Levels Of Sigma Research Articles

Appling the Lean Six Sigma Methodology in of the CostManagement & Continuous Improvement of Performance (Applied Research in the Blood Laboratory of the Hussein Teaching Hospital)

Purpose: This study aims to enable measuring performance and its quality and seeks to improve performance, is that the research problem emerged in the emergence of poor results for patients' blood test cases due to delayed results of the analysis. Theoretical framework: This study adopted the deductive approach by relying on sources and references related to the literature of the topic directly, to benefit from them in enriching the theoretical aspect of the research. Design/methodology/approach: The inductive approach through the application of the applied analytical aspect, where the financial data for the year 2021 was obtained through financial accounts, production reports, inventory cards, quality reports, observations, and personal interviews, study sample at the Hussein Teaching Hospital's blood laboratory in the service sector represented by the Lean Six Sigma methodology. Findings: The blood laboratory at Al-Hussein Teaching Hospital did not rely on modern quality systems, including the agile Six Sigma methodology. There is a lot of wastage and loss in the specified standard time, which causes a lot of delay in the time of analysis, and this indicates the discrepancy and decentralization of the process. The laboratory testing equipment breaks down many times because of its old age (these devices were obtained from the Japanese grant during the year 2003), and this causes two or three re-examinations, which drains additional time. As well, the small area of the laboratory compared to the number of visitors. Research, Practical & Social implications: The research enables measuring performance and its quality and seeks to improve performance, is that the research problem emerged in the emergence of poor results for patients' blood test cases due to delayed results of the analysis. The Hussein Teaching Hospital's blood laboratory is the service sector represented by the Lean Six Sigma methodology. In order to implement continuous improvement in the area of medical staff quality and to simultaneously provide the right environment for continuous improvement, Originality/value: This study is one of the very few studies that focus on the concept of Six Sigma as one of the tools of the modern quality management system, which is concerned with diagnosing and treating deviations to improve the performance of the blood test process, an attempt to manage cost through the use of accounting and statistical concepts, to achieve the highest levels of sigma.

GST transcriptional changes induced by a toxic Microcystis aeruginosa strain in two bivalve species during exposure and recovery phases.

Previous studies have demonstrated the modulation of glutathione transferases (GSTs) induced by microcystin (MC) alone or in combination with other cyanobacterial secondary metabolites in bivalves. However, interspecies information about which and how GST isoforms are affected by these secondary metabolites is still scarce, especially considering the dynamic process involving their uptake and elimination routes. In this context, the role of GSTs gene expression changes in response to a toxic Microcystis aeruginosa extract were examined for Mytilus galloprovincialis and Ruditapes philippinarum during exposure and recovery phases. The expression levels of sigma 1, sigma 2, pi and mu-class GST genes were analyzed in the hepatopancreas of both bivalve species during cyanobacteria extract exposure (24 h) and post-exposure (24 and 72 h). Only a significant induction of sigma 1-class GST expression was observed for R. philippinarum upon 24-hour exposure of both bivalve species to Microcystis extract. During the recovery phase, GST transcriptional changes for M. galloprovincialis were characterized by an early induction (24 h) of sigma 1 and sigma 2 transcripts. On the other hand, GST transcriptional changes for R. philippinarum during post-exposure phase were characterized by an early induction (24 h) of sigma 1 and mu transcripts and a later induction (72 h) of the four analyzed GST transcripts. Such differences reflect variable GST response mechanisms to cope with MC-producing cyanobacterial blooms exposure between these two bivalve species, revealing a higher sensitivity of R. philippinarum to Microcystis-induced stress than M. galloprovincialis. The results also suggest a much higher level of activity of the GST detoxification system during the recovery phase compared to the period of the stress exposure for both bivalve species.

Six Sigma evaluation using process capability and <i><span style="text-decoration: overline">X</span></i> control chart for reducing variance in oil density characteristic: a study in Yemen

This paper presents Six Sigma evaluation based on process capability with -X control chart to reduce variations in the oil density characteristics in an oil refinery process in Aden, Yemen. Twenty-five oil density samples with normal distribution were randomly collected, and each sample consisted of four items. Analysis indicated that the levels of sigma employed in Aden refinery are less than 3 σ. The results of this study suggested that the -X chart based on six sigma estimation method is significant to reduce variance in the oil density characteristic. In addition, the sigma levels increase when the process capability increases. Thus, the increment in sigma levels leads to improvements in the performance of the process refinery of oil density characteristic. Finally, this paper provides the necessary fundamentals and knowledge for quality control researchers and engineers to reduce variances by using Six Sigma.

IMPLEMENTASI PENERAPAN METODE SIX SIGMA PADA PROSES PERAKITAN ELEKTRO MOTOR (STUDI KASUS: PT. TATUNG ELECTRIC INDONESIA)

Tatung Electric Indonesia is a peripatetic company in manufacturing industry assembling of electro motor. So that product yielded by PT. Tatung Electric Indonesia can continue to fulfill requirement of customer, company try to improve the quality of its product by minimizing defect which during the time still happened. This defect arising caused by the existence of variation at yielded product and also that happened in the process. Six Sigma represent control technique or method to reach storey or level quality of 3.4 DPMO ( Defects Per Million Opportunities), which applied by Motorola company since year 1986. Applying strategy to be used is Define-Measure-Analyze-Improve-Control (DMAIC). This Method represent recuring step and form a make-up of quality cycle. Calculations show the company is 3.641 sigma level with DPMO 15,830. The sigma levels are on the average level of sigma industries in Indonesia (3-4 sigma). Increased levels of sigma would happen if the company immediately continuous improvement over time. Conclusion of the application of Six Sigma is the DMAIC method in the assembly process of electro motors can analyze and reduce disability that occurs that will increase the quality of products.

AtSIG6 and other members of the sigma gene family jointly but differentially determine plastid target gene expression in Arabidopsis thaliana.

Plants contain a nuclear gene family for plastid sigma factors, i.e., proteins that associate with the “bacterial-type” organellar RNA polymerase and confer the ability for correct promoter binding and transcription initiation. Questions that are still unresolved relate to the “division of labor” among members of the sigma family, both in terms of their range of target genes and their temporal and spatial activity during development. Clues to the in vivo role of individual sigma genes have mainly come from studies of sigma knockout lines. Despite its obvious strengths, however, this strategy does not necessarily trace-down causal relationships between mutant phenotype and a single sigma gene, if other family members act in a redundant and/or compensatory manner. We made efforts to reduce the complexity by genetic crosses of Arabidopsis single mutants (with focus on a chlorophyll-deficient sig6 line) to generate double knockout lines. The latter typically had a similar visible phenotype as the parental lines, but tended to be more strongly affected in the transcript patterns of both plastid and sigma genes. Because triple mutants were lethal under our growth conditions, we exploited a strategy of transformation of single and double mutants with RNAi constructs that contained sequences from the unconserved sigma region (UCR). These RNAi/knockout lines phenotypically resembled their parental lines, but were even more strongly affected in their plastid transcript patterns. Expression patterns of sigma genes revealed both similarities and differences compared to the parental lines, with transcripts at reduced or unchanged amounts and others that were found to be present in higher (perhaps compensatory) amounts. Together, our results reveal considerable flexibility of gene activity at the levels of both sigma and plastid gene expression. A (still viable) “basal state” seems to be reached, if 2–3 of the 6 Arabidopsis sigma genes are functionally compromised.

Environmental Assisted Cracking Behavior of Sigmatized Super Duplex Stainless Steel in Oilfield Production Brine

The slow strain rate test (SSRT) technique was used to investigate the stress corrosion cracking (SCC) susceptibility of sigmatized super duplex stainless steel (SDSS) grade UNS S32760, exposed to a simulated oilfield brine containing carbon dioxide/hydrogen sulfide (CO2/H2S) at 80°C. The sigma phase was precipitated by heat treatment (HT) at 850°C for various durations (0 to 12 min). Results show that low levels (less than 2%) of sigma have no profound impact on the mechanical and corrosion properties of SDSS. However, at higher levels (greater than 2%) of sigma phase, there is severe loss in ductility and increased corrosion rate. Fractographic observations using scanning electron microscopy (SEM) show that the fracture mode shifts from ductile to transgranular cracking with increasing levels of sigma phase. Electron-backscattered diffraction (EBSD) results show that crack nucleation occurs in the sigma grains. The cracking mechanism involves brittle fracture of sigma phase and slip-assisted anodic diss...

Building Bridges for Spinal Cord Repair

For nearly a century, neuroscientists have sought to restore neurological function across spinal cord lesions. Lu etal. now present significant progress toward this goal, showing in rats that transplanted neural stem cells establish a functional bridge across completely transected spinal cords.

The roles of neuronal and glial precursors in overcoming chondroitin sulfate proteoglycan inhibition

The extension of axons through the major inhibitory component of the glial scar, chondroitin sulfate proteoglycans (CSPGs), remains a key obstacle for regeneration following spinal cord injury (SCI). We have previously shown that transplants composed of neuronal and glial restricted precursors (NRP and GRP respectively) promote regeneration and connectivity in the injured spinal cord (Bonner et al., 2010, 2011), however, little is known about the properties of these precursors at a cellular level. We now report that NRP-derived neurons, in contrast to dorsal root ganglion (DRG) neurons, have the ability to extend axons and cross over from a permissive substratum (laminin) onto inhibitory CSPG in vitro. Growth cones of neurons derived from NRP, compared to DRG, exhibit significantly lower levels of the CSPG receptors protein tyrosine phosphatase sigma (PTPσ) and leukocyte common antigen-related phosphatase (LAR). GRP-conditioned medium prepared from the same cell densities did not affect the response of primary sensory neurons to CSPG confirming that the ability of NRP-derived neurons to cross onto CSPG is determined intrinsically. However, GRP-conditioned medium collected from high density cultures increased the probability of DRG axons to cross from LN onto CSPG and increased the length of DRG axons extending on CSPG. Collectively, these results suggest that (1) neurons derived from NRPs are intrinsically insensitive to CSPGs due to low levels of receptor expression, and (2) high levels of factors secreted by GRP can reduce the inhibitory effects of CSPG and promote axonal growth. These observations provide mechanistic insights into the specific roles of NRPs and GRPs in promoting regeneration and repair following SCI.

Haloperidol increases expression of the inositol 1,4,5-trisphosphate receptors in rat cardiac atria, but not in ventricles

Numerous ligands of sigma receptors are known to prolong the QT interval and therefore cause a variety of arrhythmias. High affinity binding sites for the prototypical sigma ligand haloperidol were found in membranes of cardiac myocytes from adult rats. Activation of sigma 1 receptor leads to a release of calcium from the endoplasmic reticulum that follows increased synthesis of inositol 1,4,5-trisphosphate (IP3). We studied the effect of long-term haloperidol treatment on the expression of sigma 1 receptors, IP3 receptors of type 1 and 2 in the individual parts of the rat heart, in isolated rat cardiomyocytes and in PC12 cells. We have found that prolonged treatment with haloperidol significantly increased mRNA levels of sigma 1 receptors in both atria and ventricles. Sigma 1 receptor's mRNA was increased also in isolated cardiomyocytes. Haloperidol treatment affects the expression of IP3 receptors of type 1 and 2 in cardiac atria, but not in cardiac ventricles. We observed increase in IP3 receptors in differentiated PC12 cells, but not in isolated cardiomyocytes. We propose that this increase might participate in triggering cardiac arrhythmias during haloperidol treatment, which has to be further verified.

Dynamical insights into π1σ∗ state mediated photodissociation of aniline

This article reports a comprehensive study of the mechanisms of H atom loss in aniline (C(6)H(5)NH(2)) following ultraviolet excitation, using H (Rydberg) atom photofragment translational spectroscopy. N-H bond fission via the low lying (1)pi sigma(*) electronic state of aniline is experimentally demonstrated. The (1)pi sigma(*) potential energy surface (PES) of this prototypical aromatic amine is essentially repulsive along the N-H stretch coordinate, but possesses a shallow potential well in the vertical Franck-Condon region, supporting quasibound vibrational levels. Photoexcitation at wavelengths (lambda(phot)) in the range 293.859 nm > or = lambda(phot) > or = 193.3 nm yields H atom loss via a range of mechanisms. With lambda(phot) resonant with the 1(1)pi pi(*) <-- S(0) origin (293.859 nm), H atom loss proceeds via, predominantly, multiphoton excitation processes, resonantly enhanced at the one photon energy by the first (1)pi pi(*) excited state (the 1(1)pi pi(*) state). Direct excitation to the first few quasibound vibrational levels of the (1)pi sigma(*) state (at wavelengths in the range 269.513 nm > or = lambda(phot) > or = 260 nm) induces N-H bond fission via H atom tunneling through an exit barrier into the repulsive region of the (1)pi sigma(*) PES, forming anilino (C(6)H(5)NH) radical products in their ground electronic state, and with very limited vibrational excitation; the photo-prepared vibrational mode in the (1)pi sigma(*) state generally evolves adiabatically into the corresponding mode of the anilino radical upon dissociation. However, as the excitation wavelength is reduced (lambda(phot) < 260 nm), N-H bond fission yields fragments with substantially greater vibrational excitation, rationalized in terms of direct excitation to 1(1)pi pi(*) levels, followed by coupling to the (1)pi sigma(*) PES via a 1(1)pi pi(*)/(1)pi sigma(*) conical intersection. Changes in product kinetic energy disposal once lambda(phot) approaches approximately 230 nm likely indicate that the photodissociation pathways of aniline proceed via direct excitation to the (higher) 2(1)pi pi(*) state. Analysis of the anilino fragment vibrational energy disposal-and thus the concomitant dynamics of (1)pi sigma(*) state mediated photodissociation-provides a particularly interesting study of competing sigma(*) <-- pi and pi(*) <-- pi absorption processes and develops our appreciation of the photochemistry of aromatic amines. It also allows revealing comparisons with simple amines (such as ammonia and methylamine) as well as the isoelectronic species, phenol. This study yields a value for the N-H bond strength in aniline, D(0)(H-anilino) = 31630+/-40 cm(-1).

Elevated levels of σ S inhibit biofilm formation in Escherichia coli: a role for the Rcs phosphorelay

The Rcs phosphorelay is composed of RcsC, RcsD and the response regulator RcsB, and this signalling pathway has been implicated in virulence and biofilm formation in many enteric bacteria. It was previously shown that a mutation in rcsC resulted in defective biofilm formation in Escherichia coli [Ferrières, L. & Clarke, D. J. (2003) Mol Microbiol 50, 1665-1682]. To identify the molecular mechanisms underlying the observed biofilm defect we carried out a screen looking for suppressor mutants that restored biofilm formation in the rcsC mutant background. One of the mutants was identified to be in rprA, a gene encoding a small RNA molecule that is involved in the post-transcriptional control of the alternative sigma factor, sigma(S). The expression of rprA is regulated by the Rcs phosphorelay, and there are elevated sigma(S) levels present in the rcsC mutant due to the overexpression of rprA in this background. Using different approaches, we have established that the increase in sigma(S) levels is responsible for the biofilm defect. Therefore, the Rcs phosphorelay is involved in maintaining appropriate levels of sigma(S) during biofilm formation in E. coli.

Dissection of σE‐dependent cell lysis in Escherichia coli: roles of RpoE regulators RseA, RseB and periplasmic folding catalyst PpiD

To understand the mechanism of sigma(E)-dependent cell lysis, we examined the consequences of deletion derivatives of rpoE regulators rseA, rseB and rseC on sigma(E) transcription, on levels of free versus membrane-bound sigma(E) and on OMP-biogenesis limiting factor(s) that could impact cell lysis. RT-PCR showed that individual nonpolar DeltarseA and DeltarseB increased the rpoE expression to varying extents, with pronounced induction in DeltarseA. Significantly the ratio of soluble (free) versus membrane-bound form of RpoE increased in DeltarseA, however without increase of its total amount, unraveling furthermore complexity in RpoE regulation. Significant characteristics of cell lysis, accompanied by a severe reduction in the levels of periplasmic OMP-folding factor (PpiD), were observed in DeltarseA. The cell-lysis phenotype of DeltarseA was suppressed by either rseA or ppiD plasmids, but neither by rseB nor by rseC clones. However, the cell lysis of the wild-type strain was almost completely repressed not only by the rseA clone but also by the rseB clone, suggesting RseB might be limiting in vivo. Thus, increase in the ratio of free sigma(E) in rseA mutants with a concomitant reduction in PpiD levels can account for sigma(E)-dependent lysis in concert with a potential role of small RNAs on the lysis process.

Polycyclic Aromatic Hydrocarbons and Phthalic Acid Esters in Vegetables from Nine Farms of the Pearl River Delta, South China

This study investigated the occurrence of 16 polycyclic aromatic hydrocarbons (PAHs) and 6 phthalic acid esters (PAEs) in 11 vegetable species collected from nine farms of the Pearl River Delta, South China. Twelve PAH compounds and all PAE compounds were detected by gas chromatography coupled with mass spectrometry (GC-MS) in vegetables. The total concentrations of PAHs (Sigma(PAHs)) and PAEs (Sigma(PAEs)) ranged from 7.0 to 5353 microg kg(-1) dry weight (d.w.), with a mean value of 1173 microg kg(-1) d.w., and from 0.073 to 11.2 mg kg(-1) d.w., with a mean value of 3.2 mg kg(-1) (d.w.), respectively. The highest levels of Sigma(PAHs) and Sigma(PAEs) were found in Brassica juncea and Brassica parachinensis, respectively. For the same vegetable, the bioconcentration factors (BCFs; the ratio of contaminant concentration in plant tissue to the soil concentration) of PAHs (between 0.0037 and 5.5) are generally higher than those of PAEs (between <0.0001 and 0.61). It was also noted that there were great variations of organic contaminant levels, BCFs, and benzo[a]pyrene equivalent concentrations, which depend on the various contaminants, sampling locations, and vegetable species. The occurrences of PAHs and PAEs in this study are compared with those in other studies and their sources are discussed.

Oscillator strengths and line widths of dipole-allowed transitions in N214 between 89.7 and 93.5nm

Line oscillator strengths in the 20 electric dipole-allowed bands of (14)N(2) in the 89.7-93.5 nm (111480-106950 cm(-1)) region are reported from photoabsorption measurements at an instrumental resolution of approximately 6 mA (0.7 cm(-1)) full width at half maximum. The absorption spectrum comprises transitions to vibrational levels of the 3p sigma(u) c(4)' (1)Sigma(u)(+), 3p pi(u) c(3) (1)Pi(u), and 3s sigma(g) o(3) (1)Pi(u) Rydberg states and of the b' (1)Sigma(u)(+) and b (1)Pi(u) valence states. The J dependences of band f values derived from the experimental line f values are reported as polynomials in J'(J'+1) and are extrapolated to J'=0 in order to facilitate comparisons with results of coupled Schrodinger-equation calculations. Most bands in this study are characterized by a strong J dependence of the band f values and display anomalous P-, Q-, and R-branch intensity patterns. Predissociation line widths, which are reported for 11 bands, also exhibit strong J dependences. The f value and line width patterns can inform current efforts to develop comprehensive spectroscopic models that incorporate rotational effects and predissociation mechanisms, and they are critical for the construction of realistic atmospheric radiative-transfer models.

Congener‐specific distribution of polybrominated diphenyl ethers in fish of China: Implication for input sources

A total of 390 individual fish, including seven species of freshwater-farmed fish, three species of seawater-farmed fish, and three species of wild marine fish, from China were analyzed to determine the concentrations and congener distribution of polybrominated diphenyl ethers (PBDEs). The levels of Sigma(10)PBDEs (sum of BDEs 28, 47, 66, 100, 99, 85, 154, 153, 138, and 183) in seawater-farmed fish (median, 13.6 ng/g) were significantly higher than those in freshwater-farmed fish (median, 10.1 ng/g) and in wild marine fish (median, 4.5 ng/g) on a lipid-weight basis. Lipid contents positively correlated with Sigma(10)PBDEs concentration (wet wt) in all fish species except for tilapia, bighead carp, bluntsnout bream, and common mullet; however, no significant relationship was found between fish length or weight and Sigma(10)PBDEs concentration except for one fish species. In general, BDE 47 was the dominant congener, with a mean contribution of 53.2%. Brominated diphenyl ether 209 was only detected in 14 samples, reflecting its low bioavailability or high degree of debromination. In addition, the ratios of BDE 47 to BDE 99 and of BDE 100 to BDE 99 in freshwater-farmed fish were significantly higher than those in seawater-farmed and wild marine fish. Freshwater-farmed fish had a relatively higher proportion of BDE 28, whereas seawater-farmed and wild marine fish contained more abundant BDE 66, BDE 100, and BDE 99. The difference in congener distribution might be indicative of different exposure pathways for PBDEs in three types of fish.

Alkaline phosphatase as a reporter of σS levels and rpoS polymorphisms in different E. coli strains

sigma(S) is responsible for the transcriptional regulation of genes related to protection against stresses and bacterial survival and it accumulates in the cell under conditions of stress, such as nutrient limitation. An increase in the levels of sigma(S) causes a reduction in the expression of genes that are transcribed by RNA polymerase associated with the principal sigma factor, sigma(70). phoA, that encodes alkaline phosphatase (AP) is expressed under phosphate shortage conditions, and is also repressed by sigma(S). Here we show that in a Pi-limited chemostat, accumulation of rpoS mutations is proportional to the intrinsic level of sigma(S) in the cells. Acquisition of mutations in rpoS relieves repression of the PHO genes. We also devised a non-destructive method based on the rpoS effect on AP that differentiates between rpoS (+) and rpoS mutants, as well as between high and low-sigma(S) producers. Using this method, we provide evidence that sigma(S) contributes to the repression of AP under conditions of Pi excess and that AP variation among different strains is at least partly due to intrinsic variation in sigma(S) levels. Consequently, a simple and non-destructive AP assay can be employed to differentiate between strains expressing different levels of sigma(S) on agar plates.

Hydroxy-PCBs, PBDEs, and HBCDDs in Serum from an Elderly Population of Swedish Fishermen's Wives and Associations with Bone Density

Lack of human exposure data is frequently reported as a critical gap in risk assessments of environmental pollutants, especially regarding "new" pollutants. The objectives of this study were to assess serum levels of the persistent 2,2',4,4',5,5'-hexachlorobiphenyl (CB-153), hydroxylated polychlorinated biphenyl metabolites (OH-PCBs), polybrominated diphenyl ethers (PBDEs), and hexabromocyclodode-canes (HBCDDs) in a group of Swedish middle-aged and elderly women expected to be relatively highly exposed, and to evaluate the impact of potential determinants (e.g., fish intake, age) for the inter-individual variation, as well as to investigate the association between these pollutants and bone density. No associations were found between bone mineral density or biochemical markers of bone metabolism and the analyzed environmental pollutants. Relatively high levels of CB-153 (median 260 ng/g fat) and sigma 3-OH-PCBs (median 1.7 ng/mL serum), and low concentrations of sigma6PBDEs (median 3.6 ng/g fat) were determined. Total level of HBCDDs in serum was quantified by gas chromatography with mass spectrometric detection (median 0.5 ng/g fat). HBCDD diastereomeric and enantiomeric patterns were determined by liquid chromatography with mass spectrometric detection. The dominating stereoisomer was (-)alpha-HBCDD, but 1-3% of gamma-HBCDD was also detected in the serum samples.

SpoIVH (ykvV), a requisite cortex formation gene, is expressed in both sporulating compartments of Bacillus subtilis.

It is well known that the ykvU-ykvV operon is under the regulation of the sigma(E)-associated RNA polymerase (Esigma(E)). In our study, we observed that ykvV is transcribed together with the upstream ykvU gene by Esigma(E) in the mother cell and monocistronically under Esigma(G) control in the forespore. Interestingly, alternatively expressed ykvV in either the forespore or the mother cell increased the sporulation efficiency in the ykvV background. Studies show that the YkvV protein is a member of the thioredoxin superfamily and also contains a putative Sec-type secretion signal at the N terminus. We observed efficient sporulation in a mutant strain obtained by replacing the putative signal peptide of YkvV with the secretion signal sequence of SleB, indicating that the putative signal sequence is essential for spore formation. These results suggest that YkvV is capable of being transported by the putative Sec-type signal sequence into the space between the double membranes surrounding the forespore. The ability of ykvV expression in either compartment to complement is indeed intriguing and further introduces a new dimension to the genetics of B. subtilis spore formation. Furthermore, electron microscopic observation revealed a defective cortex in the ykvV disruptant. In addition, the expression levels of sigma(K)-directed genes significantly decreased despite normal sigma(G) activity in the ykvV mutant. However, immunoblotting with the anti-sigma(K) antibody showed that pro-sigma(K) was normally processed in the ykvV mutant, indicating that YkvV plays an important role in cortex formation, consistent with recent reports. We therefore propose that ykvV should be renamed spoIVH.

Ribosomal protein S1 specifically binds to the 5' untranslated region of the Pseudomonas aeruginosa stationary-phase sigma factor rpoS mRNA in the logarithmic phase of growth.

The rpoS gene encodes the stationary-phase sigma factor (RpoS or sigma(s)), which was identified in several gram-negative bacteria as a central regulator controlling the expression of genes involved in cell survival in response to cessation of growth (stationary phase) and providing cross-protection against various stresses. In Pseudomonas aeruginosa, the levels of sigma(s) increase dramatically at the onset of the stationary phase and are regulated at the transcriptional and posttranscriptional levels. The P. aeruginosa rpoS gene is transcribed as a monocistronic rpoS mRNA transcript comprised of an unusually long 373-bp 5' untranslated region (5' UTR). In this study, the 5' UTR and total protein extracts from P. aeruginosa logarithmic and stationary phases of growth were used in order to investigate the protein-RNA interactions that may modulate the translational process. It was observed that a 69-kDa protein, which corresponded to ribosomal protein S1, preferentially binds the 5' UTR of the rpoS mRNA in the logarithmic phase and not in the stationary phase. This is the first report of a protein-rpoS mRNA 5' UTR interaction in P. aeruginosa, and the possible involvement of protein S1 in translation regulation of rpoS is discussed.

A comparative study of variation in codon 33 of the rpoS gene in Escherichia coli K12 stocks: implications for the synthesis of sigma(s).

The Escherichia coli rpoS gene encodes an RNA polymerase sigma factor (sigma S or sigma(S)) required for the expression of stationary-phase genes. In the first published rpoS sequence from E. coli K-12 codon 33 is given as CAG. However, several subsequent independent studies found the amber codon TAG at this position ( rpoSAm). Besides this amber codon, other codons such as TAT have also been found at this location in rpoS. Comparative genome analysis now leads us to propose TAG as the parental codon 33 in rpoS in E. coli K-12. Five different stocks of the strain W3110, which differ in the levels of sigma(S) protein they express, were investigated. We sequenced the rpoS gene from these, and found a T at nucleotide position 97 in four out of the five stocks and a G at position 99 in three out of the five. W1485, a parental strain of W3110, and W3350, a derivative of W3110, are also rpoSAm mutants. Such rpoSAm mutants would be expected to show no RpoS activity. The retention of partial or intermediate sigma(S) activity by suppressor-free rpoSAm mutants is therefore puzzling. We propose that a functional, N-terminally truncated, sigma(S) (Delta1-53sigma(S)) can be translated from a Secondary Translation Initiation Region (STIR) located downstream of the amber codon 33. It has recently been reported that a fragment of RpoS (Delta1-53sigma(S)) that lacks the first 53 amino acids is functional when synthesized in vivo. Taken together, our results support the hypothesis that the original codon 33 of the rpoS gene in E. coli K-12 strains is the amber codon TAG.