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To better understand its mechanism of activity towards higher alcohols, we overexpressed and purified new Geotrichum candidum GDH (GcGDH). The purified GcGDH (50.27 kDa) was then crystallized, and the crystal diffracted to a resolution of 2.3 Å using X-ray diffraction. We found that the GcGDH crystal structure belonged to space group P212121 and was comprised of two hexamers organized into an asymmetric unit, with each subunit consisting of 452 amino acid residues. The binding sites between higher alcohols or L-glutamic acid and GcGDH were consistent. The optimal reaction conditions for GcGDH and hexanol were a pH of 4.0 and temperature of 30 °C, and those for GcGDH and monosodium glutamate (MSG) were a pH of 8.0 and temperature of 20 °C. The Km values for hexanol and MSG were found to be 74.78 mM and 0.018 mM, respectively. Mutating GcGDH Lys 113 to either Ala or Gly caused a dramatic reduction in its catalytic efficiency towards both MSG and hexanol, suggesting that Lys 113 is essential to the active site of GcGDH.

This study identified the SQUAMOSA Promoter Binding Proteins transcription factor SBP from the whole genome of Quinoa by analyzing the phylogenetic relationship, gene structure, chromosome location and gene replication, upstream cis-regulatory element, tissue expression, and construction of the SBP protein interaction network, as well as analyzing the characteristics of the SBP tissue expression pattern under salt stress. It serves as the foundation for the study and application of Quinoa SBP. The results showed that 23 genes of the CqSBP family were identified by bioinformatics analysis, and their amino acid physical and chemical properties showed diversity. The family proteins were hydrophilic, and the subcellular localization showed that 23 CqSBPs were located in the nucleus. The SBP family genes are unevenly distributed on the 12 chromosomes of Quinoa; the upstream cis-acting element analysis shows that there are 49 elements with plant hormones, stress, light response, and tissue-specific expression, and all CqSBPs contain one or more TATA box elements. Protein interaction network analysis shows that all CqSBP proteins appear in the known interaction network of Arabidopsis; the expression of different SBP genes in different organs and periods of Quinoa is different, and the expression of SBP genes has certain tissue specificity. The expression of CqSBP showed many changes under salt stress.

Human rhinoviruses (HRVs) are the predominant infectious agents for the common cold worldwide. The HRV-C species cause severe illnesses in children and are closely related to acute exacerbations of asthma. 3C protease, a highly conserved enzyme, cleaves the viral polyprotein during replication and assists the virus in escaping the host immune system. These key roles make 3C protease an important drug target. A few structures of 3Cs complexed with an irreversible inhibitor rupintrivir have been determined. These structures shed light on the determinants of drug specificity. Here we describe the structures of HRV-C15 3C in free and inhibitor-bound forms. The volume-decreased S1’ subsite and half-closed S2 subsite, which were thought to be unique features of enterovirus A 3C proteases, appear in the HRV-C 3C protease. Rupintrivir assumes an “intermediate” conformation in the complex, which might open up additional avenues for the design of potent antiviral inhibitors. Analysis of the features of the three-dimensional structures and the amino acid sequences of 3C proteases suggest new applications for existing drugs.

In recent years, with the rapidly increasing demand for pure protein products in various fields (biomedicines, biochemical reagents, food industries, etc.), the need for low-cost, high-quality prot...

The early Albian Oceanic Anoxic Event 1b (OAE 1b) is well documented in the Tethys, Pacific and North Atlantic, but few studies have evaluated whether or not terrestrial records of OAE 1b exist. In order to identify terrestrial records of the early Albian OAE 1b and to infer possible driving mechanisms, an integrated multi-proxy study from the late Aptian to Albian in the Fuxin lacustrine basin was conducted, including thick, organic-rich black mudstones, total organic carbon (TOC), organic carbon isotopes (d13Corg), mercury concentration (Hg) and results from pyrolysis analyses (S2, Tmax and HI). Results show three distinct short-term negative d13Corg excursions corresponding with relatively high TOC values, which could be counterparts of the Kilian, Paquier and Leenhardt sub-events of the early Albian OAE 1b. Atmospheric CO2 concentration (pCO2) recovered from C3 plant d13Corg compositions indicates an increasing trend in Unit C during the early Albian, and there are three short-term increases of pCO2 corresponding to the three sub-events of OAE 1b at this time interval. We infer that a trend of increasing pCO2 during the Kilian sub-event in the study area is closely related to volcanism. Continental weathering calculated using chemical weathering indices (CIA, WIP and MIA(O)) show an increasing trend during the OAE 1b interval, likely resulting from warmer and more humid conditions. Mixed sources of terrestrial plants and lacustrine plankton demonstrated by pyrolysis analyses (HI vs. Tmax and S2 vs. TOC), indicate a terrestrial contribution to the organic-rich sediments of the Kilian, Paquier and Leenhardt sub-events of OAE 1b. We suggest that a CO2-forced greenhouse effect during the early Albian might have triggered the relatively warm and humid palaeoclimatic conditions, and intensified chemical weathering that combined to create high nutrient and organic matter levels that were flushed into lakes contributing to eutrophication and anoxia in lacustrine and in contemporaneous oceanic systems. (c) 2021 China University of Geosciences (Beijing) and Peking University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/ licenses/by-nc-nd/4.0/). China University of Mining and Technology (Beijing) , China; National Natural Science Foundation of China [41572090, 42002128]; Shandong Key Laboratory of Deposi-tional Mineralization & Sedimentary Mineral, Shandong University of Science and Technology, China [DMSM20190015] Published version We thank Jason Hilton for discussion, Stephen Grasby and the other anonymous reviewers for constructive and helpful reviews of the manuscript. This study was supported by the Yue Qi Scholar Project of China University of Mining and Technology (Beijing) , China; the National Natural Science Foundation of China (41572090; 42002128) , and Shandong Key Laboratory of Deposi-tional Mineralization & Sedimentary Mineral, Shandong University of Science and Technology, China (DMSM20190015) .

BACKGROUND: Predictors of recurrence in patients with lymph node-negative gastric cancer (GC) who have undergone curative resection have been widely investigated, but not the effects of predictors on timing of recurrence. OBJECTIVE: Determine the factors associated with early and late recurrence in patients with node-negative GC. DESIGN: Retrospective cohort. SETTING: Academic tertiary care center. PATIENTS AND METHODS: The study included patients with node-negative GC after curative resection between 2008 and 2018 at two institutions. Early and late recurrences were determined using a minimum P value approach to evaluate the optimal cutoff for recurrence-free survival (RFS). A competing risk model and landmark analysis were used to analyze factors associated with early and late recurrences. MAIN OUTCOME MEASURES: Recurrence-free survival and factors associated with survival. SAMPLE SIZE: 606. RESULTS: After a median follow-up of 70 months, 50 (8.3%) patients experienced recurrent disease. The optimal length of RFS for distinguishing between early (n=26) and late recurrence (n=24) was 24 months ( P =.0013). The median RFS in the early and late recurrence groups was 11 and 32 months, respectively. Diffuse tumors (hazard ratio 3.358, P =.014), advanced T stage (HR 8.804, P =.003), perineural invasion (HR 10.955, P P =.018) were independent predictors of early recurrence. Mixed tumor location (HR 5.586, P =.002), advanced T stage (HR 5.066, P P P CONCLUSIONS: Individualized therapeutic and follow-up strategies should be considered in future studies because of distinct patterns in predictors of early and late recurrence. LIMITATIONS: Retrospective design, small sample size. CONFLICT OF INTEREST: None.

Obtaining good quality protein crystals is still desirable for high resolution structural determination of the proteins using crystallography. The crystal quality is affected by the growth environment and growth methods. As frequently used methods in practical protein crystallization, sitting and hanging-drop methods are seldom compared in terms of crystal quality in the literatures. We performed a systematic comparison on the quality of the protein crystals grown in sitting and hanging-drop methods using different crystallization plates: cross-diffusion microbatch plate (CDM SD: CDM plate, sitting drop method; CDM HD: CDM plate, hanging drop method), traditional sitting and hanging-drop vapor diffusion plates (T SD: traditional plate, sitting drop method; and T HD: traditional plate, hanging drop method). Five different proteins, proteinase K (prk), lysozyme (lys), concanavalin A VI (con), catalase (cat) and α-Chymotrypsinogen A II (chy), were used. The crystal quality was compared in terms of the resolution limit, mosaicity and Wilson plot B factor. It was found that the crystals grown in CDM plate using the hanging drop method (CDM HD) exhibited the best morphology and the best crystal quality. X-ray diffraction tests showed that the CDM plate using hanging drop method is indeed a practical and useful method for obtaining high-quality protein crystals.

Monochamus saltuarius (Coleoptera: Cerambycidae) is an important native pest in the pine forests of northeast China and a dispersing vector of an invasive species Bursaphelenchus xylophilus. To investigate the bacterial gut diversity of M. saltuarius larvae in different host species, and infer the role of symbiotic bacteria in host adaptation, we used 16S rRNA gene Illumina sequencing and liquid chromatography-mass spectrometry metabolomics processing to obtain and compare the composition of the bacterial community and metabolites in the midguts of larvae feeding on three host tree species: Pinus koraiensis, Pinus sylvestris var. mongolica, and Pinus tabuliformis. Metabolomics in xylem samples from the three aforementioned hosts were also performed. Proteobacteria and Firmicutes were the predominant bacterial phyla in the larval gut. At the genus level, Klebsiella, unclassified_f__Enterobacteriaceae, Lactococcus, and Burkholderia–Caballeronia–Paraburkholderia were most dominant in P. koraiensis and P. sylvestris var. mongolica feeders, while Burkholderia–Caballeronia–Paraburkholderia, Dyella, Pseudoxanthomonas, and Mycobacterium were most dominant in P. tabuliformis feeders. Bacterial communities were similar in diversity in P. koraiensis and P. sylvestris var. mongolica feeders, while communities were highly diverse in P. tabuliformis feeders. Compared with the other two tree species, P. tabuliformis xylems had more diverse and abundant secondary metabolites, while larvae feeding on these trees had a stronger metabolic capacity for secondary metabolites than the other two host feeders. Correlation analysis of the association of microorganisms with metabolic features showed that dominant bacterial genera in P. tabuliformis feeders were more negatively correlated with plant secondary metabolites than those of other host tree feeders.

This study focuses on Early Cretaceous mudstones from the Shahai and Fuxin formations in the Fuxin continental basin. We analyse chemical weathering, land surface temperatures and palaeoclimates based on chemical weathering indices, and emphasize the implications of continental chemical weathering on nutrient fluxes into lakes and oceans. According to Cr and Ni abundance, Al2O3-TiO2, La/Sc-Th/Co and V-Ni-Th×10 plots, as well as rare earth element (REE) analysis, mudstone samples from the Shahai and Fuxin formations were derived from the same type of provenance comprising mainly felsic igneous rocks. Chemical weathering trends reflected by the Chemical Index of Alteration (CIA), Weathering Index of Parker (WIP) and the Mafic Index of Alteration for Oxidative weathering environments (MIA(O)) are consistent with each other and allow the geological succession to be divided into four stages. Land surface temperatures of the Shahai and Fuxin formations are estimated based on the linear relationship of CIA to temperature, and also can be divided into four stages consistent with those determined from chemical weathering trends. During Stage A (early part of the late Aptian) chemical weathering and land surface temperatures were relatively low and showed characteristic high fluctuations, while Stage B (latest Aptian) represented a transitional period where weathering rates and temperatures increased, and high amplitude fluctuations continued. Conditions changed markedly in Stage C (early Albian) with very high and stable weathering, and warm, humid climates, while in Stage D (middle and late Albian) conditions returned to low chemical weathering and land surface temperatures. These stages of chemical weathering and land surface temperature fluctuations represent responses to global climate fluctuations during the Early Cretaceous, with the early Albian high weathering intensities and warm, humid climates combining to create high nutrient levels that would have flushed through rivers into lakes and ultimately oceans. This correlates stratigraphically with the development of Early Cretaceous black shales during Ocean Anoxic Event 1b, showing the importance of continental weathering regimes as a causal mechanism for lake and ocean anoxia.

Deciphering the activity-conformation relationship of PTPase is of great interest to understand how PTPase activity is determined by its conformation. Here we studied the activity and conformational transitions of PTPase from thermus thermophilus HB27 in the presence of sodium dodecyl sulfate (SDS). Activity assays showed the inactivation of PTPase induced by SDS was in a concentration-dependent manner. Fluorescence and circular dichroism spectra suggested SDS induced significant conformational transitions of PTPase, which resulted in the inactivation of PTPase, and the changes of α-helical structure and tertiary structure of PTPase. Structural analysis revealed a number of hydrophobic and charged residues around the active sites of PTPase may be involved in the hydrophobic and ionic bonds interactions of PTPase and SDS, which are suggested to be the major driving force to result in PTPase inactivation and conformational transitions induced by SDS. Our results suggested the hydrophobic and charged residues around the active sites were essential for the activity and conformation of PTPase. Our study promotes a better understanding of the activity and conformation of PTPase.

The majority of outbreaks of the common cold are caused by rhinoviruses. The 2A protease (2Apro) of human rhinoviruses (HRVs) is known to play important roles in the propagation of the virus and the modulation of host signal pathways to facilitate viral replication. The 2Aprofrom human rhinovirus C15 (HRV-C15) has been expressed inEscherichia coliand purified by affinity chromatography, ion-exchange chromatography and gel-filtration chromatography. The crystals diffracted to 2.6 Å resolution. The structure was solved by molecular replacement using the structure of 2Aprofrom coxsackievirus A16 (CVA16) as the search model. The structure contains a conserved His–Asp–Cys catalytic triad and a Zn2+-binding site. Comparison with other 2Aprostructures from enteroviruses reveals that the substrate-binding cleft of 2Aprofrom HRV-C15 exhibits a more open conformation, which presumably favours substrate binding.

Protein crystallization is of great importance because protein crystals have a number of different important applications, including large-scale purification of proteins, determination of protein structure, nanoparticle preparation, and theoretical studies of crystallization. An approach often used to efficiently crystallize proteins is the use of nucleants or seeds (small fragments of protein crystals) that can help increase the probability of protein crystallization. Due to the very positive effect that seeding has on protein crystallization, seeds are now widely accepted and utilized in practical protein crystallization. Here, we show that cross-linked protein crystals (CLPCs), which retain the crystal structure but are much more stable than non-cross-linked crystals, can also be used as a new type of seed for promoting protein crystallization. Seeding with CLPCs has effects on both the reproducibility and screening of protein crystals and could improve the optical perfection (well-defined facets) of p...

The wells of the novel plate are pre-embedded with cross-linked protein microcrystals which can act as the seeds for protein crystallization. The results of crystallization screening experiments showed that the novel plate exhibited dramatically improved performance in finding crystallization conditions as compared with the original CDM plate.

We presented a systematic method that combined the cross-diffusion microbatch method (CDM) with the seeding technique to enhance protein crystal reproducibility based on a “common dispersing agent”. The common dispersing agent is used to dilute the crystal seeds. Seven proteins were screened, and more crystallization conditions were found. X-ray diffraction showed that the combined method is practical and useful for obtaining high-quality protein crystals.

We present a systematic quality comparison of protein crystals obtained with and without cross-linked protein crystal (CLPC) seeds. Four proteins were used to conduct the experiments, and the results showed that crystals obtained in the presence of CLPC seeds exhibited a better morphology. In addition, the X-ray diffraction data showed that the CLPC seeds method is a powerful tool to obtain high-quality protein crystals. Therefore, we recommend the use of CLPC seeds in preparing high-quality diffracting protein crystals.

Straw returning to soil can supplement soil nutrients required for crop growth, fertilize soil, and improve soil quality. To explore the long-term effect of straw returning on soil total nitrogen and its composition, herein, five treatments including no rice straw + no wheat straw returning (NRW), no rice straw + all wheat straw returning (W), all rice straw + no wheat straw returning (R), half rice straw + half wheat straw returning (HRW), and all rice straw + all wheat straw returning (ARW) were conducted in triplicate in Taihu Lake region, China. The effects of both straw amount and type were examined. Compared with the results obtained in 2007, the results herein obtained in 2017 showed that after 10 years of straw returning, soil total nitrogen and heavy fraction nitrogen increased, while light fraction organic matter decreased. Among the five treatments, ARW had the largest decrease in light fraction nitrogen of 8.09 g·kg秸秆还田可以补充作物生长所需的营养元素,也是提升土壤质量、构建肥沃耕层的关键措施.为揭示长期稻麦秸秆还田对土壤氮组分变化的影响,选取了江苏太湖地区稻田土壤为研究对象,设置了稻麦秸秆均不还田(NRW)、麦秆还田+稻秆不还田(W)、稻秆还田+麦秆不还田(R)、稻麦秸秆均半量还田(HRW)和稻麦秸秆均全量还田(ARW)共5个处理,研究秸秆还田量和秸秆类型对土壤全氮及氮组分构成变化的影响.结果表明: 与2007年相比,2017年除NRW处理外,其他处理土壤全氮和重组氮都增加,稻秆还田处理土壤全氮和重组氮含量最高;所有处理土壤轻组分有机质中氮含量都下降,ARW处理土壤轻组分有机质中氮含量下降最多,为8.09 g·kg

Most organisms contain glutamate dehydrogenase (E.C. 1.4.1.2-1.4.1.4). In eukaryotes, the enzyme is mainly present in mitochondria. This enzyme plays a vital role in the metabolism of nitrogen and carbon and the signaling pathway. Studies have found that glutamate dehydrogenase has a certain relationship with the occurrence and development of tumors, which is significant for tumor research, but reviews on its relationship with human tumors are rare. This review summarized the relationship between glutamate dehydrogenase and breast cancer, glioma, colorectal cancer and ovarian cancer, etc, thus providing assistance for related research.大多数生物体中都含有谷氨酸脱氢酶(Glutamate dehydrogenase, GDH) (E.C. 1.4.1.2–1.4.1.4)。在真核生物中，该酶主要存在于线粒体中，并在氮和碳的代谢以及信号通路中起着至关重要的作用。研究发现谷氨酸脱氢酶与肿瘤发生及发展有一定的关系，对于肿瘤研究具有一定意义，但是关于其与人类肿瘤的关系方面的综述很少见。文中对谷氨酸脱氢酶与乳腺癌、胶质瘤、结直肠癌以及卵巢癌等的关系进行了归纳和总结，希望可以为相关研究提供帮助。.

Phosphorus (P) is an important macronutrient for plant but can also cause potential environmental risk. In this paper, we studied the long-term fertilizer experiment (started 1980) to assess the soil P dynamic, balance, critical P value and the crop yield response in Taihu Lake region, China. To avoid the effect of nitrogen (N) and potassium (K), only the following treatments were chosen for subsequent discussion, including: C0 (control treatment without any fertilizer or organic manure), CNK treatment (mineral N and K only), CNPK (balanced fertilization with mineral N, P and K), MNK (integrated organic manure and mineral N and K), and MNPK (organic manure plus balanced fertilization). The results revealed that the response of wheat yield was more sensitive than rice, and no significant differences of crop yield had been detected among MNK, CNPK and MNPK until 2013. Dynamic and balance of soil total P (TP) and Olsen-P showed soil TP pool was enlarged significantly over consistent fertilization. However, the diminishing marginal utility of soil Olsen-P was also found, indicating that high-level P application in the present condition could not increase soil Olsen-P contents anymore. Linear-linear and Mitscherlich models were used to estimate the critical value of Olsen-P for crops. The average critical P value for rice and wheat was 3.40 and 4.08 mg kg −1 , respectively. The smaller critical P value than in uplands indicated a stronger ability of P supply for crops in this paddy soil. We concluded that no more mineral P should be applied in rice-wheat system in Taihu Lake region if soil Olsen-P is higher than the critical P value. The agricultural technique and management referring to activate the plant-available P pool are also considerable, such as integrated use of low-P organic manure with mineral N and K.

Acquired immune deficiency syndrome (AIDS) is mainly caused by the human immunodeficiency virus type-1 (HIV-1). To our knowledge, this is the first review focusing on the vital role of Pin1 in the infection of HIV-1 and the development of AIDS. We and others have demonstrated that Pin1, the only known cis-to-trans isomerase recognizing the pThr/pSer-Pro motifs in proteins, plays striking roles in several human diseases. Interestingly, recent evidence gradually indicates that Pin1 regulates several key steps of the life cycle of HIV-1, including the uncoating of the HIV-1 core, the reverse transcription of the RNA genome of HIV-1, and the integration of the HIV-1 cDNA into human chromosomes. Whereas inhibiting Pin1 suppresses all of these key steps and attenuates the replication of HIV-1, at the same time different PIN1 gene variants are correlated with the susceptibility to HIV-1 infection. Furthermore, Pin1 potentially promotes HIV-1 infection by activating multiple oncogenes and inactivating multiple tumor suppressors, extending the life span of HIV-infected cells. These descriptions suggest Pin1 as a promising therapeutic target for the prevention of HIV-1 and highlight the possibility of blocking the development of AIDS by Pin1 inhibitors.

Aldo-keto reductases (AKRs) are a superfamily of NAD(P)H-dependent oxidoreductases that catalyse the asymmetric reduction of aldehydes and ketones to chiral alcohols in various organisms. The novel aldo-keto reductase Tm1743 fromThermotoga maritimawas identified to have a broad substrate specificity and high thermostability, serving as an important enzyme in biocatalysis and fine-chemical synthesis. In this study, Tm1743 was overexpressed inEscherichia coliBL21(DE3) cells with an N-terminal His6tag and was purified by Ni2+-chelating affinity and size-exclusion chromatography. Purified recombinant enzyme was incubated with its cofactor NADP+and its substrate ethyl 2-oxo-4-phenylbutyrate (EOPB) for crystallization. Two X-ray diffraction data sets were collected at 2.0 and 1.7 Å resolution from dodecahedral crystals grown from samples containing Tm1743–NADP+–EOPB and Tm1743–NADP+, respectively. Both crystals belonged to space groupP3121, with similar unit-cell parameters. However, in the refined structure model only NADP+was observed in the active site of the full-length Tm1743 enzyme. Degradation of the N-terminal vector-derived amino acids during crystallization was confirmed by Western blot and mass-spectrometric analyses.

We present a new method for the realization of high-throughput protein crystallization screening using an array of 96 capillaries aligned in a circle. In this method, each capillary represents a single crystallization condition, and all capillaries experience identical magnetic field conditions. After crystallization, the crystals in the capillary can be directly diffracted without harvesting. This method proved easy to perform and is applicable for use in magnetic fields and may be further extended for use in other circumstances, for example, under space microgravity conditions.

We presented a systematic quality comparison of protein crystals grown using the cross-diffusion microbatch (CDM) and standard sitting-drop vapor diffusion methods. Eleven proteins were screened and it was found that crystals grown using CDM exhibited a better morphology. X-ray diffraction showed that the CDM method is practical and useful for obtaining high-quality protein crystals.

During the last two decades, C-type lectin receptors (CLRs) have been demonstrated to play key roles in initiating the host immune response against fungal infection. It is well established that CLRs, such as Dectin-1, Dectin-2, Dectin-3 and Mincle recognize the cell wall component from the infected microorganisms by using their carbohydrate recognition domain (CRD). Upon stimulation, CLRs induce multiple signal transduction cascades through their own immunereceptor tyrosine-based activation motifs (ITAMs) or interacting with ITAM-containing adaptor proteins such as FcRγ, which then lead to the activation of nuclear factor kappa B (NF-κB) through Syk- and CARD9-dependent pathway. Dissecting CLR signal cascades and their effects on host immune cells is essential to understand the molecular mechanisms in regulating host antifungal immunity. Recently, the activated CLRs including Dectin-1 and Dectin-2 are reported to undergo lysome-mediated degradation by an E3 ubiquitin ligase CBL-b. Moreover, structural analysis will help understand the molecular mechanism of these CLRs and provide clues to rational design for effective anti-fungal drugs. Overall, we summarize the current knowledge on activating and inhibitory CLRs and discuss how to boost host immune system to fight against invasive fungal infection.

Small molecule drugs obtained from synthetic compounds or natural products show their therapeutic effects by specifically binding to one or a few target proteins and modulating their functions. In contrast, undesirable drug-protein interactions may provoke harmful side effects. Furthermore, drug-protein interactions also play roles in drug activation, transport, metabolism and regulation of drug resistance. Therefore, systematically identifying binding proteins of a drug molecule is critical for understanding the mechanism of action of the drug at molecular level. In this review, we summarize current widely-used experimental approaches for proteome-wide target identification of small molecule drugs, including affinity purification, chemical proteomics and protein thermal stability based methods. The advance of these methods will quicken the pace of target deconvolution of small molecule drugs and hold promise for drug repositioning research. Moreover, these approaches also provide a powerful arsenal for exploring the signaling pathways of small molecule second messengers, signaling lipids and other regulatory metabolites.

Elodea nuttallii is widely used in Chinese mitten crab (CMC) rearing practice, but it is not a native aquatic plant and cannot endure high temperature. Thus, large E. nuttallii mortality and water deterioration events could occur during high-temperature seasons. The aim of this study was to identify the use of local macrophytes in CMC rearing practice, including Ipomoea aquatic and Oryza sativa. A completely randomized field experiment was conducted to investigate the crab yield, water quality, bacterioplankton community and functions in the three different systems (E. nuttallii, I. aquatic, and O. sativa). Average crab yields in the different macrophyte systems did not differ significantly. The I. aquatic and O. sativa systems significantly decreased the total nitrogen and nitrate-N quantities in the outflow waters during the rearing period compared to the E. nuttallii system, and the I. aquatic and O. sativa plants assimilated more nitrogen than the E. nuttallii plant. Moreover, the significant changes of bacterioplankton abundances and biodiversity in the three systems implied that cleanliness of rearing waters was concomitantly attributed to the differential microbial community and functions. In addition, principle component analysis successfully differentiated the bacterioplankton communities of the three macrophytes systems. Environmental factor fitting and the co-occurrence network analyses indicated that pH was the driver of bacterioplankton community structure. Functional predictions using PICRUSt (v.1.1.3) software based on evolutionary modeling indicated a higher potential for microbial denitrification in the I. aquatic and O. sativa systems. Notably, the O. sativa plants stopped growing in the middle of the rearing period. Thus, the I. aquatic system rather than the O. sativa system could be a feasible and environmental-friendly alternative to the E. nuttallii system in CMC rearing practice.

Potent and selective chemical probes are valuable tools for discovery of novel treatments for human diseases. NF-κB-inducing kinase (NIK) is a key trigger in the development of liver injury and fibrosis. Whether inhibition of NIK activity by chemical probes ameliorates liver inflammation and injury is largely unknown. In this study, a small-molecule inhibitor of NIK, B022, was found to be a potent and selective chemical probe for liver inflammation and injury. B022 inhibited the NIK signaling pathway, including NIK-induced p100-to-p52 processing and inflammatory gene expression, both in vitro and in vivo Furthermore, in vivo administration of B022 protected against not only NIK but also CCl4-induced liver inflammation and injury. Our data suggest that inhibition of NIK is a novel strategy for treatment of liver inflammation, oxidative stress, and injury.-Ren, X., Li, X., Jia, L., Chen, D., Hou, H., Rui, L., Zhao, Y., Chen, Z. A small-molecule inhibitor of NF-κB-inducing kinase (NIK) protects liver from toxin-induced inflammation, oxidative stress, and injury.

Alternative splicing (AS) contributes to the complexity of the mammalian proteome and plays an important role in diseases, including infectious diseases. The differential AS patterns of these transcript sequences between the healthy (HS3A) and mastitic (HS8A) cows naturally infected by Staphylococcus aureus were compared to understand the molecular mechanisms underlying mastitis resistance and susceptibility. In this study, using the Illumina paired-end RNA sequencing method, 1352 differentially expressed genes (DEGs) with higher than twofold changes were found in the HS3A and HS8A mammary gland tissues. Gene ontology and KEGG pathway analyses revealed that the cytokine-cytokine receptor interaction pathway is the most significantly enriched pathway. Approximately 16k annotated unigenes were respectively identified in two libraries, based on the bovine Bos taurus UMD3.1 sequence assembly and search. A total of 52.62% and 51.24% annotated unigenes were alternatively spliced in term of exon skipping, intron retention, alternative 5' splicing and alternative 3' splicing. Additionally, 1,317 AS unigenes were HS3A-specific, whereas 1,093 AS unigenes were HS8A-specific. Some immune-related genes, such as ITGB6, MYD88, ADA, ACKR1, and TNFRSF1B, and their potential relationships with mastitis were highlighted. From Chromosome 2, 4, 6, 7, 10, 13, 14, 17, and 20, 3.66% (HS3A) and 5.4% (HS8A) novel transcripts, which harbor known quantitative trait locus associated with clinical mastitis, were identified. Many DEGs in the healthy and mastitic mammary glands are involved in immune, defense, and inflammation responses. These DEGs, which exhibit diverse and specific splicing patterns and events, can endow dairy cattle with the potential complex genetic resistance against mastitis.

To explore the association between single nucleotide polymorphisms (SNPs) in the promoter region of the inner centromere protein (INCENP) gene and bovine semen quality, the haplotypes in 250 Chinese Holstein bulls were detected using PCR-RFLP method in this study. Two SNPs (g.-556 GT, rs 136823901 and g.-692 CT, rs 211010999) and three haplotypes (CG, TT, TG) were identified in the promoter region of INCENP. The genotype frequency and allele frequency of these two SNPs as well as the correlation between different SNP haplotype combinations and bovine semen quality were then analyzed. Our results showed that fresh sperm motility of the GT genotype was significantly higher than that of the GG genotype (P0.05) at the SNP site g.-556 GT, while fresh and frozen-thawed sperm motilities of the haplotype combinations H1H1(CCGG), H1H3(CTGT), H2H3(TTGT) and H3H3(TTTT) were significantly higher than that of H1H2 (P0.05). To further study the possible mechanisms by which g.-556 GT and g.-692 CT affect semen quality, three haplotype plasmids were respectively transfected into MLTC-1 cells. The TG haplotype demonstrated the highest luciferase activity, suggesting that g.-556 GT and g.-692 CT are functional mutations which could regulate INCENP gene expression by affecting promoter activity and thus affect semen quality.

Correction for ‘A high-performance protein crystallization plate pre-embedded with crosslinked protein microcrystals as seeds’ by Hai Hou et al., CrystEngComm, 2018, 20, 4713–4718.

Protein crystallization is affected by many parameters, among which certain parameters have not been well controlled. The temperature at which the protein and precipitant solutions are mixed (i.e., the ambient temperature during mixing) is such a parameter that is typically not well controlled and is often ignored. In this paper, we show that this temperature can influence protein crystallization. The experimental results showed that both higher and lower mixing temperatures can enhance the success of crystallization, which follows a parabolic curve with an increasing ambient temperature. This work illustrates that the crystallization solution preparation temperature is also an important parameter for protein crystallization. Uncontrolled or poorly controlled room temperature may yield poor reproducibility in protein crystallization.