Vertical black lines indicate cos and attP sites respectively. Red arrows indicate tRNA genes. Pseudogenes are marked with PI3K Inhibitor Library a black asterisk. Below the scale, arrows represent homologous proteins of bacteriophages and prophages from

different hosts with S. flexneri, E. coli and Salmonella framed within a green, red or blue box, respectively. Homologs between SfI and other phages/prophages are shown in different colors with color coding corresponding to level of homology at amino acid level, with red of 100% identity and blue of > =50% identity. Phage SfI has a very narrow host range Host specificity of serotype-converting bacteriophages has long been recognized, which results in the specific lytic spectrum and serotype conversion of S. flexneri in nature [20]. The recognition between the O-antigen of host bacterium and the tail component of a phage is the key mechanism of host specificity [20]. To determine the host range of SfI, 132 S. flexneri Selleckchem Daporinad strains of 12 serotypes (1a, 1b, 2a, 2b, 3a, 3b, 4a,

4b, 5a, Y, X and Xv) were tested following the methods described in the Methods. Apart from 10 serotype Y strains, which were all converted to serotype 1a as expected, the 24 serotype X strains tested were also lysogenized, and converted to a newly named serotype 1d [16]. The serotype 1d strains were serologically characterized as reacting with both serotype 1 specific I typing sera and serotype X specific 7;8 grouping sera [16]. Interestingly, such a serotype has already appeared in natural infections in Anhui and Henan provinces, China [21]. Except for serotypes Y and X, the other serotypes could not be lysogenized by phage SfI. A possible explanation for the host range restriction of phage SfI is phage immunity due to modification of the O-antigen as phage receptors [22]. SfI uses a site-specific mechanism for DNA ALK inhibitor packaging and has the same attP core sequence as SfII, SfIV, SfV and SfX Restriction enzyme analysis revealed that phage SfI has a linear but not circular SPTLC1 genome (data not shown). Genomic comparison found that the

SfI prophage genome has similar packaging genes to that of phage SfV; and the fragments adjacent to them were also highly similar to the cohesive end site (cos) of phage SfV [9], with only one base difference at the 5′ end (T versus A). These data suggest that SfI may use the same site-specific mechanism as SfV for packaging. Direct sequencing of the putative termini of the SfI genome extracted from free phage particles and comparison of the corresponding regions with the SfI prophage genome in strain 019 revealed a 10 nucleotide (5′- TGCCCGCCCC -3′) gap in the SfI phage genome. Therefore, we conclude that SfI uses a cos mechanism for DNA packaging as postulated for phage SfV [9], and does not use a head full mechanism (pac) as for phage Sf6 and SfX [10, 12].

Subsequently, DEPs were classified according to COG function category. It is clear that the expression of proteins involved in functions such as energy production, metabolism, transcription, translation, posttranslational modification, DNA recombination and repair, cell wall biogenesis and signal transduction mechanisms changed the most (Figure 4B). The enrichment and cluster of DEPs were performed according to Gene Ontology and KEGG Pathways functional analysis. The metabolic and biosynthetic eFT-508 biological processes were found to be different in the mutant (Figure 4C). As to KEGG functions affected in the mutant, significant difference was found in the following pathways: valine, leucine

and isoleucine biosynthesis; aminoacyl-tRNA biosynthesis; pyruvate metabolism; galactose metabolism; glycolysis; pentose phosphate pathway; and microbial metabolism in diverse environments (Figure 4D). Figure 4 Comparative proteomic analysis. (A). Protein ratio distribution. The

distribution of average find more value of protein quantification in three repeated experiments is shown. Red: fold change > 1.2, Green: fold change < −1.2. (B). COG function analysis of differentially expressed proteins. (C). KEGG pathways analysis of proteins with different expression (P value <0.05). (D). Gene ontology enrichment analysis of differentially expressed proteins. GO terms of biological process were analysed and significantly enriched catalogues are shown (P-value < 0.01). Integration of transcriptomic and proteomic analysis Most previous studies suggest a weak correlation between mRNA expression and protein expression, which may be due to post-transcriptional regulation of protein synthesis, post-translational modification or experimental errors [38–40]. However, according to the

central dogma of molecular genetics, genetic information is transmitted from DNA to message RNAs that are AG-881 manufacturer subsequently translated to proteins [41, 42]. Thus, we integrated the DEFs and DEPs to identify the overlapping genes that are expressed differently in both the transcriptome these and the proteome. One-hundred and two genes were selected (Figure 5A), and those genes with either up-regulated or down-regulated expression at both the mRNA and protein levels were subjected to bioinformatic analysis. The Gene Ontology study indicated that biological processes such as metabolic processes, catabolic processes, biosynthetic processes and translation may be affected in the mutant strain (Figure 5B). Functional classification according to COG function category indicates that, except for the general function prediction catalogue and the amino acid transport and metabolism catalogue, the genes with the greatest change in expression are classified into the cell wall/membrane/envelope biogenesis and replication catalogue and the recombination and repair catalogue (Figure 5C). Interestingly, the genetic comparison revealed that gene mutations were identified in dprA and arpU.

The age difference in having reluctance against employer interference deserves further attention. In a systematic review, no difference

in participation in WHP was found between younger and older workers (Robroek et al. 2009). However, for older workers, the situation of selleck kinase inhibitor health checks and the focus on lifestyle in the work setting may be new, while the younger workers have never known otherwise. When WHP is aimed at keeping an aging workforce healthy, special attention is needed to content and delivery of WHP and involvement of older workers in design and implementation may support better acceptance and participation. Although not statistically significant, all associations between lifestyle factors and agreeing with the statement that employer interference with employees health is a violation of privacy were in the same direction, indicating that workers with an unhealthy lifestyle GSK126 or poor health are more likely to have reluctance against this employer interference. This may be related with the potential danger of “blaming the victim”. Although it was communicated that all information would not be reported to their supervisor or employer, employees with an unhealthy lifestyle may fear potential consequences of CB-839 participation. Several studies showed that health promotion

in the workplace setting might have beneficial effects on employee lifestyle and health, as well as on reducing sick leave (Groeneveld et al. 2010; Pronk 2009). Therefore, both employee and employer might benefit from WHP. However, our results suggest that moral considerations toward health promotion program at the workplace should not be neglected and in the communication, design, and implementation of

a program deserve special attention. The main limitation in this study was the low response among non-participants, which might induce selection bias. As described in the “Methods”, due to privacy regulations, we only send out the questionnaire once without any reminders. Furthermore, it should be noted that the design and implementation of WHP across companies and countries will differ, and Tolmetin opinions of employees concerning employer involvement may also differ between cultures and countries. More research on this topic is needed in order to get insight into their potential influence on the effectiveness of WHP. This study showed that employees support the importance of health promotion in the workplace setting, but in a modest group of employees, moral considerations may play a role in their decision not to participate in workplace health promotion. Older workers were more likely to resist employer interference with their health. Therefore, special attention on such moral considerations may be needed in the communication, design, and implementation of workplace health promotion programs. Acknowledgments This work was supported by ZonMw, The Netherlands Organization for Health Research and Development (grant number 62300039).