Identification and analysis of the amylase-binding protein B (AbpB) and gene (abpB) from Streptococcus gordonii

The binding of salivary amylase to Streptococcus gordonii has previously been shown to involve a 20-kDa amylase-binding protein (AbpA). S. gordonii also releases an 82-kDa protein into the supernatant that binds amylase. To study this 82-kDa component, proteins were precipitated from bacterial culture supernatants by the addition of acetone or purified amylase. Precipitated proteins were separated by SDS-PAGE and transferred to a sequencing membrane. The P2 kDa band was then sequenced, yielding a 25 N-terminal amino acid sequence, CGFIFGRQLTADGSTMFGPTEDYP. Primers derived from this sequence were used in an inverse PCR strategy to clone the full-length gene from S. gordonii chromosomal DNA. An open reading frame of 1959 bp was noted that encoded a 652 amino acid protein having a predicted molecular mass of 80 kDa. The first 24 amino acid residues were consistent with a hydrophobic signal peptide, followed by a 25 amino acid N-terminal sequence that shared identity (24 of 25 residues) with the amino acid sequence of purified AbpB. The abpB gene from strains of S. gordonii was interrupted by allelic exchange with a 420-bp fragment of the abpB gene linked to an erythromycin cassette. The 82-kDa protein was not detected in supernatants from these mutants. These abpB mutants retained the ability to bind soluble amylase. Thus, AbpA, but not AbpB, appears sufficient to be the major receptor for amylase binding to the streptococcal surface. The role of AbpB in bacterial colonization remains to be elucidated.     

Modeling CADASIL vascular pathologies with patient-derived induced pluripotent stem cells

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy(CADASIL)is a rare hereditary cerebrovascular disease caused by a NOTCH3 mutation.However,the underlying cellular and molecular mechanisms remain unidentified.Here,we generated non-integrative induced pluripotent stem cells(iPSCs)from fibroblasts of a CADASIL patient harboring a heterozygous NOTCH3 mutation(c.3226C>T,p.R1076C).Vascular smooth muscle cells(VSMCs)differentiated from CADASIL-specific iPSCs showed gene expression changes associated with disease phenotypes,including activation of the NOTCH and NF-kB signaling pathway,cytoskeleton disorganization,and excessive cell proliferation.In comparison,these abnormalities were not observed in vascular endothelial cells(VECs)derived from the patients iPSCs.Importantly,the abnormal upregulation of NF-kB target genes in CADASIL VSMCs was diminished by a NOTCH pathway inhibitor,providing a potential therapeutic strategy for CADASIL.Overall,using this iPSCbased disease model,our study identified clues for studying the pathogenic mechanisms of CADASIL and developing treatment strategies for this disease.     

Regulatory action of prolactin on the in vitro growth of CD34+ve human hemopoietic progenitor cells

The pituitary hormone prolactin (Prl) is known to act as a local regulator of immune cell function, and Prl-binding receptors (Prl-R) have been described to share distinctive features with the members of the newly described cytokine/hemopoietin receptor superfamily. Here we show that the hormone can functionally interact with lineage-specific hemopoietic factors. When highly purified progenitor cells (CD34+ve) were seeded in semisolid methylcellulose cultures in the presence of interleukin (IL)-3, granulocyte-macrophage colony stimulating factor (GM-CSF), and erythropoietin (Epo), a selective enhancing effect of Prl on the formation of colony forming unit-granulocyte (CFU-G) and burst forming unit-erythroid (BFU-E) colonies was observed. The effect of the hormone was plotted as a bell shaped curve, with the optimal response at the supraphysiological concentration of 50 ng/ml. Limiting dilution analysis showed that Prl acted directly on hemopoietic progenitors. This was confirmed by the observation on the CD34+ve cells of Prl-binding sites reacting with the specific monoclonal antibodies (mAbs), U5 and PrR-7A. Immunoprecipitation of the metabolically labeled CD34+ve cells with the PrR-7A mAb revealed a structure of 43 kD under reducing conditions. Analysis of the early events associated with the Prl/Prl-R interaction showed an increased number of cells engaged in DNA and hemoglobin synthesis. Enhanced erythroid differentiation of CD34+ve cells in the presence of Prl was secondary to upmodulation of receptors for the lineage-specific factor Epo. Together these data demonstrate the existence of a functional interplay between Prl. and hemopoietic factors. © 1995 Wiley-Liss, Inc.     

基于环境库兹涅茨曲线的我国大气污染防治重点区域环境空气质量与经济增长关系研究

改革开放以来,中国经济迅猛发展,但大气污染等环境问题日益突出。进入21世纪,我国通过颁布实施多项大气污染防治政策,将京津冀及周边地区、长三角地区、珠三角地区等大气污染较严重的区域划定为重点区域,针对性制定治污措施和实施减排工程,努力推动区域环境空气质量改善。基于2000-2019年我国31个省（自治区、直辖市）（以下简称31个省份）GDP,以及SO₂、PM₁₀、NO₂三项大气污染物浓度数据,利用环境库兹涅茨曲线（EKC,Environmental Kuznets Curve）模型,对31个省份和京津冀及周边地区、长三角地区、珠三角地区的经济增长情况、大气污染物浓度演变以及二者之间的关系进行了系统全面的分析评估。研究结果显示：（1）近年来实施的各项大气污染防治政策,特别是2013年以来颁布实施的《大气污染防治行动计划》《打赢蓝天保卫战三年行动计划》,推动环境空气质量改善的同时,促进了经济发展与环境保护长期关系协调性逐步增强,除NO₂浓度呈U型外,31个省份SO₂浓度、PM₁₀浓度与人均GDP的EKC曲线呈倒U型和倒N型,并处于快速下降阶段。（2）京津冀及周边地区SO₂浓度与人均GDP呈倒U型,且处于快速下降阶段;PM₁₀和NO₂浓度均呈现U型关系,且均处于上升期。（3）长三角地区SO₂、PM₁₀浓度与人均GDP呈现倒U型和U型,但均处于下降阶段;NO₂浓度与人均GDP无相关关系。（4）珠三角地区SO₂、PM₁₀和NO₂浓度与人均GDP均呈现倒U型关系,且均处于下降阶段。为此,建议"十四五"期间我国政府要继续实施新一轮的大气污染防治行动计划,聚焦机动车NO_x污染管控,大力推动NO₂浓度稳步下降,以实现我国环境空气质量持续改善,为统筹经济高质量发展和生态环境高水平保护奠定坚实基础。     

Sphingosine kinase: biochemical and cellular regulation and role in disease

Sphingolipids have emerged as molecules whose metabolism is regulated leading to generation of bioactive products including ceramide, sphingosine, and sphingosine-1-phosphate. The balance between cellular levels of these bioactive products is increasingly recognized to be critical to cell regulation; whereby, ceramide and sphingosine cause apoptosis and growth arrest phenotypes, and sphingosine-1-phosphate mediates proliferative and angiogenic responses. Sphingosine kinase is a key enzyme in modulating the levels of these lipids and is emerging as an important and regulated enzyme. This review is geared at mechanisms of regulation of sphingosine kinase and the coming to light of its role in disease.     

Prion protein attenuates excitotoxicity by inhibiting NMDA receptors

It is well established that misfolded forms of cellular prion protein (PrP [PrP(C)]) are crucial in the genesis and progression of transmissible spongiform encephalitis, whereas the function of native PrP(C) remains incompletely understood. To determine the physiological role of PrP(C), we examine the neurophysiological properties of hippocampal neurons isolated from PrP-null mice. We show that PrP-null mouse neurons exhibit enhanced and drastically prolonged N-methyl-d-aspartate (NMDA)-evoked currents as a result of a functional upregulation of NMDA receptors (NMDARs) containing NR2D subunits. These effects are phenocopied by RNA interference and are rescued upon the overexpression of exogenous PrP(C). The enhanced NMDAR activity results in an increase in neuronal excitability as well as enhanced glutamate excitotoxicity both in vitro and in vivo. Thus, native PrP(C) mediates an important neuroprotective role by virtue of its ability to inhibit NR2D subunits.     

Microcella flavibacter sp. nov., isolated from marine sediment,and reclassification of Chryseoglobus frigidaquae,Chryseoglobus indicus,and Yonghaparkia alkaliphila as Microcella frigidaquae comb. nov., Microcella indica nom. nov., and Microcella alkalica nom. nov.

A novel Gram-staining positive, aerobic, rod-shaped, non-motile and yellow-pigmented actinobacterium, designated strain WY83^T, was isolated from a marine sediment of Indian Ocean. Strain WY83^T grew optimally at 30–35 °C, pH 7–8 and with 0–3% (w/v) NaCl. The predominant menaquinones were MK-10, MK-11 and MK-12, and the major fatty acids were C_19:1 ω9c/C19:1 ω11c, anteiso-C_15:0, C_17:0 3OH, and iso-C_16:0. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol and one unidentified glycolipid. The cell-wall peptidoglycan contained lysine as a diamino acid. The DNA G?+?C content was 72.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences and ninety-two bacterial core genes indicated that strain WY83^T formed an evolutionary lineage with Chryseoglobus frigidaquae JCM 14730^T, Chryseoglobus indicus CTD02-10-2^T, Yonghaparkia alkaliphila JCM 15138^T, Microcella alkaliphila DSM 18851^T and Microcella putealis DSM 19627^T within the radiation enclosing members of the family Microbacteriaceae. All pairwise percentage of conserved proteins between strain WY83^T and the closely related phylogenetic neighbors were greater than 65%. The average nucleotide identity and in silico DNA–DNA hybridization values were both below the thresholds used for the delineation of a new species. On the basis of the evidence presented, strains WY83^T, Y. alkaliphila JCM 15138^T, C. frigidaquae JCM 14730^T, M. alkaliphila DSM 18851^T and M. putealis DSM 19627^T should belong to different species of the same genus. Strain WY83^T represents a novel species of the genus Microcella, for which the name Microcella flavibacter sp. nov. is proposed. The type strain is WY83^T (=?KCTC 39637^T?=?MCCC 1A07099^T). Furthermore, Chryseoglobus frigidaquae, Chryseoglobus indicus, and Yonghaparkia alkaliphila were reclassified as Microcella frigidaquae comb. nov., Microcella indica nom. nov., and Microcella alkalica nom. nov., respectively.

     

Simultaneous binding of Guidance Cues NET1 and RGM blocks extracellular NEO1 signaling

1. Download : Download high-res image (287KB)
2. Download : Download full-size image

     

Single Cell Analysis of Lymph Node Tissue from HIV-1 Infected Patients Reveals that the Majority of CD4+ T-cells Contain One HIV-1 DNA Molecule

Genetic recombination contributes to the diversity of human immunodeficiency virus (HIV-1). Productive HIV-1 recombination is, however, dependent on both the number of HIV-1 genomes per infected cell and the genetic relationship between these viral genomes. A detailed analysis of the number of proviruses and their genetic relationship in infected cells isolated from peripheral blood and tissue compartments is therefore important for understanding HIV-1 recombination, genetic diversity and the dynamics of HIV-1 infection. To address these issues, we used a previously developed single-cell sequencing technique to quantify and genetically characterize individual HIV-1 DNA molecules from single cells in lymph node tissue and peripheral blood. Analysis of memory and naïve CD4⁺ T cells from paired lymph node and peripheral blood samples from five untreated chronically infected patients revealed that the majority of these HIV-1-infected cells (>90%) contain only one copy of HIV-1 DNA, implying a limited potential for productive recombination in virus produced by these cells in these two compartments. Phylogenetic analysis revealed genetic similarity of HIV-1 DNA in memory and naïve CD4⁺ T-cells from lymph node, peripheral blood and HIV-1 RNA from plasma, implying exchange of virus and/or infected cells between these compartments in untreated chronic infection.     

Potential for Biocontrol of Lasiodiplodia theobromae (Pat.) Griff. & Maubl. in Banana Fruits by Trichoderma Species

Fifteen Trichoderma isolates were tested for their antagonistic ability against Lasiodiplodia theobromae. Trichoderma harzianum exhibited the greatest inhibition in dual culture. Microscopic investigation demonstrated direct parasitism and coiling of T. harzianum and T. viride around hyphae of L. theobromae, causing swollen, deformed, shortened, or rounded cells of the pathogen. Granulation of cytoplasm and disintegration of the hyphal walls of L. theobromae also were noted in dual culture. Trichoderma viride reduced rotting by 29.07 to 65.06% in artificially inoculated banana fruits. Treatment of banana fruits with T. viride 4 h prior to inoculation with L. theobromae provided better protection than simultaneous application or treatment 4 h after inoculation.