene is a late gene, and the pUL21 localizes to the nucleus and cytoplasm. DEV UL21 is a virion component. In addition, pUL21 can interacts with pUL16. These ﬁndings provide insight into the characteristics of UL21 and the interaction between pUL21 and binding partner pUL16. Our study enhances understanding of DEV pUL21.
Proteomic Study of Hypothalamus in Pigs Exposed to Heat Stress
Authors: Yu Tianyue , Yong Yan-Hong , Li Jun-yu , Fang Biao , Hu Can-ying , Wu Lian-yun , Gooneratne Ravi , Patil Yadnyavalkya , Ju Xianghong 
Published: November 2019

Abstract
Background: With evidence of warming climates, it is important to understand the effects of heat stress in farm animals in order to minimize production losses. Study of changes in the brain proteome induced by heat stress may aid in understanding how heat stress impacts brain function. The hypothalamus is a key region in the brain that controls the pituitary gland, which is responsible for the secretion of several important hormones. Result: In this study, we examined the hypothalamic protein profile of ten pigs (30–40 kg body weight), 5 of which were subjected to heat stress (35 ± 1°C; relative humidity = 90%) and 5 acting as controls (28 ± 3°C; RH = 90%). The isobaric tags for relative and absolute quantification (iTRAQ) analysis of the hypothalamus identified 1710 peptides corresponding to 360 proteins, and 295 deferentially expressed proteins (DEPs), 148 of which were up-regulated and 147 down-regulated in heat-stressed animals. Ingenuity Pathway Analysis software predicted 30 canonical pathways, 4 functional groups, and 4 regulatory networks of interest and the DEPs mainly concentrated in the cytoskeleton of the pig hypothalamus during heat stress. Conclusions: The upstream regulators of these 295 DEPs in the hypothalamus of the pig under HS are mainly transcriptional regulators, chemical drugs, and sRNA. This study provides reference data for further study of the mechanism of HS on hypothalamic physiology and metabolism(Illustration 1).
Unilateral visual impairment in a patient undergoing chemotherapy: a case report and clinical findings
Authors: Yuan Xia , Feng Yuliang , Li Dan , Li Mei 
Published: November 2019

Abstract
Visual impairment occurred as an infrequent form of chemotherapeutic toxicity and was often underestimated despite of several reports. We described a case of acute unilateral visual impairment after one cycle of intravenous chemotherapy of a normal dose, aiming at raising attention to chemotherapy-induced ocular toxicity. Case presentation: The patient developed a progressive vision loss in the right eye during the chemotherapy. After one cycle of intravenous chemotherapy, her visual acuity decreased by 0.6 in the right eye (VOD=0.4) compared to the previous value of 1.0 (VOD=1.0). No evidence of ocular infiltration was observed from the cerebral magnetic resonance imaging (MRI). During her follow-up period, we documented the ophthalmologic examinations including visual acuity, visual field (VF), visual evoked potential (VEP), electroretinogram (ERG), fundus photograph (FP), fundus fluorescein angiography (FFA) and optical coherence tomography (OCT). Ophthalmoscope examination and fundus photograph showed optic disc edema, fuzzy boundary and linear hemorrhages in her right eye. Fundus fluorescein angiography (FFA) revealed capillary underdevelopment at the nasal and superior temporal area of the optic disc in the early phase and capillary fluorescein leakage in the late phase. The result of VEP test suggested the impaired function of the optic nerve. Thus, a diagnosis of nonarteritic anterior ischemic optic neuropathy (NAION) was made by the ophthalmologist according to these results. The patient was prescribed prednisone combined with neuroprotective drugs, which did not work. After the cessation of chemotherapy, her impaired vision gradually recovered. Conclusions: This is the first reported case of acute visual impairment in a patient who underwent chemotherapy of a normal dose. It is indicated that while receiving benefits from chemotherapy, cancer patients simultaneously suffer from the risk of vision loss. Key words: Vision loss, Chemotherapy, Nonarteritic anterior ischemic optic neuropathy
Full-length transcriptome sequences of Agropyron cristatum facilitate the prediction of putative genes for thousand-grain weight in a wheat-A. cristatum translocation line
Authors: Author) Shenghui Zhou(Former Corresponding , Zhang Jinpeng , Han Haiming , Zhang Jing , Huihui Ma , Zhang Zhi , Lu Yuqing , Liu Weihua , Yang Xinming , Li Xiuquan , Author) Lihui Li(New Corresponding 
Published: November 2019

Abstract
Agropyron cristatum (L.) Gaertn. (2n = 4x = 28; genomes PPPP) is a wild relative of common wheat (Triticum aestivum L.) and provides many desirable genetic resources for wheat improvement. However, there is still a lack of reference genome and transcriptome information for A. cristatum, which severely impedes functional and molecular breeding studies.Results Single-molecule long-read sequencing technology from Pacific Biosciences (PacBio) was used to sequence full-length cDNA from a mixture of leaves, roots, stems and caryopses and constructed the first full-length transcriptome dataset of A. cristatum, which comprised 44,372 transcripts. As expected, the PacBio transcripts were generally longer and more complete than the transcripts assembled via the Illumina sequencing platform in previous studies. By analyzing RNA-Seq data, we identified tissue-enriched transcripts and assessed their GO term enrichment; the results indicated that tissue-enriched transcripts were enriched for particular molecular functions that varied by tissue. We identified 3,398 novel and 1,352 A. cristatum-specific transcripts compared with the wheat gene model set. To better apply this A. cristatum transcriptome, the A. cristatum transcripts were integrated with the wheat genome as a reference sequence to try to identify candidate A. cristatum transcripts associated with thousand-grain weight in a wheat-A. cristatum translocation line, Pubing 3035.Conclusions Full-length transcriptome sequences were used in our study. The present study not only provides comprehensive transcriptomic insights and information for A. cristatum but also proposes a new method for exploring the functional genes of wheat relatives under a wheat genetic background. The sequence data have been deposited in the NCBI under BioProject accession number PRJNA534411.