Mitogenomes provide new insights of evolutionary history of Boreheptagyiini and Diamesini (Diptera: Chironomidae: Diamesinae)

Mitogenomes have been widely used for phylogenetic reconstruction of various Dipteran groups, but specifically for chironomid, they have not been carried out to resolve the relationships. Diamesinae (Diptera: Chironomidae) are important bioindicators for freshwater ecosystem monitoring, but its evolutionary history remains uncertain for lack of information. Here, coupled with one previously published and 30 new mitogenomes of Diamesinae, we carried out comparative mitogenomic analysis and phylogenetic analysis. Mitogenomes of Diamesinae were conserved in structure, and all genes arranged in the same order as the ancestral insect mitogenome. All protein‐coding genes in Diamesinae were under stronger purifying selection than those of other nonbiting midge species, which may exhibit signs of adaptation to life at cold living conditions. Phylogenetic analyses strongly supported the monophyly of Diamesinae, with Boreheptagyiini deeply nested within Diamesini. In addition, phylogenetic relationship of selected six genera was resolved, except Sympotthastia remained unstable. Our study revealed that the mitogenomes of Diamesinae are highly conserved, and they are practically useful for phylogenetic inference.     

Oxidized high-density lipoprotein promotes CD36 palmitoylation and increases lipid uptake in macrophages

Oxidized high-density lipoprotein (oxHDL) reduces the ability of cells to mediate reverse cholesterol transport and also shows atherogenic properties. Palmitoylation of cluster of differentiation 36 (CD36), an important receptor mediating lipoprotein uptake, is required for fatty acid endocytosis. However, the relationship between oxHDL and CD36 has not been described in mechanistic detail. Here, we demonstrate using acyl-biotin exchange analysis that oxHDL activates CD36 by increasing CD36 palmitoylation, which promotes efficient uptake in macrophages. This modification increased CD36 incorporation into plasma lipid rafts and activated downstream signaling mediators, such as Lyn, Fyn, and c-Jun N-terminal kinase, which elicited enhanced oxHDL uptake and foam cell formation. Furthermore, blocking CD36 palmitoylation with the pharmacological inhibitor 2-bromopalmitate decreased cell surface translocation and lowered oxHDL uptake in oxHDL-treated macrophages. We verified these results by transfecting oxHDL-induced macrophages with vectors expressing wildtype or mutant CD36 (mCD36) in which the cytoplasmic palmitoylated cysteine residues were replaced. We show that cells containing mCD36 exhibited less palmitoylated CD36, disrupted plasma membrane trafficking, and reduced protein stability. Moreover, in ApoE^−/−CD36^−/− mice, lipid accumulation at the aortic root in mice receiving the mCD36 vector was decreased, suggesting that CD36 palmitoylation is responsible for lipid uptake in vivo. Finally, our data indicated that palmitoylation of CD36 was dependent on DHHC6 (Asp-His-His-Cys) acyltransferase and its cofactor selenoprotein K, which increased the CD36/caveolin-1 interaction and membrane targeting in cells exposed to oxHDL. Altogether, our study uncovers a causal link between oxHDL and CD36 palmitoylation and provides insight into foam cell formation and atherogenesis.     

Intravital lipid droplet labeling and imaging reveals the phenotypes and functions of individual macrophages in vivo

Macrophages play pivotal roles in the maintenance of tissue homeostasis. However, the reactivation of macrophages toward proinflammatory states correlates with a plethora of inflammatory diseases, including atherosclerosis, obesity, neurodegeneration, and bone marrow (BM) failure syndromes. The lack of methods to reveal macrophage phenotype and function in vivo impedes the translational research of these diseases. Here, we found that proinflammatory macrophages accumulate intracellular lipid droplets (LDs) relative to resting or noninflammatory macrophages both in vitro and in vivo, indicating that LD accumulation serves as a structural biomarker for macrophage phenotyping. To realize the staining and imaging of macrophage LDs in vivo, we developed a fluorescent fatty acid analog-loaded poly(lactic-co-glycolic acid) nanoparticle to label macrophages in mice with high efficiency and specificity. Using these novel nanoparticles, we achieved in situ functional identification of single macrophages in BM, liver, lung, and adipose tissues under conditions of acute or chronic inflammation. Moreover, with this intravital imaging platform, we further realized in vivo phenotyping of individual macrophages in the calvarial BM of mice under systemic inflammation. In conclusion, we established an efficient in vivo LD labeling and imaging system for single macrophage phenotyping, which will aid in the development of diagnostics and therapeutic monitoring. Moreover, this method also provides new avenues for the study of lipid trafficking and dynamics in vivo.Supplementary key words: macrophage, inflammation, lipid droplet, nanoparticle delivery, in vivo imaging, fatty acid analog, bone marrow, systemic inflammation, lipid trafficking, biomarker

Macrophages, a type of immune cells, almost reside in all tissues of body, from the skin to the bone marrow (BM) (1). Macrophages have remarkable plasticity, and they can be activated into specific subtypes by modifying their physiology and functions in response to local environmental cues. Activated macrophages are commonly divided into proinflammatory killing subtype and anti-inflammatory repairing subtype. Proinflammatory macrophages responding to bacteria, IFN-γ, and lipopolysaccharide (LPS) are involved in host defense and inflammation, whereas anti-inflammatory macrophages responding to interleukin-4 (IL-4), IL-10, and IL-13 play a pivotal role in tissue homeostasis and remodeling (2). Increasing evidence indicates that the reactivation of macrophages toward proinflammatory states under diverse kinds of stress is correlated with a plethora of inflammatory diseases, such as atherosclerosis, diabetes, obesity, rheumatoid arthritis, neurodegeneration, and BM failure syndromes (3, 4). Thus, characterization of macrophage activation status and the underlying molecular mechanism in situ will help elucidate their functions in these diseases; however, in vivo analysis of the macrophage activation status in their native multicellular microenvironment is challenging.Although lipid droplets (LDs) have been initially described as intracellular fat storage organelles in adipocytes, increasing studies indicate that myeloid cells also form LDs under inflammation and stress (5, 6). Macrophages, as the effector cells of innate immunity, are found to form LDs to support their host defense when exposed to pathogens, such as parasites, bacteria, and viruses (7, 8, 9, 10, 11). However, abnormal LD accumulation in tissue-resident macrophages correlates with the pathogenesis of various inflammatory diseases. For instance, foam cells in atherosclerotic lesions can maintain the local inflammatory response by secreting proinflammatory cytokines (12, 13, 14). Moreover, LD-accumulating microglia contribute to neurodegeneration by producing high levels of reactive oxygen species (ROS) and secreting proinflammatory cytokines (15). These findings indicate that LD accumulation might be a hallmark of macrophages with proinflammatory functions.In this study, based on the typical activation of in vitro BM-derived macrophages, we find that proinflammatory M_{(LPS + IFN-γ)} macrophages are characterized by LD accumulation, whereas resting macrophages and anti-inflammatory M_(IL-4) and M_(IL-10) macrophages do not contain any LDs. These features also hold for Matrigel plug-recruited macrophages and tissue-resident macrophages in mice. These findings demonstrate that LD accumulation could serve as a morphological index to distinguish proinflammatory macrophages from others.It is feasible to distinguish LD-containing cells using imaging techniques, which has translational potential for identification of proinflammatory macrophages in vivo. However, current techniques for LD visualization are traditional in vitro staining method, and in vivo staining and imaging of LD in individual macrophages remains a challenge. Through nanocarrier screening, we selected the poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) as nanocarrier to deliver the lipophilic carbocyanine dye (DiI_C18(5) solid (1,1''-dioctadecyl-3,3,3'',3''-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt) [DiD]) and lipid staining dye (C1-BODIPY 500/510-C12) into macrophages. Using these dual fluorescence-labeled PLGA NPs, we achieved in situ and in vivo functional identification of single macrophages in various tissues under systemic or local inflammatory stress. Collectively, this study establishes an efficient in vivo labeling and imaging system of intracellular LDs for phenotyping the activation status and functions of individual macrophages in their dynamic niche, which is pivotal for disease diagnosis and preclinical research.     

太行山南麓山区栓皮栎-扁担杆生态系统水分利用策略

分析了太行山南麓低丘山区降水、泉水、地下水、土壤水以及栓皮栎、扁担杆的氢氧稳定同位素特征,结合Iso Source模型确定了栓皮栎和扁担杆水分来源的季节性差异,并对栓皮栎和扁担杆水分利用策略进行分析。结果表明,同一生态系统中的栓皮栎和扁担杆枝条水的δ18O和δD值差别明显。雨季中栓皮栎和扁担杆水分来源较浅,以0—20 cm土壤水分为主,但旱季中栓皮栎和扁担杆水分主要来源均比雨季明显加深,其中栓皮栎主要利用40—60 cm土壤水分,扁担杆则主要利用20—40 cm土壤水分。此外,旱季后期栓皮栎还利用部分泉水,其比例达到了19.6%。二者水分来源的不同,使得栓皮栎与扁担杆在旱季期间能避开用水冲突。旱季中生长在生态系统上层的栓皮栎中午部分气孔关闭,蒸腾速率下降,生长在生态系统下层的扁担杆日均蒸腾速率、气孔导度则分别比栓皮栎下降了46.94%和30.58%。栓皮栎和扁担杆分别采取了深水源及部分气孔关闭和浅水源及低蒸腾耗散的水分利用策略来利用旱季中有限的水分,因而其组成的生态系统表现出较强地适合太行山南麓脆弱环境的生态适应性。     

天山林区六种灌木生物量的建模及其器官分配的适应性

灌木全株生物量估算模型的构建仍存在一定困难,对灌木生物量在器官分配上所体现的适应性研究也不够充分。以天山林区6种常见灌木为研究对象,在天山的东段、中段、西段林区分别设置样地进行群落调查,由此以全株收获法取得6种常见灌木若干标准株的全株、根、枝、叶及各径级根的生物量,将D~2H(地径平方与高度的乘积)与V(冠幅面积与高度的乘积)分别选为估测模型的自变量,通过回归分析法建立了各种灌木全株生物量的最优估算模型,然后比较了此6种灌木全株生物量在营养器官上分配差异以及根系生物量在径级上的分配差异。结果表明:(1)天山林区6种常见灌木中,小檗(Berberis heteropoda Schrenk)、忍冬(Lonicera hispida Pall.ex Roem.et Schuet.)、栒子(Cotoneaster melanocarpus Lodd.)的全株生物量约为8.48—9.01 kg,蔷薇(Rosa spinosissima L.)、绣线菊(Spiraea hypericifolia L.)、方枝柏(Juniperus pseudosabina Fisch.et Mey.)的全株生物量约为2.71—3.20 kg;(2)蔷薇、绣线菊、栒子的全株生物量最优估测模型是以V为自变量的函数,小檗、忍冬、方枝柏的全株生物量最优估测模型是以D~2H为自变量的函数,各模型R~2值均在0.850以上,且在P0.05水平上达到显著,模型模拟结果达到了较高的准确度;(3)6种灌木全株生物量在根、枝上的分配比重差异不显著,仅在叶上的分配比重有差异(P0.05);根系生物量在径级上的分配均呈现随根系径级下降而减少的规律,6种灌木在径级大于2 mm根上的分配比重存在差异(P0.05,径级大于20 mm根为P0.01水平);(4)6种灌木全株生物量在营养器官上的分配差异以及根系生物量在径级上的分配差异均体现了各物种对其生境选择的适应策略。     

气候变化对滇西北碧塔海流域景观演变的影响

气候变化是景观演变的重要驱动力,高海拔地带生态系统对气候变化的响应较为敏感和迅速。选择地处青藏高原东南缘,受人为干扰相对较小的碧塔海流域为研究对象,利用1958—2011年气象资料以及1955、1974、1981、1994、2005和2011年6个年代的景观类型面积,采用气候倾向率估计、Mann-Kendall突变检验方法和Pearson相关系数等统计方法,研究了香格里拉县气候变化背景下碧塔海流域景观演变特征。结果表明:(1)1958—2011年来,滇西北香格里拉县多年平均气温为5.9℃,多年平均活动积温为2146.1℃,多年平均年极端高温为24.3℃,多年平均年极端低温为-18.6℃,多年平均降水量为631.7mm。气温、活动积温和年极端低温呈显著上升趋势,近54年来气温平均上升了1.94℃,升温速率远高于云南和全国水平。年极端高温和降水量呈增加趋势,但增加不明显。滇西北高原碧塔海流域背景气候呈现显著的变暖趋势。(2)1955—2011年来,碧塔海流域自然景观演变规律为草甸和灌草丛向有林地演变,气温、活动积温和年极端低温升高趋势与有林地景观面积的增加趋势大体一致,与草甸和灌草丛变化趋势相反。(3)气温、活动积温和年极端低温与碧塔海流域草甸、灌草丛和有林地的景观面积变化具有显著的相关性,是流域景观演变的主导气候要素,在草甸和灌草丛演变成有林地的过程中发挥着重要作用。极端高温和降水量对流域景观演变的作用相对较弱。     

硝酸盐型厌氧铁氧化菌的种类、分布和特性

硝酸盐型厌氧铁氧化(NAFO)是指微生物在厌氧条件下利用硝酸盐或亚硝酸盐作为电子受体,将低价铁(二价铁或零价铁)氧化为高价铁(三价铁)的过程。具有NAFO代谢能力的微生物称为硝酸盐型厌氧铁氧化菌(NAFOM)。NAFO是微生物领域的重大发现,也是环境领域开发新型脱氮技术和地学领域研究铁、氮循环的理论依据。整理文献报道的NAFOM资料,分析NAFOM系统发育性状,探讨典型NAFOM的生态分布及其营养、代谢特性,以期为NAFOM菌种资源的开发、地球铁素和氮素循环的研究、NAFO过程的优化提供借鉴。     

SARS-CoV-2 envelope protein causes acute respiratory distress syndrome (ARDS)-like pathological damages and constitutes an antiviral target

Cytokine storm and multi-organ failure are the main causes of SARS-CoV-2-related death. However, the origin of excessive damages caused by SARS-CoV-2 remains largely unknown. Here we show that the SARS-CoV-2 envelope (2-E) protein alone is able to cause acute respiratory distress syndrome (ARDS)-like damages in vitro and in vivo. 2-E proteins were found to form a type of pH-sensitive cation channels in bilayer lipid membranes. As observed in SARS-CoV-2-infected cells, heterologous expression of 2-E channels induced rapid cell death in various susceptible cell types and robust secretion of cytokines and chemokines in macrophages. Intravenous administration of purified 2-E protein into mice caused ARDS-like pathological damages in lung and spleen. A dominant negative mutation lowering 2-E channel activity attenuated cell death and SARS-CoV-2 production. Newly identified channel inhibitors exhibited potent anti-SARS-CoV-2 activity and excellent cell protective activity in vitro and these activities were positively correlated with inhibition of 2-E channel. Importantly, prophylactic and therapeutic administration of the channel inhibitor effectively reduced both the viral load and secretion of inflammation cytokines in lungs of SARS-CoV-2-infected transgenic mice expressing human angiotensin-converting enzyme 2 (hACE-2). Our study supports that 2-E is a promising drug target against SARS-CoV-2.Subject terms: Cell death, Molecular biology     

TEB/POLQ plays dual roles in protecting Arabidopsis from NO-induced DNA damage

Nitric oxide (NO) is a key player in numerous physiological processes. Excessive NO induces DNA damage, but how plants respond to this damage remains unclear. We screened and identified an Arabidopsis NO hypersensitive mutant and found it to be allelic to TEBICHI/POLQ, encoding DNA polymerase θ. The teb mutant plants were preferentially sensitive to NO- and its derivative peroxynitrite-induced DNA damage and subsequent double-strand breaks (DSBs). Inactivation of TEB caused the accumulation of spontaneous DSBs largely attributed to endogenous NO and was synergistic to DSB repair pathway mutations with respect to growth. These effects were manifested in the presence of NO-inducing agents and relieved by NO scavengers. NO induced G2/M cell cycle arrest in the teb mutant, indicative of stalled replication forks. Genetic analyses indicate that Polθ is required for translesion DNA synthesis across NO-induced lesions, but not oxidation-induced lesions. Whole-genome sequencing revealed that Polθ bypasses NO-induced base adducts in an error-free manner and generates mutations characteristic of Polθ-mediated end joining. Our experimental data collectively suggests that Polθ plays dual roles in protecting plants from NO-induced DNA damage. Since Polθ is conserved in higher eukaryotes, mammalian Polθ may also be required for balancing NO physiological signaling and genotoxicity.