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Sinopoda yichangensis Zhu, Zhong & Yang, 2020 was described from a single male from Qiaoliao Village, Hubei Province, China. To date, no additional specimens have been recorded. The female is reported for the first time from the type locality. Detailed morphological descriptions of the female, with photographs of living specimens and copulatory organs, are provided.

In this study, we aim to identify the clinical significance of basonuclin 1 ( BNC1) expression in ovarian carcinoma (OV) and to explore its latent mechanisms. Via integrating in-house tissue microarrays, gene chips, and RNA-sequencing data, we explored the expression and clinical value of BNC1 in OV. Immunohistochemical staining was utilized to confirm the protein expression status of BNC1. A combined SMD of –2.339 (95% CI: –3.649 to –1.028, P < 0.001) identified that BNC1 was downregulated based on 1346 samples, and the sROC (AUC = 0.93) showed a favorable discriminatory ability of BNC1 in OV patients. We used univariate and multivariate Cox regulation to evaluate the prognostic role of BNC1 for OV patients, and a combined hazard ratio of 0.717 (95% CI: 0.445–0.989, P < 0.001) revealed that BNC1 was a protective factor for OV. Furthermore, the fraction of infiltrating naive B cells, memory B cells, and other immune cells showed statistical differences between the high- and low- BNC1 expression groups through cell-type identification by estimating relative subsets of RNA transcripts (CIBERSORT) algorithm. Enrichment analysis showed that BNC1 may have a relationship with immune-related items in OV. By predicting the potential regulatory transcription factors (TFs) of BNC1, friend leukemia virus integration 1 ( FLI1) may be a potential upstream TF of BNC1. Corporately, a decreasing trend of BNC1 may serve as a tumor suppressor and prognostic biomarker in OV patients. Moreover, BNC1 may take part in immune-related pathways and influence the fraction of tumor-infiltrating immune cells.

Cytochrome P450s (P450s) are the most versatile catalysts utilized by plants to produce structurally and functionally diverse metabolites. Given the high degree of gene redundancy and challenge to functionally characterize plant P450s, protein engineering is used as a complementary strategy to study the mechanisms of P450-mediated reactions, or to alter their functions. We previously proposed an approach of engineering plant P450s based on combining high-accuracy homology models generated by Rosetta combined with data-driven design using evolutionary information of these enzymes. With this strategy, we repurposed a multi-functional P450 (CYP87D20) into a monooxygenase after redesigning its active site. Since most plant P450s are membrane-anchored proteins that are adapted to the micro-environments of plant cells, expressing them in heterologous hosts usually results in problems of expression or activity. Here, we applied computational design to tackle these issues by simultaneous optimization of the protein surface and active site. After screening 17 variants, effective substitutions of surface residues were observed to improve both expression and activity of CYP87D20. In addition, the identified substitutions were additive and by combining them a highly efficient C11 hydroxylase of cucurbitadienol was created to participate in the mogrol biosynthesis. This study shows the importance of considering the interplay between surface and active site residues for P450 engineering. Our integrated strategy also opens an avenue to create more tailoring enzymes with desired functions for the metabolic engineering of high-valued compounds like mogrol, the precursor of natural sweetener mogrosides.The online version contains supplementary material available at 10.1007/s42994-021-00056-z.

Background Traditional soy protein isolate (SPI)-based gel products, such as tofu, are generally produced by heating and by addition of metal salt ions to adjust the hydrophobicity and electrostatic force of soybean protein to facilitate the formation of a uniform network structure. However, the gelation rate of the soy protein gel network structure is difficult to control. Theoretically, epigallocatechin-3-gallate (EGCG) could be used to alter the surface hydrophobicity of thermally-induced SPI to improve their gelation rate and form a more uniform network structure, thus improving SPI-based gel properties (hardness, water-holding capacity and rheological properties). Results An SPI-EGCG complex (SPIE) was prepared, and properties of the resulting gel, following induction of transglutaminase (TG), were evaluated. Results showed that EGCG are bound to thermally-induced SPI primarily via hydrophobic and hydrogen bonding, thus altering the secondary structure composition and reducing surface hydrophobicity of proteins in thermally-induced SPI. Furthermore, the optimum amount of EGCG required to improve the gel strength, water-holding capacity and rheological properties was ≤0.04:1 (SPI 1 g L -1 , EGCG:SPI, w/w). Thermal stability analysis further indicated that EGCG in SPIE was more stable than free EGCG after heating. Conclusion This study demonstrated that EGCG can improve the gel properties of TG-cross-linked SPIE, while EGCG in SPIE exhibits enhanced thermal stability. Additionally, the results of this study provide a novel strategy for the development of SPI-based gel foods with improved gel properties and that are enriched with bioactive compounds. This article is protected by copyright. All rights reserved.

ObjectivesTo determine the prevalence and genetic characteristics of ESBL-producing Escherichia coli in retail raw meats from Singapore markets.MethodsA total of 634 raw meat (chicken, pork and beef) samples were collected from markets in Singapore during June 2017–October 2018. The samples were enriched overnight and then incubated on Brilliance™ ESBL Agar. Presumptive ESBL isolates were confirmed using the double-disc synergy test. Confirmed ESBL-producing E. coli were sent for WGS and bioinformatic analysis was performed.ResultsThe prevalence of ESBL-producing E. coli in chicken, pork and beef meats was 51.2% (109/213), 26.9% (58/216) and 7.3% (15/205), respectively. A total of 225 ESBL-producing E. coli were isolated from 184 samples. β-Lactam resistance genes were detected in all isolates. After β-lactam resistance genes, the most common antimicrobial resistance genes detected were aminoglycoside resistance genes (92.4%). One hundred and seventy-two (76.4%), 102 (45.3%) and 52 (23.1%) isolates carried blaCTX-M genes, blaTEM genes and blaSHV genes, respectively. blaCTX-M-55 (57/225, 25.3%) and blaCTX-M-65 (40/225, 17.8%) were the most frequent ESBL genes. Colistin resistance genes (including mcr-1, mcr-3 and mcr-5) were found in 15.6% of all isolates.ConclusionsThis study indicates that ESBL-producing E. coli are widely found in retail raw meats, especially chicken, in Singapore. Occurrence of MDR (resistance to at least three classes of antimicrobial) and colistin resistance genes in retail raw meat suggests potential food safety and public health risks.

Background Spiders are the most dominant predatory natural enemies of insect pests in the tea plantation ecosystem. There has been a large amount of literature published about the investigation of spider species in Chinese tea plantations from 1982 to 2020. Here, the spider species in Chinese tea plantations has been summarised and the dominant spider species in each regional tea plantation recorded. To date, there were 535 spider species from 40 families reported in Chinese tea plantations. New information There are 245 spider species from 13 families now being added to the checklist. A total of 89 spider species from 19 families were the dominant species, amongst them, Agelena labyrinthica , Allagelena difficilis , Neoscona theisi , Clubiona deletrix , Clubiona japonicola , Hylyphantes graminicola , Pardosa laura , Oxyopes sertatus , Evarcha albaria , Plexippus paykulli , Coleosoma octomaculatum , Ebrechtella tricuspidata and Xysticus ephippiatus were recorded in many tea plantations. The checklist will provide important data for the biodiversity and distribution of spiders in tea plantations of China.

Pseudopoda taibaischana Jäger, 2001 (Sparassidae) is redescribed based on new material from the type locality in Taibaishan Nation Forest Park of Shaanxi Province, China. The female is described and illustrated for the first time, and a redescription is provided for the male.

Two new species of the genus Thelcticopis Karsch, 1884, T. dahanensis Zhu & Zhong, sp. nov. (♂) and T. unciformis Zhu & Zhong, sp. nov. (♂), are described and figured from Taiwan Island. Thelcticopis severa (L. Koch, 1875) is recorded from Guangdong and Fujian provinces for the first time. So far, Thelcticopis, including four species from China, is mainly distributed in the tropical or subtropical areas of China (Hainan, Taiwan, Yunnan, Guangdong, Fujian).

Background Recently, abundant evidence indicated proinflammatory cytokines might play a crucial role in pathophysiology and treatment of depression. According to our preclinical research, we propose glycyrrhizic acid (GZA) for an adjunctive treatment owing to its safety, economical and anti-inflammatory profile. Methods Eligible participants were recruited and randomly allocated into independent treatment groups of SSRI+GZA (n = 30) and SSRI+PBO (placebo, n = 26). Depressive symptoms and specific serum biomarkers were detected during the 4-week treatment course. Afterward, the relationships between biomarkers and clinical effects were explored. Results Depressive symptoms relieved more in SSRI+GZA than SSRI+PBO, both at week 2 (P = 0.003) and week 4 (P = 0.016). Meanwhile, at week 4, both response rate (P = 0.035) and remission rate (P = 0.031) acutely became higher in SSRI+GZA compared with SSRI+PBO. Mediation analysis further demonstrated that TNF-α reduction mediated the association between GZA treatment and clinical improvement, the indirect effect lay between 0.124 and 3.514 (95% CI). The exploratory analysis also suggested that the symptomatic improvement existed in patients with high-inflammation (baseline CRP > 3 mg/L) rather than those with low-inflammation (baseline CRP ≤ 3 mg/L). Limitations The sample size in this study was not large enough and the follow-up duration was relatively short. Conclusions This study offers a novel strategy for the diagnosis, categorization, individualization and prognosis regarding upgrading traditional antidepressant therapy, which is from biomarkers to diagnostic indicator and therapeutic target. Patients are necessary to be classified according to the inflammatory state, those with high levels of baseline inflammation should receive combined treatment with anti-inflammatory agents like GZA.

The exact mechanisms associated with secondary brain damage following traumatic brain injury (TBI) remain unclear; therefore, identifying the critical molecular mechanisms involved in TBI is essential. The mRNA expression microarray GSE2871 was downloaded from the Gene Expression Omnibus (GEO) repository. GSE2871 comprises a total of 31 cerebral cortex samples, including two post-TBI time points. The microarray features eight control and seven TBI samples, from 4 hours post-TBI, and eight control and eight TBI samples from 24 hours post-TBI. In this bioinformatics-based study, 109 and 66 differentially expressed genes (DEGs) were identified in a Sprague-Dawley (SD) rat TBI model, 4 and 24 hours post-TBI, respectively. Functional enrichment analysis showed that the identified DEGs were significantly enriched in several terms, such as positive regulation of nuclear factor-κB transcription factor activity, mitogen-activated protein kinase signaling pathway, negative regulation of apoptotic process, and tumor necrosis factor signaling pathway. Moreover, the hub genes with high connectivity degrees were primarily related to inflammatory mediators. To validate the top five hub genes, a rat model of TBI was established using the weight-drop method, and real-time quantitative polymerase chain reaction analysis of the cerebral cortex was performed. The results showed that compared with control rats, Tnf-α, c-Myc, Spp1, Cxcl10, Ptprc, Egf, Mmp9, and Lcn2 were upregulated, and Fn1 was downregulated in TBI rats. Among these hub genes, Fn1, c-Myc, and Ptprc may represent novel biomarkers or therapeutic targets for TBI. These identified pathways and key genes may provide insights into the molecular mechanisms of TBI and provide potential treatment targets for patients with TBI. This study was approved by the Experimental Animal Ethics Committee of the First Affiliated Hospital of Nanchang University, China (approval No. 003) in January 2016.

Thitarodes (Lepidoptera, Hepialidae) is the only genus that hosts to the Ophiocordyceps sinensis, a traditional Chinese medicine considered as a powerful medicinal supplement. In this study, the complete mitochondrial genomes (mitogenomes) of two species, T. damxungensis and T. pui, have been sequenced, which are 15,928 bp and 15,362 bp in size respectively, and both contain 13 protein-coding genes (PCGs), 2 rRNAs, 22 tRNAs and an AT-rich region. Like other hepialoids, the gene arrangement of the mitogenomes of T. damxungensis and T. pui is identical to the ancestral arrangement but differs from those of other lepidopteran species on account of the different arrangements of trnM, trnI, and trnQ. The size of AT-rich region is 545 bp in T. damxungensis and 1030 bp in T. pui. Tandem repetition in the AT-rich region is responsible for the length difference of the A + T-rich region in both species. In Hepialidae, the phylogenetic study based on the dataset of the sequences that combined the protein-coding genes and RNA genes suggested that the species T. yunnanensis should still belong to the genus Thitarodes rather than Ahamns, which is different from the results based on the traditional phylogeny.

Introduction: As of June 7, 2021, the outbreak of Coronavirus Disease 2019 (COVID-19) has spread to more than 200 countries. The global number of reported cases is more than 172.9 million, with more than 3.7 million deaths, and the number of infected individuals is still growing rapidly. Consequently, events and activities around the world were canceled or postponed, and the preparation for sporting events were greatly challenged. Under such circumstances, about 11,000 athletes from ~206 countries are arriving in Tokyo for the 32nd Summer Olympic Games. Therefore, it is urgently necessary to assess the occurrence and spread risk of COVID-19 for the Games.Objectives: To explore effective prevention and control measures for COVID-19 in large international events through simulations of different interventions according to risk assessment.Methods: We used a random model to calculate the number of initial infected patients and used Poisson distribution to determine the number of initial infected patients based on the number of countries involved. Furthermore, to simulate the COVID-19 transmission, the susceptible-exposed-symptomatic-asymptomatic-recovered-hospitalized (SEIARH) model was established based on the susceptible-exposed-infectious-recovered (SEIR) mathematical model of epidemic diseases. According to risk assessment indicators produced by different scenarios of the simulated interventions, the risk of COVID-19 transmission in Tokyo Olympic Games was assessed.Results: The current COVID-19 prevention measures proposed by the Japan Olympic Committee need to be enhanced. And large-scale vaccination will effectively control the spread of COVID-19. When the protective efficacy of vaccines is 78.1% or 89.8%, and if the vaccination rate of athletes reaches 80%, an epidemic prevention barrier can be established.

ObjectiveEscherichia coli ST410 with blaNDM-5 has been increasingly detected as multidrug resistance pathogens globally, even though there are very few reports of infections caused by blaNDM-5 producing E. coli in Singapore[1]. And significantly limit sequencing information of blaNDM-5 carried E. coli strain from Singapore. In 2018, our group obtained a carbapenem resistance E. coli ST410 strain SrichA-1 isolated from reservoir water in Singapore, determined to harbor the NDM-5 gene. (BioSample Accession: SAMN18579051).MethodsThe susceptibility test to antimicrobials was performed with microdilution minimum inhibitory concentration (MIC) test and interpreted according to the Clinical And Laboratory Standards Institute (CLSI) -M100 standards. The genomic DNA of this strain was extracted and send for Whole-genome sequencing(WGS) with the Illumina platform. The WGS analysis was processed with the Center for Genomic Epidemiology (CGE, DTU) server.ResultsDuring the minimum inhibitory concentration (MIC) test, SrichA-1 has shown strong resistance to all the beta-lactams, including cephalosporin and carbapenem, which can not be inhibited by the clavulanic acid. Further whole genome sequencing analysis has shown that the strain harboring five beta-lactamase genes covers all class A to D, including the carbapenemase genes as blaNDM-5.ConclusionHere, we reported the complete chromosome sequence of this isolate as well as the sequence of a cycler plasmid. The pSGNDM-5 plasmid was furtherly identified to carry four beta-lactamase genes, including blaNDM-5, blaCTX-M-15, blaTEM-1B, blaOXA-1, while a blaCMY-2 was detected to be located on the chromosome.

As the role of the aquatic environment in the “One Health” approach has called increasing attention, the studies of Antimicrobial resistance (AMR) spreading in the water bodies have been reported worldwide. However, there are still limited studies on the AMR carrier in the reservoir water in Singapore. Since 2018, our group has collect water samples from six reservoirs in Singapore and isolated the beta-lactam resistant bacteria from them. We then characterized the isolates with Whole-genome sequencing (WGS) and successfully identified ESBL-producing bacteria from three sampling reservoirs, and confirmed their resistance with both phenotypic and sequencing methods. To better understand the AMR spreading locally, we compared our isolates with isolates from other WGS studies in Singapore covered humans, food, and the enviroment. From there, we noticed the same sequence type (ST) as ST10, ST23, and ST38 has been shared among the environment, food, and humans, as well as the same beta-lactamase genes, are widely distributed among multiple sources. Further genetic environment comparison of beta-lactamase has suggested their spreading as conserved gene clusters among different species and sources. And this hypothesis has been supported by the successful conjugation of blaCTX-M-15 from Klebsiella pneumonia to Escherichia coli (E .coli). We also applied the shotgun metagenomic sequencing to understand the community of bacteria in reservoir water and detect the AMR genes. The composition of bacteria has shown different diversity among different samples. Besides, different beta-lactamase genes have been identified compared to culture depended methods. Here, we suggest that sequencing analysis has great potential in understanding AMR spreading in the “One-Health” approach. A genetic-based AMR risk assessment is in urgent need in Singapore.

Due to its special ways of hiding and lifestyle, Thelcticopis is a type of spider that is very difficult to collect. In 2018, we collected two huntsman spiders in Guangxi. After comparison with other Thelcticopis species, such as anterior median eye larger than other eyes, they were found to belong to the genus of Thelcticopis. Currently, four Thelcticopis species are reported from China, T. severa (L. Koch, 1875), T. zhengi Liu, Li & Jäger, 2010, T. dahanensis Zhu & Zhong, 2020 and T. unciformis Zhu & Zhong, 2020. They are mainly distributed in the tropical or subtropical areas of China (Fujian, Guangdong, Guangxi, Hainan, Hongkong, Hunan, Taiwan, Yunnan and Zhejiang). In this paper, we diagnose and describe a new species, Thelcticopis pinmini sp. nov., which was collected from Guangxi Province, China.

Arabidopsis thaliana, one of the most important model plants, has played an essential role in every biological field including evolutionary biology. Recent population genomic studies have gradually clarified the origin and evolution of this species. Nevertheless, incongruent patterns among gene trees based on cytogenetic data have highlighted the importance of understanding the life history evolution and landscape biogeography of extant A. thaliana populations. Here, we focus on the maternally inherited chloroplast genome in A. thaliana and carry out phylogeographic analyses and coalescent time estimations. The maternal lineage of A. thaliana originated in the European to West and Central Asian regions in the Early Pleistocene. Relicts, the ancient lineages suggested by population genomic data, are not ancestral maternal lineages, but are derived from the European population. Part of the European population then dispersed eastward and spread to the Indian region, and finally extended to the Yangtze River region. The branching patterns and evolutionary time scales of the maternal genealogy are significantly different from those estimated from analyses of autosomal genes, and these cannot be explained by incomplete lineage sorting of the ancestral polymorphisms during the coalescent process due to large differences in the evolutionary time scale involved.

Functional manipulation of biosynthetic enzymes such as cytochrome P450s (or P450s) has attracted great interest in metabolic engineering of plant natural products. Cucurbitacins and mogrosides are plant triterpenoids that share the same backbone but display contrasting bioactivities. This structural and functional diversity of the two metabolites can be manipulated by engineering P450s. However, the functional redesign of P450s through directed evolution (DE) or structure-guided protein engineering is time consuming and challenging, often because of a lack of high-throughput screening methods and crystal structures of P450s. In this study, we used an integrated approach combining computational protein design, evolutionary information, and experimental data-driven optimization to alter the substrate specificity of a multifunctional P450 (CYP87D20) from cucumber. After three rounds of iterative design and evaluation of 96 protein variants, CYP87D20, which is involved in the cucurbitacin C biosynthetic pathway, was successfully transformed into a P450 mono-oxygenase that performs a single specific hydroxylation at C11 of cucurbitadienol. This integrated P450-engineering approach can be further applied to create a de novo pathway to produce mogrol, the precursor of the natural sweetener mogroside, or to alter the structural diversity of plant triterpenoids by functionally manipulating other P450s.

To evaluate the efficacy and safety of antithrombotic prophylaxis and to explore potential risk factors for thrombotic/bleeding events in patients with positive antiphospholipid (aPL) antibodies receiving invasive procedures. All aPL-positive patients who underwent invasive procedures in Peking Union Medical College Hospital, from January 2002 to April 2018, were retrospectively enrolled. Demographic features, clinical features, antiphospholipid antibody profiles, types of invasive procedures, and antithrombotic management, as well as complications and outcomes, were systematically reviewed and recorded. A total of 111 aPL-positive patients with 130 invasive procedures were enrolled. One hundred nine (83.8%) cases were on regular antithrombotic therapy which started at least 1 month prior to the invasive procedures, with 58 (44.6%) receiving anticoagulation therapy, 27 (20.8%) receiving antiplatelet therapy, and 24 (18.5%) receiving both. During the periprocedural period, the median time free of antithrombotic therapy was 2.5 days (interquartile range 1.5–6.0 days). Two (1.5%) periprocedural thrombotic events and 18 (13.8%) bleeding events were identified. Large open/laparoscopic surgeries of the thorax and abdomen were associated with a higher risk of bleeding (OR 3.46, 95% CI 1.24–9.67, p = 0.014). All bleeding events were manageable and not life-threatening. Aggressive antithrombotic therapy was associated with fewer thrombotic events in aPL-positive patients receiving invasive procedures, but might contribute to an increased bleeding rate, especially in large open surgeries. This study justifies more caution in prophylactic antithrombotic therapy in periprocedural aPL-positive patients.

The taxonomic status of Pseudopodaaureola (He & Hu, 2000) is revised based on the re-examination of the type specimen and fresh material collected from the type locality. The cheliceral dentition, with a large denticle field between two anterior and three posterior teeth, the male palps with simple RTA arising distally, narrow, not filiform embolus and sheet-like membranous conductor, and the female epigyne with a visible median septum indicate that the species does not belong to Pseudopoda or to the originally assigned genus, Heteropoda. Based on these characters, the species is here transferred to Gnathopalystes. The male is redescribed and the female is described and illustrated for the first time.

Hepatocellular carcinoma (HCC) is the most common type of liver cancer with high mortality. Identifying key molecules involved in the regulation of HCC development is of great clinical significance. SOX11 is a transcription factor belonging to group C of Sry-related high mobility group box family whose abnormal expression is frequently seen in many kinds of human cancers. Here, we noted that the expression of SOX11 was decreased in human HCC tumors compared with that in matched normal tissues. Overexpression of SOX11 promoted growth inhibition and apoptosis in HCC cell line HuH-7. Mechanistically, SOX11 enhanced the expression of nemo-like kinase and the phosphorylation of TCF4, thereby blunting the activation of oncogenic Wnt/β-catenin signaling pathway in HuH-7 cells. Finally, SOX11 was also found to sensitize HuH-7 cells to chemotherapy drugs cisplatin and 5-fluorouraci. Therefore, our study identifies SOX11 as a potential tumor suppressor in HCC and may hopefully be beneficial for the clinical diagnosis or treatment of HCC.

Many plant species that occur at the high-altitudes such as Qinghai-Tibet Plateau (QTP) under extreme climate conditions have potential medicinal functions. To explore the potential use of such plants, we extracted the essential oil from Artemisia dracunculus var. qinghaiensis Y. R. Ling collected from QTP at the flower-buds stage (0.67% v/w) by the hydro-distillation method. The chemical compositions and antimicrobial activity of the essential oil were determined using the gas chromatography/mass spectrometry (GC/MS) and agar disk-diffusion methods, respectively. Thirty-seven compounds accounting for 60.88% of the total detected components, with a high proportion of monoterpenes (42.12%) were identified in the essential oil. Of these, sabinene (19.19%), β-terpinene (8.94%), terpinen-4-ol (3.83%), and α-pinene (3.08%) were identified as the major compounds. The essential oil showed inhibitory activity against seven microbial strains (Staphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella paratyphi, and Saccharomyces cerevisiae) at the minimum concentrations (MICs) of 1.25–5.0 μl/ml. Our results suggest that the essential oil of the QTP A. dracunculus var. qinghaiensis can be used as a potential biomedical resource.

The effects of partial enzymatic hydrolysis of soymilk on the characteristics of transglutaminase (TG)-crosslinked tofu gel were studied. SDS-PAGE showed that the molecular weight of the partially hydrolyzed soybean protein was reduced to that of a digested peptide (less than 43.0 kDa) when papain was added at more than 50 μL/100 mL soymilk. The content of free sulfhydryls, β-sheets, and random coils in papain-treated soymilk increased. When TG was added to soy milk after papain treatment and tofu gel was formed, its storage modulus increased from 957.44 to 1241.39 Pa. The gel strength, water-holding capacity, and nonfreezing water content of the tofu gel were greater than those without enzyme treatment. Scanning electron microscopy revealed that limited papain hydrolysis stimulated TG-catalyzed cross-linking of soymilk to form a dense gel network structure, whereas an extended enzymatic hydrolysis of soymilk did not promote crosslinking by TG. PRACTICAL APPLICATION: This work investigated the effect of partial hydrolysis on TG cross-linked tofu gel. Partial hydrolysis of soybean protein with papain can promote TG cross-linking reaction, thus form a dense network structure, increase gel strength, and water-holding capacity. Therefore, it can be used to produce a good gel product with higher gel strength, springiness, water-holding capacity, and a more dense microstructure.; © 2018 Institute of Food Technologists®.

The Clubiona zilla-group is a relatively small species group, distributed exclusively in East Asia, with only three species clearly documented so far. Clubiona hooda Dong & Zhang, 2016, which was previously placed in the C. trivialis-group, is assigned to the C. zilla-group in the present paper. A new spider of the C. zilla-group from Jiugong Mountain in China is described under the name of C. jiugong sp. nov. Detailed descriptions and photographs of the new species are provided.

Burkholderia mallei, the causative agent of glanders, is a gram-negative intracellular bacterium. Depending on different routes of infection, the disease is manifested by pneumonia, septicemia, and chronic infections of the skin. B. mallei poses a serious biological threat due to its ability to infect via aerosol route, resistance to multiple antibiotics and to date there are no US Food and Drug Administration (FDA) approved vaccines available. Induction of innate immunity, inflammatory cytokines and chemokines following B. mallei infection, have been observed in in vitro and small rodent models; however, a global characterization of host responses has never been systematically investigated using a non-human primate (NHP) model. Here, using a liquid chromatography-tandem mass spectrometry (LC-MS/MS) approach, we identified alterations in expression levels of host proteins in peripheral blood mononuclear cells (PBMCs) originating from naïve rhesus macaques (Macaca mulatta), African green monkeys (Chlorocebus sabaeus), and cynomolgus macaques (Macaca fascicularis) exposed to aerosolized B. mallei. Gene ontology (GO) analysis identified several statistically significant overrepresented biological annotations including complement and coagulation cascade, nucleoside metabolic process, vesicle-mediated transport, intracellular signal transduction and cytoskeletal protein binding. By integrating an LC-MS/MS derived proteomics dataset with a previously published B. mallei host-pathogen interaction dataset, a statistically significant predictive protein-protein interaction (PPI) network was constructed. Pharmacological perturbation of one component of the PPI network, specifically ezrin, reduced B. mallei mediated interleukin-1β (IL-1β). On the contrary, the expression of IL-1β receptor antagonist (IL-1Ra) was upregulated upon pretreatment with the ezrin inhibitor. Taken together, inflammasome activation as demonstrated by IL-1β production and the homeostasis of inflammatory response is critical during the pathogenesis of glanders. Furthermore, the topology of the network reflects the underlying molecular mechanism of B. mallei infections in the NHP model.

Background: The fast-spreading of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli (ESBL-producing E. coli) and ESBL genes has become a big challenge to public health. The risk of spreading ESBL genes and pathogens in the environment and community has raised public health concern. The characterizing and whole-genome sequencing studies of ESBL-producing bacteria from reservoir water in Singapore is still limited. Materials and methods: The reservoir water sample was taken from two randomly selected sampling points of the Chinese Garden (Jurong river reservoir), which is a popular reservoir park in Singapore. The bacteria of the water sample were collected with 0.45 &micro, m filter membranes and enriched before processing with ESBL-producing E. coli screening. The collected ESBL positive isolates were further characterized by both phenotypic tests including disc diffusion and microdilution Minimum Inhibitory Concentration (MIC) test, and also genotypic test as whole-genome sequencing analysis. Besides, to investigate the transferability of the resistance gene, a conjugation test was performed with the J53 E. coli strain as the gene receptor. Result: Nine ESBL-producing E. coli isolates were collected and confirmed as ESBL-producing with both phenotypic and genotypic tests. A potential pathogen as ST131 clade A isolate was identified, and all isolates were determined to harbor a blaCTX-M gene. Among them, strain J1E4 was resistant to polymyxin E and confirmed to harboring a conjugatable mcr-1 gene. Further genetic environment analysis has reflected a conversed gene cluster formed by insert sequence (IS), blaCTX-M-15, and WbuC family cupin-fold metalloprotein, which may potentially jump from the plasmids to the chromosome. Conclusion: The first time we reported the whole genome sequencing (WGS) data of ESBL-producing E. coli including potential pathogen (ST131) present in reservoir water in Singapore. The ESBL-producing E. coli from reservoir water also carrying conjugatable colistin resistance genes which may become a risk to human health.

BackgroundAstragalus membranaceus Bge. var. mongolicus (Bge.) Hsiaois one of the most common herbs widely used in South and East Asia, to enhance people’s health and reinforce vital energy. Despite its prevalence, however, the knowledge about phytochemical compositions and metabolite biosynthesis inAstragalus membranaceus Bge. var. mongolicus (Bge.) Hsiaois very limited.ResultsAn integrated metabolomics and transcriptomics analysis using state-of-the-art UPLC-Q-Orbitrap mass spectrometer and advanced bioinformatics pipeline were conducted to study global metabolic profiles and phytochemical ingredients/biosynthesis inAstragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. A total of 5435 metabolites were detected, from which 2190 were annotated, representing an order of magnitude increase over previously known. Metabolic profiling ofAstragalus membranaceus Bge. var. mongolicus (Bge.) Hsiaotissues found contents and synthetic enzymes for phytochemicals were significantly higher in leaf and stem in general, whereas the contents of the main bioactive ingredients were significantly enriched in root, underlying the value of root in herbal remedies. Using integrated metabolomics and transcriptomics data, we illustrated the complete pathways of phenylpropanoid biosynthesis, flavonoid biosynthesis, and isoflavonoid biosynthesis, in which some were first reported in the herb. More importantly, we discovered novel flavonoid derivatives using informatics method for neutral loss scan, in addition to inferring their likely synthesis pathways inAstragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao.ConclusionsThe current study represents the most comprehensive metabolomics and transcriptomics analysis on traditional herbAstragalus membranaceus Bge. var. mongolicus (Bge.) Hsiao. We demonstrated our integrated metabolomics and transcriptomics approach offers great potentials in discovering novel metabolite structure and associated synthesis pathways. This study provides novel insights into the phytochemical ingredients, metabolite biosynthesis, and complex metabolic network in herbs, highlighting the rich natural resource and nutritional value of traditional herbal plants.

In the past year, Prof. Jian Chen conducted several spider collections in Hubei Province. Almost 1000 spiders were collected. After diagnosis, two of them were found to belong to the genus Sinopoda Jäger, 1999. Two Sinopoda Jäger, 1999 species, both from Hubei Province, including one new species, are treated in the current paper: S. angulata Jäger, Gao & Fei, 2002 and S. yichangensis sp. n. (♂). The male of S. angulata is described for the first time from Enshi Tujia and Miao Autonomous Prefecture, Hubei Province. New geographic records are provided as well as photos of the copulatory organs and habitus.

The species of Prosoponoides Millidge & Russell-Smith, 1992 from China are studied, including one new species, Prosoponoides youyiensis Liu & Chen n. sp., and two known species, Prosoponoides hamatum Millidge & Russell-Smith, 1992 and Prosoponoides sinense (Chen, 1991). Morphological descriptions and illustrations of P. hamatum and P. youyiensis Liu & Chen n. sp. are given, and digital photos of P. sinense are provided.

Nonsynonymous-synonymous substitution rate ratio (d N/d S) is an important measure for evaluating selective pressure based on the protein-coding sequences. Maximum likelihood (ML) method with codon-substitution models is a powerful statistic tool for detecting amino acid sites under positive selection and adaptive evolution. We analyzed the hepatitis C virus (HCV) envelope protein-coding sequences from 18 general geno/subtypes worldwide, and found 4 amino acid sites under positive selection. Since these sites are located in different immune epitopes, it is reasonable to anticipate that our study would have potential values in biomedicine. It also suggests that the ML method is an effective way to detect adaptive evolution in virus proteins with relatively high genetic diversity.

Objectives The objective of this study was to address the biological function of miR-7 in an animal model of systemic lupus erythematosus. Methods MRLlpr/lpr lupus mice were administrated antagomiR-7 or a scramble control by tail vein for 5weeks. Three groups of animals’ tissues were assessed for lupus manifestations by immunofluorescence and immunohistochemistry, and serum was examined for levels of autoantibodies and inflammatory cytokines. Splenic B cell subsets were assessed for intracellular expression of PI3K signaling by FACS. Finally, the ability of the miR-7 antagomir to regulate the expansion of T follicular helper (Tfh) cells and B cell hyperresponsiveness was further explored. Results We found that miR-7 was up-regulated in MRLlpr/lpr lupus mice and directly targeted PTEN mRNA in B cells. Up-regulated miR-7 in MRLlpr/lpr lupus B cells was negatively correlated with PTEN expression. Notably, miR-7 antagomir treatment reduced lupus manifestations in MRLlpr/lpr lupus mice. miR-7-mediated down-regulation of PTEN/AKT signaling promoted B cell differentiation into plasmablasts/plasma cells and spontaneous germinal center (GC) formation, whereas miR-7 antagomir normalized splenic B cell subtypes. Besides suppressing the activation of B cells, miR-7 antagomir intervention also down-regulated STAT3 phosphorylation and production of IL-21 and reduced Tfh expansion. Conclusion The above data have demonstrated the critical roles of miR-7 not only in regulating PTEN expression and also B cell and Tfh cell function in lupus-prone MRLlpr/lpr lupus mice. Furthermore, the disease manifestations in MRLlpr/lpr lupus mice are efficiently improved by miR-7 antagomir, indicating miR-7 as a potential treatment strategy in SLE.

Sinopoda spiders are a diverse group with limited dispersal ability. They are remarkably sympatric among related species, which often results in misidentification and incorrect matching of sexes. In order to understand the evolutionary relationships and revise the taxonomy problems in this genus, we offer the first molecular phylogeny of Sinopoda. Our results strongly support the monophyly of Sinopoda and its sister relationship with Spariolenus and reject the monophyly of the S. okinawana species group. We establish three new species groups based on both molecular and morphological data. Our phylogeny also illuminates some taxonomic issues and clarifies some species limits: (1) Supporting the newly revised matching of sexes in S. longiducta and S. yaanensis by Zhong et al. (2019). (2) The original description of S. campanacea was based on mismatched sexes. S. changde is proposed as a junior synonymy of S. campanacea, while the original female ‘S. campanacea’ is here described as a new species: S. papilionaceous Liu sp. nov. (3) The type series of S. serpentembolus contains mismatched sexes. The female is considered as S. campanacea, while we here report the correctly matched females of S. serpentembolus. (4) We describe one additional new species: S. wuyiensis Liu sp. nov. Our first molecular phylogeny of Sinopoda provides a tool for comparative analyses and a solid base for the future biodiversity and taxonomic work on the genus.

Recent studies have shown that bacteria can also undergo apoptosis, which has gradually attracted researchers' attention. Cisplatin is a first-line drug to treat several cancers, but it can damage beneficial bacteria. Hence it is very important to explore the damage mechanism of cisplatin on beneficial bacteria. In this study, Lactobacillus paracasei, one kind of beneficial bacteria, was used as the model to investigate cisplatin damage. Conventional detection showed that cisplatin induced the apoptosis of Lactobacillus paracasei. Then Fourier transform infrared (FTIR) microspectroscopy was used to detect biomacromolecular changes in Lactobacillus paracasei apoptosis, and the following results were obtained: ① Second derivative IR spectra showed the changes of DNA, proteins, polysaccharides and lipids; ② Peak-area ratios suggested the changes of the protein and lipid structure and the decrease of DNA content; ③ Principal component analysis (PCA) further revealed significant changes in the DNA and protein content/structure. This study may have a new insight into the adverse reaction mechanism of cisplatin on Lactobacillus, moreover, it suggests that FTIR microspectroscopy may be a useful supplementary tool for investigating bacterial apoptosis.

In this study, we elucidated the microbial transglutaminase-induced gelation properties and thermal gelling ability of soy protein isolate (SPI) and wheat gluten (WG) mixture following ultrahigh pressure (UHP, 100–400 MPa) pretreatment. UHP treatment induced unfolding and aggregation within SPI/WG protein molecules, which led to increases in free sulfhydryl group content and surface hydrophobicity. However, the transglutaminase cross-linking reaction facilitated the formation of hydrophobic interactions and disulfide bonds and thus resulted in higher gel strength, water holding capacity, and denser and more homogeneous gel networks of transglutaminase cross-linked SPI/WG gels. Rheological measurements revealed that the addition of UHP steps might generate a higher storage modulus (G′) value of MTGase-induced SPI/WG gelation during the heating-cooling cycle (25 °C → 95 °C → 25 °C). Our results indicated that various chemical interactions including covalent interactions (i.e., e-(γ-glutamyl)lysine bonds and disulfide bonds) and non-covalent interactions (i.e., electrostatic forces and hydrophobic interactions) were involved in SPI/WG gel network structures. Hydrophobic interactions and disulfide bonds are significantly increased with the pressure level (100–400 MPa) compared with that of the unpressurized control. Furthermore, UHP treatment reduced the α-helix and β-turn content but increased the β-sheet and random coil structures. Thus, UHP treatment may be considered as a novel technique to expand the utilization of SPI/WG mixture in the food protein gelation industry.

Colistin is regarded as one of the last-resort antimicrobials for treatment of Gram-negative bacterial infections (1). Several cases of plasmid-borne colistin resistance genes mcr-1, mcr-3-like, and mcr-4.2 in clinical Enterobacteriaceae isolates, including Escherichia coli, have been reported in Singapore (2-4). However, the mcr-5 gene has not been reported in clinical isolates in Singapore. Previously, we reported the antimicrobial resistance (AMR) genotype and phenotype of E. coli SGEHI2010ENV103 isolates from ready-to-eat food in Singapore and documented the first isolate carrying mcr-5.1 in Singapore (5). In this study, we further analyzed this isolate. Here, we report the first complete nucleotide sequence of a transferable plasmid harboring mcr-5.1 in E. coli isolated from ready-to-eat chicken rice in Singapore. (Chicken rice is a common dish in Singapore which is composed of cooked chicken and seasoned rice, served with sauce and cucumber garnishes). Published version

As an important member of cytochrome P450 (CYP) enzymes, human CYP1A2 is associated with the metabolism of caffeine and melatonin and the activation of precarcinogens. Besides, this CYP protein also involves in metabolizing 5-10% of clinical medicines. Some peripheral mutations in CYP1A2 (P42R, I386F, R431W, and R456H) significantly decrease the enzyme activities, resulting in a vital reduction in substrate metabolisms. To explore the effects of these peripheral mutations, we constructed a membrane-binding model for the full-length human CYP1A2 and studied their dynamic behaviors on lipid membranes. Free energy calculations indicate that the peripheral mutations donot influence substrate binding. P42R is located in the N-terminal anchor, and its positive charged sidechain is adverse to membrane binding. I386F enhances the van der Waals contacts of the water channel bottleneck and R456H breaks the hydrogen bonding interactions that function to position the BC loop, both of which result in a significant inhibition on the water channel. R431W causes a sidechain conformational rearrangement for aromatic residues around the substrate channel, making it in a closed state in most cases. Our computational simulations demonstrate that pi-pi stacking interactions are essential for substrate binding and channel opening. We hope that these findings may be of general relevance to the mutation-induced activity changes for CYP proteins, providing useful information for understanding the CYP-mediated drug metabolism.

Traumatic brain injury (TBI) and status epilepticus (SE) have both been linked to development of human epilepsy. Although distinct etiologies, current research has suggested the convergence of molecular mechanisms underlying epileptogenesis following these insults. One such mechanism involves the neurotrophin brain-derived neurotrophic factor (BDNF) and its high-affinity receptor, tropomyosin related kinase B (TrkB). In this review, we focus on currently available data regarding the pathophysiologic role of BDNF/TrkB signaling in epilepsy development. We specifically examine the axonal injury and SE epilepsy models, two animal models that recapitulate many aspects of TBI- and SE-induced epilepsy in humans respectively. Thereafter, we discuss aspiring strategies for targeting BDNF/TrkB signaling so as to prevent epilepsy following an insult or suppress its expression once developed. This article is part of the special issue entitled 'New Epilepsy Therapies for the 21st Century - From Antiseizure Drugs to Prevention, Modification and Cure of Epilepsy'.

The spider genus Sinopoda Jager, 1999 in China is studied. Seventeen new species are described: S. abstrusa sp. nov. (♂♀, Xizang), S. aequalis sp. nov. (♂♀, Henan), S. apiculiformis sp. nov. (♂♀, Hunan), S. brevis sp. nov. (♂♀, Liaoning), S. changde sp. nov. (♂♀, Hunan, Shaanxi), S. columnaris sp. nov. (♀, Guizhou), S. curva sp. nov. (♂, Guangxi), S. dehiscens sp. nov. (♀, Hunan), S. erromena sp. nov. (♀, Hunan), S. improcera sp. nov. (♂♀, Yunnan), S. lata sp. nov. (♀, Yunnan), S. luyui sp. nov. (♂♀, Sichuan, Gansu), S. ovata sp. nov. (♂♀, Chongqing), S. pyramidalis sp. nov. (♂♀, Sichuan), S. tumefacta sp. nov. (♂♀, Yunnan), S. yaanensis sp. nov. (♂♀, Sichuan, Yunnan), S. yanlingensis sp. nov. (♂♀, Hunan). Five known species are redescribed or diagnosed and differentiated from related species: the males of S. liui Zhong, Cao & Liu, 2017 and S. longshan Yin, Peng, Yan & Bao, 2000 and the female of S. pengi Song & Zhu, 1999 are reported for the first time; the female of S. longiducta Zhang, Zhang & Zhang, 2015 is considered mismatched and transferred to S. yaanensis sp. nov. ; photographs and new records are provided for S. globosa Zhang, Zhang & Zhang, 2015.

Despite essential roles played by long noncoding RNAs (lncRNAs) in development and disease, methods to determine lncRNA cis-elements are lacking. Here, we developed a screening method named “Tiling CRISPR” to identify lncRNA functional domains. Using this approach, we identified Xist A-Repeats as the silencing domain, an observation in agreement with published work, suggesting Tiling CRISPR feasibility. Mechanistic analysis suggested a novel function for Xist A-repeats in promoting Xist transcription. Overall, our method allows mapping of lncRNA functional domains in an unbiased and potentially high-throughput manner to facilitate the understanding of lncRNA functions.

Melon is an economically important fruit crop that has been cultivated for thousands of years; however, the genetic basis and history of its domestication still remain largely unknown. Here we report a comprehensive map of the genomic variation in melon derived from the resequencing of 1,175 accessions, which represent the global diversity of the species. Our results suggest that three independent domestication events occurred in melon, two in India and one in Africa. We detected two independent sets of domestication sweeps, resulting in diverse characteristics of the two subspecies melo and agrestis during melon breeding. Genome-wide association studies for 16 agronomic traits identified 208 loci significantly associated with fruit mass, quality and morphological characters. This study sheds light on the domestication history of melon and provides a valuable resource for genomics-assisted breeding of this important crop., This work was supported by funding from the Agricultural Science and Technology Innovation Program (to Yongyang Xu, S.H., Z.Z. and H.W.), the China Agriculture Research System (CARS-25 to Yongyang Xu and H.W.), the Leading Talents of Guangdong Province Program (00201515 to S.H.), the Shenzhen Municipal (The Peacock Plan KQTD2016113010482651 to S.H.), the Dapeng district government, National Natural Science Foundation of China (31772304 to Z.Z.), the Science and Technology Program of Guangdong (2018B020202007 to S.H.), the National Natural Science Foundation of China (31530066 to S.H.), the National Key R&D Program of China (2016YFD0101007 to S.H.), USDA National Institute of Food and Agriculture Specialty Crop Research Initiative (2015-51181-24285 to Z.F.), the European Research Council (ERC-SEXYPARTH to A.B.), the Spanish Ministry of Economy and Competitiveness (AGL2015–64625-C2-1-R to J.G.-M.), Severo Ochoa Programme for Centres of Excellence in R&D 2016–2010 (SEV-2015–0533 to J.G.-M.), the CERCA Programme/Generalitat de Catalunya to J.G.-M. and the German Science Foundation (SPP1991 Taxon-OMICS to H.S.).

I'm sorry, we can't use the final publisher's pdf for this journal (http://www.sherpa.ac.uk/romeo/issn/2095-9273/), but we can use the post-print (ie final draft post-refereeing) if you have it? RVO 16/1/18

To discover leads for next-generation chemoprotective antimalarial drugs,we tested more than 500,000 compounds for their ability to inhibit liver-stage development of luciferase-expressing Plasmodium spp. parasites (681 compounds showed a half-maximal inhibitory concentration of less than 1micromolar).Cluster analysis identified potent and previously unreported scaffold families as well as other series previously associated with chemoprophylaxis. Further testing through multiple phenotypic assays that predict stage-specific and multispecies antimalarial activity distinguished compound classes that are likely to provide symptomatic relief by reducing asexual blood-stage parasitemia from those which are likely to only prevent malaria. Target identification by using functional assays, in vitro evolution, or metabolic profiling revealed 58 mitochondrial inhibitors but also many chemotypes possibly with previously unidentified mechanisms of action. INTRODUCTION Malaria remains a devastating disease, affecting 216 million people annually, with 445,000 deaths occurring primarily in children under 5 years old. Malaria treatment relies primarily on drugs that target the diseasecausing asexual blood stages (ABS) of Plasmodium parasites, the organisms responsible for human malaria. Whereas travelers may rely on shortterm daily chemoprotective drugs, those living in endemic regions require long-termmalaria protection such as insecticide-treated nets (ITNs) and vector control. However, ITNs do not fully shield individuals from malaria, may lose potency with time, and can be bulky and difficult to use. Another concern is that mosquitosmay become resistant to the active insecticides that are used in ITNs and vector control. RATIONALE As the possibility of malaria elimination becomesmore tangible, the ideal antimalarial medicine profile should include chemoprotection. Chemoprotectivemedicines typically work against the exoerythrocytic parasite forms that invade and develop in the liver and are responsible for the earliest asymptomatic stage of the infection. Such medicines could be formulated to provide long-acting prophylaxis, safeguarding individuals that are living near or traveling to areas that have been cleared of parasites. Long-acting chemoprotection in endemic regions could also greatly reduce circulating parasite numbersandpotentially replace a vaccine in an elimination campaign. Although millions of compounds have been screened for activity against parasiteABS, and some have been subsequently tested for potential prophylactic activity, large-scale searches that beginwith prophylactic activity have not been performed because of the complexity of the assay: This assay requires the production of infected laboratory-rearedmosquitoes and hand-dissection of the sporozoiteinfected salivary glands frommosquito thoraxes. A Plasmodium vivax liver-stage schizont on a lawn of hepatocytes. The parasite schizont has been stained with antibodies to parasite HSP70 (red) and UIS4 (yellow). Cell (parasite and hepatoma) nuclei are shown in blue. This study identifies compounds that can prevent the development of these liver-stage parasites and may function as chemoprotective drugs for malaria. RESULTS To discover leads for next-generation chemoprotective antimalarial drugs, we used luciferase-expressing Plasmodium spp. parasites, dissected from more than a million mosquitoes over a 2-year period, to test more than 500,000 compounds for their ability to inhibit liver-stage development of malaria (681 compounds showed a half-maximal inhibitory concentration of

The objective of the present study was to explore the molecular mechanism with which a single nucleotide polymorphism (rs61764370) interferes with the interaction between the 3'-untranslated region (3'-UTR) of Kirsten rat sarcoma viral oncogene homolog (KRAS) and let-7a, and its association with the metastasis of osteosarcoma (OS). In this study, we confirmed that KRAS is a target of let-7a in OS cells, and the introduction of rs61764370 minor allele into KRAS 3'-UTR significantly compromised the microRNA (miRNA)/mRNA interaction using a luciferase reporter system. Additionally, a total of 36 OS tissue samples of three different genotypes (TT,22; TG,10; GG,4) were obtained, and the expression of let-7a and KRAS was determined. We showed that let-7a mRNA expression was similar between each group whereas the mRNA and protein expression of KRAS in the TT genotype group was significantly lower than that in the GT or GG genotype groups. Moreover, we identified a negative regulatory relationship between let-7a and KRAS. Furthermore, we demonstrated that let-7a and KRAS interfered with the viability, invasiveness and migration of OS cells genotyped as TT. In the OS cells genotyped as TG, let-7a exerted minimal effects, and the effect of KRAS siRNA remained. Taken together, the findings of the present study demonstrated that the KRAS 3'-UTR rs61764370 polymorphism interfered with miRNA/mRNA interaction, and showed that the minor allele was associated with an elevated risk of developing metastatic disease in OS.

Soy protein isolate (SPI) and wheat gluten (WG) are widely used in commercial food applications in Asia for their nutritional value and functional properties. However, individually each exhibits poor gelation. In this study, we examined the microbial transglutaminase (MTGase)-induced gelation properties of SPI and WG mixtures with high intensity ultrasonic pretreatment. Ultrasonic treatment reduced the particle size of SPI/WG molecules, which led to improvements in surface hydrophobicity (Ho) and free sulfhydryl (SH) group content. However, MTGase crosslinking facilitated the formation of disulfide bonds, markedly decreasing the content of free SH groups. Ultrasonic treatment improved the gel strength, water holding capacity, and storage modulus and resulted in denser and more homogeneous networks of MTGase-induced SPI/WG gels. In addition, ultrasonic treatment changed the secondary structure of the gel samples as determined by Fourier transform infrared spectroscopic analysis, with a reduction in α-helices and β-turns and an increase in β-sheets and random coils. Thus, ultrasound is useful in facilitating the gelation properties of MTGase-induced SPI/WG gels and might expand their utilization in the food protein gelation industry.

Background The integration of soybean protein isolate (SPI) with wheat gluten (WG) crosslinked via microbial transglutaminase (MTGase) may enhance the formation of ϵ-(γ-glutamyl)lysine covalent bonds, because SPI is rich in lysine and WG contains more glutamine. Microwave pretreatment may accelerate enzymatic reactions. In this study, we aimed to elucidate the effects of microwave pretreatment on the gelation properties of SPI and WG crosslinked with MTGase. Results Interestingly, the gel strength, water-holding capacity (WHC) and storage modulus (G') values of MTGase-induced SPI/WG gels were dramatically improved with increasing microwave power. Moreover, the MTGase crosslinking reaction promoted the formation of disulfide bonds, markedly reducing the free SH group and soluble protein content of gels. Fourier transform infrared spectroscopic analysis of SPI/WG gels showed that microwave pretreatment increased the proportion of α-helices and β-turns and decreased the proportion of β-sheets. Results from scanning electron microscopy indicated that the MTGase-induced SPI/WG gels had denser and more homogeneous microstructures after microwave pretreatment. Conclusion The effect of microwave pretreatment is useful in advancing gelation characters of MTGase-induced SPI/WG gels and provides the possibility for expanding the application of food protein. © 2015 Society of Chemical Industry.

The giant rhinoceros beetles (Dynastini, Scarabaeidae, Coleoptera) are distributed in tropical and temperate regions in Asia, America and Africa. Recent molecular phylogenetic studies have revealed that the giant rhinoceros beetles can be divided into three clades representing Asia, America and Africa. Although a correlation between their evolution and the continental drift during the Pangean breakup was suggested, there is no accurate divergence time estimation among the three clades based on molecular data. Moreover, there is a long chronological gap between the timing of the Pangean breakup (Cretaceous: 110-148 Ma) and the emergence of the oldest fossil record (Oligocene: 33 Ma). In this study, we estimated their divergence times based on molecular data, using several combinations of fossil calibration sets, and obtained robust estimates. The inter-continental divergence events among the clades were estimated to have occurred about 99 Ma (Asian clade and others) and 78 Ma (American clade and African clade), both of which are after the Pangean breakup. These estimates suggest their inter-continental divergences occurred by overseas sweepstakes dispersal, rather than by vicariances of the population caused by the Pangean breakup.

Emodinol, 1β,3β,23-trihydroxyolean-12-en-28-acid, as the main active ingredient firstly extracted from the rhizomes of Elaeagus pungens by our research group, has been demonstrated to exhibit uricosuric activity by our previous study. The aim of this study was to evaluate the uricosuric and nephroprotective effects of emodinol and explore its possible mechanisms in potassium oxonate-induced hyperuricemic mice with renal dysfunction. Mice were orally administrated 250 mg/kg of potassium oxonate once daily for 7 consecutive days to induce hyperuricemia with renal dysfunction. Emodinol was given at doses of 25, 50, and 100 mg/kg on the same day 1 h after oxonate treatment, and allopurinol (10 mg/kg) was given as a positive control. After 1 week, serum uric acid, serum creatinine, urine uric acid, urine creatinine, blood urea nitrogen, and hepatic xanthine oxidase activity were determined. The mRNA and protein levels of urate transporter 1, glucose transporter 9, ATP-binding cassette subfamily G member 2, organic anion transporter 1, oncoprotein-induced transcript 3, and organic cation/carnitine transporters in the kidney were detected by real-time polymerase chain reaction and Western blot analysis. In addition, urinary and renal Tamm-Horsfall glycoprotein concentrations were examined by ELISA assays. Emodinol significantly reduced serum urate levels, increased urinary urate levels and fractional excretion of uric acid, and inhibited hepatic xanthine oxidase activity in hyperuricemic mice. Moreover, potassium oxonate administration led to dys expressions of renal urate transporter 1, glucose transporter 9, ATP-binding cassette subfamily G member 2, organic anion transporter 1, and oncoprotein-induced transcript 3 as well as alternations of uromodulin concentrations, which could be reversed by emodinol. On the other hand, treatment of emodinol caused upregulated expressions of organic cation/carnitine transporters, resulting in an improvement of renal function characterized by decreased serum creatinine and blood urea nitrogen levels. Emodinol exhibited hypouricemic and nephroprotective actions by inhibiting xanthine oxidase activity and regulating renal ion transporters and oncoprotein-induced transcript 3, which may be a potential therapeutic agent in hyperuricemia and renal dysfunction.

Pieridae is a butterfly family whose higher classification has not yet been settled. In this research, we used completed mitochondrial genome sequence to identify and analyze the relationship of three subfamilies of Pieridae (Dismorphiinae, Coliadinae and Pierinae). We have sequenced the complete mitogenomes of Aporia crataegi, Gonepteryx rhamni, and Appias remedios, which are 15,148 bp, 15,118 bp and 15,223 bp in length, respectively. These three mitogenomes have a typical set of 37 genes and an A+T-rich region. The protein-coding genes, excluding the cox1 gene, in the three mitogenomes are initiated by the typical ATN codons. The putative initiation codon for the cox1 gene in the three species is CGA. tRNA genes in the three Pieridae mitogenomes show typical clover leaf structures, except tRNASer(AGN) which lacks the dihydrouridine (DHU) stem. The length of A+T-rich region varies in the three species, which leads to the variation in the mitochondrial genome size. The characterizations of the three mitogenomes enrich our knowledge on the Lepidopteran mitogenome and provide us genetic information to reconstruct the phylogenetic tree. Finally, the phylogenetic analyses using mitogenome data showed that the relationship among three subfamilies of Pieridae is (Dismorphiinae + (Coliadinae + Pierinae)).