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PurposeCXC chemokines are mediators which mediate immune cells migration to sites of inflammation and injury. Chemokine C-X-C motif ligand 16 (CXCL16) plays an important role in the occurrence and development of sepsis through leukocyte chemotaxis, leukocyte adhesion and endotoxin clearance. In this study, we selected a set of tagging single nucleotide polymorphisms (tag SNPs) in the CXCL16 gene and investigated their clinical relevance to the development of sepsis and multiple organ dysfunction syndrome (MODS) in patients with major trauma in three independent Chinese Han populations.MethodsA total of 1,620 major trauma patients were enrolled in this study. Among these patients, 920 came from Chongqing in western China, 350 came from Zhejiang Province in eastern China, and 350 came from Guizhou Province in southwestern China. The improved multiplex ligation detection reaction (iMLDR) method was employed in the genotyping and genetic association analyses to determine the associations between CXCL16 haplotypes and sepsis morbidity rate and higher MOD scores in three cohorts.ResultsOnly CXCL16 T123V181 haplotype was associated with an increased risk for sepsis morbidity and higher MOD scores in the three cohorts (OR = 1.89,P= 0.001 for the Chongqing cohort; OR = 1.76,P= 0.004 for the Zhejiang cohort; OR = 1.55,P= 0.012 for the Guizhou cohort). The effect of T123V181 haplotype on the chemotaxis, migration and endotoxin clearance of immune cells were further observed. Protein modeling analysis showed that T123 and V181 might alter the structure of the CXCL16 active center. Thus it enhanced the chemotaxis and adhesion ability of immunocytes.ConclusionWe demonstrate the mechanism of CXCL16 T123V181 haplotype which regulates the sepsis morbidity rate and thus provide a new biomarker for early diagnosis of sepsis and MODS.Clinical Trial Registrationwww.ClinicalTrials.gov, identifier NCT01713205 (https://www.clinicaltrials.gov/ct2/results?cond=&term=+NCT01713205&cntry=&state=&city=&dist=).

miR-608 has been indicated to play an important role in the pathogenesis of various inflammation-related diseases, including sepsis and several types of cancers. However, there is little information about the underlying mechanism, especially in inflammatory cells. In this study, an hsa-miR-608-inhibition cell model was constructed in U937 cells using a lentivirus, and gene expression profiles were determined by a cDNA microarray. Altogether, 682 genes showed a difference greater than 1.2-fold, including 184 genes downregulated and 498 genes upregulated. Among these genes, one potential miR-608-target gene, ELANE, was further investigated. A positive relationship between the expression of miR-608 and that of ELANE was found both in vivo and in vitro. In addition, decreased expression of miR-608 resulted in overexpression of ELANE at both the mRNA and protein levels. Cotransfection of HEK293T cells with a miR-608 mimic inhibited reporter activity, and mutation of the miRNA seed sequences abolished the repression of reporter activity. These results suggest that miR-608 is an important posttranscriptional regulator of ELANE expression in human monocytes and may play an important role in the process of inflammation. miR-608 and neutrophil elastase may be novel targets for the diagnosis or treatment of sepsis.

Background Genetic backgrounds have been recognized as significant determinants of susceptibility to sepsis. CXC chemokines play a significant role in innate immunity against infectious diseases. Genetic polymorphisms of CXC chemokine genes have been widely studied in inflammatory and infectious diseases but not in sepsis. Thus, we aimed to investigate the clinical relevance of CXC chemokine gene polymorphisms and susceptibility to sepsis in a traumatically injured population. Methods Thirteen tag single nucleotide polymorphisms were selected from CXC chemokine genes using a multimarker tagging algorithm in the Tagger software. Three independent cohorts of injured patients (n = 1700) were prospectively recruited. Selected single nucleotide polymorphisms were genotyped using an improved multiplex ligation detection reaction method. Cytokine production in lipopolysaccharide-stimulated whole blood was measured using an enzyme-linked immunosorbent assay. Results Among the 13 tag single nucleotide polymorphisms, four single nucleotide polymorphisms (rs1429638, rs266087, rs2297630, and rs2839693) were significantly associated with the susceptibility to sepsis, and three (rs3117604, rs1429638, and rs4074) were significantly associated with an increased multiple organ dysfunction score in the derivation cohort. However, only the clinical relevance of rs1429638 and rs266087 was confirmed in the validation cohorts. In addition, rs2297630 was significantly associated with interleukin 6 production. Conclusion The rs1429638 polymorphism in the CXCL1 gene and the rs2297630 polymorphism in the CXCL12 gene were associated with altered susceptibility to sepsis and might be used as important genetic markers to assess the risks of sepsis in trauma patients. Level of evidence Prognostic and epidemiologic study, level II.

Tremella fuciformis is an important edible mushroom that has been widely cultivated and used as food and medicinal ingredient in traditional Chinese medicine. In the past decades, many researchers have reported that T. fuciformis polysaccharides (TPS) possess various bioactivities, including anti-tumor, immunomodulatory, anti-oxidation, anti-aging, repairing brain memory impairment, anti-inflammatory, hypoglycemic and hypocholesterolemic. The structural characteristic of TPS has also been extensively investigated using advanced modern analytical technologies such as NMR, GC-MS, LC-MS and FT-IR to dissect the structure-activity relationship (SAR) of the TPS biomacromolecule. This article reviews the recent progress in the extraction, purification, structural characterization and applications of TPS.

Sepsis arises from an aberrant and excessive host response to infection. Long noncoding RNAs (lncRNAs) are involved in multiple cellular functions, including inflammation and immunity. However, to date there has been no systematic attempt to identify lncRNAs whose expression is changed after the induction of the innate immune response. In this study, we profiled global lncRNA and mRNA expression changes in peripheral blood mononuclear cells (PBMCs) treated with lipopolysaccharide (LPS) using a microarray platform. Of the 40 914 lncRNAs screened, 596 were significantly upregulated and 250 were significantly downregulated (corrected P

Polymeric quaternary ammonium salts (PQASs) exhibit antibacterial action and are less toxic, less stimulatory to the human body and have easier-to-modify functionalities than small molecular antibacterial agents. However, few studies on the structure-activity relationship and toxicity mechanism of PQASs against fungi have been reported. We previously described the synthesis of a novel PQAS, namely, a homopolymer of (2-methacrylamido)propyltetrabenzyldimethylammonium chloride (PQD-BC), and discovered that the polymer exhibits antifungal activities not only against Fusarium oxysporum f. sp. Cubense tropical race 4 (Foc 4), the pathogen of banana wilt, but also against Rhizoctonia solani (R. solani), the pathogen of rice sheath blight (RShB). Furthermore, we studied the mechanism of action of PQD-BC against Foc 4, which is markedly different from R. solani in morphology and life cycle. Therefore, the structure-antifungal activity relationship and toxicity mechanism of PQD-BC against R. solani were extensively studied in this work and compared with those of the low-molecular-weight quaternary ammonium salt benzyldimethyldodecylammonium chloride (BC), and the results play an important role in identifying long-term and low-toxicity fungicides that can suppress the sclerotia of R. solani. The results showed that PQD-BC and BC can destroy the structural integrity and morphology of a cell, such as by loss of the cell wall and plasma membrane integrity, leading to the release of intracellular contents and can induce mitochondrial dysfunction and interference with genomic DNA and inhibit the formation of sclerotia. However, PQD-BC showed a special mechanism for causing the lipid peroxidation of the cell membrane; this mechanism was not observed with BC. The newly elucidated mechanism accounts for differences between polymers and small-molecule compounds and provides a theoretical basis for further application of PQAS against fungi and sclerotia.

A series of acrylate polymers containing quaternary ammonium salts (PDMAEMA-BC) having tunable molecular weights were synthesized via atom transfer radical polymerization (ATRP), and the effect of the degree of polymerization (DP) on the antimicrobial activity against bacteria ( Escherichia coli , Staphylococcus albus ), pathogenic fungi ( Candida albicans ) and phytopathogenic fungi ( Rhizoctonia solani and Fusarium oxysporum f. sp. cubense race 4) was systematically assessed. The antimicrobial properties against E. coli , S. albus and C. albicans were characterized using the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) or minimum bactericidal concentration (MBC) values, whereas the antimicrobial activities against R. solani and Foc4 were evaluated using the effective concentration (EC 50 and EC 90 ), MIC and MFC values. The results indicated that the PDMAEMA-BC homopolymers showed better antimicrobial activities than the corresponding monomer, i.e., the acrylate quaternary ammonium salt monomer (DMAEMA-BC), and the optimal antimicrobial activities were obtained for moderate PDMAEMA-BC chain lengths. These results help to understand the antimicrobial mechanism of polymeric quaternary ammonium salts and highlight their potential application as fungicidal agents for controlling both human and plant diseases.

Polymeric quaternary ammonium salts (PQAS) have been widely used to prevent microbial contamination, but little is known about their activity against phytopathogenic fungi. Our previous report described the synthesis of two novel PQAS, namely a homopolymer of (2-methacrylamido) propyltetrabenzyldimethylammonium chloride (PQD-BC) and dimethylaminopropyl benzyl chloride-grafted polysiloxanes (PDMS-g-BC); we demonstrated their structure-activity relationship against phytopathogenic fungi such as R. solani and Fusarium oxysporum f. sp. cubense tropical race 4 (Foc4). Here, we analyzed the structure-activity relationship and toxicity mechanism of these compounds at the molecular level against Foc4 compared with the low molecular-weight quaternary ammonium salt benzalkonium chloride (BC). The results revealed that PQD-BC and PDMS-g-BC application inhibited the growth of Foc4 in a concentration-dependent manner and that PDMS-g-BC exhibited higher activity than PQD-BC. In addition, these polymers were found to induce cell death in Foc4 by disrupting the cellular structure integrity, such as the loss of the cell wall and plasma membrane integrity and oxidative stress (lipid peroxidation), leading to the release of intracellular contents and inducing mitochondrial dysfunction and interference with genomic DNA. The newly elucidated mechanism provides possible applications in which PQAS can be used against phytopathogenic fungi.

Tau aggregates into paired helical filaments within neurons, a pathological hallmark of Alzheimer's disease. Heparin promotes tau aggregation and recently has been shown to be involved in the cellular uptake of tau aggregates. Although the tau-heparin interaction has been extensively studied, little is known about the glycan determinants of this interaction. Here, we used surface plasmon resonance (SPR) and NMR spectroscopy to characterize the interaction between two tau fragments, K18 and K19, and several polysaccharides, including heparin, heparin oligosaccharides, chemically modified heparin, and related glycans. Using a heparin-immobilized chip, SPR revealed that tau K18 and K19 bind heparin with a K-D of 0.2 and 70 mu M, respectively. In SPR competition experiments, N-desulfation and 2-O-desulfation had no effect on heparin binding to K18, whereas 6-O-desulfation severely reduced binding, suggesting a critical role for 6-O-sulfation in the tau-heparin interaction. The tau-heparin interaction became stronger with longer-chain heparin oligosaccharides. As expected for an electrostatics-driven interaction, a moderate amount of salt (0.3 M NaCl) abolished binding. NMR showed the largest chemical-shift perturbation (CSP) in R2 in tau K18, which was absent in K19, revealing differential binding sites in K18 and K19 to heparin. Dermatan sulfate binding produced minimal CSP, whereas dermatan disulfate, with the additional 6-O-sulfo group, induced much larger CSP. 2-O-desulfated heparin induced much larger CSP in K18 than 6-O-desulfated heparin. Our data demonstrate a crucial role for the 6-O-sulfo group in the tau-heparin interaction, which to our knowledge has not been reported before.

A series of linear hydrophilic cationic polymers with different charge density and molecular weights were synthesized by one-step polymerization process. The effect of the hydrophobicity and molecular weights on the antifungal activity against Rhizoctonia solani (R. solani) and Fusarium oxysporum f. sp. cubense race 4 (Foc4) was assessed. The biotoxicity of the cationic polymers were evaluated based on their median lethal concentration (LC50) for zebrafish and silkworm and median lethal dose (LD50) for Kunming mice. The results indicated that the balance between antifungal activity and biotoxicity could be well tuned by controlling the hydrophobic-hydrophilic balance. The minimum inhibitory concentration (MIC) of PEPB10 and PEPB25 against R. solani were 160 μg/mL and 80 μg/mL, respectively. And the LD50 for Kunming mice of PEPB10 and PEPB25 were more than 5000 mg/kg, which mean that PEPB10 and PEPB25 with high hydrophilicity show low toxicity and better selectivity for R. solani. The cationic polymers can kill the R. solani by damaging their membranes and exchanging the Ca2+ or/and Mg2+ cations of their membranes or cell wall. These results help to understand the antifungal mechanism of low-toxic polymeric quaternary ammonium salts and highlight their potential application as highly selective fungicidal agents for controlling plant diseases.

Ganoderma lucidum, commonly known as "Lingzhi" in Chinese, are well-known medicinal mushrooms. Lingzhi has been used in traditional Chinese herbal medicines for more than two thousand years. G. lucidum polysaccharides (GLPs) are present at high levels in G. lucidum cells and GLPs have molecular weights ranging from thousands to millions. GLPs have been widely studied for their various biological activities, such as antioxidant, antitumor, anti-inflammatory, antiviral, anti-diabetes, and immunomodulatory activities. The methods for GLPs extraction and characterization are mature, but the comprehensive research on the relationship between GLPs structure (i.e., molecular weight, tertiary structure, branching, substituents, and monosaccharide composition) and function is still quite limited. The aim of this review is to update and summarize the mechanisms of the various bioactive polysaccharides extracted from G. lucidum. The information presented on these bio-mechanisms should be valuable in the research and development of GLPs-derived therapeutics.

Antrodia camphorata (AC) is a precious medicinal mushroom native to Taiwan and famous for its excellent pharmacological activity. A ball mill assisted mechanochemical extraction method was applied in the extraction of triterpenoids from Antrodia camphorata. Compared with the ethanol hot thermal reflux method, mechanochemical-assisted extraction afforded an increased yield of triterpenoids to 1.82 &plusmn, 0.04% under conditions of mixing with 10 WT% NaHCO3, milling for 20 min, and extracting with water and chloroform. Triterpenoids from Antrodia camphorata extracted by the mechanochemical-assisted extraction method (TAEM) resulted in stronger pharmacological activity as compared to that extracted by ethanol (TAEE). HPLC and LC-MS/MS results showed that the mechanochemical method could extract triterpenoids which were barely extracted by ethanol extraction. The results of this study could provide valuable ideas and a basis for the application of the mechanochemical-assisted extraction method in the extraction of triterpenoids from AC.

Pleurotus eryngii (also known as king trumpet mushroom or king oyster mushroom) is an edible mushroom cultivated widely in many regions of the world. Polysaccharides from P. eryngii have a variety of biological activities, including anti-oxidant, anti-hyperlipidemic, anti-tumor, immunoregulatory and bacteriostatic. This paper reviews the extraction/purification, structural analysis and pharmacological activities of polysaccharides from this mushroom and provides updated research progress in areas important for the processing and product development of P. eryngii derived agents.

In cancer therapy, there still remains great challenges such as controlling cancer metastasis and the side effects from chemotherapy. Many anti-cancer drugs have cytotoxicity to normal cell with a high concentration, on the other hand, the anti-cancer effect will not be ideal if it didn’t reach the effective concentrations. So there is a way to make balance between the cytotoxicity and effective concentrations, sustained release can not only prolong the duration of the effective concentration but also reduce the time of peak concentration. Herein, we designed a sandwich structured antitumor composite with a three-layer film including: i) the top and bottom layers of ethyl cellulose (EC) containing Ganoderma lucidum triterpenes (GLT); ii) the middle layer of polyvinyl alcohol (PVA) containing G. lucidum polysaccharides (GLP). The films were fabricated layer-by-layer using electrospinning with the aim of providing the sustained release of the antitumor activity of GLT and GLP for use in cancer therapy. This sandwich structured material has potential for sustained antitumor release, high loading capacity (1.6% for GLT and 7.9% for GLP) and large surface area. The morphology, structure, biocompatibility, and the release profile of the encapsulated GLP and GLT in this fabricated composite were characterized. The in vitro antitumor effects of this drug film were studied using carcinoma cells, as SGC-7901, A549, Hela and Caco-2, with the IC50 of 51.2, 90.7, 93.0 and 21.7 μg/mL, respectively. In brief, this medical film achieved a good antitumor effect at the cellular level.

Fusarium oxysporum f. sp. cubense race 4 (Foc4) has significant impacts on banana production. Foc4 conidia can survive for many years in the soil, making control difficult. Thus, the inhibition of the conidia that serve as a continual source of inoculum in soil should be an effective way to control banana Fusarium wilt. In this paper, we first investigated the activities of polydimethylsiloxane-polymethacrylate block copolymers containing quaternary ammonium salts (PDMS-b-QPDMAEMA, labeled SixQ5) against Foc4 with microbroth dilution and toxicity plate assays. Then, the environmental behavior of these compounds, i.e., their adsorption and migration in soils, was further studied. Later, the effect and duration of SixQ5 on Foc4 in soil and the effects of SixQ5 on soil microbial diversity were evaluated. In addition, we conducted pot experiments to study the effect of Si5Q5 on banana Fusarium wilt. The results showed that SixQ5 showed a good inhibitory effect on Foc4, was easily adsorbed by the soil and was relatively immobile in soil. The inhibitory effect of SixQ5 on Foc4 was maintained for 30 d under continual inoculation of Foc4 into the soil (Foc4 addition every 2 d), and SixQ5 remained active in the soil for 90 d with Foc4 addition every 10 d. Soil microbial population analysis revealed that the culturable total fungal and bacterial populations increased and the culturable actinobacterial population decreased SixQ5 with addition. Pot experiments showed that Si5Q5 can reduce Foc4 infection in banana seedlings and does not harm banana seedlings. The stability and inhibitory effects on Foc4 demonstrate the potential application of SixQ5 in banana fields to prevent Foc4 conidial infection in the long term.

The interactions between carboxybetaine-functionalized polydimethylsiloxanes (PDMS-g-CB) and bovine serum albumin (BSA) in water solution were investigated from varied aspects based on different techniques, including fluorescence spectroscopy, UV-Vis spectroscopy, circular dichroism spectroscopy, isothermal titration calorimetry, and atomic force microscopy. Due to the weak interaction between PDMS-g-CB and BSA, another polymer with similar structure as PDMS-g-CB, i.e., quaternary ammonium salt-functionalized polydimethylsiloxanes (PDMS-g-QAS), was synthesized and chosen as the positive control, and the results indicated that the free PDMS-g-CB in solution could exhibit good resistance against proteins, which was mainly due to the zwitterions-side groups’ binding ability between PDMS chain and protein. Both the characterization methods and the results could help us to understand the interaction between free water-soluble polymers and protein, and extend the application of PDMS-g-CB and design new antifouling materials.

Three series of acrylamide monomers/polymers containing quaternary ammonium salts (QASs), i.e., acrylamide QAS monomers (QDs), homopolymers of QDs (PQDs) and copolymers of QDs with acrylate monomer (PQDCs), were synthesized and employed in antimicrobial tests against both bacteria ( Escherichia coli ( E . coli ) and Staphylococcus albus ( S . albus )) and phytopathogenic fungi ( Rhizoctonia solani ( R . solani ) and Fusarium oxysporum f . sp. cubense race 4 ( Foc 4)). The antibacterial activity of the QASs was evaluated by determining the minimum inhibitory concentration (MIC) against E . coli and S . albus by the TTC coloration method, and the antifungal activity was measured by mycelia growth inhibition as well as MIC and the minimum fungicidal concentration (MFC) values. The results indicated that PQD homopolymers and PQDC copolymers showed far better antimicrobial activities than QD monomers. PQDC copolymers by incorporating hydrophobic acrylate units into the main chain of polyacrylamide backbone displayed even better antimicrobial activities, depending on QAS structure and hydrophobic content. Moreover, polymers with benzyl group attached to nitrogen atom showed better inhibitory effect on bacteria and phytopathogenic fungi. The results could assist understanding and development of future design of antimicrobial polymers as potential fungicide agents to control plant disease.

A series of dimethylaminopropyl benzyl chloride grafted polysiloxanes (PDMS-g-BCs) with tunable molecular weights and cationic content were synthesized, and the effect of polymer structure on the antimicrobial activities against bacteria ( Escherichia coli ( E . coli ) and Staphylococcus albus ( S . albus )) and phytopathogenic fungi ( Rhizoctonia solani ( R . solani ) and Fusarium oxysporum f . sp . cubense race 4 ( Foc 4)) were systematically assessed. The antibacterial activity was evaluated by determining the minimum inhibitory concentration (MIC) against E . coli and S . albus by the broth dilution method, and the antifungal activity was measured by mycelia growth inhibition as well as by the MIC and minimum fungicidal concentration (MFC) values. The molecular weight and cationic content were major determinants of the activities of PDMS-g-BCs; when the molecular weight was approximately 2500 g/mol and the cationic content was approximately 20 mol% of the total siloxane units, PDMS-g-BCs gained strong antimicrobial activities toward both E . coli and Foc 4, comparable to those of a commonly used broad-spectrum microbicide benzalkonium chloride, making PDMS-g-BCs promising fungicidal agents for plant disease control.

An edible fungal polysaccharide termed as ABP was obtained by extraction with hot water, and followed successive chromatographic purification using DEAE-Sepharose Fast Flow column and Sephacryl S-300 High-Resolution column. A symmetrical peak was obtained on high-performance size-exclusion chromatography with an average molecular weight of 5.17 × 104 Da, which was named ABP, and its main components were ᴅ-glucose and ᴅ-mannose. Based on the study of methylation analysis, along with FT-IR, GC, GC-MS, 1D 1H and 13C NMR and 2D NMR (H-HCOSY, TOCSY, HMQC, and NOESY), its chemical structure was featured with a repeating unit (1→6) linking β-ᴅ-Glcp as the main backbone with (1→4)-linked α-ᴅ-Manp units. The structure of the mainly repeating units of ABP was established as: →6) − β − D − Glucp − (1→4) − α− D −Manp(1→6) − β − D − Glucp − (1→6) − β − D − Glucp − (1→

A water-soluble polysaccharide containing 3-O-methyl galactose (PCP60W) was isolated from fruiting bodies of Pleurotus citrinopileatus and purified by anion-exchange and gel column chromatography. This polysaccharide has an average molecular weight of 2.74 × 104 Da and its structure was elucidated using monosaccharide composition and methylation analysis combined with one- and two-dimensional (COSY, TOCSY, NOESY, HMQC and HMBC) NMR spectroscopy. PCP60W was shown to be a linear partially 3-O-methylated α-galactopyranan comprised of 6-linked galactose, 6-linked 3-O-methyl galactose and 4-linked glucose in a ratio of 3.0:1.0:0.6. This work provides additional evidence for the view that 3-O-methyl galactose is common to the genus Pleurotus.

Polyporus umbellatus is a medicinal fungus, has been used in traditional Chinese medicine for thousands years for treatment of edema, scanty urine, vaginal discharge, jaundice and diarrhea. The structure of a soluble polysaccharide (named PUP80S1), purified from the sclerotia of Polyporus umbellatus was elucidated by gas chromatography (GC), GC-mass spectrometry and nuclear magnetic resonance spectroscopy. PUP80S1 is a branched polysaccharide containing approximately 8.5% uronic acid and having an average molecular weight of 8.8kDa. Atomic force microscopy of PUP80S1 reveals a globular chain conformation in water. Antioxidant tests, Oxygen radical absorption capacity and 2,2-diphenyl-1-picrylhydrazyl radical scavenging assays indicate that PUP80S1 possesses significant antioxidant activity. But the related polysaccharide, PUP60S2, which contains more uronic acid residues and a higher level of branching, shows better antioxidant activity. These results suggest that structure features of polysaccharides play an important role in their physiological functions.

Boletus edulis is a well-known delicious mushroom. In this study, three crude polysaccharides (BEPF30, BEPF60 and BEPF80) were isolated from the fruiting bodies of B. edulis with boiling water. Chemical and physical characteristics of the three crude polysaccharides were investigated by the combination of chemical and instrumental analysis methods. Their antioxidant activities were investigated in vitro systems including hydroxyl assay, superoxide radical assay, reducing power and chelating activity. Among these three polysaccharides, BEPF60 showed more significant reducing power and chelating activity; and highest inhibitory effects on superoxide radical and hydroxyl radical. These results indicated that polysaccharides extracted from B. edulis might be employed as ingredients in healthy and functional food to alleviate the oxidative stress.

Background Previous epidemiological studies have presented conflicting evidence regarding associations between interleukin-1 (IL-1) polymorphisms and sepsis susceptibility. We have performed a meta-analysis to evaluate possible associations between IL-1 polymorphisms and sepsis risk. Methods Eligible literature was retrieved from PubMed, Embase and Web of Knowledge databases until Jun 15, 2013. The pooled odds ratio (OR) and 95% confidence interval (CI) were calculated using random-effects model in the overall and subgroup analysis based on ethnicity, sepsis severity and quality score. Results Eighteen studies addressing five IL-1 polymorphisms were included in this meta-analysis. For IL-1A-889 (rs1800587) polymorphism, significant association was observed in overall comparison for allelic effect (OR = 1.47, 95% CI = 1.01-2.13, P = 0.04). There were no significant associations between either IL-1B-511 (rs16944) or IL-1B-31 (rs1143627) and sepsis susceptibility in overall or subgroup analyses. For IL-1B + 3594 (rs143634) polymorphism, genotype TT decreased sepsis risk in overall analysis (OR = 0.59, 95% CI = 0.36-0.97, P = 0.04), as well as in Caucasian (OR = 0.57, 95% CI = 0.34-0.95, P = 0.03) and sepsis (OR = 0.55, 95% CI = 0.31-0.97, P = 0.04) subgroup analysis. For IL-1RN VNTR polymorphism, significant association was observed in overall comparison for allelic effect (OR = 1.40, 95% CI = 1.01-1.95, P = 0.04). Furthermore, the effect sizes of IL-1RN VNTR on sepsis risk increased with disease severity (septic shock OR > severe sepsis OR > sepsis OR). Conclusions Our meta-analysis indicated that IL-1A-889, IL-1B + 3954 and IL-1RN VNTR might be associated with sepsis susceptibility. However, further studies with larger sample sizes and from homogenous populations would be necessary to validate these findings.

We described a triplex polymerase chain reaction (PCR) and triplex pyrosequencing assay which allowed a simultaneous determination of three tag single nuleotide polymorphisms (tag SNPs) in the lipopolysaccharide-binding protein (LBP) gene: rs1780623, rs11536972 and rs2232618. This method enables a fast and cost-effective genotyping and a simultaneous determination of the three tag SNPs.

This paper describes the design and construction of chimeric human rhinoviruses that contain immunogenic regions from other pathogens as part of their surface coat proteins. Segments encoding the poliovirus 3 Sabin VP1 and VP2 proteins, the influenza hemagglutinin (HA) glycoprotein, and the human immunodeficiency virus gp120 surface and gp41 transmembrane glycoproteins were inserted into a full-length clone of human rhinovirus 14 (HRV14) at regions corresponding to neutralizing immunogenic sites IA (NIm-IA) and II (NIm-II). Of 12 chimeric constructs described, 3 produced viable virus. An HRV14 chimeric virus containing five amino acids of influenza HA (corresponding to 300 A2 of solvent-accessible surface area) had wild-type HRV14 growth characteristics and was neutralized by four of four anti-influenza HA antisera with reciprocal neutralizing titers ranging from 180 to 330. However, antisera raised in two guinea pigs against the HRV14:influenza HA chimera did not show significant neutralization of relevant strains of influenza. These results are the first to demonstrate the feasibility of making viable chimeras of human rhinoviruses displaying heterologous immunogens.

To determine the hypothesis that genetic variations of the lipopolysaccharide-binding protein (LBP) gene influence risk for the development of sepsis and multiple organ dysfunction (MOD) in patients with major trauma.Lipopolysaccharide-binding protein plays a central role in innate immune response as the first line of defense and directing the microbial-induced activation of the inflammatory host response. Although a total of 112 single nucleotide polymorphisms (SNPs) have been identified so far within the entire LBP gene, only a few SNPs have been studied.Nine haplotype tagging SNPs (htSNPs) were selected from 51 SNPs with a minor allele frequency of ≥5% using the HapMap database for the Chinese Han population. Two independent cohorts of major trauma patients were recruited. The 9 htSNPs were genotyped using pyrosequencing method and analyzed in relation to the risk of development of sepsis and MOD, LBP production, and lipopolysaccharide (LPS)-induced activation of peripheral blood leukocytes. Moreover, the functionality of the rs2232618 polymorphism was assessed by the observation of its effects on the binding and activation of LPS and the LBP-CD14 interaction.Among the 9 htSNPs, only the rs2232618 was significantly associated with higher susceptibility to sepsis and MOD in the 2 independent cohorts of major trauma patients recruited from southwest and eastern China. This SNP was also significantly associated with LPS-induced activation of peripheral blood leukocytes. In addition, the rs2232618 polymorphism could enhance LBP protein activities, showing significant increases in LPS binding to macrophages, LPS-induced cellular activation, and LBP-CD14 interaction at the presence of the variant LBP protein.The rs2232618 polymorphism is a functional SNP and confers host susceptibility to sepsis and multiple organ dysfunction in patients with major trauma.

The activity of a new water-soluble molecule, SCH 38057, against picornaviruses is described. SCH 38057 inhibited plaque formation of selected entero- and rhinoviruses in a range of 10.2 to 29.1 microM (50% endpoint) and had a therapeutic index of 10 against poliovirus type 2 (polio 2) in HeLa cells. When administered orally or subcutaneously, SCH 38057 protected mice infected with either coxsackievirus B3 (CVB3) or echovirus-9 from mortality. The molecule provided a low level of protection against thermal inactivation of virus, indicating that SCH 38057 interacts with the picornavirus capsid. Binding studies with [3H]SCH 38057 revealed that the molecule binds to CVB3 and human rhinovirus 14 (HRV14) in a ratio of 29 and 19 molecules per viral particle, respectively. The affinity constant for SCH 38057 binding to CVB3 was 7.0 x 10(-4) M. When added to cultures of infected cells at 3 h after infection, SCH 38057 markedly inhibited viral RNA synthesis. This finding with lack of inhibition of attachment and loss of infectious virus after attachment were interpreted to indicate that, although SCH 38057 binds to the viral capsid, the molecule exerts its antiviral effect after the initial stage of picornavirus uncoating, i.e., after conversion of the 156S infectious viral particle to smaller subviral species.

SCH 38057 (1-[6-(2-chloro-4-methoxyphenoxy)-hexyl]imidazole hydrochloride) is a new, water-soluble antiviral compound that has inhibitory activities against a number of picornavirus infections. The structure of the human rhinovirus 14 (HRV14) complex with SCH 38057 was determined at 3.0 A resolution by single-crystal diffraction techniques using synchrotron X-radiation. SCH 38057 was found to bind at the innermost end of the hydrophobic pocket within the capsid protein VP1, a locus of binding of other antipicornaviral agents; however, the complex differs from previously reported complexes in two important aspects. It leaves a considerable volume near the entrance to the binding pocket unoccupied. In addition, the alterations in the conformation of the VP1 polypeptide are similar to, but more extensive than those observed in HRV14 complexes with other antiviral agents. Although only 9 amino acids of VP1 have close contacts with the SCH 38057 molecule (within 3.6 A), at least 36 amino acids from both VP1 and VP3 have significantly altered conformations (C alpha movement0.5 A versus native). The structures of complexes of HRV14 with SCH 38057 and WIN 51711 are compared. Aromatic ring interactions between picornavirus capsid residues and antiviral inhibitors are proposed to be among the major determinants for positioning of these compounds.

AIDS, caused by human immunodeficiency virus (HIV), is one of the world's most serious health problems, with current protocols being inadequate for either prevention or successful long-term treatment. In retroviruses such as HIV, the enzyme reverse transcriptase copies the single-stranded RNA genome into double-stranded DNA that is then integrated into the chromosomes of infected cells. Reverse transcriptase is the target of the most widely used treatments for AIDS, 3'-azido-3'-deoxythymidine (AZT) and 2',3'-dideoxyinosine (ddI), but resistant strains of HIV-1 arise in patients after a relatively short time. There are several nonnucleoside inhibitors of HIV-1 reverse transcriptase, but resistance to such agents also develops rapidly. We report here the structure at 7 A resolution of a ternary complex of the HIV-1 reverse transcriptase heterodimer, a monoclonal antibody Fab fragment, and a duplex DNA template-primer. The double-stranded DNA binds in a groove on the surface of the enzyme. The electron density near one end of the DNA matches well with the known structure of the HIV-1 reverse transcriptase RNase H domain. At the opposite end of the DNA, a mercurated derivative of UTP has been localized by difference Fourier methods, allowing tentative identification of the polymerase nucleoside triphosphate binding site. We also determined the structure of the reverse transcriptase/Fab complex in the absence of template-primer to compare the bound and free forms of the enzyme. The presence of DNA correlates with movement of protein electron density in the vicinity of the putative template-primer binding groove. These results have important implications for developing improved inhibitors of reverse transcriptase for the treatment of AIDS.

Background Recent studies on the association between CD14-159C/T polymorphism and sepsis showed inconclusive results. Accordingly, we conducted a comprehensive literature search and a meta-analysis to determine whether the CD14-159C/T polymorphism conferred susceptibility to sepsis or was associated with increased risk of death from sepsis. Methodology Data were collected from the following electronic databases: PubMed, Embase, Medline, Web of Knowledge, and HuGE Navigator, with the last report up to June 15, 2012. The odds ratio (OR) and 95% confidence interval (CI) were used to assess the strength of association. We summarized the data on the association between CD14-159C/T polymorphism and sepsis in the overall population and subgroup by ethnicity and sepsis subtype. Principal Findings A total of 16 studies on sepsis morbidity (1369 cases and 2382 controls) and 4 studies on sepsis mortality (731 sepsis patients) met the inclusion criteria for meta-analysis. Overall analysis showed no strong evidences of association with sepsis susceptibility under any genetic model. However, slight associations were found in Asian populations (dominant model: OR = 1.38, 95%CI = 0.96–1.98, P = 0.08) and septic shock patients (dominant model: OR = 1.72, 95%CI 1.05–2.83, P = 0.03; allelic model: OR = 1.52, 95%CI 1.09–2.12, P = 0.01) in the stratified analysis. Moreover, there was borderline association between CD14-159C/T and sepsis mortality under the dominant genetic model (OR = 1.44, 95%CI = 0.98–2.11, P = 0.06). Conclusions/Significance This meta-analysis suggests that the CD14-159C/T polymorphism may not be a significant susceptibility factor in the risk of sepsis and mortality. Only weak associations were observed in Asian populations and septic shock patients. More studies based on larger sample sizes and homogeneous sepsis patients are needed to confirm these findings.

Random systematic mutagenesis was used to generate a library of human rhinovirus 14 chimeras that each display a segment from the V3 loop of human immunodeficiency virus type 1. The sequence XXIGPGRAXX, where X could be any of the 20 amino acids, was inserted at the neutralizing immunogenic site II of human rhinovirus 14 between VP2 residues 159 and 160. Twenty-five unique chimeric viruses were isolated, and the identity of their randomized residues was determined. A nonrandom amino acid distribution that may reflect structural requirements for viability was observed at the randomized positions. Fifteen of 25 chimeras were neutralized by one or more of a panel of four anti-human immunodeficiency virus type 1 V3 loop antibody preparations, indicating that antigenicity had been successfully transplanted. Libraries of chimeric viruses produced by using the techniques described may be a source of vaccines and other immunotherapeutic reagents. The random systematic mutagenesis methodology described should be generally useful for the rapid transplantation of foreign sequences into viral coat and other proteins to produce libraries containing members with the desired properties.

Preparations of coxsackievirus B1 (CVB1) derived from an infectious cDNA clone have been crystallized in multiple crystal forms. Using high intensity synchrotron radiation, an orthorhombic form of the crystals was shown to diffract X-rays to at least 2.9 A resolution. The unit cell has a primitive lattice with dimensions a = 323 A, b = 450 A, and c = 522 A. A crystallographic asymmetric unit of these CVB1 crystals probably contains an entire virus particle, implying the presence of 60-fold non-crystallographic redundancy. This CVB1 crystal form appears to be suitable for high-resolution structure determination by X-ray crystallography.

A chimeric virus library was designed whereby sequences corresponding to the V3 loop of human immunodeficiency virus type 1 (HIV-1) were presented on the surface of human rhinovirus 14. The V3 loop sequences consisted of a relatively conserved segment of seven amino acids and five adjacent residues that were allowed to vary in proportion to their seroprevalence among HIV-1 isolates of North America and Europe. A technique called random systematic mutagenesis was used to incorporate the composite V3 loop sequences flanked by zero to two randomized amino acids. This library could contain 2.7 x 10(8) members having diverse sequences and conformations. Immunoselection of a portion of this library by using two neutralizing V3 loop-directed monoclonal antibodies followed by selection for desirable growth and purification characteristics yielded a set of chimeric rhinoviruses, five of which are described. The inserted sequences in the five chimeras do not match those of any known isolate of HIV-1. Nonetheless, all five chimeras were neutralized by antibodies directed against different strains of HIV-1 and were able to elicit the production of antibodies that bind V3 loop peptides from diverse HIV-1 isolates. Moreover, antisera derived from four of the five chimeras were capable of neutralizing one or more strains of HIV-1 in cell culture. This study demonstrates that random systematic mutagenesis in conjunction with antibody screening is a powerful and efficient means to obtain antigenic chimeras with relevant immunogenic properties.