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Arctium lappa, commonly called burdock, has a long medicinal and edible history. It has recently gained increasing attention because of its economic value. In this study, we obtained the complete chloroplast genome of A. lappa by Illumina Hiseq. The complete chloroplast genome of A. lappa is a typical circular structure with 152 708 bp in length. The GC content in the whole chloroplast genome of A. lappa is 37.7%. A total of 37 tRNA genes, 8 rRNA genes, and 87 protein-coding genes were successfully annotated. And the chloroplast genome contains 113 unique genes, 19 of which are duplicated in the inverted repeat. The distribution of 39 simple sequence repeats was analysed, and most of them are in the large single-copy (LSC) sequence. An inversion comprising 16 genes was found in the LSC region, which is 26 283 bp long. We performed multiple sequence alignments using 72 common protein-coding genes of 29 species and constructed a Maximum Parsimony (MP) tree. The MP phylogenetic result shows that A. lappa grouped together with Carthamus tinctorius, Centaurea diffusa, and Saussurea involucrata. The chloroplast genome of A. lappa is a valuable resource for further studies in Asteraceae. Key words: Arctium lappa, chloroplast genome, molecular structure, phylogenetic relationship. L'Arctium lappa, ou grande bardane, est connue depuis longtemps comme plante medicinale et comestible. Elle a recemment suscite une attention croissante en raison de sa valeur economique. Dans cette etude, les auteurs ont decode le genome chloroplastique complet de l'A lappa par sequencage Illumina Hiseq. Le genome chloroplastique complet de l A. lappa presente une structure circulaire typique comprenant 152 708 pb. Le contenu en GC est de 37,7 %. Au total, 37 genes d'ARNt, 8 genes d'ARNr et 87 genes codant pour des proteines ont ete annotes. Le genome chloroplastique compte 113 genes uniques, dont 19 sont dupliques au sein de la region inversee repetee. La distribution de 39 microsatellites a ete analysee et la plupart se trouvent au sein de la grande region a simple copie (LSC). Une inversion comprenant 16 genes et mesurant 26 283 pb a ete detectee au sein de la region LSC. Les auteurs ont realise des alignements multiples sur 72 genes communs codant pour des proteines chez 29 especes et produit un arbre phylogenetique par une approche de parcimonie maximale (MP). Le resultat de cette analyse phylogenetique a montre que l'A. lappa appartenait au meme groupe que le Carthamus tinctorius, le Centaurea diffusa et le Saussurea involucrata. Le genome chloroplastique de l'A. lappa constitue une ressource utile pour de futures etudes chez les Asteraceae. [Traduit par la Redaction] Mots-cles : Arctium lappa, genome chloroplastique, structure moleculaire, relation phylogenetique., Introduction Arctium lappa, a biennial herb of Asteraceae, is commonly known as a medicinal plant in many parts of the world, and it has been used as a therapeutic plant [...]

Yttria-stabilized zirconia (YSZ, Y2O3-ZrO2) ceramic with titanium dioxide nanoparticle (nanoTiO2) is prepared using conventional pressureless and two-step sintering (TSS) methods. The microstructure, phase composition, and mechanical properties of TSS-TiO2-ZrO2-Y2O3 nanocomposite ceramics are compared to those of the ceramics sintered using conventional sintering. The effect of TSS and nanoparticles on the sintering densification of the YSZ ceramics is investigated. X-ray diffractometry revealed that nanoTiO2 destabilizes c-ZrO2 and stabilizes t-ZrO2 in the YSZ. In addition, TSS is more conducive for grain refinement and sintering densification of the TiO2-ZrO2-Y2O3 nanocomposite ceramics; the average grain size of the samples decreased from 1.446 (conventional sintering) to 0.492 μm (TSS). Meanwhile, the relative density and sintering shrinkage of the TSS-1450 sample reached the maximum (98.23% and 22.92%, respectively) at a nanoTiO2 fraction of 7 wt%. The maximum microhardness and bending strength of the TSS-1450 ceramic matrix with 7-wt% nanoTiO2 were 1823 HV and 505 MPa, respectively. The nanoparticles distributed in the grain boundary induce the crack to the matrix, which induces the transgranular fracture, and nanoTiO2 improves the densification and mechanical properties of YSZ ceramics more than titanium dioxide microparticles. [ABSTRACT FROM AUTHOR]

After solutionizing and aging at 980°C for various durations, the compressive deformation behaviour and mechanisms of the directionally solidified nickel-based superalloy CM247LC are investigated at room temperature. Experimental results show that the yield strength decreases gradually from 991 ± 1.2–672 ± 7.5 MPa with increasing the γ′ precipitate size from 145 ± 2–604 ± 19 nm. Transmission electron microscope observations on the slightly deformed specimens reveal that, in contrast to previous researches, plastic deformation of all the specimens is controlled by strongly coupled dislocations cutting through γ′ precipitates. Based on these experimental results, the relationship between the yield strength and precipitate size is discussed. [ABSTRACT FROM AUTHOR]

Bidens plants are annuals or perennials of Asteraceae and usually used as medicinal materials in China. They are difficult to identify by using traditional identification methods because they have similar morphologies and chemical components. Universal DNA barcodes also cannot identify Bidens species effectively. This situation seriously hinders the development of medicinal Bidens plants. Therefore, developing an accurate and effective method for identifying medicinal Bidens plants is urgently needed. The present study aims to use phylogenomic approaches based on organelle genomes to address the confusing relationships of medicinal Bidens plants. Illumina sequencing was used to sequence 12 chloroplast and eight mitochondrial genomes of five species and one variety of Bidens. The complete organelle genomes were assembled, annotated and analysed. Phylogenetic trees were constructed on the basis of the organelle genomes and highly variable regions. The organelle genomes of these Bidens species had a conserved gene content and codon usage. The 12 chloroplast genomes of the Bidens species were 150,489 bp to 151,635 bp in length. The lengths of the eight mitochondrial genomes varied from each other. Bioinformatics analysis revealed the presence of 50–71 simple sequence repeats and 46–181 long repeats in the organelle genomes. By combining the results of mVISTA and nucleotide diversity analyses, seven candidate highly variable regions in the chloroplast genomes were screened for species identification and relationship studies. Comparison with the complete mitochondrial genomes and common protein-coding genes shared by each organelle genome revealed that the complete chloroplast genomes had the highest discriminatory power for Bidens species and thus could be used as a super barcode to authenticate Bidens species accurately. In addition, the screened highly variable region trnS-GGA-rps4 could be also used as a potential specific barcode to identify Bidens species. [ABSTRACT FROM AUTHOR]

Small-conductance [Ca.sup.2+]-activated [K.sup.+] channels (SK channels, [K.sub.Ca], channels) have been reported in excitable cells, where they aid in integrating changes in intracellular [Ca.sup.2+] with membrane potential. We recently reported for the first time the functional existence of SK2 ([K.sub.Ca]2.2) channels in human and mouse cardiac myocytes. Here, we report cloning of SK1 ([K.sub.Ca]2.1) and SK3 ([K.sub.Ca]2.3) channels from mouse atria and ventricles using RT-PCR. Full-length transcripts and their variants were detected for both SK1 and SK3 channels. Variants of mouse SK1 channel (mSK1) differ mainly in the COOH-terminal structure, affecting a portion of the sixth transmembrane segment (S6) and the calmodulin binding domain (CaMBD). Mouse SK3 channel (mSK3) differs not only in the number of polyglutamine repeats in the N[H.sub.2] terminus but also in the intervening sequences between the polyglutamine repeats. Full-length cardiac mSK1 and mSK3 show 99 and 91% nucleotide identity with those of mouse colon SK1 and SK3, respectively. Quantification of SK1, SK2, and SK3 transcripts between atria and ventricles was performed using real-time quantitative RT-PCR from single, isolated cardiomyocytes. SKI transcript was found to be more abundant in atria compared with ventricles, similar to the previously reported finding for SK2 channel. In contrast, SK3 showed similar levels of expression in atria and ventricles. Together, our data are the first to indicate the presence of the three different isoforms of SK channels in heart and the differential expression of SK1 and SK2 in mouse atria and ventricles. Because of the marked differential expression of SK channel isoforms in heart, specific ligands for [Ca.sup.2+]-activated [K.sup.+] currents may offer a unique therapeutic opportunity to modify atrial cells without interfering with ventricular myocytes. calcium-activated potassium current; mouse cardiac myocyte

Titanium alloy is an important metal material with excellent specific strength, which is widely used in aerospace field, nuclear industry, chemical medicine, and military industry. In order to investigate the connection conditions of TC4 titanium alloy and 316L stainless steel at different temperatures, the braze welding measurement with Ti57Zr13Cu21Ni9 filler metal was conducted in vacuum. The microstructure, morphology and phase of the joint were characterized by SEM (scanning electron microscope), EDS (Energy Dispersive Spectrometer) and XRD (X-ray diffraction), respectively. Microhardness and shear strength of the joint at room temperature and the bonding mechanism of TC4 and 316L were also investigated. The obtained results revealed that the main phases in the diffusion layer were Ti-based solid solution and Ti-Fe (TiFe and TiFe2) intermetallic compoundsands (IMCs) the center of the braze was mainly composed of Ti-Fe IMCs, (Ti, Zr)2(Ni, Cu), Ti-based solid solution. Additionally, the increase of brazing temperature firstly increased and then decreased the average shear strength with the maximum value of 133.9 MPa at 960 °C. [ABSTRACT FROM AUTHOR]

Vacuum brazing of TC4 titanium alloy and 316L stainless steel was conducted using Ag70.5Cu27.5Ti2 filler metal, the effects of microstructure and mechanical properties of brazing temperature on interface were investigated, and the bonding mechanism of brazed joint was also established. The results show that the filler metal and the base metal on both sides were completely diffused and bonded well. The microstructure of brazed joint interface is as follows: TC4 titanium alloy/diffusion layer(α-Ti + Ti2Cu)/reaction layer (including TiCu and Ti2Cu)/joint center area/TiFe layer/316L stainless steel. When the holding time lasted for 15 min, the shear strength of brazed joint first increased and then decreased as the brazing temperature rose. When the brazing temperature upped to 870 °C, the maximum of shear strength reached 190 MPa, and then the fracture occurred in the center of the brazed joint. The hardness of the joints gradually increased as the temperature rose, and the mean value reached maximum of 323 HV at 930 °C. [ABSTRACT FROM AUTHOR]

Abstract Background Cancers arise through an evolutionary process in which cell populations are subjected to selection; however, to date, the process of bladder cancer, which is one of the most common cancers in the world, remains unknown at a single-cell level. Results We carried out single-cell exome sequencing of 66 individual tumor cells from a muscle-invasive bladder transitional cell carcinoma (TCC). Analyses of the somatic mutant allele frequency spectrum and clonal structure revealed that the tumor cells were derived from a single ancestral cell, but that subsequent evolution occurred, leading to two distinct tumor cell subpopulations. By analyzing recurrently mutant genes in an additional cohort of 99 TCC tumors, we identified genes that might play roles in the maintenance of the ancestral clone and in the muscle-invasive capability of subclones of this bladder cancer, respectively. Conclusions This work provides a new approach of investigating the genetic details of bladder tumoral changes at the single-cell level and a new method for assessing bladder cancer evolution at a cell-population level.

Abstract Background Earlier studies have reported the production of histamine in colorectal cancers (CRCs). The effect of histamine is largely determined locally by the histamine receptor expression pattern. Recent evidence suggests that the expression level of histamine receptor H4 (HRH4) is abnormal in colorectal cancer tissues. However, the role of HRH4 in CRC progression and its clinical relevance is not well understood. The aim of this study is to evaluate the clinical and molecular phenotypes of colorectal tumors with abnormal HRH4 expression. Methods Immunoblotting, real-time PCR, immunofluorescence and immunohistochemistry assays were adopted to examine HRH4 expression in case-matched CRC samples (n = 107) and adjacent normal tissues (ANTs). To assess the functions of HRH4 in CRC cells, we established stable HRH4-transfected colorectal cells and examined cell proliferation, colony formation, cell cycle and apoptosis in these cells. Results The protein levels of HRH4 were reduced in most of the human CRC samples regardless of grade or Dukes classification. mRNA levels of HRH4 were also reduced in both early-stage and advanced CRC samples. In vitro studies showed that HRH4 over-expression caused growth arrest and induced expression of cell cycle proteins in CRC cells upon exposure to histamine through a cAMP -dependent pathway. Furthermore, HRH4 stimulation promoted the 5-Fu-induced cell apoptosis in HRH4-positive colorectal cells. Conclusion The results from the current study supported previous findings of HRH4 abnormalities in CRCs. Expression levels of HRH4 could influence the histamine-mediated growth regulation in CRC cells. These findings suggested a potential role of abnormal HRH4 expression in the progression of CRCs and provided some new clues for the application of HRH4-specific agonist or antagonist in the molecular therapy of CRCs.

The protein phosphatase 2C (PP2C) gene family, which is known to participate in cellular processes, is one of the most conserved plant-specific gene families that regulate signal transduction in eukaryotic organisms. However, the PP2C gene family of Glycyrrhiza uralensis Fisch. has not yet been systematically characterized. In this study, 76 PP2C genes of G. uralensis (GuPP2Cs) were identified. Phylogenetic analysis divided these GuPP2Cs into 12 subfamilies. The gene structure and protein motifs of the GuPP2Cs were then analysed. The analysis of the transcriptomic data of the GuPP2Cs revealed that nine genes were highly sensitive to abscisic acid (ABA) stress and might respond to ABA treatment. These nine genes were validated via quantitative real-time polymerase chain reaction. This study provides a foundation for future studies on the functional PP2C genes of G. uralensis and the improvement of the cultivation and quality of G. uralensis. [ABSTRACT FROM AUTHOR]

Ceramic coatings containing calcium and phosphorus were prepared on the surface of Ti-3Zr-2Sn-3Mo-25Nb by micro-arc oxidation. The effects of Ca/P ratio, oxidation time, current density and duty ratio on the coating were studied by orthogonal experiment under a phosphate electrolyte to determine the optimal process parameters. To analyse the effects of NaF additions, 1 g L−1, 2 g L−1, 3 g L−1, 4 g L−1 and 5 g L−1 NaF were added into the electrolyte, respectively. From analysis, the authors draw the following conclusions: NaF can improve the conductivity of electrolyte and facilitate the densification of coating; the hardness, thickness and roughness of the coating have been improved to some extent; the most obvious improvement to the coating is effected by adding 3 g L−1 NaF into the electrolyte. Furthermore, passivation has occurred on the MAO-NaF-coatings immersed in simulated body fluids. The most improved corrosion resistance of the MAO-oxidation coating is gained by adding 3 g L−1. [ABSTRACT FROM AUTHOR]

Analysis of the relationships among wild species of section Moutan in the plant genus Paeonia has traditionally been problematic. Interspecies relationships cannot be effectively determined using phenotypic traits alone or through analysis of nuclear or chloroplast DNA fragments. Elucidation of complete chloroplast genome sequences will aid the identification and phylogeny of these species. In this study, the complete chloroplast genomes of three sect. Moutan plants were sequenced and analyzed. Comparative and phylogenetic analyses of the complete chloroplast genomes of all eight species of sect. Moutan were then conducted. The three complete chloroplast genomes gained in this study showed four-part annular structures, and the genome length, structure, GC content, codon usage, and gene distribution were highly similar. There was greater variation in the noncoding regions of the sequences than in the conserved protein-coding regions. Sequence variations in the small single copy (SSC) regions and large single copy (LSC) regions were considerably greater than those in the inverted repeat (IR) regions. Phylogenetic analysis revealed that the species of sect. Moutan clustered in one branch and then subdivided into smaller branches. As for the three complete chloroplast genome sequences obtained in this study, Paeonia jishanensis clustered with another P. jishanensis sequence from the GenBank database, Paeonia qiui clustered with Paeonia rockii , and Paeonia delavayi var. lutea clustered with Paeonia ludlowii. It was also found that the complete chloroplast genomes, LSC regions, and SSC regions all showed great abilities in identification and phylogenetic analysis of the species of sect. Moutan , while IRs regions and highly variable regions were not suitable for the species of sect. Moutan. [ABSTRACT FROM AUTHOR]

Ephedrae Herba and Ephedrae Radix et Rhizoma (Mahuang) have been used as Chinese herbal medicines. Ephedra plants mainly live in deserts and have good governance of desertification. Despite their important medicinal and environmental protection value, dietary supplements containing ephedrine from Ephedra species may threaten the health of people. Morphological resemblance amongst species causes difficulty in identifying the original species of Ephedra herbs. Chloroplast (CP) genome shows good prospects in identification and phylogenetic analysis. This study introduced the structures of the CP genomes of three Ephedra species and analysed their phylogenetic relationships. Three complete CP genomes of Ephedra showed four-part annular structures, namely, two single-copy regions and two inverted repeat regions. The entire CP genomes of three Ephedra species in terms of size were 109,550 bp (E. sinica), 109,667 bp (E. intermedia), and 109,558 bp (E. equisetina). Each CP genome of the three Ephedra species encoded 118 genes, including 73 protein-coding genes, 37 tRNA genes and 8 ribosomal RNA genes. Eleven high-variation regions were screened through mVISTA to be potential specific DNA barcodes for identifying Ephedra species. Maximum likelihood and maximum parsimony trees showed that CP genomes could be used to identify Ephedra species. The Ephedra species had a close phylogenetic relationship with Gnetum species and Welwitschia mirabilis. This research provided valuable information for the identification and phylogenetic analysis of gymnosperms and drug safety of Ephedra. [ABSTRACT FROM AUTHOR]

The present study aimed to observe the identification of biomarkers of silicosis based on the differentially expressed serum proteins between normal healthy individuals and patients with silicosis fibrosis. A total number of 20 patients with clinically diagnosed silicosis were screened, which were designated as the foundation treatment group. In addition, 20 age-matched healthy patients attending a check-up at the physical examination department were selected. Serum samples were obtained and a combined protein chip with surface-enhanced laser desorption ionization flight mass spectrometry was applied to perform serum analysis. Data preprocessing, screening differences in peak, hierarchical cluster analysis, Principal Component Analysis, construction of a decision tree model, and prediction based on the differences between peaks corresponding to proteins were performed to analyze the data. The results revealed differences in the proteins in serum between the normal group and the group prior to foundation treatment prediction. The corresponding names of the protein peak, predicted protein, and gene name were as follows: M1948_00, complement c3 frag, C3; M2017_02, amyloid-βa4 protein, APP; and M2879_56, hepcidin, HAMP. Differentially expressed serum proteins in the normal group and the basis treatment group were predicted, including M2017_02, amyloid-βa4 protein, APP; M2879_56, hepcidin, HAMP; and M3224_97, fibrinogen-α chain frags, FGA. The differentially expressed serum proteins in the group prior to basis treatment and the group following basis treatment were predicted, including M2001_69, amyloid-βa4 protein, APP; M2017_02, amyloid-βa4 protein, APP, M4144_81, plasma protease c1 inhibitor frag, and SERPING1. In conclusion, there were differences in the proteins in serum between the patients with silicosis fibrosis and healthy individuals. [ABSTRACT FROM AUTHOR]

Objective. To observe the curative effect of VAWI on Xinjiang Uygur patients with silicosis fibrosis. Methods. After we diagnosed the 40 patients with the first phase of silicosis, we randomly divided them into two groups: the basic treatment group (group A, n=20) and the VAWI group (group B, n=20). At the same time, we selected the age-matched healthy patients (n=20). We applied the combined protein chip with SELDI-TOF-MS to carry out the serum analysis. The data were analyzed throughout data preprocessing, difference in PEAK screening, hierarchical cluster analysis, and Principal Component Analysis (PCA). We built decision tree model and predict the difference between the PEAK corresponding proteins. Results. The proteins peaks corresponding to name, predicted protein, and gene name were as follows: M2001_69, amyloid beta a4 protein, APP, and M2017_02, amyloid beta a4 protein, APP. The different expression of proteins in patients with silicosis was found before and after with VAWI treatment. The predicted proteins were as follows: M1982_50, amyloid beta a4 protein, APP; M3164_50, fibrinogen alpha chain frag, FGA; M3379_28, fibrinogen alpha chain frag, FGA; and so on. Conclusion. VAWI presented curative effect on patients with silicosis fibrosis via the alternation of proteins expression in serum. [ABSTRACT FROM AUTHOR]

Human CMTM3 has been proposed as a putative tumor suppressor gene. The loss of CMTM3 has been found in several carcinomas. However, the regulation of CMTM3 expression and its function in tumor progression remain largely unknown. Here, we investigated the regulation of CMTM3 expression, function and molecular mechanism in human testicular cancer cells. CMTM3 was frequently downregulated or silenced in testicular cancer cell lines and tumor tissues but highly expressed in normal testis tissues. The re-expression of CMTM3 significantly suppressed the colony formation, proliferation, and migration capacity of testicular cancer cells by inducing a G2 cell cycle arrest and apoptosis. Moreover, the re-expression of CMTM3 activated the transcription of p53, induced p53 accumulation, up-regulated the expression of p21, and increased the cleavage of caspase 9, 8, 3, and PARP. The downregulation of CMTM3 in clinical tumor tissues was associated with the methylation of a single CpG site located within the Sp1/Sp3-responsive region of the core promoter. These results indicate that CMTM3 can function as tumor suppressor through the induction of a G2 cell cycle arrest and apoptosis. CMTM3 is thus involved in testicular cancer pathogenesis, and it is frequently at least partially silenced by the methylation of a single, specific CpG site in tumor tissues. [ABSTRACT FROM AUTHOR]

We reported the complete chloroplast genome sequences of Rhus potaninii which was characterized by de novo assembly with Illumina sequencing data. The size of R. potaninii complete chloroplast genome is 159,620 bp in length and includes a large single copy region of 87,722 bp, a small single copy region of 18,948 bp, and a pair of inverted repeats of 26,475 bp. Its GC content is 37.9%. A total of 133 genes were predicted, including 86 protein-coding genes, 8 rRNA genes, 37 tRNA genes, and 2 pseudogenes. Maximum-likelihood (ML) phylogenetic tree indicates that R. potaninii is sister to R. chinensis. [ABSTRACT FROM AUTHOR]

Loss-of-function mutations in the KCNQ4 channel cause DFNA2, a subtype of autosomal dominant non-syndromic deafness that is characterized by progressive sensorineural hearing loss. Previous studies have demonstrated that the majority of the pathogenic KCNQ4 mutations lead to trafficking deficiency and loss of KCNQ4 currents. Over the last two decades, various strategies have been developed to rescue trafficking deficiency of pathogenic mutants; the most exciting advances have been made by manipulating activities of molecular chaperones involved in the biogenesis and quality control of the target protein. However, such strategies have not been established for KCNQ4 mutants and little is known about the molecular chaperones governing the KCNQ4 biogenesis. To identify KCNQ4-associated molecular chaperones, a proteomic approach was used in this study. As a result, two major molecular chaperones, HSP70 and HSP90, were identified and then confirmed by reciprocal co-immunoprecipitation assays, suggesting that the HSP90 chaperone pathway might be involved in the KCNQ4 biogenesis. Manipulating chaperone expression further revealed that two different isoforms of HSP90, the inducible HSP90α and the constitutive HSP90β, had opposite effects on the cellular level of the KCNQ4 channel; that HSP40, HSP70, and HOP, three key components of the HSP90 chaperone pathway, were crucial in facilitating KCNQ4 biogenesis. In contrast, CHIP, a major E3 ubiquitin ligase, had an opposite effect. Collectively, our data suggest that HSP90α and HSP90β play key roles in controlling KCNQ4 homeostasis via the HSP40-HSP70-HOP-HSP90 chaperone pathway and the ubiquitin-proteasome pathway. Most importantly, we found that over-expression of HSP90β significantly improved cell surface expression of the trafficking-deficient, pathogenic KCNQ4 mutants L274H and W276S. KCNQ4 surface expression was restored by HSP90β in cells mimicking heterozygous conditions of the DFNA2 patients, even though it was not sufficient to rescue the function of KCNQ4 channels. [ABSTRACT FROM AUTHOR]

A systematic review of the Chinese literature was conducted from 1956 to 2005. The survey included both online and manual searching, as well as expert discussions aimed at providing insight into factors affecting benzene exposure levels in paint/coatings industries. Data extracted from 204 papers included: (1) year of occurrence, (2) type of paint/coatings products, (3) type of industries where the products were used or produced, (4) job titles and work activities, (5) type of literature searched, (6) working conditions whenever data were available, and (7) exposure levels. Most benzene measurements were short-term samples for comparison with the Chinese maximum allowable concentration standard. The accuracy and precision of the sampling and analytical methods were not reported. The distribution of benzene concentrations was tested and found to fit neither normal nor lognormal distributions. Analysis of variance (comparison for more than two groups) and t-test (comparison for two groups) were conducted on Blom-transformed benzene concentration data. The overall median benzene exposure levels were 215, 82, 31, and 6 mg/m3 during the periods 1956-1978, 1979-1989, 1990-2001, and 2002-2005, respectively. Mean benzene exposure was significantly lower for paint manufacturing than paint spraying. No significant difference was found among paint types and benzene exposure for paint application. Benzene exposure was significantly higher in workplaces judged to have poor ventilation. No significant differences were found in benzene exposure as a function of industry type. Even though substantially lower when compared with levels in the past, recent benzene exposure measurements suggested that many facilities in the paint/coatings industries in China still have benzene concentrations that are above the current China occupational exposure limit for benzene (6 mg/m3 as a time-weighted average). Benzene concentrations from the present exercise, while not directly supporting quantitative retrospective exposure estimating, provide insight on relative benzene exposure for painting tasks in the reported industries over time. [Supplemental materials are available for this article. Go to the publisher's online edition of the Journal of Occupational and Environmental Hygiene for the following free supplemental resource: tables detailing benzene concentration for different types of painting and other tasks in Chinese industry.] [ABSTRACT FROM AUTHOR]

An outcome of the intricate K+ regulation in the cochlear duct is the endocochlear potential (EP), ∼80 mV, the "battery" that runs hair-cell transduction; however, the detailed molecular mechanisms for the generation of the EP remain unclear. We provide strong evidence indicating that the intermediate cells (ICs) of the stria vascularis (StV) express outward K+ current that rectifies inwardly at positive potentials. The channel belongs to the ether-a-go-go-related gene (erg) family of K+ channels. We cloned an ERG1a channel in the mouse inner ear (MERG1a). The cellular distribution of MERG1a in the cochlea displayed the highest levels of immunoreactivity in the ICs and modest reactivity in the marginal cells as well as in several extrastrial cells (e.g., hair cells). Functional expression of the StV-specific MERG1a channel reveals a current that activates at relatively negative potentials (approximately -50 mV) and shows rapid inactivation reflected as inward rectification at depolarized potentials. The current was sensitive to the methanesulfonanilide drug E-4031 (IC50, ∼165 nM) and the recombinant peptide rBeKm-1 (IC50, ∼16 nM), and the single-channel conductance in symmetrical K+ was ∼14 pS. The site of expression of MERG1a and its functional phenotype (e.g., modulation of the current by external K+) make it one of the most likely candidates for establishing the high throughput of K+ ions across ICs to generate EP. In addition, the property of the channel that produces marked K+ extrusion in increased external K+ may be important in shaping the dynamics of K+ cycling in the inner ear. [ABSTRACT FROM AUTHOR]

Multiple Ca2+ channels confer diverse functions to hair cells of the auditory and vestibular organs in the mammalian inner ear. We used gene-targeting technology to generate alpha1D Ca2+ channel-deficient mice to determine the physiological role of these Ca2+ channels in hearing and balance. Analyses of auditory-evoked brainstem recordings confirmed that alpha1D-/- mice were deaf and revealed that heterozygous (alpha1D+/-) mice have increased hearing thresholds. However, hearing deficits in alpha1D+/- mice were manifested mainly by the increase in threshold of low-frequency sounds. In contrast to impaired hearing, alpha1D-/- mice have balance performances equivalent to their wild-type littermates. Light and electron microscope analyses of the inner ear revealed outer hair cell loss at the apical cochlea, but no apparent abnormality at the basal cochlea and the vestibule. We determined the mechanisms underlying the auditory function defects and the normal vestibular functions by examining the Ba2+ currents in cochlear inner and outer hair cells versus utricular hair cells in alpha1D+/- mice. Whereas the whole-cell Ba2+ currents in inner hair cells consist mainly of the nimodipine-sensitive current (approximately 85%), the utricular hair cells express only approximately 50% of this channel subtype. Thus, differential expression of alpha1D channels in the cochlear and utricular hair cells confers the phenotype of the alpha1D null mutant mice. Because vestibular and cochlear hair cells share common features and null deletion of several genes have yielded both deafness and imbalance in mice, alpha1D null mutant mice may serve as a model to disentangle vestibular from auditory-specific functions. [ABSTRACT FROM AUTHOR]

Functional interactions between ligand-gated, voltage-, and Ca(2+)-activated ion channels are essential to the properties of excitable cells and thus to the working of the nervous system. The outer hair cells in the mammalian cochlea receive efferent inputs from the brain stem through cholinergic nerve fibers that form synapses at their base. The acetylcholine released from these efferent fibers activates fast inhibitory postsynaptic currents mediated, to some extent, by small-conductance Ca(2+)-activated K+ channels (SK) that had not been cloned. Here we report the cloning, characterization, and expression of a complete SK2 cDNA from the mouse cochlea. The cDNAs of the mouse cochlea alpha9 and alpha10 acetylcholine receptors were also obtained, sequenced, and coexpressed with the SK2 channels. Human cultured cell lines transfected with SK2 yielded Ca(2+)-sensitive K+ current that was blocked by dequalinium chloride and apamin, known blockers of SK channels. Xenopus oocytes injected with SK2 in vitro transcribed RNA, under conditions where only outward K+ currents could be recorded, expressed an outward current that was sensitive to EGTA, dequalinium chloride, and apamin. In HEK-293 cells cotransfected with cochlear SK2 plus alpha9/alpha10 receptors, acetylcholine induced an inward current followed by a robust outward current. The results indicate that SK2 and the alpha9/alpha10 acetylcholine receptors are sufficient to partly recapitulate the native hair cell efferent synaptic response. [ABSTRACT FROM AUTHOR]

A partial DNA library of Streptomyces ansochromogenes 7100 was constructed by using plasmid pl J702 as vector and white mutant W19 as recipient. About 3 000 clones were obtained, two of which gave rise to the grey phenotype as wild type 7100. The plasmids were isolated from two transformants. The result indicated that the 5.2 kb and 5.8 kb DNA fragments were inserted into plJ702. The resulting recombinant plasmids were designated as pNL-1 and pNL-2 respectively. The 1.25 kb Pstl I - Apa I DNA fragment from pNL-1 was recognized as its complementarity to W19 strain. The nucleotide sequence of the 3.0 kb Pst I DNA fragment including 1.25 kb was determined and analyzed. The result indicated that this DNA fragment contains one complete open reading frame (ORF1) which encodes a protein with 295 amino acid residues, and this gene was designated as sawB. The deduced protein has 81% amino acid identities in comparison with that encoded by whiH in Streptomyces coelicolor. The function of sawB gene was studied by using strategy of gene disruption, and the resulting sawB mutant failed to form spores and produced loosely coiled aerial hyphal. The result showed that sawB is closely related to hyphal coiling and sporulation in S. ansochromogenes, and also indicated that the sawB can complement whiH mutant (C119) to restore the grey phenotype of Streptomyces coelicolor J 1501 (wild type). [ABSTRACT FROM AUTHOR]

Several genetically stable mutants blocked in nikkomycin biosynthesis were obtained after the slightly germinated spores of Streptomyces ansochromogenes, a nikkomycin producer, were treated with ultra violet radiation. One of the mutants is the same in morpholotical differentiation as the wild type strain and is designated as NBB19. A DNA library was constructed using plasmid pIJ702 as cloning vector, NBB19 as cloning recipient. A 6 kb DNA fragment which can genetically complement NBB19 was cloned when screening the library for antifungal activity. Sequence analysis showed that the 3 kb Bgl II - Sal I fragment contains one complete ORF (ORF1) and one partial ORF (ORF2). ORF1 is designated as sanA. sanA is 1 365 bp, encoding a protein consisting of 454 amino acid residues. Database searching indicated that sanA is homologous to the hypothetical methyltransferase in Pyrococcus horikoshii with 25% identities and 41% positives. Disruptant of sanA lost the ability to synthesize nikkomycin. It indicated that sanA is a novel gene which is essential for nikkomycin biosynthesis. [ABSTRACT FROM AUTHOR]