Phosphorylated LuxO activates transcription of five regulatory sRNAs (Qrr1-5), four of which, together with the chaperone Hfq, destabilize the mRNA for the master regulator LuxR. (C) In the presence of AIs, LuxO is dephosphorylated, and LuxR is produced. LuxR activates genes responsible for bioluminescence, biofilm formation and exoproteolytic activity,

and represses genes involved in type III secretion and siderophore production V. harveyi is an opportunistic pathogen mainly for shrimps, but also for fish, squids and lobsters [25–27] and causes #Emricasan ic50 randurls[1|1|,|CHEM1|]# major losses in shrimp aquaculture [28]. The response to QS signals is of interest in this context, because genes regulated by QS encode proteins required for biofilm formation [3]

and virulence factors, such as siderophores [29], type III secretion (e.g. vscP) [30] and exoproteolytic activity (e.g. vhp) [17, 31], in addition to bioluminescence (using the lux system) [32]. Here we focused on the single cell analysis of fluorescent reporter strains bearing plasmids containing promoter::gfp fusions, which allowed us to simultaneously monitor the expression of two AI-regulated genes in single cells. Results AI-regulated bioluminescence correlates well with the activity of the corresponding promoter::gfp fusion To expand our previous findings on heterogeneous behavior of a V. harveyi population found for bioluminescence [3] to other AI-regulated genes, we decided to construct promoter::gfp fusions. It was important to use a wild type DNA-PK inhibitor genetic background to monitor bioluminescence as a marker for an intact QS cascade in each strain. Therefore, all promoter::gfp fusions are plasmid based. To set up the reporter system we tested first a plasmid containing a promoter::gfp fusion of the constitutively expressed housekeeping gene recA to estimate the degree of heterogeneity

in the expression of this gene [33]. Wild type cells conjugated with this plasmid were grown to the exponential growth phase, stained with propidium iodide to identify dead cells (about 5%), and single cells in the same field of view were analyzed in phase contrast and fluorescence Glycogen branching enzyme modes. Images were analyzed using ImageJ. Luminescence and fluorescence intensities of each living cell are expressed as intensity values per cell after normalization to the same cell size. All living cells were fluorescent, indicating expression of recA in all cells. Fluorescence intensities were determined in about 1,400 cells. The average fluorescence intensity was calculated to be 1,017 a.u./cell [(a.u.) arbitrary units] with a standard deviation of 9.9% (data not shown). For comparison all living cells of strain BB120gfp containing a chromosomal encoded gfp were fluorescent and showed an average fluorescence intensity of 1,085 a.u./cell with a standard deviation of 10.5% (data not shown).

Next, we aligned all hits with MAFFT [43] and discarded those without sequence information for the YCYL or PAAP region and removed 100% identical sequences using Jalview [44], leaving us with a set of 286 WNV sequences for which we calculated the respective motif occurrences. The strain designations as listed in the alignment were taken from the NCBI taxonomy on West Nile viruses: http://​www.​ncbi.​nlm.​nih.​gov/​Taxonomy/​Browser/​wwwtax.​cgi?​id=​11082.

selleck chemical Several of these strains like selleck chemicals Sarafend belong to the pathogenic lineage 2. These are: West Nile virus H442, West Nile virus SA381/00, West Nile virus SA93/01, West Nile virus SPU116/89. Please note that the Kunjin virus has been recognized as WNV strain which is also visible by the identical sequences in the 2 displayed patterns. Acknowledgements We would like to thank Dr. Robert B. Tesh (University of Texas Medical Branch, Galveston) for kindly providing the WNV serum, Dr. Ted Pierson (NIAID) for the WNV constructs and the NIH AIDS research and reference reagent program for providing the HIV-Ig. References 1. Brinton MA: The molecular biology of West SN-38 purchase Nile Virus: a new invader of the western hemisphere. Annu Rev Microbiol 2002, 56:371–402.PubMedCrossRef 2. Lindenbach BD, Thiel HJ, Rice CM: Flaviviridae:

the viruses and their replication. Philadelphia, PA: Fields virology Lippincott William & Wilkins; 2007:1101–1152. 3. Calvert AE, Huang CY, Blair CD, Roehrig JT: Mutations in the West Nile prM protein affect VLP and virion secretion in vitro. Virology 2012, 433:35–44.PubMedCrossRef 4. Setoh YX, Prow NA, Hobson-Peters J, Lobigs M, Young PR, Khromykh AA, Hall RA: Identification of residues in West Nile virus

pre-membrane protein that influence viral particle secretion and virulence. J Gen Virol 2012, 93:1965–1975.PubMedCrossRef 5. Li J, Bhuvanakantham R, Howe J, Ng ML: Identifying the region influencing the cis-mode of maturation of West Nile (Sarafend) virus using chimeric infectious clones. Biochem Biophys Res Commun 2005, 334:714–720.PubMedCrossRef 6. Mackenzie JM, Westaway EG: Assembly and maturation of the flavivirus Kunjin virus appear Cetuximab molecular weight to occur in the rough endoplasmic reticulum and along the secretory pathway, respectively. J Virol 2001, 75:10787–10799.PubMedCrossRef 7. Mason PW: Maturation of Japanese encephalitis virus glycoproteins produced by infected mammalian and mosquito cells. Virology 1989, 169:354–364.PubMedCrossRef 8. Nowak T, Farber PM, Wengler G: Analyses of the terminal sequences of West Nile virus structural proteins and of the in vitro translation of these proteins allow the proposal of a complete scheme of the proteolytic cleavages involved in their synthesis. Virology 1989, 169:365–376.PubMedCrossRef 9. Garrus JE, von Schwedler UK, Pornillos OW, Morham SG, Zavitz KH, Wang HE, Wettstein DA, Stray KM, Cote M, Rich RL, et al.

coli strain DH5α by introduction of pMS2KI (lane 4 and lane 5). The presence of a 259-bp amplicon showed that caroS2I was transcribed constitutively (panel caroS2I in Figure 3). The caroS2I gene was transcribed unexpectedly in mutant strain TF1-2 even though the plasmid pMS2KI was introduced (lane 3). This demonstrated that caroS2I is expressed constitutively regardless of whether the gene caros2K is transcribed. Possibly an individual promoter for caroS2I gene

is located behind the Tn5 insertion site in the caroS2K gene. CaroS2I transcripts were detected in strain SP33 with plasmid pGS2I (lanes 6 and 7). Although both the SP33 strains (with or without pGEM T-easy) were susceptible to Carocin S2, SP33/pGS2I appeared to grow in the presence of CaroS2K 3-deazaneplanocin A mw (Figure 4B). Figure 4 Recovery and immunity activity of carocin S2. (A) Antibacterial activity of carocin S2 from different strains. The indicator was Pcc strain SP33. Strain number: 1, F-rif-18; 2, TF1-2; 3, TF1-2/pMS2KI; 4,

DH5α/pMS2KI; 5, DH5α. (B) Assay for caroS2I. The colony and inoculated strains were F-rif-18. The indicator strains were: 1, SP33; 2, this website SP33/pGEM-T easy; 3, SP33/pGS2I. To prove that pMS2KI contained the gene for Carocin S2, pMS2KI was introduced into TF1-2 and E. coli DH5α. Both TF1-2/pMS2KI and DH5α/pMS2KI had ability to express the activity of Carocin S2 (Figure 4A). The size of inhibition zone around strain TF1-2/pMS2KI was equal to that around DH5α/pMS2KI but still smaller than that around the wild-type strain F-rif-18. On the other hand, the quantity of transcripts expressed in vivo and in vitrodid not usually correspond. p53 activator Deduction of the amino acid sequence of PD-1 antibody Carocin S2 The carocin S2 gene consists of two ORFs (Additional file 1, Figure S7): one containing the 2352-bp caroS2K gene and the other containing the 273-bp caroS2I gene. The stop codon (TGA) of caroS2K overlaps the first start codon of caroS2I by 4-bp (ATGA). The amino sequences were deduced from the nucleotide sequence of the carocin S2 gene using DNASIS-Mac software (HITACHI, Japan) and compared to other analogous

proteins using the BLAST and FASTA search tools. ORF1 was found to encode a 783-amino acid protein with a high degree of homology to Pcc21 carocin D, Escherichia coli colicin D and Klebsiella oxytoca klebicin D (Figure 5); ORF2 was found to encode a 90-amino acid protein that shows homology to the immunity proteins of colicin D and klebicin D (Figure 5). Thus, caroS2K produces an antibiotic with a deduced molecular mass of 85 kDa. CaroS2I (a 10-kDa protein of 90 amino acids) was shown to confer resistance to CaroS2K. It is particularly noteworthy that the homology between CaroS2K and Colicin D and Klebicin D is at the C-terminal end of these proteins where the catalytic center of a ribonuclease is located.

iniae vaccine component. check details Conclusions In summary, this study buy GSK461364 presents MtsA as a novel solute-binding protein that can contribute to iron transport. This is the first ABC transporter member to be identified from S. iniae. We have shown that MtsA is a lipoprotein which can bind to heme, and is expressed in vivo during Kunming mice infection by S. iniae HD-1. More

importantly, this is the first report on the cloning of ABC transporter lipoprotein from S. iniae genomic DNA, and its immunogenicity is indicative of its possible use as an S. iniae subunit vaccine. Methods Bacterial strains and growth conditions Streptococcus iniae HD-1 was isolated from Threeband sweetlips (Plectorhynchus cinctus) from Guangdong province, PRC. The microorganism was stored in our lab and cultured according to the methods described by Zhou et al [45]. Briefly, S. iniae isolate HD-1 cells were grown in brain heart infusion broth (BHI, Oxoid Ltd.), and BHI broth with 1.5% agar (Guangdong Huankai Microbial Sci. & Tech, Co., Ltd.) was used as Blebbistatin molecular weight the solid medium. Escherichia coli DH5α and BL21 (DE3) strains (Beijing Newprobe Biotechnology Co., Ltd.) were used for gene

cloning and protein expression, respectively. Cloning and reverse transcription analysis of mtsABC Genomic DNA was extracted from the S. iniae HD-1 strain using the Wizard genomic DNA purification kit (Promega Co., Ltd.), as recommended by the manufacturer, Amylase and the material was quantified by measuring the absorbance at 260 nm. PCR was carried out with 1 μg of DNA using the primers listed in Additional file 1, Table S6. The primers were designed based on the conserved regions of the published amino acid sequence of metal ABC transporter (Additional file 1, Table S6-1), and the full-length product was obtained by SiteFinding-PCR (Additional file 1, Table S6-2, 6-3), as described by Tai et al [46]. The PCR products were sequenced

to rule out spurious mutations (Invitrogen Co., Ltd.). S. iniae HD-1 cells grow to the logarithmic phase were harvested by centrifugation, and total RNA was extracted by the Pure Yield™ RNA midiprep system (Promega, USA, Co., Ltd.). Total RNA was then incubated with RNase I at 37°C for 30 min to remove the contaminating genomic DNA. The material was quantified spectrophotometrically by ultraviolet absorption spectrometry (CE2302, Gene Quest), and its integrity was verified on a 0.8% agarose gel. First-strand cDNA was synthesized from 1 μg total RNA using the first-strand cDNA synthesis kit with ReverTra Ace-α-reverse transcriptase (Toyobo Co., Ltd.). The cDNA synthesized above was used as the template to amplify genes using the ORF-specific primers listed in Additional file 1, Table S7, and the PCR products were sequenced at Invitrogen Corporation to confirm their specificity. Expression of recombinant MtsA The genomic DNA of S.

Bronchoalveolar lavages Bronchoalveolar lavage (BAL) fluid was harvested as previously described [20]. Mice were euthanized by 4SC-202 in vitro injection of Pentobarbital (Sanofi Santé Animale, Libourne, France) and the respiratory

tract was exposed by dissection. A small incision was made near the top of the trachea, and a blunt-end 20-gauge needle was inserted and tied in place with surgical thread around the trachea. BAL www.selleckchem.com/products/jq-ez-05-jqez5.html fluid was obtained by 10 rounds of filling the lungs with 0.7 ml PBS and withdrawing as much of the liquid as possible. The samples were centrifuged to collect BAL fluid cells. BAL fluid cells were washed and resuspended in 1 ml PBS and aliquots were removed for counting with a hemocytometer and for cytospin centrifugation on a microscope slide, followed by DNA staining with Hoechst 33342 for identification of cell types. To determine the numbers of macrophages and neutrophils in the samples, 100 cells from several microscopy fields

were identified. Flow cytometry using macrophage buy Lenvatinib marker antibodies F4/80 (Miltenyi-Biotec, Bergisch Gladbach, Germany) and Gr-1 (Biolegend, San diego CA USA) was used to verify the extent of macrophage depletion within the BAL of clodrolip treated animals. Cell viability was evaluated using the trypan dye exclusion (Sigma-Aldrich). In vivo and in vitro imaging of bioluminescence Non-specific serine/threonine protein kinase Images were acquired using an IVIS 100 system according to the manufacturer’s instructions and as previously described [16]. In brief, 100 μl of PBS containing 3.33 mg D-luciferin was intraperitoneally injected in mice before each measurement. Mice were anesthetized using a constant flow of 2.5% isofluorane mixed with oxygen using an XGI-8 gas anesthesia system (Xenogen

Corporation). Images from mice were acquired 10 min after luciferin injection. Acquisition and quantification were performed using Living Image software version 3.1 (Xenogen Corporation). Quantification of photons per second emitted by each organ was performed by defining regions of interest corresponding to the respective organ of interest. The presence of A. fumigatus within the different organs was confirmed by histopathological analysis. For in vitro measurement of fungal germination within the BAL, D-luciferin in a final concentration of 10 mM was added directly to cells pelleted at the surface of chamber slides. The reaction was pre-incubated for 10 min at room temperature and measurement was performed with the IVIS 100 system. Determination of fungal DNA from infected lungs by quantitative real-time PCR A quantitative real-time PCR approach was selected to determine the fungal burden by quantification of the amount of fungal DNA among the total DNA isolated from lung tissues. The lung of a mouse not infected with A. fumigatus served as negative control.

Vector differences greater than 2 represent proteins with the highest change in expression, while vector differences less than 0.5 represent proteins with little statistical change Captisol mw in expression. This calculation allowed us to eliminate values of high change between exponential and stationary phase samples when variation between Selleckchem AZD4547 replicates was higher than that of the change in exponential vs stationary

phase samples. We propose that a vector difference of ≥ 0.5 as a confident change in expression between exponential and stationary phase proteins. Changes in protein expression levels were manually verified. Differences in protein expression between stationary and exponential phase cell-free extracts of core metabolic proteins Selleck RepSox are summarized in Table  1. A total of 166 of 252 encoded core metabolic proteins were detected using a combination of both shotgun and 4-plex acquisition methods. Twenty-four percent (24%) of proteins detected using 4-plex 2D-HPLC-MS/MS had a change in expression with a V diff greater than 0.5. Nineteen percent (19%) of these proteins increased during the transition

from exponential to stationary phase, while only 4% decreased in stationary phase, and 15% of these differentially expressed proteins changed by a magnitude greater than 1. Table 1 Protein detection using shotgun (single-plex) and iTRAQ labelled 4-plex 2D-HPLC-MS/MS and relative changes in protein expression levels Core metabolic protein categories Total genes Proteins detected Changes in protein levels (Stat/Exp)   1-Plex 4-Plex Total V diff  ≥ 0.5         Increased Decreased Non-catalytic cellulosomal proteins 8 5 6 7 0 0 Cellulosomal glycosidase 73 29 26 31 2 1 Non-cellulosomal glycosidases 35 17 13 19 3 0 RsgI-like σ-factors and anti-σI factors 9 3 2 3 0 0 Cello-oligosaccharide ABC transporters 14 9 8 10 2 1 Glycolysis 20 15 15 15 3 1 Pentose phosphate pathway 6 4 3 5 1 0 Energy storage 13 11 11 13 3 0 Pyruvate formation

from phosphoenolpyruvate 8 8 8 8 0 2 End-product synthesis from pyruvate 49 39 38 41 12 0 Energy generation 17 14 14 14 2 1 Total 252 154 144 166 28 6 Core metabolic proteins MycoClean Mycoplasma Removal Kit were classified into functional categories. The total number of protein encoding genes in each category and the number of corresponding proteins detected are provided. The number of proteins that changed during transition from exponential to stationary phase were listed only when their vector difference (V diff ) was greater than 0.5. Proteins detected can be viewed in Additional files 3 and 4. Central carbohydrate metabolism Global proteomic analysis is fundamental in verifying carbon utilization and end-product synthesis pathways. While mRNA expression profiles provide a great wealth of information with regards to transcriptional patterns, proteomics can rectify the discrepancy between transcription and translation.

Second (or step 2), a negative pulse is applied to create the conducting filament at LRS (approximately 20 kΩ). A negative forming voltage, which determines the conducting filament size, is reduced RepSox order from 2.6 to 1.1 V with a 100-ns pulse width. However, a conventional negative forming voltage (-2.6 V) is shown in blue line, this changes HRS (approximately 15 MΩ) to LRS (approximately 10 kΩ). Quantum-size effect and percolation models of RESET for different

switching materials have been explained to understand the conducting filaments [135, 136]. Another method of reducing CC can be used to control the conducting filament size, which can be achieved by adjusting the resistivity of the bulk TaO x layer. The resistivity can reduce the forming current by controlling the oxygen content of TaO x [120]. In this case, the conducting filament size becomes smaller and oxygen vacancy becomes larger when the oxygen content is increased. The observed switching is due to the change of barrier KU-57788 mw height on the application of voltage. When positive voltage was applied, O2- ions migrate from bulk and accumulate near the TE. Oxidation reaction increases the barrier height and device comes to the HRS. On the other hand, when negative voltage was applied on the TE, O2- ions move away from TE and reduction reaction lowers the barrier height which brings the device into LRS. Hence, the barrier height change

on the application of bias voltage due to redox reaction is responsible for the observed switching.

Several kinds of electrode materials were examined and found that the materials having high work function show stable resistance switching behavior. The significant check details improvement in the retention characteristics at 150°C under the small current operation of 80 μA by two-step forming are obtained as compared to single-step forming. Two-step electroforming process is very critical to have controlled conducting filament diameter as well as the RRAM could be Nepicastat chemical structure operated as low current at 80 μA. The W/TiO x /TaO x /W memory device showed good bipolar resistive switching characteristics with different CCs from 10 to 100 μA (Figure 12[41]). The low-resistance state decreases with increasing CCs from 10 to 100 μA (Figure 12a,b), which will be useful for multi-level data storage applications. As the filament diameter increases with higher CCs, the low-resistance state decreases, and the value of RESET voltage increases. The RESET current can be scaled down to 23 μA at a low CC of 10 μA. Figure 13a,b shows the device-to-device uniformity of LRS/HRS and SET/RESET voltage, respectively. The cumulative probability distribution is small for both LRS/HRS as well as set/reset voltage. The resistance ratio of HRS/LRS is >100, and the device can be operated below ±5 V. The device can be switched more than 104 AC cycles with stable LRS, as shown in Figure 14a.

Direct costs for internal procedures are mainly related to the gafchromic film. On average, direct and Pictilisib price indirect costs are 0,23 and 0,65 € per bag, respectively. The cost for personnel involved are; IRE technicians approx. 42 € per hour and Medical Physicist approx. 67 € per hour (data provided by the IRE Administration). The cost of internal

dosimetric verification is 1,00 €/bag. The list of costs for external and internal procedures is reported in Table 3 per bag. Table 3 Comparison of costs/bag irradiated with external and internal procedures   COSTS for External procedures (€/bag) COSTS for Internal procedures (€/bag) Indirect cost (§) 8 0,65 Direct cost (°) – 0,23 Technician (Transfusion Dep.) (°°) 20,44 8,54 Wortmannin Technician (Radiotherapy Dep.) (°°) – 0,63 Dosimetric verification (°°) – 1,00 Cost for one irradiation to be corresponded to External Institute 38 – Total cost for blood selleck products bag 66,44 11,05 Note: (§) assuming also the cost of LINAC

depreciation (100 €/h), the scanner depreciation (2 €/h); (°) including the cost of gafchromic films; (°°) see Table 1 and 2 for the time. The cost of the implementation of the internal procedure was 144,24 € and included the cost of the box and the treatment planning study. One thousand nine hundred and ninety six blood components were irradiated internally in the first year, so the overall savings to IFO was about € 110.558,44. All the blood component bags were transfused.

Discussion The procedure was developed, verified and has since been successfully implemented in the Transfusion, Fossariinae Medical Physics and Radiotherapy Departments, irradiating about two thousand blood components internally in the first year. The one-field irradiation procedure is much more easy to perform and time saving compared to other techniques reported in literature and based on LINAC [11–13]. There is no allowance for set-up error and the entire dose delivery procedure lasts only 3 minutes/box. The blood components are irradiated at the request of the Transfusion Department. The procedure is no longer carried out soley according to daily necessity but also on a regular weekly basis and stored for up to two weeks. The IRE procedure delivering a mean dose of 32 Gy (range: 27-35 Gy) is in accordance with the Italian Decree [14] and International Recommendations [3]. The gafchromic film, inserted into each box, is a visual reminder that the blood components have been irradiated, and the data analysis guarantees that the intended dose matches with that delivered. In fact, the gafchromic films serve multiple purposes: 1) to avoid a erroneous (no/duplicated) irradiation of the same box when multiple irradiations are programmed in the same session; 2) to measure the dose delivered to a particular reference point, close to the box top; 3) to implement a quality control programme of blood irradiation.

In other bacteria, like X. campestris, OhrR contains a second cysteine located on the COOH extremity of the OhrR protein (C127 for X. campestris). Oxidation of the protein initiates by the formation of a sulphenic derivative of the reactive cysteine (C22) followed by the formation of a disulfide

bond with C127 of the other OhrR subunit [30]. While ohr homologues are widely distributed in bacterial genomes [19], the role of ohr and ohrR was only studied in a few number of bacteria: X. campestris, B. subtilis, Agrobacterium tumefasciens, Pseudomonas aeruginosa and Streptomyces coelicolor this website [20, 31–35]. In many bacteria, peroxide stress was studied only via H2O2 stress. In S. meliloti, H2O2 resistance has been extensively studied [8, 10, 11] while OHP resistance is poorly understood. This study aims at evaluating the role of ohr and ohrR genes on OHP resistance in S. meliloti. The analysis of the biochemical properties of ohr and ohrR Pexidartinib in vivo mutants and the expression pattern suggests that this system should play an important role in sensing and protection of S. meliloti from OHPs. Results Identification of Ohr and OhrR homologues in S. meliloti Blast search of S. meliloti genome

for homologues of X. campestris Ohr protein revealed two paralogues, SMa2389 and SMc00040, showing 52 and 57% identity respectively with Ohr of X. campestris. They possess conserved active site cysteines of Ohr/OsmC proteins [19]. SMa2389 learn more is annotated as OsmC. SMc00040 has been shown to be induced by peroxide stress [11]; it is divergently located from a gene encoding a Cell Cycle inhibitor MarR family regulator that has 49 and 45% identity with the OhrR regulatory protein of X. campestris and B. subtilis respectively. SMc01945 has been previously published as OhrR like repressor since it presents 40% identity with OhrR of X. campestris [11]; the adjacent gene cpo (SMc01944) has been shown to encode a secreted peroxidase. Co-localisation on the genome of ohr and ohrR was found in all bacteria

in which these genes were investigated [20, 31, 36], suggesting that SMc00040 and SMc00098 encodes respectively Ohr and OhrR proteins. ohr mutant growth is inhibited by organic peroxides In order to investigate the role of ohr (SMc00040) and ohrR (SMc00098) in oxidative stress defence, S. meliloti strains with an ohrR deletion or carrying an insertion in ohr were constructed. The ability of these mutants to resist exposure to oxidants was evaluated; neither of the two had any growth defect when grown aerobically in complete medium LB or in minimal medium GAS. Moreover they possessed the same plating efficiency as wild type strain. The influence of organic peroxides on growth of wild type, ohr and ohrR strains was analysed by adding increasing amounts of t-butyl hydroperoxide (tBOOH) and cumene hydroperoxide (CuOOH) to LB medium and determining the maximal OD570 nm reached by the cultures.

Results As of 30 April 2006, 226 workers had died, adding 55 deaths to the earlier follow-up (see Table 1). Nine persons were lost to follow up, as they were not registered Berzosertib datasheet by the communal personal administration any more. Table 2 Cause-specific mortality in 570 workers exposed to dieldrin and aldrin stratified into three dose groups Cause of death Total group Low intake Moderate intake High intake Obs SMR (95% CI) Obs SMR (95% CI) Obs SMR (95% GS-4997 purchase CI) Flavopiridol (Alvocidib) Obs SMR (95% CI)

All causes 226 69.0* 60.3–78.7 59 75.1* 57.2–96.9 78 72.1* 57.0–90.0 89 67.0* 53.8–82.4 Neoplasms 82 76.4* 60.8–94.9 27 100.3 66.1–145.9 27 75.1 49.5–109.3 28 66.2* 44.0–95.6 Cardiovascular disease 80 59.9* 47.5–74.6 17 54.1* 31.5–86.6 30 67.6* 45.6–96.6 33 59.4* 40.9–83.4 find more Respiratory disease 20 74.3 45.4–114.7 5 87.3 28.5–204.9 5 56.0 18.2–130.7 10 84.4 40.5–155.3 Others causes 35 61.1* 42.6–85.0 7 50.2 20.2–103.4 14 76.7 42.0–128.8 14 63.0 34.4–105.7 Unknown 9     3     2     4     Neoplasms, cause specific 82     27     27     28      Oesophagus 4 159.3 43.4–407.9 2 286.5 34.7–1,035.1 1 116.6 3.0–649.4 1 107.5 2.7–599.1  Stomach and small intestine 8 96.0 41.5–189.2 5 249.3 80.9–581.7 2 75.5 9.0–269.2 1 30.0 0.8–167.1  Large intestine 7 96.7 38.9–199.2 1 54.6 1.4–304.0 2 81.9 9.9–296.0 4 139.5 38.0–357.1  Rectum 6 214.8 78.8–467.6 3 441.8 91.1–1,291.2 1 109.7 2.8–610.9 2 175.6 21.3–634.3  Liver and biliary passages 4 216.1 58.9–553.9 2 426.4 51.6–1540.5 2 322.6 39.1–1,165.3 0 0 0–414.4  Pancreas 3 66.5 13.7–194.3 1 86.4 2.2–481.6 0 0 0–197.1 2 113.0 13.7–408.2  Trachea and lung cancer 26 63.0* 41.1–92.3 7 66.7 26.8–137.1 12 85.9 44.4–150.0 7 43.3* 17.4–89.2  Skin 3 302.4 62.4–883.8 1 357.1 9.0–1,989.9 2 611.6 74.1–2,209.4 0 0 0–843.9  Kidney 2 69.8 8.5–252.2 0 0 0–392.1 0 0 0–307.9 2 184.7 22.4–667.1  Prostate cancer 5 55.3 18.0–129.2 2 102.9 12.5–371.6 1 32.8 0.8–182.