The role of leadership in implementing lean manufacturing

It is widely accepted that for the successful implementation of lean manufacturing, the senior management commitment is of great importance. However, the lean journey is usually a long one, and eventually management commitment creeps. Furthermore, the involvement of employees in daily improvements is also critical for the success of implementation. Lean leadership can be considered as a way of sustaining and improving the employee performance in lean production systems. In the present study, a thorough literature review is presented focusing in reviewing the principles of lean leadership and the practices that can lead in improving the employee performance. Furthermore, the characteristics and qualities of lean leader are discussed Paper originally presented at Manufacturing Systems 4.0 – Proceedings of the 50th CIRP Conference on Manufacturing Systems, held 3 – 5 May, 2017, Taichung City, Taiwan

A-site mRNA cleavage is not correlated with ssrA-peptide tagging activity.

<p><b>A</b>) Northern blot analysis of <i>flag-(m)ybeL-PP</i> transcripts in cells lacking 3′-to-5′ exoribonucleases. Total RNA was isolated from <i>E. coli ssrA⁻</i> cells that lack the indicated RNase genes and probed with a radiolabeled oligonucleotide that hybridizes the 5′-UTR of <i>flag-(m)ybeL-PP</i> message. The lane labeled <i>in vitro</i> contains <i>flag-(m)ybeL-PP</i> mRNA that is truncated at the stop codon. The migration positions of full-length and truncated transcripts are indicated. <b>B</b>) SsrA(DD)-peptide tagging of His₆-YbeL-PP. His₆-YbeL-PP chains were purified from cells of the indicated genetic backgrounds and resolved by SDS-PAGE and stained with Coomassie blue. <b>C</b>) Quantification of A-site mRNA cleavage and ssrA(DD) tagging efficiency. The percentage of A-site truncated mRNA was determined by quantifying northern blot hybridization signals as described in Methods. The effect of each RNase gene deletion was examined in an <i>ssrA⁻</i> background, and the data (in white bars) represent the mean ± SEM for at least three independently prepared RNA samples. Full-length and ssrA(DD)-tagged His₆-YbeL-PP chains were isolated from <i>ssrA(DD)</i> cells and quantified by densitometry. Tagging efficiency (in gray bars) is reported as the percentage of total chains that carry ssrA(DD) peptides. The presented data represent the mean ± SEM from four independent experiments.</p

Pulse-chase analysis of ribosome recycling.

<p><b>A</b>) Autoradiography of [³⁵S]-labeled peptidyl-tRNAs isolated from <i>rnb⁺</i> and Δ<i>rnb</i> cells. Cells were pulse-labeled and samples taken at the indicated times for denaturing polyacrylamide gel electrophoresis and autoradiography. The band corresponding to peptidyl prolyl-tRNA₂^Pro was identified by northern blot hybridization (not shown). <b>B)</b> Double exponential equation fits to experimental data. Because there are at least two distinct pathways for the turnover of peptidyl-tRNA (i.e. release factor mediated termination and tmRNA-SmpB mediated rescue) double exponential decay equations were fitted to the experimental data. Representative fits for <i>rnb⁺</i> and Δ<i>rnb</i> cells are shown. All experiments were conducted twice and the reported values correspond to the mean ± SEM.</p

YafO does not mediate +12 processing during translational arrest.

<p>Northern blot analysis of <i>flag-(m)ybeL-PP</i> transcripts. Total RNA was isolated from <i>E. coli ssrA⁻</i> cells with the indicated genotypes and probed with an oligonucleotide that hybridizes to the 5′-UTR of <i>flag-(m)ybeL-PP</i> message. The lane labeled <i>in vitro</i> contains <i>flag-(m)ybeL-PP</i> mRNA that is truncated at the stop codon. The migration positions of full-length and truncated transcripts are indicated.</p

Peptidyl-tRNA half-lives.^a

a<p><i>T</i>_1/2 values were determined from double-exponential decay equations fitted to data as described in Methods. Reported values are the mean ± standard error of the mean (SEM).</p

Effect of RNA helicase deletions on mRNA processing and ssrA(DD)-peptide tagging.

<p><b>A</b>) Northern blot analysis of <i>flag-(m)ybeL-PP</i> transcripts. Total RNA was isolated from <i>E. coli ssrA⁻</i> cells with the indicated genotypes and probed with an oligonucleotide that hybridizes to the 5′-UTR of <i>flag-(m)ybeL-PP</i> message. The lanes labeled <i>in vitro</i> contain <i>flag-(m)ybeL-PP</i> mRNA that is truncated at the stop codon. The migration positions of full-length and truncated transcripts are indicated. <b>B</b>) Quantification of ssrA(DD) tagging efficiency. Full-length and ssrA(DD)-tagged His₆-YbeL-PP chains were quantified by densitometry and tagging efficiency reported as the percentage of total chains that carry ssrA(DD) peptides. Reported values represent the mean ± SEM from two independent experiments.</p

Suppression of A-site mRNA cleavage.

<p><b>A</b>) The <i>flag-(m)ybeL-PP</i> transcript encodes an N-terminal FLAG epitope fused to the C-terminal 49 residues of <i>E. coli</i> YbeL-PP. The position of the P-site and A-site codons during ribosome pausing is indicated by boxed P and A, respectively. Northern blots were hybridized with an oligonucleotide probe that binds both messages just upstream of the initiation codon as indicated. The downward pointing arrows show A-site and +12 cleavage sites as mapped in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0081319#pone.0081319-GarzaSnchez3" target="_blank">[29]</a>. <b>B</b>) Northern blot analysis of <i>flag-(m)ybeL-PP</i> transcripts. Total RNA isolated from cells with the indicated <i>ssrA</i> and <i>rnb</i> genotypes was probed with a radiolabeled oligonucleotide that hybridizes the 5′-UTR of <i>flag-(m)ybeL-PP</i> message. Control transcripts that are truncated at the stop codon were produced by <i>in vitro</i> transcription and run as a gel-migration marker in the lane labeled <i>in vitro</i>. The migration positions of full-length and truncated transcripts are indicated. <b>C</b>) Quantification of A-site truncation products. The percentage of A-site truncated mRNA in each genetic background was determined by quantifying northern blot hybridization signals as described in Methods. Reported values represent the mean ± SEM for at least three independently prepared RNA samples.</p

CdiA^UPEC536 and HA-CdiA^UPEC536 are exported to the cell surface.

<p>A) Schematic of the CdiA^UPEC536 exoprotein depicting the locations of the inserted N-terminal hemagglutinin (HA) epitope tag and the CdiA-CT region (residues Val3016– Ile3242) used to generate anti-CdiA-CT^UPEC536 polyclonal antibodies. Regions corresponding to the secretion signal sequence and the toxic tRNase domain are indicated. The vertical VENN sequence demarcates the N-terminal margin of the variable CdiA-CT sequence. Residue numbers are shown by the scale bar below. B) Whole-cell immunoblot analysis of <i>E. coli</i> cells expressing CdiA^UPEC536 and HA-CdiA^UPEC536. Cells expressing HA-CdiA^UPEC536 (HA-CdiA), CdiA^UPEC536 (CdiA) or no effector protein (CDI^-) were fixed without permeabilization and stained with anti-HA or anti-CdiA-CT^UPEC536 (anti-CdiA-CT) antibodies. Where indicated, cells were treated with proteinase K (proK) prior to fixation. Stained cells were spotted onto nitrocellulose membrane and analyzed with an Odyssey infrared imager.</p

CdiA-CT^UPEC536 is delivered into target cells.

<p>A) Anti-CdiA-CT^UPEC536 immunofluorescence microscopy of CDI^UPEC536 co-cultures. Non-fluorescent inhibitor cells expressing either CdiA^UPEC536 (CdiA) or HA-CdiA^UPEC536 (HA-CdiA) were mixed with red fluorescent target cells (2∶1 inhibitor-to-target ratio) for 1 h. Cells were then fixed and permeabilized for fluorescence microscopy using anti-CdiA-CT^UPEC536 (anti-CdiA-CT) antibodies as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057609#s4" target="_blank">Methods</a>. Green fluorescence is indicative of anti-CdiA-CT immunostaining. Where indicated (+), cells were treated with proteinase K prior to fixation. The histogram quantifies the percentage of target cells (average ± SEM) with surface and internal CdiA-CT^UPEC536 antigen staining. At least 150 target cells from two independent experiments were scored for the quantification of CdiA delivery. B) Anti-HA epitope immunofluorescence microscopy of CDI^UPEC536 co-cultures. Co-culture conditions and sample preparation was as described in panel A except that anti-HA antibodies were used for immunofluorescence. Green fluorescence is indicative of anti-HA immunostaining. Where indicated (+), cells were treated with proteinase K prior to fixation.</p

CdiA^UPEC536 is transferred to the surface of target cells.

<p>A) Immunofluorescence microscopy of CDI^UPEC536 co-cultures. Non-fluorescent inhibitor cells (expressing either CdiA^UPEC536 or HA-CdiA^UPEC536) were mixed with red fluorescent target cells (2∶1 inhibitor-to-target ratio) for 1 h, then analyzed by fluorescence microscopy using anti-CdiA-CT^UPEC536 (anti-CdiA-CT) or anti-HA antibodies as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057609#s4" target="_blank">Methods</a>. Green fluorescence is indicative of anti-CdiA-CT and anti-HA staining of cell surfaces. Inhibitor cells in the CDI^- panel carry an empty vector without a CDI system. Where indicated (+), samples were treated with proteinase K prior to fixation, but cells were not permeabilized. B) Transfer of CdiA-CT^UPEC536 antigen to target cells. The percentage of <i>bamA</i>⁺ and <i>bamA101</i> target cells with CdiA-CT^UPEC536 antigen was quantified after one and two hours of co-culture with CdiA^UPEC536 inhibitor cells. The <i>bamA101</i> allele was complemented with plasmid pBamA (corresponding to pDAL950 in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0057609#pone-0057609-t001" target="_blank">Table 1</a>). C) Transfer of HA antigen to target cells. Quantifications in panels B and C were determined by analysis of 150 target cells from two independent experiments. The reported values represent the average ± SEM.</p