Chemiluminescence of 4‐styrylphthal‐hydrazides with crown ether as ionophore

Two 4‐styrylphthalhydrazides bearing crown ether moieties as ionophores were prepared and their chemiluminescence (CL) behaviour was investigated. Their aerobic oxidation in the presence of excess tert‐BuOK in DMSO provided the CL with much shorter emission wavelengths than those of the corresponding phthalate dianions, indicating that the emission arose from the excited phthalates incorporated with potassium ions at the ionophore sites. Such incorporation of potassium ions with the crown sites also reduced the fluorescence intensities of the phthalate dianions which caused an energy transfer CL, resulting in the additional emissions of the phthalhydrazide monoanions in much longer wavelength regions. On the other hand, only the emission from the excited phthalate dianion was detected under aqueous conditions using aqueous hydrogen peroxide in acetonitrile as the oxidant, but no meaningful difference in the CL intensity depending on the kind of metal cations was observed. Copyright © 2008 John Wiley & Sons, Ltd.     

Cytokinin biosynthesis in crown gall tissue of Vinca rosea: Metabolism of isopentenyladenine

When isopentenyl[8-¹⁴C]adenine was incubated with crown gall tumour tissue of Vinca rosea, it was stereospecifically hydroxylated to trans-zeatin and its derivatives, which are the endogenous free cytokinins in this tissue. Adenine, adenosine and adenine nucleotides were the major degradation products.     

Analysis of potential virulence genes and competence to transformation in Haemophilus influenzae biotype aegyptius associated with Brazilian Purpuric Fever

Brazilian Purpuric Fever (BPF) is a hemorrhagic pediatric illness caused by Haemophilus influenzae biogroup aegyptius (Hae), a bacterium that was formerly associated with self-limited purulent conjunctivitis. BPF is assumed to be eradicated. However, the virulence mechanisms inherent to Hae strains associated with BPF is still a mystery and deficient in studies. Here, we aim to analyze the role of the autotransporter genes related to adherence and colonization las, tabA1, and hadA genes through RT-qPCR expression profiling and knockout mutants. Relative quantification by real-time PCR after infection in human cells and infant rat model suggests that las was initially downregulated probably duo to immune evasion, tabA1, and hadA were overexpressed in general, suggesting an active role of TabA1 and HadA1 adhesins in Hae in vitro and in vivo. Transformation attempts were unsuccessful despite the use of multiple technical approaches and in silico analysis revealed that Hae lacks genes related to competence in Haemophilus, which could be part of the elucidation of the difficulty of genetically manipulating Hae strains.     

Real‐time PCR quantification of Bemisia tabaci (Homoptera: Aleyrodidae) B‐biotype remains in predator guts

The cotton whitefly, Bemisia tabaci (Gennadius) B‐biotype, is fed on by a wide variety of generalist predators, but there is little information on these predator–prey interactions, especially under field conditions. In this study, a real‐time polymerase chain reaction (PCR) assay was developed to quantify B. tabaci B‐biotype remains in predator gut. The B. tabaci B‐biotype genomic DNA copy number was referred to the actual amount of BT1 isolate, the B. tabaci B‐biotype specific DNA fragment. The numbers of BT1 isolate in one B. tabaci B‐biotype egg, individual adult and a single red‐eyed nymph were 2.56 × 10³, 2.56 × 10⁴, and 1.29 × 10⁴ copies, respectively. When Propylaea japonica adults fed on one, two, four, eight or 16 red‐eyed nymphs, the detected numbers of BT1 isolate ranged from 2.77 × 10⁴ to 4.05 × 10⁵ copies, forming a strong linear relationship (R² = 0.9899). Following the consumption of two red‐eyed nymphs, prey DNA was detectable in 100% of P. japonica at t = 0, decreasing to 80.0% and 60.0% after 1–4 h and 8 h of digestion, respectively, with 3.36 × 10⁴–1.25 × 10³ BT1 isolate copies. The predation by field‐collected predators, 26 larvae of P. japonica, and of Harmonia axyridis each, Chrysopa spp. larvae (Chrysopa pallens and C. formosa, 18 individuals in total), and a single adult of Scymnus hoffmanni, 19 adults of Orius sauteri and nine adult spiders (Erigonnidium graminicolum and Neoscona doenitzi), on B. tabaci B‐biotype were quantified. Of the 99 analysed predator individuals, 3.65 × 10²–4.60 × 10⁵ copies of BT1 isolate, equivalent to 0.8–18.8 red‐eyed nymphs were detected. These results suggest that TaqMan real‐time PCR technology may provide a rapid and sensitive method for quantifying B. tabaci B‐biotype remains in predator guts and will be invaluable in assessing the food web relationship between prey and arthropod predators.     

Biosynthesis of N2-(1,3-dicarboxypropyl) ornithine [nopalinic acid] in crown gall tumour tissue

Protein extract from crown gall tumour tissue, induced on Nicotiana tabacum by Agrobacterium tumefaciens strain T37, synthesized nopalinic acid [N²-(1,3-dicarboxypropyl)ornithine] from l-ornithine and α-ketoglutarate in the presence of NADPH. Label was incorporated into nopalinic acid from both l-ornithine-[¹⁴C] and α-ketoglutarate-[¹⁴C] in vivo. Nopaline [N²-(1,3-dicarboxypropyl)arginine] did not appear to be metabolized to nopalinic acid in vivo.     

Modeling population size independent tissue epigenomes by ChIL‐seq with single thin sections

Recent advances in genome‐wide technologies have enabled analyses using small cell numbers of even single cells. However, obtaining tissue epigenomes with cell‐type resolution from large organs and tissues still remains challenging, especially when the available material is limited. Here, we present a ChIL‐based approach for analyzing the diverse cellular dynamics at the tissue level using high‐depth epigenomic data. “ChIL for tissues” allows the analysis of a single tissue section and can reproducibly generate epigenomic profiles from several tissue types, based on the distribution of target epigenomic states, tissue morphology, and number of cells. The proposed method enabled the independent evaluation of changes in cell populations and gene activation in cells from regenerating skeletal muscle tissues, using a statistical model of RNA polymerase II distribution on gene loci. Thus, the integrative analyses performed using ChIL can elucidate in vivo cell‐type dynamics of tissues.     

西帕依固龈液联合西吡氯铵治疗牙龈炎的疗效分析

目的:探讨西帕依固龈液联合西吡氯铵治疗牙龈炎的临床疗效。方法:选择我院于2013年10月-2015年2月收治的130例牙龈炎患者,将其随机分成实验组和对照组各65例,对照组给予西吡氯铵治疗,实验组给予西帕依固龈液联合西吡氯铵治疗,比较两组患者的菌斑指数(PLI)、牙龈指数(GI)、龈沟出血指数(SBI)以及治疗疗效。结果:治疗8天后,实验组和对照组患者的治疗总有效率分别为96.92%和81.54%,组间比较差异具有统计学意义(P0.05)。两组患者治疗前PLI、GI、SBI比较无显著差异(P0.05);治疗后,两组患者各项指标均较治疗前有所改善,与各自治疗前比较差异具有统计学意义(P0.05),但两组治疗后的指标差异无统计学意义(P0.05)。两组患者不良反应的发生率分别为10.77%和9.23%,差异无统计学意义(x~2=0.0855,P0.05)。结论:西帕依固龈液与西吡氯铵联合使用治疗牙龈炎具有确切的疗效,同时具有较高的安全性。     

NCAPG is differentially expressed during longissimus muscle development and is associated with growth traits in Chinese Qinchuan beef cattle

Based on RNA-seq analysis, we recently found that the bovine NCAPG(non-SMC condensin I complex, subunit G) gene is differentially expressed duringdevelopment of the longissimus muscle. In the present study, we validated this resultand, using quantitative real-time PCR analysis, identified two adjacent genes, LCORLand DCAF16, that are more abundant in fetal muscle tissue; further analysis oftissue-specific expression patterns indicated high abundance ofNCAPG in muscle. Since no polymorphisms were detected in aprevious study of Qinchuan cattle, we extended our investigation to examine theoccurrence of single-nucleotide polymorphisms (SNPs) in the NCAPGgene. Three SNPs, i.e., one located in the intron region (g47747: T> G), a synonymous mutation (g52535: A > G) and a missense mutation (g53208: T> G) that leads to a change in the amino acid of interest (pIle442Met), weredetected in a population of Qinchuan beef cattle (n = 300). Association analysisshowed that these SNPs were significantly associated with the growth traits ofQinchuan beef cattle. Our results indicate that the bovine NCAPGgene may be involved in the development of the longissimus muscle. Thesepolymorphisms in the NCAPG gene may be useful for marker-assistedselection of optimal body size in Qinchuan beef cattle.     

Identification of novel candidate circulating biomarkers for malignant soft tissue sarcomas: Correlation with metastatic progression

Soft tissue sarcomas (STS) are a heterogeneous group of rare tumors for which identification and validation of biological markers may improve clinical management. The fraction of low‐molecular‐weight (LMW) circulating proteins and fragments of proteins is a rich source of new potential biomarkers. To identify circulating biomarkers useful for STS early diagnosis and prognosis, we analyzed 53 high‐grade STS sera using hydrogel core‐shell nanoparticles that selectively entrap LMW proteins by size exclusion and affinity chromatography, protect them from degradation and amplify their concentration for mass spectrometry detection. Twenty‐two analytes mostly involved in inflammatory and immunological response, showed a progressive increase from benign to malignant STS with a relative difference in abundance, more than 50% when compared to healthy control. 16 of these were higher in metastatic compared to non‐metastatic tumors. Cox‘s regression analysis revealed a statistical significant association between the abundance of lactotransferrin (LTF) and complement factor H‐related 5 (CFHR5) and risk of metastasis. In particular, CFHR5 was associated with the risk of metastasis. The role of circulating proteins involved in metastatic progression will be crucial for a better understanding of STS biology and patient management.     

Synthesis of d‐mannitol‐based crown ethers and their application as catalyst in asymmetric phase transfer reactions

A few new d ‐mannitol‐based monoaza‐15‐crown‐5 type chiral lariat ethers and 18‐crown‐6 type macrocycles were synthesized. These crown compounds were used as phase transfer catalysts in asymmetric Michael addititons and in a Darzens condensation under mild conditions to afford the corresponding products in a few cases in good to excellent enantioselectivities. In the Michael addition of diethyl acetoxymalonate to trans‐chalcone, in the addition of diethyl acetamidomalonate to ß‐nitrostyrene, in the reaction of diethyl bromomalonate with benzylidene malononitriles, in the cyclopropanation reaction of diethyl bromomalonate and 2‐benzylidene‐1,3‐indandione, and in the Darzens condensation of α‐chloroacetophenone with benzaldehyde, maximum enantioselectivities of 39%, 65%, 99%, 56%, and 62%, respectively, were obtained in the presence of the d ‐mannitol‐based macrocycles as the catalysts.     

Synthesis of l‐threitol‐based crown ethers and their application as enantioselective phase transfer catalyst in Michael additions

A few new l ‐threitol‐based lariat ethers incorporating a monoaza‐15‐crown‐5 unit were synthesized starting from diethyl l ‐tartrate. These macrocycles were used as phase transfer catalysts in asymmetric Michael addition reactions under mild conditions to afford the adducts in a few cases in good to excellent enantioselectivities. The addition of 2‐nitropropane to trans ‐chalcone, and the reaction of diethyl acetamidomalonate with β‐nitrostyrene resulted in the chiral Michael adducts in good enantioselectivities (90% and 95%, respectively). The substituents of chalcone had a significant impact on the yield and enantioselectivity in the reaction of diethyl acetoxymalonate. The highest enantiomeric excess (ee ) values (99% ee ) were measured in the case of 4‐chloro‐ and 4‐methoxychalcone. The phase transfer catalyzed cyclopropanation reaction of chalcone and benzylidene‐malononitriles using diethyl bromomalonate as the nucleophile (MIRC reaction) was also developed. The corresponding chiral cyclopropane diesters were obtained in moderate to good (up to 99%) enantioselectivities in the presence of the threitol‐based crown ethers.     

N- 乙酰转移酶NAT10 在软组织肿瘤中的表达及意义

目的：观察N-乙酰转移酶NAT10蛋白在软组织肉瘤中的表达及与类型、分级的关系。方法：通过原核表达NAT10蛋白免疫制备特异性多克隆抗体,并经免疫印迹鉴定;以组织芯片一免疫组化检测166例软组织肉瘤和28例良性肿瘤及瘤样病变中NAT10蛋白的表达。结果：制备多克隆抗体经Western印迹鉴定与NAT10具有特异结合性。免疫组化显示166例软组织肉瘤中NAT10蛋白阳性95例,阳性率为57％（95／166）,28例良性肿瘤及瘤样病变中4例阳性14％（4／28）。两者间有显著性差异（P〈0．05）。NAT10表达的主要分布为：滑膜肉瘤76％（13／17）、恶性纤维组织细胞瘤75％（15／20）、原始神经外胚叶瘤（PNET）70％（16／23）、横纹肌肉瘤70％（7／10）、恶性外周神经鞘膜瘤50％（11,／22）、隆突性皮肤纤维肉瘤50％（7／14）、平滑肌肉瘤43％（6／14）、脂肪肉瘤42％（8／19）、黏液性纤维肉瘤38％（6／16）。统计比较显示：滑膜肉瘤与黏液性纤维肉瘤和脂肪肉瘤,以及恶性纤维组织细胞瘤与黏液性纤维肉瘤之间NAT10表达具有显著性差异（P〈0．05）;而其它各组间无明显差异（P〉0．05）。同时,NAT10蛋白强阳性表达（≥＋＋）多存在于滑膜肉瘤（53％,9／17）、横纹肌肉瘤（40％,4／10）及恶性纤维组织细胞瘤（40％,8／20）。在FNCLGC分级中,19例I级肉瘤中NAT10阳性表达率为42％（8／19）,44例Ⅱ级肉瘤为43％（19／44）,70例Ⅲ级肉瘤为73％（51／70）。Ⅲ级NAT10阳性率显著高于Ⅱ级组和Ⅰ级组（均为P〈0．05）。结论：研究表明N-乙酰转移酶NAT10表达于多种人软组织肉瘤,尤其在高度恶性肉瘤,因此有可能为软组织肉瘤的分级及预后因子。     

Updating the geographical distribution and frequency of Aedes albopictus in Brazil with remarks regarding its range in the Americas

The geographical distribution of Aedes albopictus in Brazil was updated according to the data recorded across the country over the last eight years. Countrywide house indexes (HI) for Ae. albopictus in urban and suburban areas were described for the first time using a sample of Brazilian municipalities. This mosquito is currently present in at least 59% of the Brazilian municipalities and in 24 of the 27 federal units (i.e., 26 states and the Federal District). In 34 Brazilian municipalities, the HI values for Ae. albopictus were higher than those recorded for Ae. aegypti, reaching figures as high as HI = 7.72 in the Southeast Region. Remarks regarding the current range of this mosquito species in the Americas are also presented. Nineteen American countries are currently infested and few mainland American countries have not confirmed the occurrence of Ae. albopictus. The large distribution and high frequency of Ae. albopictus in the Americas may become a critical factor in the spread of arboviruses like chikungunya in the new world.     

An integrated approach with Trichoderma harzianum DGA01 and hot water treatment on control of crown rot disease and retention of overall quality in banana

A biological control of crown rot disease of banana fruit was analysed using an integrated approach combining hot water treatment and Trichoderma harzianum strain DGA01. Treated fruits were stored at 22–25 °C and 90–95% relative humidity for 2 weeks. The bioefficacy of fungal antagonist in vitro towards crown rot-causing pathogens, namely Lasiodiplodia theobromae, Thielaviopsis paradoxa, Colletotrichum musae and Fusarium verticillioides, was enhanced by 11.41% following hot water treatment (50 °C, 20 min). DGA01 germinated on the fruit 48 h after inoculation and parasitised the pathogen. Postharvest application showed that hot water treatment and conidial suspension of DGA01 (10⁶ ml^?1) applied singly performed significantly better than the untreated control in reducing the incidence of crown rot, but were not as effective as the fungicide. The combination of hot water treatment and DGA01 gave 93% control of fruit decay which was comparable with fungicide treatment of 95%. The quality of fruit was markedly improved in hot water treatment + DGA01 as compared to those dipped in fungicide solution. The inconsistencies of single treatments, by DGA01 or hot water dips, in controlling crown rot such as variation in severity of disease among treatments and within a treatment, were lessened by dipping the fruit in DGA01 conidial suspension following hot water treatment.     

Association of TYK2 polymorphisms with autoimmune diseases: A comprehensive and updated systematic review with meta-analysis

Autoimmune diseases are characterized by the loss of self-tolerance, leading to immune-mediated tissue destruction and chronic inflammation. Tyrosine kinase 2 (TYK2) protein plays a key role in immunity and apoptosis pathways. Studies have reported associations between single nucleotide polymorphisms (SNPs) in the TYK2 gene and autoimmune diseases; however, results are still inconclusive. Thus, we conducted a systematic review followed by meta-analysis. A literature search was performed to find studies that investigated associations between TYK2 SNPs and autoimmune diseases (multiple sclerosis, systemic lupus erythematosus, Crohn’s disease, ulcerative colitis, psoriasis, rheumatoid arthritis, type 1 diabetes, and inflammatory bowel disease). Pooled odds ratios (OR) with 95 % CI were calculated using random (REM) or fixed (FEM) effects models in the Stata 11.0 Software. Thirty-four articles were eligible for inclusion in the meta-analyses, comprising 9 different SNPs: rs280496, rs280500, rs280523, rs280519, rs2304256, rs12720270, rs12720356, rs34536443, and rs35018800. Meta-analysis results showed the minor alleles of rs2304256, rs12720270, rs12720356, rs34536443, and rs35018800 SNPs were associated with protection against autoimmune diseases. Moreover, the A allele of the rs280519 SNP was associated with risk for systemic lupus erythematosus. Our meta-analyses demonstrated that the rs2304256, rs12720270, rs12720356, rs34536443, rs35018800, and rs280519 SNPs in the TYK2 gene are associated with different autoimmune diseases.     

Bridging of partially negative atoms by hydrogen bonds from main‐chain NH groups in proteins: The crown motif

The backbone NH groups of proteins can form N1N3‐bridges to δ‐ve or anionic acceptor atoms when the tripeptide in which they occur orients them appropriately, as in the RL and LR nest motifs, which have dihedral angles 1,2‐α_Rα_L and 1,2‐α_Lα_R, respectively. We searched a protein database for structures with backbone N1N3‐bridging to anionic atoms of the polypeptide chain and found that RL and LR nests together accounted for 92% of examples found (88% RL nests, 4% LR nests). Almost all the remaining 8% of N1N3‐bridges were found within a third tripeptide motif which has not been described previously. We term this a “crown,” because of the disposition of the tripeptide CO groups relative to the three NH groups and the acceptor oxygen anion, and the crown together with its bridged anion we term a “crown bridge.” At position 2 of these structures the dihedral angles have a tight α_R distribution, but at position 1 they have a wider distribution, with ? and ψ values generally being lower than those at position 1. Over half of crown bridges involve the backbone CO group three residues N‐terminal to the tripeptide, the remainder being to other main‐chain or side‐chain carbonyl groups. As with nests, bridging of crowns to oxygen atoms within ligands was observed, as was bridging to the sulfur atom of an iron‐sulfur cluster. This latter property may be of significance for protein evolution. Proteins 2015; 83:2067–2076. © 2015 Wiley Periodicals, Inc.     

A functional promoter polymorphism −607G>C of WNT10B is associated with abdominal fat in Korean female subjects

WNT10B has been implicated as a potential regulator of adipogenesis in cellular and animal models of obesity. In this study, we attempted to characterize the associations between common genetic polymorphisms of WNT10B and fat accumulation in a sample of 1029 Korean female subjects. Direct sequencing of genomic DNAs of 45 subjects identified six common single-nucleotide polymorphisms (SNPs) of WNT10B, which were in almost complete linkage disequilibrium. Among the six SNPs, −607G>C (rs833840) showed differential nuclear factor binding in an electrophoretic mobility shift assay and differential promoter activity in a reporter assay, implicating it as a functional regulatory SNP. When body compositions of the subjects determined using bio-impedance analysis were compared according to their −607G>C genotype, only body fat mass showed a significant association. Body masses of protein, mineral and water showed no association. For more accurate evaluation of the effects of −607G>C genotype on body fat, cross-sectional fat areas of the subjects measured by abdominal computed tomography were compared. Genotype of −607G>C was significantly associated with abdominal total fat and abdominal subcutaneous fat areas (P=.009 and P=.007 in recessive model, respectively). Of the 1029 subjects, 576 were treated with a 1 month very low calorie diet and changes of body weight and composition were compared with −607G>C genotype. No significant associations were evident. This study is the first report of the association of common genetic polymorphism of WNT10B with human fat accumulation.     

Correlation of Edaphic Factors with Plant-parasitic Nematode Population Densities in a Forage Field

Two hundred soil samples from the A_p horizon of a reed canarygrass field overlaying several different but related soils in northern Minnesota were analyzed for plant-parasitic nematodes and 22 edaphic factors. Pratylenchus penetrans was the predominant nematode taxon. Others were Aglenchus agricola, Tylenchorhynchus spp., Heterodera trifolii, Paratylenchus spp., Tylenchus maius, and Criconemella sp. Five nematode taxa, P. penetrans, A, agricola, Tylenchorhynchus spp., H. trifolii, and Paratylenchus spp., were correlated with particle size, Tylenchus maius and Criconemella sp. were correlated with effective cation exchange capacity. Nematode field spatial arrangements were related to a combination of statistically significant positive and negative soil factor effects on the nematode populations. Contour maps derived by geostatistical techniques were used to visually validate statistically significant correlations of nematode and soil data. Contour mapping to supplement traditional statistical techniques can be used to achieve a more holistic approach to studies of nematode-soil interrelationships.     

Evaluation of a xenogeneic VEGF vaccine in dogs with soft tissue sarcoma

Active immunization against pro-angiogenic growth factors or their receptors is an emerging strategy for controlling tumor growth and angiogenesis. Previous studies in rodent tumor models have indicated that immunization against xenogeneic growth factors is more likely to induce effective anti-tumor responses than immunization against the autologous growth factor. However, the effectiveness or safety of the xenogeneic vaccination approach has not been previously assessed in a clinically relevant outbred, spontaneous tumor model. Therefore, we investigated the safety and anti-tumor and anti-angiogenic effects of a xenogeneic vascular endothelial cell growth factor (VEGF) vaccine in pet dogs with spontaneous cancer. Nine dogs with soft tissue sarcoma were immunized with a recombinant human VEGF vaccine over a 16-week period. The effects of immunization on antibodies to human and canine VEGF, circulating VEGF concentrations, tumor microvessel density (MVD), and tumor growth were assessed. The xenogeneic VEGF vaccine was well-tolerated by all dogs and resulted in induction of humoral responses against both human and canine VEGF in animals that remained in the study long enough to receive multiple immunizations. Three of five multiply immunized dogs also experienced sustained decreases in circulating plasma VEGF concentrations and two dogs had a significant decrease in tumor MVD. The overall tumor response rate was 30% for all treated dogs in the study. We conclude therefore that a xenogeneic VEGF vaccine may be a safe and effective alternative means of controlling tumor growth and angiogenesis.