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Carbon reserves in rice straw (stem and sheath) before flowering contribute to a significant portion of grain filling. However, the molecular mechanism of carbon reserve remobilization from straw to grains remains unclear. In this study, super rice LYP9 and conventional rice 9311 showed different carbon reserve remobilization behaviors. The transcriptomic profiles of straws of LYP9 and 9311 were analyzed at three stages of grain filling. Among the differentially expressed genes (DGs), 5,733 genes were uniquely up- or down-regulated at 30 days after anthesis (DAA) between LYP9 and 9311 in comparison with 681 at 10 DAA and 495 at 20 DAA, suggesting that the gene expression profile of LYP9 was very different from that of 9311 at the late stage of grain filling. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis and Gene Ontology (GO) classification of DGs both showed that the carbohydrate catabolic pathway, plant hormone signal transduction and photosynthesis pathway were enriched in DGs, suggesting their roles in carbon reserve remobilization, which explains to a certain extent the difference in non-structural carbohydrate content, photosynthesis and ABA content between the two cultivars during grain filling. Further comparative analysis and confirmation by quantitative real-time PCR and enzyme assays suggest that genes involved in trehalose synthesis (trehalose-phosphate phosphatase and trehalose 6-phosphate synthase/phosphatase), starch degradation (β-amylase) and sucrose synthesis (sucrose-phosphate synthase and sucrose synthase) were important for carbon reserve remobilization, whereas ABA content was determined by the counteraction of NCED1 and ABA8ox1 genes. The higher expression level of all these genes and ABA content in 9311 resulted in better efficiency of carbon reserve remobilization in 9311 than in LYP9. [ABSTRACT FROM AUTHOR]

Sucralose is the most widely used artificial sweetener, and its health effects have been highly debated over the years. In particular, previous studies have shown that sucralose consumption can alter the gut microbiota. The gut microbiome plays a key role in processes related to host health, such as food digestion and fermentation, immune cell development, and enteric nervous system regulation. Inflammation is one of the most common effects associated with gut microbiome dysbiosis, which has been linked to a series of human diseases, such as diabetes and obesity. The aim of this study was to investigate the structural and functional effects of sucralose on the gut microbiota and associated inflammation in the host. In this study, C57BL/6 male mice received sucralose in their drinking water for 6 months. The difference in gut microbiota composition and metabolites between control and sucralose-treated mice was determined using 16S rRNA gene sequencing, functional gene enrichment analysis and metabolomics. Inflammatory gene expression in tissues was analyzed by RT-PCR. Alterations in bacterial genera showed that sucralose affects the gut microbiota and its developmental dynamics. Enrichment of bacterial pro-inflammatory genes and disruption in fecal metabolites suggest that 6-month sucralose consumption at the human acceptable daily intake (ADI) may increase the risk of developing tissue inflammation by disrupting the gut microbiota, which is supported by elevated pro-inflammatory gene expression in the liver of sucralose-treated mice. Our results highlight the role of sucralose-gut microbiome interaction in regulating host health-related processes, particularly chronic inflammation. [ABSTRACT FROM AUTHOR]

Introduction: Nogo-A is an important neurite growth-regulatory protein in the adult and developing nervous system. Recently, increasing evidence has shown that Nogo-A plays important roles in cardiac development and may act as a potential indicator for heart failure. In addition, increased oxidative stress has been found in individuals with cardiovascular diseases. However, not much is known regarding the expression levels of Nogo-A and reactive oxygen species (ROS) in patients with coronary heart disease (CHD). Therefore, we sought to investigate the relationship between Nogo-A, ROS levels and CHD.Material and Methods: The plasma Nogo-A and ROS concentrations of 122 acute coronary syndrome (ACS), 101 unstable angina pectoris (UAP), and 21 acute myocardial infarction (AMI) patients and 56 healthy controls were measured by enzyme-linked immunosorbent assay (ELISA). We further generated a receiver operating characteristic (ROC) curve to assess the diagnostic accuracy of Nogo-A and ROS in CHD.Results: The Nogo-A and ROS levels were significantly higher in patients with CHD than those in healthy controls. In addition, multivariate logistic regression analysis revealed that the level of Nogo-A (odds ratio (OR) = 1.624, 95% confidence interval: 1.125-2.293, p = 0.009) is a risk factor for prediction of CHD. Nogo-A has diagnostic value, with an optimal threshold of 5.466 ng/ml for maximized diagnostic performance (59% sensitivity and 78.6% specificity, area under curve, p < 0.05). However, ROS concentration is not a risk factor for prediction of CHD (OR = 0.999, 95% confidence interval: 0.997-1.001, p = 0.320).Conclusions: Increased plasma Nogo-A level may be associated with CHD. [ABSTRACT FROM AUTHOR]

Background: Environmental chemical-induced perturbations of gut microbiome are associated with a series of adverse health outcomes. The effects of triclosan on human health have been controversial in recent years. The purpose of this study is to investigate the functional impact of triclosan on the mouse gut microbiome and the link between triclosan exposure and resistomes in gut bacteria. Methods: We combined 16S rRNA gene sequencing and shotgun metagenomics sequencing to examine the compositional and functional impact of triclosan exposure on the gut microbiota of C57BL/6 mice. Results: 16S rRNA sequencing results revealed that 13-week triclosan exposure in drinking water induced significant perturbations in mouse gut bacterial assemblages with distinct trajectories compared to controls. Metagenomics sequencing results indicated a remarkable enrichment of gut bacterial genes related to triclosan resistance, stress response, antibiotic resistance and heavy metal resistance. Conclusions: Triclosan exposure has a profound impact on the mouse gut microbiome by inducing perturbations at both compositional and functional levels. To our best knowledge, this is the first evidence regarding the functional alterations of gut microbiome induced by triclosan exposure, which may provide novel mechanistic insights into triclosan exposure and associated diseases. [ABSTRACT FROM AUTHOR]

The gut microbiome plays a key role in energy production, immune system development, and host resistance against invading pathogens, etc. Disruption of gut bacterial homeostasis is associated with a number of human diseases. Several environmental chemicals have been reported to induce alterations of the gut microbiome. Diazinon, one of important organophosphate insecticides, has been widely used in agriculture. Diazinon and its metabolites are readily detected in different environmental settings and human urine. The toxicity of organophosphates has been a long-standing public health concern. We recently demonstrated that organophosphate insecticide diazinon perturbed the gut microbiome composition of mice. However, the functional impact of exposure on the gut microbiome has not been adequately assessed yet. In particular, the molecular mechanism responsible for exposure-induced microbial profile and community structure changes has not been identified. Therefore, in this study, we used metatranscriptomics to examine the effects of diazinon exposure on the gut metatranscriptome in C57BL/6 mice. Herein, we demonstrated for the first time that organophosphate diazinon modulated quorum sensing, which may serve as a key mechanism to regulate bacterial population, composition, and more importantly, their functional genes. In addition, we also found that diazinon exposure activated diverse stress response pathways and profoundly impaired energy metabolismof gut bacteria. These findings provide new understandings of the functional interplay between the gut microbiome and environmental chemicals, such as organophosphates. [ABSTRACT FROM AUTHOR]

Background: α-Amylase plays a pivotal role in a broad range of industrial processes. To meet increasing demands of biocatalytic tasks, considerable efforts have been made to isolate enzymes produced by extremophiles. However, the relevant data of α-amylases from cold-adapted fungi are still insufficient. In addition, bread quality presents a particular interest due to its high consummation. Thus developing amylases to improve textural properties could combine health benefits with good sensory properties. Furthermore, iron oxide nanoparticles provide an economical and convenient method for separation of biomacromolecules. In order to maximize the catalytic efficiency of α-amylase and support further applications, a comprehensive characterization of magnetic immobilization of α-amylase is crucial and needed. Results: A novel α-amylase (AmyA1) containing an open reading frame of 1482 bp was cloned from Antarctic psychrotolerant fungus G. pannorum and then expressed in the newly constructed Aspergillus oryzae system. The purified recombinant AmyA1 was approximate 52 kDa. AmyA1 was optimally active at pH 5.0 and 40 °C, and retained over 20% of maximal activity at 0-20 °C. The Km and Vmax values toward soluble starch were 2.51 mg/mL and 8.24 × 10-2 mg/(mL min) respectively, with specific activity of 12.8 × 103 U/mg. AmyA1 presented broad substrate specificity, and the main hydrolysis products were glucose, maltose, and maltotetraose. The influence of AmyA1 on the quality of bread was further investigated. The application study shows a 26% increase in specific volume, 14.5% increase in cohesiveness and 14.1% decrease in gumminess in comparison with the control. AmyA1 was immobilized on magnetic nanoparticles and characterized. The immobilized enzyme showed improved thermostability and enhanced pH tolerance under neutral conditions. Also, magnetically immobilized AmyA1 can be easily recovered and reused for maximum utilization. Conclusions: A novel α-amylase (AmyA1) from Antarctic psychrotolerant fungus was cloned, heterologous expression in Aspergillus oryzae, and characterized. The detailed report of the enzymatic properties of AmyA1 gives new insights into fungal cold-adapted amylase. Application study showed potential value of AmyA1 in the food and starch fields. In addition, AmyA1 was immobilized on magnetic nanoparticles and characterized. The improved stability and longer service life of AmyA1 could potentially benefit industrial applications. [ABSTRACT FROM AUTHOR]

Background: Marine mud is an abundant and largely unexplored source of enzymes with unique properties that may be useful for industrial and biotechnological purposes. However, since most microbes cannot be cultured in the laboratory, a cultivation-independent metagenomic approach would be advantageous for the identification of novel enzymes. Therefore, with the objective of screening novel lipolytic enzymes, a metagenomic library was constructed using the total genomic DNA extracted from marine mud. Results: Based on functional heterologous expression, 34 clones that showed lipolytic activity were isolated. The five clones with the largest halos were identified, and the corresponding genes were successfully overexpressed in Escherichia coli. Molecular analysis revealed that these encoded proteins showed 48-79 % similarity with other proteins in the GenBank database. Multiple sequence alignment and phylogenetic tree analysis classified these five protein sequences as new members of known families of bacterial lipolytic enzymes. Among them, EST4, which has 316 amino acids with a predicted molecular weight of 33.8 kDa, was further studied in detail due to its strong hydrolytic activity. Characterization of EST4 indicated that it is an alkaline esterase that exhibits highest hydrolytic activity towards p-nitrophenyl butyrate (specific activity: 1389 U mg-1) at 45 °C and pH 8.0. The half-life of EST4 is 55 and 46 h at 40 and 45 °C, respectively, indicating a relatively high thermostability. EST4 also showed remarkable stability in organic solvents, retaining 90 % of its initial activity when incubated for 12 h in the presence of hydrophobic alkanes. Furthermore, EST4 was used as an efficient whole-cell biocatalyst for the synthesis of short-chain flavor esters, showing high conversion rate and good tolerance for high substrate concentrations (up to 3.0 M). These results demonstrate a promising potential for industrial scaling-up to produce short-chain flavor esters at high substrate concentrations in non-aqueous media. Conclusions: This manuscript reports unprecedented alcohol tolerance and conversion of an esterase biocatalyst identified from a marine mud metagenomic library. The high organic solvent tolerance and thermostability of EST4 suggest that it has great potential as a biocatalyst. [ABSTRACT FROM AUTHOR]

Background: Everolimus -eluting stent (EES) is common used in patients undergoing percutaneous coronary interventions (PCI). Our purpose is to evaluate long-term clinical outcomes of everolimus -eluting stent (EES) versus paclitaxel-eluting stent (PES) in patients undergoing percutaneous coronaryinterventions (PCI) in randomized controlled trials (RCTs).Methods: We searched Medline, EMBASE, Cochrane Library, CNKI, VIP and relevant websites ( https://scholar-google-com.ezproxy.lib.usf.edu/ ) for articles to compare outcomes between everolimus-eluting stent and paclitaxel-eluting stent without language or date restriction. RCTs that compared the use of everolimus -eluting stent and paclitaxel-eluting stent in PCI were included. Variables relating to patient, study characteristics, and clinical endpoints were extracted. Meta-analysis was performed using RevMan 5.2 software.Results: We identified 6 published studies (from three randomized trials) more on everolimus-eluting stent (n = 3352) than paclitaxel-eluting (n = 1639), with follow-up duration ranging from 3, 4 and 5 years. Three-year outcomes of everolimus-eluting stent compared to paclitaxel-eluting were as following: the everolimus-eluting stent significantly reduced all-cause death (relative risk [RR]:0.63; 95% confidence interval [CI]: 0.46. to 0.82), MACE (RR: 0.56; 95% CI: 0.41 to 0.77), MI (RR: 0.64; 95% CI: 0.48 to 0.86), TLR (RR: 0.72; 95% CI: 0.59 to 0.88), ID-TLR (RR: 0.74; 95% CI: 0.59 to 0.92) and ST (RR: 0.54; 95% CI: 0.32 to 0.90). There was no difference in TVR between the everolimus-eluting and paclitaxel-eluting (RR: 0.76; 95% CI: 0.58 to 1.10); Four-year outcomes of everolimus-eluting compared to paclitaxel-eluting: the everolimus-eluting significantly reduced MACE (RR: 0.44; 95% CI: 0.18 to 0.98) and ID-TLR (RR: 0.47; 95 % CI: 0.23 to 0.97). There was no difference in MI (RR: 0.48; 95% CI: 0.16 to 1.46), TLR (RR: 0.46; 95% CI: 0.20 to 1.04) and ST ((RR: 0.34; 95% CI: 0.05 to 2.39). Five-year outcomes of everolimus-eluting stent compared to paclitaxel-eluting: There was no difference in ID-TLR (RR: 0.67; 95% CI: 0.45 to 1.02) and ST (RR: 0.71; 95% CI: 0.28 to 1.80).Conclusions: In the present meta-analysis, everolimus-eluting appeared to be safe and clinically effective in patients undergoing PCI in comparison to PES in 3-year clinical outcomes; there was similar no difference in reduction of ST between EES and PES in long-term(≥ 4 years) clinical follow-ups. Everolimus-eluting is more safety than paclitaxel-eluting in long-term clinical follow-ups, whether these effects can be applied to different patient subgroups warrants further investigation. [ABSTRACT FROM AUTHOR]

A DNA segment encoding a novel a-amylase was cloned from the Antarctic psychrotolerant fungi Geomyces pannorum. The gene has an open reading frame of 1761 bp, deduced 586 amino acids, revealing a relatively low homology to a-amylases fromdiverse fungal genera. The a-amylase gene was overexpressed in Pichia pastoris, and the recombinanta-amylasewas purified and characterized. Enzymatic activity was optimal at pH 6.0 and 70°C, and the specific activity was 9.78⨰103 U/mg when using soluble starch as substrate. Moreover, the enzyme retained more than 90% of its maximum activity over a wide pH range (6.0-9.0). Exceptionally, the main hydrolysis products of soluble starch catalyzed by the enzyme were glucose and maltose, of which glucose reached a concentration of 68.8% (w/w). This is the first report on identifying thermophilic fungal amylase from psychrotolerant fungi. Also, the preliminary study on the high-glucose-producing a-amylase with detailed enzymatic properties shows a potential application prospect in syrup industry. [ABSTRACT FROM AUTHOR]

Background: The desiccation-tolerant moss Bryum argenteum is an important component of the Biological Soil Crusts (BSCs) found in the Gurbantunggut desert. Desiccation tolerance is defined as the ability to revive from the air dried state. To elucidate the molecular mechanisms related to desiccation tolerance, we employed RNA-Seq and digital gene expression (DGE) technologies to study the genome-wide expression profiles of the dehydration and rehydration processes in this important desert plant. Results: We applied a two-step approach to investigate the gene expression profile upon rehydration in the moss Bryum argenteum using Illumina HiSeq2000 sequencing platform. First, a total of 57,247 transcript assembly contigs (TACs) were obtained from 54.79 million reads by de novo assembly, with an average length of 863 bp and N50 of 1,372 bp. Among the reconstructed TACs, 36,916 (64.5%) revealed similarity with existing protein sequences in the public databases. 23,509 and 21,607 TACs were assigned GO and KEGG annotation information, respectively. Second, samples were taken from 3 hydration stages: desiccated (Dry), rehydrated 2 h (R2) and rehydrated 24 h (R24), and DEG libraries were constructed for Differentially Expressed Genes (DEGs) discovery. 4,081 and 6,709 DEGs were identified in R2 and R24, compared with Dry, respectively. Compared to the desiccated sample, up-regulated genes after two hours of hydration are primarily related to stress responses. GO function enrichment network, EKGG metabolic pathway and MapMan analysis supports the idea of the rapid recovery of photosynthesis after 24 h of rehydration. We identified 770 transcription factors (TFs) which were classified into 50 TF families. 142 TF transcripts were up-regulated upon rehydration including 23 members of the ERF family. Conclusions: In this study, we constructed a pioneering, high-quality reference transcriptome in B. argenteum and generated three DGE libraries to elucidate the changes of gene expression upon rehydration. Expression profiles consistent with the rapid recovery of photosynthesis (at R2) and the re-establishment of a positive carbon balance following rehydration (at R24) were observed. Our study will extend our knowledge of bryophyte transcriptomes and provide further insight into the molecular mechanisms related to rehydration and desiccation-tolerance. [ABSTRACT FROM AUTHOR]

Syntrichia caninervis is the dominant bryophyte of the biological soil crusts found in the Gurbantunggut desert. The extreme desert environment is characterized by prolonged drought, temperature extremes, high radiation and frequent cycles of hydration and dehydration. S. caninervis is an ideal organism for the identification and characterization of genes related to abiotic stress tolerance. Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) expression analysis is a powerful analytical technique that requires the use of stable reference genes. Using available S. caninervis transcriptome data, we selected 15 candidate reference genes and analyzed their relative expression stabilities in S. caninervis gametophores exposed to a range of abiotic stresses or a hydration-desiccation-rehydration cycle. The programs geNorm, NormFinder, and RefFinder were used to assess and rank the expression stability of the 15 candidate genes. The stability ranking results of reference genes under each specific experimental condition showed high consistency using different algorithms. For abiotic stress treatments, the combination of two genes (α-TUB2 and CDPK) were sufficient for accurate normalization. For the hydration-desiccation-rehydration process, the combination of two genes (α-TUB1 and CDPK) were sufficient for accurate normalization. 18S was among the least stable genes in all of the experimental sets and was unsuitable as reference gene in S. caninervis. This is the first systematic investigation and comparison of reference gene selection for RT-qPCR work in S. caninervis. This research will facilitate gene expression studies in S. caninervis, related moss species from the Syntrichia complex and other mosses. [ABSTRACT FROM AUTHOR]

Background Fungal amylase, mainly constitute of fungal a-amylase and fungal glucoamylase, are utilized in a broad range of industries, such as starch hydrolysis, food and brewing. Although various a-amylase and glucoamylase have been found in various fungi, the amylases from Aspergillus dominate the commercial application. One of the main problems exist with regard to these commercial use of amylases is the relatively low thermal and acid stability. In order to maximize the efficiency of starch processing operations, the operating range of glucoamylase and α-amylase, in terms of pH, temperature and compatibility with other enzymes should be improved. So developing fungal amylases with increased thermostability and acid stability which could be used in commercial starch process has been attracting researchers? interest continually. Besides, synergetic action of glucoamylase and a-amylase could facilitate the degradation of starch. And co-expressing glucoamylase with a-amylase in one host could avoid the need to ferment repeatedly and improves cost-effectiveness of the process. Results A novel fungal glucoamylase (RpGla) gene encoding a putative protein of 512 amino acid residues was cloned from Rhizomucor pusillus. BLAST analysis revealed that RpGla shared the highest identity of 51% with the Rhizopus oryzae glucoamylase (ABB77799.1). The fungal glucoamylase RpGla was expressed in Pichia pastoris (resulting in KM71/9KGla) with maximum activity of 1237 U ml-1. The optimum pH and temperature of RpGla were pH 4.0 and 70C, respectively. Fungal a-amylase (RpAmy) gene was also cloned from R. pusillus and transformed into KM71/9KGla, resulted in recombinant yeast KM71/9KGla-ZaAmy, which harboring the RpGla and RpAmy genes simultaneously. The maximum saccharogenic activity of KM71/9KGla-ZaAmy was 2218 U mml-1, which improved 79% compared to KM71/9KGla. Soluble starch hydrolyzed by purified RpGla achieved 43% glucose and 34% maltose. Higher productivity was achieved with a final yield of 48% glucose and 47% maltose catalyzed by purified enzyme preparation produced by KM71/9KGla-ZαAmy. Conclusions A novel fungal glucoamylase and fungal a-amylase genes were cloned from Rhizomucor pusillus. The two enzymes showed good thermostability and acid stability, and the similar biochemical properties facilitated synergetic action of the two enzymes. A dramatic improvement was seen in amylase activity through co-expressing RpGla with RpAmy in Pichia pastoris. This is the first report of improving activity through co-expression glucoamylase with a-amylase in P. pastoris. Besides, fungal glucoamylase and a-amylase from R. pusillus were shown as promising candidates for further application in starch hydrolysis. [ABSTRACT FROM AUTHOR]

Background: Syntrichia caninervis is a desiccation-tolerant moss and the dominant bryophyte of the Biological Soil Crusts (BSCs) found in the Mojave and Gurbantunggut deserts. Next generation high throughput sequencing technologies offer an efficient and economic choice for characterizing non-model organism transcriptomes with little or no prior molecular information available. Results: In this study, we employed next generation, high-throughput, Illumina RNA-Seq to analyze the poly-(A) + mRNA from hydrated, dehydrating and desiccated S. caninervis gametophores. Approximately 58.0 million paired-end short reads were obtained and 92,240 unigenes were assembled with an average size of 493 bp, N50 value of 662 bp and a total size of 45.48 Mbp. Sequence similarity searches against five public databases (NR, Swiss-Prot, COSMOSS, KEGG and COG) found 54,125 unigenes (58.7%) with significant similarity to an existing sequence (E-value ≤ 1e-5) and could be annotated. Gene Ontology (GO) annotation assigned 24,183 unigenes to the three GO terms: Biological Process, Cellular Component or Molecular Function. GO comparison between P. patens and S. caninervis demonstrated similar sequence enrichment across all three GO categories. 29,370 deduced polypeptide sequences were assigned Pfam domain information and categorized into 4,212 Pfam domains/families. Using the PlantTFDB, 778 unigenes were predicted to be involved in the regulation of transcription and were classified into 49 transcription factor families. Annotated unigenes were mapped to the KEGG pathways and further annotated using MapMan. Comparative genomics revealed that 44% of protein families are shared in common by S. caninervis, P. patens and Arabidopsis thaliana and that 80% are shared by both moss species. Conclusions: This study is one of the first comprehensive transcriptome analyses of the moss S. caninervis. Our data extends our knowledge of bryophyte transcriptomes, provides an insight to plants adapted to the arid regions of central Asia, and continues the development of S. caninervis as a model for understanding the molecular aspects of desiccation-tolerance. [ABSTRACT FROM AUTHOR]

New recycling technologies have been developed lately to enhance the value of the fiberglass powder-resin powder fraction (FRP) from waste printed circuit boards. The definite aim of the present paper is to present some novel methods that use the image forces for the separation of the resin powder and fiberglass powder generated from FRP during the corona electrostatic separating process. The particle shape charactization and particle trajectory simulation were performed on samples of mixed non-metallic particles. The simulation results pointed out that particles of resin powder and particles of fiberglass powder had different detach trajectories at the conditions of the same size and certain device parameters. An experiment carried out using a corona electrostatic separator validated the possibility of sorting these particles based on the differences in their shape characteristics. The differences in the physical properties of the different types of particles provided the technical basis for the development of electrostatic separation technologies for the recycling industry. [ABSTRACT FROM AUTHOR]

Serine protease inhibitors, or serpins, are a group of widely distributed proteins with similar structures that use conformational change to inhibit proteases. Antithrombin (AT) is a member of the serine protease inhibitor superfamily and a major coagulation inhibitor in all vertebrates, but its evolutionary origin remains elusive. In this study we isolated for the first time a cDNA encoding an antithrombin homolog, BjATl, from the protochordate Branchiostoma japonicum. The deduced protein BjATl consisted of 338 amino acids sharing 36.7% to 41.1% identity to known vertebrate ATs. BjATl contains a potential N-linked glycosylation site, two potential heparin binding sites and the reactive center loop with the absolutely conserved sequence Gly-Arg-Ser; all of these are features characteristic of ATs. All three phylogenetic trees constructed using Neighbor-Joining, Maximum-Likelihood and Bayesian-Inference methods also placed BjATl together with ATs. Moreover, BjATl expressed in yeast cells was able to inhibit bovine thrombin activity by forming a SDS-stable BjATlthrombin complex. It also displays a concentration-dependent inhibition of thrombin that is accelerated by heparin. Furthermore, BjATl was predominantly expressed in the hepatic caecum and hind-gut, agreeing with the expression pattern of AT in mammalian species. All these data clearly demonstrate that BjATl is an ortholog of vertebrate ATs, suggesting that a primitive coagulation system emerged in the protochordate. [ABSTRACT FROM AUTHOR]

Background: Bacterial plant pathogens are very harmful to their host plants, which can cause devastating agricultural losses in the world. With the development of microbial genome sequencing, many strains of phytopathogens have been sequenced. However, some misannotations exist in these phytopathogen genomes. Our objective is to improve these annotations and store them in a central database DIGAP. Description: DIGAP includes the following improved information on phytopathogen genomes. (i) All the 'hypothetical proteins' were checked, and non-coding ORFs recognized by the Z curve method were removed. (ii) The translation initiation sites (TISs) of 20% ~ 25% of all the protein-coding genes have been corrected based on the NCBI RefSeq, ProTISA database and an ab initio program, GS-Finder. (iii) Potential functions of about 10% 'hypothetical proteins' have been predicted using sequence alignment tools. (iv) Two theoretical gene expression indices, the codon adaptation index (CAI) and the E(g) index, were calculated to predict the gene expression levels. (v) Potential agricultural bactericide targets and their homology-modeled 3D structures are provided in the database, which is of significance for agricultural antibiotic discovery. Conclusion: The results in DIGAP provide useful information for understanding the pathogenetic mechanisms of phytopathogens and for finding agricultural bactericides. DIGAP is freely available at http://ibi.hzau.edu.cn/digap/. [ABSTRACT FROM AUTHOR]

Headspace solid-phase microextraction has been applied to the analysis of volatile components in longan ( Dimocarpus longan Lour.). Silica fiber coated with divinylbenzene–carboxen–polydimethylsiloxane (DVB–CAR–PDMS) was found to be more efficient for extracting volatile compounds than other fibers such as those coated with polydimethylsiloxane, polyacrylate, and carboxen–polydimethylsiloxane. The SPME headspace volatiles were collected using a DVB–CAR–PDMS fiber. The extraction conditions were optimized using a response surface experimental design to analyze the effect of three factors: extraction temperature, equilibrium time and extraction time. The best response was obtained when the extraction temperature was around 50 °C, equilibrium time near 15 min and extraction time close to 25 min. The volatile extracts were analyzed using gas chromatograph/mass spectrometry system. Sixty-two different volatile compounds from the headspace of longan were identified. The linearity was good in the considered concentration ranges ( R2 ≥ 0.965). Average recoveries ranged from 78.6 to 108.3% and showed good accuracy. The precision of the present method was found to be good (RSD < 13.59%). Therefore, this method can be applied to determine the quality of the longan or other volatile components as an alternative to full-scale headspace analysis. [ABSTRACT FROM AUTHOR]

Background: For industrial bioconversion processes, the utilization of surface-displayed lipase in the form of whole-cell biocatalysts is more advantageous, because the enzymes are displayed on the cell surface spontaneously, regarded as immobilized enzymes. Results: Two Pichia pastoris cell surface display vectors based on the flocculation functional domain of FLO with its own secretion signal sequence or the α-factor secretion signal sequence were constructed respectively. The lipase gene lipB52 fused with the FLO gene was successfully transformed into Pichia pastoris KM71. The lipase LipB52 was expressed under the control of the AOX1 promoter and displayed on Pichia pastoris KM71 cell surface with the two Pichia pastoris cell surface display vectors. Localization of the displayed LipB52 on the cell surface was confirmed by the confocal laser scanning microscopy (CLSM). The LipB52 displayed on the Pichia pastoris cell surface exhibited activity toward p-nitrophenol ester with carbon chain length ranging from C10 to C18, and the optimum substrate was p-nitrophenol-caprate (C10), which was consistent with it displayed on the Saccharomyces cerevisiae EBY100 cell surface. The hydrolysis activity of lipase LipB52 displayed on Pichia pastoris KM71-pLHJ047 and KM71-pLHJ048 cell surface reached 94 and 91 U/g dry cell, respectively. The optimum temperature of the displayed lipases was 40°C at pH8.0, they retained over 90% activity after incubation at 60°C for 2 hours at pH 7.0, and still retained 85% activity after incubation for 3 hours. Conclusion: The LipB52 displayed on the Pichia pastoris cell surface exhibited better stability than the lipase LipB52 displayed on Saccharomyces cerevisiae cell surface. The displayed lipases exhibited similar transesterification activity. But the Pichia pastoris dry cell weight per liter (DCW/L) ferment culture was about 5 times than Saccharomyces cerevisiae, the lipase displayed on Pichia pastoris are more suitable for whole-cell biocatalysts than that displayed on Saccharomyces cerevisiae cell surface. [ABSTRACT FROM AUTHOR]

Background: To explore the correlation between serum uric acid (SUA) and prognosis in patients with chronic heart failure (CHF) after revascularization. Methods: A total of 126 patients with CHF undergoing revascularization [coronary artery intervention (PCI) or coronary artery bypass grafting (CABG)J in the hospital were enrolled as CHF group between December 2021 and October 2022, while 126 healthy controls during the same period were enrolled as healthy control group. The levels of SUA, inflammatory factors and cardiac function in the two groups were detected. The correlation between SUA level and inflammatory factors, cardiac function levels was analyzed. All patients in CHF group were followed up for 6 months to observe prognosis. The differences in the above indexes among patients with different prognosis were compared. The risk factors of prognosis were analyzed by multivariate Logistic regression analysis, and their predictive value for prognosis was evaluated by ROC curves analysis. Results: SUA, IL-6, TNF-a, NT-proBNP and LVEDD in CHF group were higher than those in healthy control group, while LVEF was lower than that in healthy control group (P<0.05). Among the 126 patients within 6 months after surgery, there were 24 cases (19.05%) with adverse cardiac events (poor prognosis). SUA, IL-6, TNF-a, NT-proBNP and LVEDD in patients with poor prognosis were higher than those with good prognosis, while LVEF was lower than that with good prognosis (P<0.05). SUA level was positively correlated with IL-6, TNF-a, NT-proBNP and LVEDD (r=0.283, 0.292, 0.322, 0.355, P<0.05), while negatively correlated with LVEF (r=-0.368, P<0.05). High level of SUA and low LVEF were independent risk factors of prognosis (OR=1,486, 0.678, P<0.05). Both SUA and LVEF were of prognostic value (AUC=0.805, 0.809, P<0.05). Conclusion: SUA level is significantly increased in CHF patients, which is an independent risk factor of poor prognosis in CHF patients after revascularization, with high prognostic value. [ABSTRACT FROM AUTHOR]

Platelets from patients with myeloproliferative neoplasms (MPNs) exhibit a hyperreactive phenotype. Here, we found elevated P-selectin exposure and platelet-leukocyte aggregates indicating activation of platelets from essential thrombocythemia (ET) patients. Single-cell RNA-seq analysis of primary samples revealed significant enrichment of transcripts related to platelet activation, mTOR, and oxidative phosphorylation in ET patient platelets. These observations were validated via proteomic profiling. Platelet metabolomics revealed distinct metabolic phenotypes consisting of elevated ATP generation accompanied by increases in the levels of multiple intermediates of the tricarboxylic acid cycle, but lower a-ketoglutarate (a-KG) in MPN patients. Inhibition of PI3K/AKT/mTOR signaling significantly reduced metabolic responses and hyperreactivity in MPN patient platelets, while a-KG supplementation markedly reduced oxygen consumption and ATP generation. Ex vivo incubation of platelets from both MPN patients and Jak2 V617F-knockin mice with a-KG supplementation significantly reduced platelet activation responses. Oral a-KG supplementation of Jak2 V617F mice decreased splenomegaly and reduced hematocrit, monocyte, and platelet counts. Finally, a-KG treatment significantly decreased proinflammatory cytokine secretion from MPN CD14+ monocytes. Our results reveal a previously unrecognized metabolic disorder in conjunction with aberrant PI3K/AKT/mTOR signaling that contributes to platelet hyperreactivity in MPN patients. [ABSTRACT FROM AUTHOR]

In order to investigate the effect of ultrasound-assisted fermentation on its flavor and quality, mackerel enzymatic hydrolysate was, fermented with Saccharomyces longosporus to enhance the flavor, and flavor and quality changes during different fermentation stages were monitored by measurement of total bacterial count and amino acid nitrogent content, sensory evaluation, and principal component analysis (PCA) of electronic nose data. Meanwhile, nutritional evaluation of the fermented product was carried out by analysis of free and essential amino acid composition, and the composition of flavor compounds was analyzed by gas chromatography-ion mobility spectrometry (GC-IMS). It was found that PCA effectively distinguished the flavor differences between the ultrasound and control groups during the fermentation process. The total bacterial count, amino acid nitrogen content and sensory score of the ultrasound group were higher than those of the control group during the whole fermentation process. On day 12 of fermentation, the ultrasound group had the best flavor quality with a total bacterial count of 8.90 (lg(CFU/mL)) and an amino acid nitrogen content of 0.78 g/100 g, and the fermented product had a rich umami taste and a fruity aroma. Moreover, the content of total free amino acids in the fermented product was 6.373 g/100 g, and the essential amino acid index (EAAI) value was 82.90, indicating a high nutritional value. GC-IMS results showed that the contents of off-flavor compounds such as, saturated straight-chain aldehydes gradually decreased, or even disappeared after 12 days of fermentation. In contrast, the contents of unsaturated alcohols and aroma-active substances (ketones and esters) significantly increased in the ultrasonic group compared with the control group. Thus, ultrasound-assisted treatment could improve the flavor quality of fermented mackerel enzymatic hydrolysate. This study may provide a theoretical basis for the application of ultrasonic technology in the development of seafood seasoning.

A novel Lycopodium alkaloid, lycocasine A (1), and seven known Lycopodium alkaloids (2–8), were isolated from Lycopodiastrum casuarinoides. Their structures were determined through NMR, HRESIMS, and X-ray diffraction analysis. Compound 1 features an unprecedented 5/6/6 tricyclic skeleton, highlighted by a 5-aza-tricyclic[6,3,1,02,6]dodecane motif. In bioactivity assays, compound 1 demonstrated weak inhibitory activity against acid-sensing ion channel 1a.

In this paper, a novel polysulfone (PSF)-Fe3O4 composite ultrafiltration membrane was prepared and employed to purify the flavonoids from the ginkgo biloba extraction (GBE). The mass susceptibility (w) of the ginkgo flavonoids with 98% purity was tested to be greater than 0, so they showed paramagnetism. Hysteresis curve indicated that the composite membrane have superparamagnetism for the addition of nano-sized Fe3O4 particles. The process of ultrafiltration was performed without and in the magnetic field with different intensities, respectively. The flavonoids content in the products is much superior when performed in the magnetic field than that without magnetic field. In the absence of magnetic field, the flavonoids content in the products was purified to 30.16% from 15.60% in crude GBE. Once the magnetic field with 0.2 T was added in the process, the purity was enhanced to 35.24%. Furthermore, the purity was persistently increasing with the incremental magnetic intensity. When the magnetic intensity was strengthened to 1.0 T, the purity approaches saturation, with being up to 52.87%. [ABSTRACT FROM AUTHOR]

ObjectivesOver the years, it has been found that colchicine offers substantial benefits in secondary prevention in patients with coronary artery disease (CAD). We studied the effects of colchicine timing because there are no guidelines about when to provide it during the perioperative period for patients with CAD.MethodsUp to January 1, 2023, seven electronic literature databases were screened (including three English databases and four Chinese databases). Randomized controlled trials included only treatment with colchicine in the perioperative period of CAD. The Cochrane Evaluation Tool was used to judge the risk of bias in research. Statistical analysis was performed by Stata 16.0 software.ResultsWe evaluated twelve studies that found colchicine to be effective in decreasing the occurrence of major adverse cardiac events (MACEs) (p

Background Primary thyroid lymphoma (PTL) is a rare type of extranodal lymphoma, which is prone to misdiagnosis and missed diagnosis due to atypical clinical and ultrasonic features. Objective To summarize and analyze the features of 2-D ultrasonography and contrast-enhanced ultrasonography in PTL. Methods A retrospective, comparative analysis was conducted on ultrasonographic features of 16 patients with pathologically confirmed PTL (PTL group) and 16 thyroid cancer patients (thyroid cancer group) recruited from Tangshan Gongren Hospital during December 2012 to December 2020. Results Pathological results of PTL group: 10 cases were diffuse large B-cell lymphoma, 4 cases were mucous-associated lymphoid tissue extranodular marginal zone B-cell lymphoma, 1 case was follicular lymphoma, and 1 case was Burkitt lymphoma. Pathological results of thyroid cancer group: 11 cases were papillary carcinoma, 4 cases were follicular carcinoma and 1 case was medullary carcinoma. The rate of thyroid enlargement in PTL group was higher than that in thyroid cancer group (13/16 vs 5/16) (χ2=8.127, P0.05) . PTL and thyroid cancer groups also had no statistically significant difference in the rate of cervical lymph node metastases (8/16 vs 11/16) (χ2=1.166, P>0.05) . Contrast-enhanced ultrasonography showed high enhancement during arterial and venous phases in PTL, and low enhancement at arterial and venous phases in thyroid cancer. Conclusion PTL has specific ultrasonic manifestations. A diagnosis of PTL could be made when a patient manifests as thyroid enlargement, diffuse hypoechoic, non-calcified, regular-shaped lesions with well-defined margins and posterior echo enhancement, and high enhancement at arterial and venous phases.

Genomic integration of genes and pathway-sized DNA cassettes is often an indispensable way to construct robust and productive microbial cell factories. For some uncommon microbial hosts, such as Mycolicibacterium and Mycobacterium species, however, it is a challenge. Here, we present a multiplexed integrase-assisted site-specific recombination (miSSR) method to precisely and iteratively integrate genes/pathways with controllable copies in the chromosomes of Mycolicibacteria for the purpose of developing cell factories. First, a single-step multi-copy integration method was established in M. neoaurum by a combination application of mycobacteriophage L5 integrase and two-step allelic exchange strategy, the efficiencies of which were ~100% for no more than threecopy integration events and decreased sharply to ~20% for five-copy integration events. Second, the R4, Bxb1 and ΦC31 bacteriophage Att/Int systems were selected to extend the available integration toolbox for multiplexed gene integration events. Third, a reconstructed mycolicibacterial Xer recombinases (Xer-cise) system was employed to recycle the selection marker of gene recombination to facilitate the iterative gene manipulation. As a proof of concept, the biosynthetic pathway of ergothioneine (EGT) in Mycolicibacterium neoaurum ATCC 25795 was achieved by remodeling its metabolic pathway with a miSSR system. With six copies of the biosynthetic gene clusters (BGCs) of EGT and pentose phosphate isomerase (PRT), the titer of EGT in the resulting strain in a 30 mL shake flask within 5 days was enhanced to 66 mg/L, which was 3.77 times of that in the wild strain. The improvements indicated that the miSSR system was an effective, flexible, and convenient tool to engineer the genomes of Mycolicibacteria as well as other strains in the Mycobacteriaceae due to their proximate evolutionary relationships. [ABSTRACT FROM AUTHOR]

Hydroxychloroquine (HCQ) is derivative of the heterocyclic aromatic compound quinoline, which has been used for the treatment of autoimmune diseases. The central purpose of this study was to investigate therapeutic effects and inflammatory immunological molecular mechanism of HCQ in experimental autoimmune hepatitis (AIH). Treatment with HCQ ameliorated hepatic pathologic damage, inflammatory infiltration, while promoted regulatory T cell (Treg) and down-regulated CD8+T cell differentiation in AIH mice induced by S-100 antigen. In vitro, HCQ also suppressed pro-inflammatory cytokine (IFN-γ, TNF-α, and IL-12) secretion, promoted anti-inflammatory cytokine (TGF-β1) secretion. HCQ mainly impaired T cell lipid metabolism but not glycolysis to promote Treg differentiation and function. Mechanistically, HCQ down-regulated GRK2 membrane translocation in T cells, inhibited GRK2-PI3K interaction to reduce the PI3K recruiting to the membrane, followed by suppressing the phosphorylation of PI3K-AKT-mTOR signal. Pretreating T cells with paroxetine, a GRK2 inhibitor, disturbed HCQ effect to T cells. HCQ also reversed the activation of the PI3K-AKT axis by 740 Y-P (PI3K agonist). Meanwhile, HCQ inhibited the PI3K-AKT-mTOR, JAK2-STAT3-SOCS3 and increased the AMPK signals in the liver and T cells of AIH mice. In conclusion, HCQ exhibited specific and potent therapeutic effects on AIH and attendant liver injury, which was attributed to HCQ acted on GRK2 translocation, inhibited metabolism-related PI3K-AKT and inflammation-related JAK2-STAT3 signal in T lymphocytes, thereby modulating lipid metabolism of T cell function to regulate Treg differentiation and function.

Pregna-1,4,9(11),16(17)-tetraene-3,20-dione (pregnatetraenedione) is the most useful precursor for the synthesis of corticosteroids in industry. However, the production of pregnatetraenedione is still a complex and expensive process. Here, we proposed a greener and more economical route for the synthesis of pregnatetraenedione, which was achieved via the conversion of phytosterols to an uncommon steroid compound, 9a-hydroxy-3-oxo-4,17(20)-pregna-diene-20-carboxylic acid (9-OHPDC), by an engineered Mycolicibacterium neoaurum. 9-OHPDC is a metabolite of sterols obtained by partially degrading its C17 side-chain and hitherto it is difficult to exclusively produce due to its complex metabolic process. In this study, we systematically investigated the determinants related to the generation of 9-OHPDC in the metabolic process of phytosterols and developed a 9-OHPDC-producing strain with the methyl-esterified 9-OHPDC (9-OHPDC-M) as the main product by metabolic engineering. The production of 9-OHPDC and 9-OHPDC-M respectively reached 9.86 g L-1 and 1.27 g L-1 in about 6 days and the total mole yield was more than 85.9%. Using 9-OHPDC-M as a synthon, subsequently, a four-step chemical process for the synthesis of pregnatetraenedione was designed and verified, which was composed of the following steps: hydrolysis of 9-OHPDC-M to 9-OHPDC, decarboxylation rearrangement, dehydration and A1 dehydrogenation. From phytosterols to pregnatetraenedione, the combined microbial and chemical process showed a total weight yield of up to 35.4%, significantly higher than that of the current industrial routes. In addition, compared with the current routes, the new synthesis route has the advantages of shorter reaction steps, milder reaction conditions, reduced use of organic solvents and abandonment of toxic catalysts. In a word, this study provided a green route for the production of pregnatetraenedione, which can be used for corticosteroid production in industry. [ABSTRACT FROM AUTHOR]