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Proposes, Scope, Roles and responsibility, Team, Audit program and schedule yearly bases , Procedure including check list. Example of implementation Schedule: Annual Self Inspection Schedule max end of first quarter Each department should be audited at least once a year.
Quality System Function is responsible to ensures the execution of self inspection is in accordance with predetermined schedule with tolerance plus minus one months from scheduled. Audit shall be conducted by independent individuals not having direct responsibility for the area being audited. Auditor should maintain their knowledge of internal and external regulations and develop their audit skills.
Assessment is needed each year to maintain auditor qualification status. Observer - An observer may be part of audit team upon invitation by the audit unit. The observer is not actively involved in the audit process. Open communication between the auditor and auditee should be maintained. Audit observations should be based on objective evidence, such as direct observations of on-going processes, personnel interviews or evaluation of recorded data. If during inspection a critical incompliance is observed, an immediate action must be taken.
Audit observation shall be objective, fair and balanced, recorder in adequate detail and easy to understood. Reference is made to underlying documents, if applicable. Monitored by Company management. Example implementation: Auditee should prepare CAPA form within one month after self inspection report approved. For CAPA which should be coordinated to other department, it is responsibility of auditee for the coordination process.
Conducted by outside or independent specialist or internal team designated by Management. Process Equipment Design and construction Equipment maintenance and cleaning Calibration Computerized system Documentation and records Documentation system and specification Equipment cleaning and use records Record of raw materials, intermediate, API labeling and packaging materials Master production instruction Master production and control record Laboratory control record Batch production record review.
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Material Management General control Receipt and quarantine Sampling and testing of incoming production materials Storage Re-evaluation Production and in-process control Production operation Time limit In-process sampling and control Blending batches of intermediate or API Contamination control. Validation Validation policy Validation documentation Qualification Approaches to process validation Process validation program Periodic review of validated systems Cleaning validation Validation of analytical method Change Control.
A list of approved suppliers of starting material should be established and reviewed. For new supplier, should be evaluated history and nature of the materials. And if audit is required, it should determine the suppliers ability to confirm with GMP standard. All established supplier should be evaluated regularly. This procedure is applicable to all supplier who supply products Scope and services which may affect the quality of products, as follows: Maintenance and calibration services, validation services, pest control.
Utility including suppliers form preventive maintenance. Prepare supplier qualification plan and report. Review supplier qualification plan and report. Perform qualification step as required in qualification plan.
Flag for inappropriate content. Related titles. Jump to Page. Search inside document. Monitored by Company management Example implementation: Perform qualification step as required in qualification plan C. Documents Similar To Self Qualification.
D Intracellular survival was measured using bacteria cultured to the E phase with or without theophylline. The first three genotypes from left to right on the x axis were cultured to the E phase in LB broth media without inducer. The inducer was added during culture in broth, as well as to the tissue culture medium during the infection period, for the final three genotypes depicted on the x axis. Residues within TM3 for E.
Channel-facing residues facilitate movement of protons across the IM Fig. Mutating the E. Several residues are conserved for S. Therefore, substitution mutant alleles, tolQ AV and tolQ GD , were engineered and inserted downstream of the riboswitch on the mutant genome Fig. As a control for specificity, we engineered a neutral substitution allele, tolQ GV , on the mutant genome. This indicated that the negative phenotypic effect of expressing the mutant TolQ protein was specific to the GD substitution, which likely perturbs TolQ function by imparting an electronegative charge to the channel lumen Fig.
Salmonellae encounter a variety of host-killing mechanisms during systemic pathogenesis. Therefore, we reasoned that S. Typhimurium Tol-Pal proteins might differentially promote survival during murine bacteremia. In contrast, statistically fewer surviving salmonellae were enumerated from the mice infected with each of the seven tol-pal mutant genotypes of S.
Typhimurium than those infected with the wild type. Though these mutants were not attenuated in phagocytes, statistically fewer ybgC and cpoB mutant salmonellae were recovered from the mice than the wild type.
Therefore, each Tol-Pal protein contributes to S. Tol-Pal proteins differentially promote S. The mice were euthanized at 48 h.
The spleens and livers were dissected and homogenized. In fact, the loss of each S. Typhimurium Tol-Pal protein impacted Rcs signaling activity, cell morphology, Rif resistance, and survival during bacteremia, including loss of YbgC and CpoB, whose biological relationship with Tol-Pal was unresolved Fig.
The proteins and mechanisms required for Gram-negative bacteria to transport GPLs across the periplasm have eluded scientific discovery until recently 47 , 52 , We now understand that some mechanisms involve multicomponent protein systems that span the envelope and derive energy from the IM in order to move GPLs across the periplasm.
In support of a direct binding mechanism, TolA promiscuously binds multiple OM lipoproteins, periplasmic proteins, colicins, and even phage particles The mechanism could also be indirect. For example, Tol-Pal membrane contact sites could facilitate GPL movement by influencing the activity of other GPL trafficking proteins and their mechanisms 36 , Analogously, Tol-Pal might promote formation of membrane hemifusions between the IM outer leaflet and OM inner leaflet.
These intermediate bilayers could be conduits for GPL trafficking between the periplasmic leaflets of two segregated bilayers Fig.
Future biochemical work is necessary to deduce the exact biochemical mechanism. Salmonellae maintain two to four times fewer PGls within the OM than within the IM, but the mechanism and significance are not known Fig. However, the ybgC mutant Rif sensitivity and virulence attenuation were far less severe than those for the tolQ , tolR , and tolA mutants Fig.
Typhimurium tol-pal phenotypes, and additional biochemical mechanisms of Tol-Pal are necessary to fully promote antimicrobial resistance and virulence. Regarding the biological role for S. In support of PGls acting as receptors for antibiotics, spontaneous Gram-positive resistance to daptomycin, a cyclic lipopeptide antibiotic, involves mutations that cause three biochemical phenotypes related to membrane PGl homeostasis.
These include loss of PGl synthesis, neutralization of anionic PGl head groups by the addition of lysine, and inversion of lysinylated PGl molecules into the outer leaflet of the plasma membrane 55 , By analogy, S. Palmitoylating PGl head groups might prevent cathelicidin antimicrobial peptides from inserting into the S. Typhimurium bilayer, since PagP enhances salmonella resistance to these toxins Finally, PGls are the principal donor substrates for lipoprotein biosynthesis Typhimurium morphology, Rif resistance, and survival during infection.
The interacting proteins include acyl carrier protein ACP , sn -glycerolphosphate acyltransferase PlsB , and phosphatidylserine synthase Pss The functional consequence of the YbgC interactions with these proteins is not known.
The Pss enzyme is essential for enterobacterial viability and necessary for PE synthesis The exact biochemical role of CpoB during S. Typhimurium infection is more difficult to predict given the presumed absence of catalytic activity. For E. Recently, CpoB was implicated in altering peptidoglycan cross-linking for septation This was determined using in vitro protein interaction analysis and LpoB-induced Pbp1B-murein transpeptidation reactions.
However, no clear septation or murein cross-linking defect has been demonstrated for E. Therefore, the function of CpoB and the significance of the TolA interaction have not been fully elucidated. We show that CpoB promotes reduced S. In contrast, loss of CpoB did not influence S.
We postulate that during systemic pathogenesis, CpoB executes a key biochemical mechanism. It is possible that the mechanism involves TolQRA. Alternatively, CpoB functions independently of Tol-Pal to promote survival in mice. The binding interfaces for the TolA-CpoB interaction have been reported Therefore, it should be possible to determine the functional role of the predicted S. Typhimurium CpoB-TolA interaction toward the pathogenesis phenotype.
Many types of Gram-negative bacteria harbor the Tol-Pal system. In the case of S. Typhimurium, one common biological function of the seven Tol-Pal proteins is to promote survival during systemic pathogenesis in mice. These data will provide key insight into the biochemical involvement of S.
Typhimurium Tol-Pal during infection.
For the materials used in this study, see Materials and Methods in the supplemental material. The wild-type Salmonella enterica serovar Typhimurium S. Typhimurium bacteria were derived from the s genotype and contain a chromosomally integrated wza-lacZ gene promoter fusion Table S1 E-phase bacteria were obtained by a 1: Plasmid-bearing bacteria were grown with the respective antibiotics as follows: All the deletion-insertion mutants were created using the phage lambda red recombinase system Typhimurium genome, was achieved using the pGRG37 vector Second-site TolQ complementation was achieved by adding 0.
Point mutations were placed at the att Tn7 integration site as described above. E-phase bacteria were pelleted and resuspended in Z-buffer, and OD readings were taken to quantify culture density.
Bacteria were permeabilized with chloroform and 0.
Na 2 CO 3 at 1 M was added as a stop solution, and the mixture was centrifuged. For live-cell imaging, E-phase phase bacteria were labeled with FM 0. Agarose pads were made using a 0.
FMlabeled cells were spotted onto the pads and air dried. A coverslip was placed on the slide, and the sides were sealed with a hot-glue gun. Plating efficiency on rifampin Rif was determined by first adjusting the density of E-phase bacterial cultures to an OD of 1. The cells were then serially diluted fold in PBS. For the tolQ mutant rescue experiments, 0. Triplicate wells were infected for each genotype, and surviving intracellular CFU were enumerated at 2 and 6 hpi.
For additional details, see Materials and Methods in the supplemental material. Typhimurium bacteria were back diluted 1: The cells were collected by centrifugation and resuspended in a sucrose solution to begin the osmotic spheroplasting and lysis procedure, which allows for the efficient separation of the bilayers.
The IM and OM were isolated using a discontinuous sucrose density gradient and ultracentrifugation The membranes were resuspended in 1 ml of 1 mM Tris, pH 7. We thank Samuel I.
Miller for the idea to use wza-lacZ to identify S. Typhimurium barrier loci and Daleep K. Arora for technically assisting in laboratory tasks leading to this publication.
Supplemental material for this article may be found at https: National Center for Biotechnology Information , U.
Journal List Infect Immun v.
Cpob 2006 ebook download
Infect Immun. Published online Jun Prepublished online May 7. Revathi Masilamani , a Melina B. Cian , a and Zachary D. Dalebroux a. Melina B. Zachary D.
Author information Article notes Copyright and License information Disclaimer. Corresponding author. Address correspondence to Zachary D. Dalebroux, ude. Salmonella Tol-Pal reduces outer membrane glycerophospholipid levels for envelope homeostasis and survival during bacteremia.
Infect Immun Received Apr 27; Accepted Apr This is an open-access article distributed under the terms of the Creative Commons Attribution 4. This article has been cited by other articles in PMC. Associated Data Supplementary Materials Supplemental material. ABSTRACT Salmonellae regulate membrane lipids during infection, but the exact proteins and mechanisms that promote their survival during bacteremia remain largely unknown. Open in a separate window. FIG 1. Typhimurium tol-pal loci causes Rcs signaling activity to increase.
FIG 2. FIG 3. Overnight cultures of wild-type and tolQ , tolR , and tolA mutant salmonellae were back diluted 1: The cells were collected by centrifugation and resuspended in a sucrose solution, which began the osmotic spheroplasting and lysis procedure. The membranes were isolated by discontinuous sucrose density gradient ultracentrifugation A number sign indicates a statistically significant difference for wild-type S.
Tol-Pal proteins variably promote S. Typhimurium morphology. FIG 4. Typhimurium Tol-Pal proteins variably promote Rif resistance. Select Tol-Pal proteins are necessary for S. Inducing TolQ translation, in cis , restores Rcs signaling inactivity, cellular morphology, Rif resistance, and intracellular survival to tolQ mutant S. FIG 5. FIG 6. What is the biological role of S. Typhimurium limiting OM PGl concentrations? What is the biochemical role of YbgC and CpoB during systemic infection?
Bacterial strains, growth conditions, and genetics. Rif sensitivity assay. Membrane fractionation, protein quantification, and GPL extraction.
Supplementary Material Supplemental material: Click here to view. Footnotes Supplemental material for this article may be found at https: Now you see me, now you don't: Nat Rev Microbiol Salmonellae interactions with host processes.
Exploiting host immunity: Trends Immunol Santos JC, Enninga J. At the crossroads: Cell Microbiol Salmonellae PhoPQ regulation of the outer membrane to resist innate immunity.
Curr Opin Microbiol Fortifying the barrier: Making a membrane on the other side of the wall. Biochim Biophys Acta The power of asymmetry: Annu Rev Microbiol Mechanism of assembly of the outer membrane of Salmonella typhimurium. Isolation and characterization of cytoplasmic and outer membrane.
J Biol Chem PhoPQ regulates acidic glycerophospholipid content of the Salmonella Typhimurium outer membrane. MinD and MinE interact with anionic phospholipids and regulate division plane formation in Escherichia coli. Differential affinities of MinD and MinE to anionic phospholipid influence Min patterning dynamics in vitro.
Mol Microbiol Narita SI, Tokuda H. Bacterial lipoproteins; biogenesis, sorting and quality control.
petunjuk operasional cpob 2012 pdf
Crystal structure of E. Nat Commun 7: Konovalova A, Silhavy TJ. Outer membrane lipoprotein biogenesis: Lol is not the end. Detecting envelope stress by monitoring beta-barrel assembly. Cell Transmembrane domain of surface-exposed outer membrane lipoprotein RcsF is threaded through the lumen of beta-barrel proteins. Elife 5: Laloux G, Collet JF. Major Tom to ground control: J Bacteriol Genome expression analyses revealing the modulation of the Salmonella Rcs regulon by the attenuator IgaA.
Majdalani N, Gottesman S. The Rcs phosphorelay: Activation of the Rcs signal transduction system is responsible for the thermosensitive growth defect of an Escherichia coli mutant lacking phosphatidylglycerol and cardiolipin. Exploring the relationship between lipoprotein mislocalization and activation of the Rcs signal transduction system in Escherichia coli.
Microbiology Egan AJF. Bacterial outer membrane constriction. Proteins The YbgC protein encoded by the ybgC gene of the tol-pal gene cluster of Haemophilus influenzae catalyzes acyl-coenzyme A thioester hydrolysis. FEBS Lett Transmembrane alpha-helix interactions are required for the functional assembly of the Escherichia coli Tol complex.
J Mol Biol Characterization of the tol-pal region of Escherichia coli KWe now understand that some mechanisms involve multicomponent protein systems that span the envelope and derive energy from the IM in order to move GPLs across the periplasm. Juran TQ Trilogi Kualitas 1. It was possible that the deletion-insertion alleles were causing polar effects on adjacent tol-pal genes Fig. While they are not qualified to dispense medicines, providing them information to refer patients for appropriate care remains a valuable but unrealized aim.
Rosita Andriani. John W.
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