, 1998; Holo et al., 2001; Maldonado et al., 2003; Diep et al., 2009). Strain-related differences in bactericidal activity affect the susceptibility of other microorganisms to plantaricins and organic acids (Ehrmann et al., 2000; Omar et al., 2006; Nielsen et al., 2010). None of the strains had genes for plantaricins NC8, S, or W (Table 1). With the methodology used, plantaricin A-, EF-, JK-, and N-related genes were detectable in all strains except for TO1001 (Table 1). Similar to the case of TO1001, L. plantarum strain 3.9.1, isolated from an African fermented

food, does not have any of these plantaricin genes (Omar et al., 2006). Certain L. plantarum strains show the following different types of plantaricin-related gene combinations: (1)

plnEF and plnW; (2) plnD, plnEF, plnI, and plnG; (3) plnD, plnJ, plnK, and plnG; (4) INCB018424 chemical structure plnD, plnEF, plnI, plnK, and plnG; (5) plnA, plnC, plnD, plnEF, plnI, plnJ, plnK, and plnN (Omar et al., 2006; Moghadam et al., see more 2010). Thus, the characteristics of the gene combinations carried for the production of plantaricins in TO1000, TO1002, and TO1003 are unique among the known L. plantarum strains isolated from fermented products. The synthesis of plantaricin A is observed from early exponential to early stationary phase. During stationary phase, the amount of plantaricin A strikingly declines (Diep et al., 1994). The addition of sucrose to the medium enhances production of nisin, another bacteriocin produced by Lactococcus lactis, (Devuyst & Vandamme, 1992). Thus, bacterial growth rate and available nutrients are associated with antimicrobial activity. In fact, the rates of fermentation differed among the four strains at 30 and 60 days of storage (Tables 3 and 4), suggesting that, in addition to the divergence in the available carbohydrates, the capacity for production of organic acids, and

the pH and temperature preferences for growth, antimicrobial activity may also be an important factor in the regulation of silage fermentation quality. Further 4-Aminobutyrate aminotransferase studies are needed both to elucidate the production of plantaricins by the TO strains inoculated in silage and to understand their roles in the improvement of silage quality. In conclusion, phenotypic and genotypic differences were present among LAB strains in spite of their belonging to the same species and subspecies, and the fermentation quality of silage inoculated with different conspecific strains differed significantly, supporting the idea that suitable LAB inoculants should be selected on a strain basis. Because TO1002 most effectively improved the fermentation quality in terms of pH decrease, regulation of undesirable microorganisms, and high DM recovery, this strain should be the most suitable inoculant for longer storage of paddy rice silage. The selected L. plantarum subsp.

In-depth qualitative interviews were undertaken

with 11 key MHRA members. A recorded semi-structured interview conducted within MHRA’s building, a topic guide (the role of pharmacists and GPs, which elements should be considered and how this should be communicated) was used to interview. A purposive sample of knowledgeable participants recruited thought a gatekeeper from different employment levels, including senior management, middle management, employees and senior employees, with knowledge of the counterfeiting medicines issue. University ethics committee approval for the overall project was gained. Framework learn more analysis approach was used to identify themes (2). Three main themes were identified relating to the roles of pharmacists and GPs in combating counterfeit medicines from the perspective of MHRA’s members. The first theme identified four roles for pharmacists and GPs in combating counterfeit medicines; these were: being vigilant for any suspicion of counterfeit cases; being a good source of reporting to the regulatory agency; providing check details awareness and advice for patients; as well as needing to source their medicines from a secured supply chain. The second theme related to how those roles should be communicated by the regulatory agency to pharmacists and GPs; participants recommended using media tools, working with their professional bodies and training

such as undergraduate and CPD courses. The third theme focused on what decision-makers within a regulatory agency should consider when defining those roles. Participants suggested; the regulatory agency should consider improving their communication and

speeding access to the relevant information; the need for the regulatory agency to taking patient’s confidentiality seriously in dealing with this issue; and the amount of information the agency should share with the pharmacists and GPs regarding counterfeiting medicines. This study was developed in the context of a very limited range of published Isoconazole literature. Senior and middle management MHRA managers have a clear view as to what the role of pharmacists and GPs should be in the combatting counterfeit medicines. A need to better communicate the role of pharmacists and GPs was also identified in addition to methods of delivering this. The views of the professions themselves on this are currently unknown. For the roles of pharmacists and GPs in combating counterfeit medicines to be better understood and refined, further studies are required to address the contribution and views of other stakeholders of the regulatory agency. 1. Jackson G, Patel S, Khan S. Assessing the problem of counterfeit medications in the United Kingdom. International Journal of Clinical Practice. 2012;66(3):241–250. 2. Srivastava A, Thomson SB. Framework analysis: a qualitative methodology for applied policy research. JOAAG. 2009;4(2):72–79. H. Family, E. Bell, V. Choo, S. Hassan, D.

Quantitative real time PCR was also performed to validate the corresponding rise in the transcript levels of these genes. Escherichia coli YZ2005 for Red/ET homologous recombination was kindly provided by Dr Youming Zhang (Genebridges GmbH, Germany). Escherichia coli S17-1 was used as the donor strain in intergeneric conjugation.

The spinosad-producing strain S. spinosa CCTCC M206084 was isolated by our laboratory from the south of China. For routine use, all strains of E. coli were grown in Luria–Bertani medium at 37 °C supplemented with antibiotics as required (apramycin, Am, 50 μg mL−1). Saccharopolyspora spinosa Mitomycin C purchase was grown in tryptic soy broth (TSB; Difco) at 30 °C. For fermentation, S. spinosa and its exconjugants were first grown for 2 days at 30 °C in the seed medium containing 1% glucose, 0.9% yeast extract, 0.2% MgSO4·7H2O, and 0.05% KH2PO4, followed by 10 days in production medium PM1 containing 0.1% KNO3, 0.05% K2HPO3·3H2O, 0.001% FeSO4, 0.05% MgSO4·7H2O, 0.4% yeast, and 0.4% tryptone. To improve yield further, fermentation was performed in a modified production medium GW-572016 chemical structure PM2 containing 6% glucose,

2% starch, 2% soybean meal, 1% fish meal, 1% corn syrup, 0.3% glutamine, 1% soybean oil, and 0.4% CaCO3. Plasmid pSET152 was obtained from Dr Meifeng Tao (Central China Agricultural University, China) and was used as template for PCR amplifying the linear cloning vector. The Red/ET recombination was performed as described previously (Zhang et al.,

2000). To clone the partial spinosyn biosynthetic gene cluster (c. 18 kb) directly, a 50-μL aliquot of Red/ET-competent (ET+) E. coli YZ2005 cells was co-electroporated with 0.3 μg of linear cloning vector and 5 μg genomic DNA of S. spinosa CCTCC M206084 in a Bio-Rad Gene Interleukin-2 receptor Pulser Apparatus (Bio-Rad Ltd, Richmond, CA). The linear cloning vector was amplified with primer pair P1/P2 (Supporting Information, Table S1) using pSET152 as template. Each primer P1/P2 contains a 50-bp homologous arm for the cloning of the spinosyn gene cluster. To guarantee the correction of the sequence of the homologous arms, two c. 800-bp fragments covering the homologous arms from S. spinosa CCTCC M206084 were amplified and sequenced using primer pairs P3/P4, P5/P6 designed according to the published spinosyn biosynthetic gene cluster sequence of S. spinosa NRRL 18538 (GenBank accession number: AY007564, Waldron et al., 2001). The sequencing results had 99% identities with the corresponding sequences of S. spinosa NRRL 18538. Two 50-bp regions were chosen as homologous arms. The genomic DNA was isolated according to Kieser et al. (2000) and was completely digested by Xho I (Takara, Japan) which occurs outside the c. 18-kb target genes to expose the homologous arms.

Family history was notable for malignancies including breast, nasopharyngeal and colon cancers. Physical exam disclosed hypertension, bilaterally enlarged, firm, non-tender

parotid glands, fine bibasilar crackles and bipedal edema. Anti Ro/Sjögren’s syndrome antigen A antibody was positive, with negative tests for anti La/Sjögren’s syndrome antigen B and anti-nuclear antibody (ANA). Chest radiographs showed basal infiltrates. Sjögren’s syndrome associated with glomerulonephritis and interstitial lung disease was Cabozantinib datasheet diagnosed, and she received pulse methylprednisololone followed by oral prednisone with dramatic improvement. Two months later, while on prednisone 5 mg/day, she returned to the clinic with an enlarging fixed non-tender right breast mass. She underwent modified radical mastectomy of the right breast, and pathologic report revealed diffuse, small cell, non-Hodgkin’s lymphoma of the breast; axillary lymph nodes were negative for tumor. She opted for alternative find more therapy and did not return to the clinic until

7 months later when she developed sudden monocular blindness in the right eye with no other systemic manifestations. Magnetic resonance imaging (MRI) revealed swelling and enhancement of intracanalicular and pre-chiasmatic segments of the right optic nerve and right side of the optic chiasm. Considerations were Devic’s disease versus metastases. She received pulse methylprednisolone therapy (1 g/day for 3 days) many with partial recovery of vision. She is scheduled for lymphoma chemotherapy to include rituximab. “
“The aim of this study was to assess the effects of anti-tumor necrosis factor (TNF) agents or disease-modifying antirheumatic drugs (DMARDs) on hepatitis B virus (HBV) reactivation in hepatitis B surface antigen (HBsAg)-positive patients with rheumatic diseases. Evidence of HBV reactivation after anti-TNF therapy or DMARDs

in HBsAg-positive patients with rheumatic disease was summarized by performing a systematic review. A total of 122 HBsAg-positive rheumatic disease-positive patients undergoing treatment with an anti-TNF agent or with DMARDs were identified in nine studies. In eight of the studies, the anti-TNF agents used were etanercept in 56 cases, adalimumab in 25 cases and infliximab in 14 cases. Follow-up periods ranged from 6 to 52 months. Antiviral prophylaxis was administrated in 48 of the 122 patients (39.3%). HBV reactivation in HBsAg-positive patients taking an anti-TNF agent or DMARD was reported in 15 cases (15/122 = 12.3%). Ten of the 15 patients provided individual data on HBV reactivation: four patients had rheumatoid arthritis, four had ankylosing spondylitis and two had psoriatic arthritis; four received etanercept, and two received infliximab. In one of the four etanercept-treated cases in which the patient had elevated HBV-DNA levels, antiviral prophylaxis was also administered.

To eliminate the disturbing

effect of the fusion protein (Fig. 3b), the fusion transposase producer plasmid was eliminated from five yjjY mutants and the motility of these strains was tested again. Reduced motility was observed in all cases, indicating that in (or close to) the yjjY gene, a DNA segment is located that affects motility. Because the sequence of the yjjY insertion site showed high similarity to the consensus used by the wt IS30 transposase, we tested whether the wt IS30 uses this target sequence as a hot spot. Only seven yjjY mutants were TGF-beta inhibitor found to be generated by the wt IS30 out of the 222 mutants tested. These data demonstrate that the fusion transposase has a much more pronounced target preference for the yjjY hot spot (17.3%) compared with that of the wt transposase (3.2%). In this study, we have worked out and successfully applied a novel method based on IS30-mediated site-directed mutagenesis in order to produce nonflagellated S. Enteritidis mutants. The system was constructed based on the assumption that the FljA repressor component of the fusion transposase – as a DNA-binding protein – would bind to its target (the operator of fliC), and as a consequence, insertions could be concentrated with a relatively high frequency in the flagellin operon. The system constructed on the above basis worked well

and generated insertions. It turned out that the sequenced insertion sites showed pronounced similarity to the IS30 consensus sequence Bay 11-7085 of insertions (Table 1;

Olasz et al., 1998). This buy Tanespimycin indicated that the fusion transposase retained the target recognition ability of the wt IS30 transposase. Another feature of the insertions was that four target sites – called hot spots – were utilized several times. One of these hot spots was the target sequence in the fliD gene and these insertions resulted in nonmotile phenotypes. This fact could be considered as a proof of FljA-targeted transposition, because fliD is located in close proximity to the fliC operator sequence, which is the binding site of the native FljA repressor protein. These data suggested that the fusion of the FljA repressor protein modulated the target preference of the IS30 transposase and increased the frequency of integration into a new target site not preferred by the wt transposase. This result is in good agreement with earlier observations that the target preference of IS30 transposase can be modified by fusing the enzyme to unrelated DNA-binding proteins (Szabo et al., 2003 and unpublished data). Unexpectedly, another highly preferred hot spot was identified in the putative gene yjjY. Although this target site was recognized by both the wt and the fusion transposase, the frequency of the mutations generated by the IS30–FljA transposase was almost six times higher than that of the wild type (17.3% vs. 3.2%).

The reaction was loaded onto a 12.5% SDS-PAGE gel, which was autoradiographed and analyzed by BAS1800 (Fuji film). For Western blot analysis, the cytoplasmic domain of BtkB was incubated with 0.1 mM ATP, 1 mM DTT, and 5 mM MgCl2 at 37 °C for 1 h. Also, ATPase activity was performed in 20 μL of 40 mM Tris–HCl buffer (pH 7.0), 1 mM DTT, 5 mM MgCl2, 1 mM ATP, and 2 μg BtkB at 37 °C for 60 min. Released phosphate was measured with the malachite green reagent (Enzo life sciences). Myxococcus xanthus wild-type and btkB mutant cells were grown in CYE medium and harvested in the exponential growth

phase and stationary phase. Also, developmental cells were prepared on CF agar plates or CYE medium containing 0.5 M glycerol. Approximately 2 × 107 cells were dissolved in SDS sample buffer, and denatured find more proteins were separated by SDS-polyacrylamide gel electrophoresis (PAGE). Proteins were then transferred to PVDF membranes for Western blotting. The membranes were incubated with a horseradish peroxidase–conjugated antiphosphotyrosine PY20 (Santa Cruz Biotechnology). Blots were developed with ECL reagent (GE Healthcare). Total RNA was isolated from exponential and stationary phase cells and during cell development at 24, 48, and 72 h and then treated with DNase (Promega). After inactivation of DNase, cDNA was synthesized from the RNA samples (each 0.8 μg) using PrimeScript II RTase (Takara Bio Inc.) and random

hexamers, and PCR was performed with Kapa SYBR Fast qPCR master mix (KAPABiosystems), primers (RTbtkBN and RTbtkBC, Table S1), and the synthesized cDNA using the ABI 7300 real-time cycler. The mRNA levels of a downstream gene (MXAN_1029) were also determined Selleck Sirolimus by qRT-PCR analysis using the primers (RT1029N and RT1029C, Table S1). A control without reverse transcriptase was performed to detect residual contaminating genomic DNA. Exponential phase cells (8 × 108 cells mL−1) in CYE medium were used for the assays. Cells were harvested, washed, and resuspended at approximately 5 × 108 cells mL−1 in TM buffer. A total Adenylyl cyclase of 360 μL of the cell

suspension was mixed with 40 μL dye stock solution (150 μg mL−1 Congo red and 100 μg mL−1 trypan blue). The assay was performed as previously described (Black & Yang, 2004). Cells grown in CYE medium were harvested in the exponential growth phase and stationary phase, washed three times with distilled water, and then sonicated without glass beads three times for 1 min each. Cells were also starved on CF agar and harvested at 48 and 96 h. The cells were sonicated with glass beads five times for 1 min each. The supernatant and pellet were separated by centrifugation three times at 10 000 g for 10 min. The pellet was washed three times with distilled water. The sugar contents of the supernatant and pellet suspension were determined at 490 nm by the phenol–sulfuric acid method, with glucose as the standard (Dubois et al., 1956). BtkB consists of 710 amino acid residues with a calculated molecular mass of 78.4 kDa.

The reaction was loaded onto a 12.5% SDS-PAGE gel, which was autoradiographed and analyzed by BAS1800 (Fuji film). For Western blot analysis, the cytoplasmic domain of BtkB was incubated with 0.1 mM ATP, 1 mM DTT, and 5 mM MgCl2 at 37 °C for 1 h. Also, ATPase activity was performed in 20 μL of 40 mM Tris–HCl buffer (pH 7.0), 1 mM DTT, 5 mM MgCl2, 1 mM ATP, and 2 μg BtkB at 37 °C for 60 min. Released phosphate was measured with the malachite green reagent (Enzo life sciences). Myxococcus xanthus wild-type and btkB mutant cells were grown in CYE medium and harvested in the exponential growth

phase and stationary phase. Also, developmental cells were prepared on CF agar plates or CYE medium containing 0.5 M glycerol. Approximately 2 × 107 cells were dissolved in SDS sample buffer, and denatured Selleckchem CYC202 proteins were separated by SDS-polyacrylamide gel electrophoresis (PAGE). Proteins were then transferred to PVDF membranes for Western blotting. The membranes were incubated with a horseradish peroxidase–conjugated antiphosphotyrosine PY20 (Santa Cruz Biotechnology). Blots were developed with ECL reagent (GE Healthcare). Total RNA was isolated from exponential and stationary phase cells and during cell development at 24, 48, and 72 h and then treated with DNase (Promega). After inactivation of DNase, cDNA was synthesized from the RNA samples (each 0.8 μg) using PrimeScript II RTase (Takara Bio Inc.) and random

hexamers, and PCR was performed with Kapa SYBR Fast qPCR master mix (KAPABiosystems), primers (RTbtkBN and RTbtkBC, Table S1), and the synthesized cDNA using the ABI 7300 real-time cycler. The mRNA levels of a downstream gene (MXAN_1029) were also determined Dasatinib mouse by qRT-PCR analysis using the primers (RT1029N and RT1029C, Table S1). A control without reverse transcriptase was performed to detect residual contaminating genomic DNA. Exponential phase cells (8 × 108 cells mL−1) in CYE medium were used for the assays. Cells were harvested, washed, and resuspended at approximately 5 × 108 cells mL−1 in TM buffer. A total Clomifene of 360 μL of the cell

suspension was mixed with 40 μL dye stock solution (150 μg mL−1 Congo red and 100 μg mL−1 trypan blue). The assay was performed as previously described (Black & Yang, 2004). Cells grown in CYE medium were harvested in the exponential growth phase and stationary phase, washed three times with distilled water, and then sonicated without glass beads three times for 1 min each. Cells were also starved on CF agar and harvested at 48 and 96 h. The cells were sonicated with glass beads five times for 1 min each. The supernatant and pellet were separated by centrifugation three times at 10 000 g for 10 min. The pellet was washed three times with distilled water. The sugar contents of the supernatant and pellet suspension were determined at 490 nm by the phenol–sulfuric acid method, with glucose as the standard (Dubois et al., 1956). BtkB consists of 710 amino acid residues with a calculated molecular mass of 78.4 kDa.

106 It may be that at the expense of generating mutations, mammalian cells may use transient up-regulation of Pol ι to deal with replication arrest by DNA damage for survival.107 However, continuous over-expression of such error prone DNA polymerase, for instance by chronic hypoxia, may

result in a high rate of point mutations.108 As mentioned above, germline mutations in NBS1 predispose it to the Nijmegen breakage syndrome. The NBS1 protein forms a complex with MRE11A and RAD50 called MRN, which interacts with double-strand breaks and begins the DNA damage response by recruiting the ATM protein (see above). Inactivation of NBS1 impairs the function of MRN, leading to a high sensitivity to radiation, CIN and defective cell cycle checkpoints. To et al. demonstrated that hypoxia (1% O2 for Romidepsin solubility dmso 16 h) down-regulates NBS1 expression at the mRNA and protein levels in cancer cell lines.109 They showed that this down-regulation is

HIF1 but not HIF2 dependent and is mediated by reduction of Sp1-MYC by competing Sp1-HIF1 at the promoter region of the NBS1 locus, similar to the MSH2 locus.86,109 All cancers contain a much greater number of genetic and epigenetic alterations than do corresponding PLX3397 normal cells. At nucleotide levels, these alterations include: substitutions of one base by another,

insertions or deletions of small or large segments of DNA, rearrangements, copy number increases, copy number reductions, acquisition of foreign DNA (virus) in some cases and hypermethylation Atorvastatin or hypomethylation of guanosine residue.3 The cancer genome also shows changes in numbers of whole or parts of chromosomes. It is reasonable to assume that these genetic alterations can be caused in part by exposure to environmental carcinogens. Data from the whole genome sequencing of melanoma showed clearly the contribution of UV radiation to the melanoma genome.110 Interestingly, there is a sign of the second genetic insult after UV damage is detected in the genome and this is characterized by an increase in the frequency of C > A transversions.110 It is tempting to speculate that the second event occurring in the melanoma genome may be associated with H/R. As reviewed in this article, H/R is a strong candidate for induction of genetic alterations and the DNA damage response found in cancer genomes and tissues; however, our insights into H/R on the cellular genome are all based on experiments performed in tissue culture or in animal models. The question is whether H/R really plays the same contributing role for genetic instability in human tumor tissues as observed in experimental systems.

The bacterial cells of a culture and the extracellular medium were separated in their different compartments as described in Materials and methods and CtpA was detected in the subcellular fractions by Western blot. CtpA could not be determined in fractionations prepared from the P. aeruginosa PAO1 wild-type strain after overnight culture in liquid media (data not shown). Variation of culture conditions such as different growth times and increased temperature also did not result in CtpA detection. This was probably due

to an extremely low redundancy of the protein, which may be only expressed in very low amounts or may only be present for a short time during the cell cycle in concentrations below the detection limit. Therefore,

an expression vector was constructed containing the coding sequence of the PA5134 Selleckchem ALK inhibitor ORF, without the putative promoter region but maintaining the ribosome-binding site and the signal peptide. The coding region was under transcriptional control of the lac promoter which is constitutively transcribed in P. aeruginosa resulting in a moderate overexpression (Rosenau & Jaeger, 2004). When P. aeruginosa harbouring this plasmid was grown for 4 h and fractionated afterwards, Western blot analysis showed that RG7422 in vitro CtpA was clearly detectable and resided exclusively in the periplasmic fraction (Fig. 2). The extracellular protein exotoxin A and the periplasmic protein DsbA detected with the respective specific antisera served as controls (Lory et al., 1983; Urban et al., 2001). Lane 1 of Fig. 2 represents the protein from whole cells and demonstrates that CtpA was expressed and could

be detected with the peptide-specific antibodies. CtpA showed the same distribution within the fractions as the known periplasmic protein DsbA (lane 3). Moreover, CtpA was not detected in the cytoplasmic fraction or in the membrane fraction, as lanes 4 and 5 indicate. The first localization study of Hara et al. (1991) in maxicells found that Prc was primarily present in the cytoplasmic membrane and periplasm. Silber et al. (1992) later identified Prc in the periplasm and membrane fractions. In our experiments, P. aeruginosa CtpA was present only in the periplasm, indicating that the presence of Prc found in the nonperiplasmic fraction of the former studies is likely due Carnitine palmitoyltransferase II to the artificial amounts of the protease present due to overexpression, as suggested by the authors. Lane 5 shows that no extracellular CtpA was present, whereas exotoxin A in lane 5 was solely detected in the extracellular fraction, as expected. A CTP from C. trachomatis, Tsp, was able to interfere with the NF-κB pathway by cleaving the p65 protein of the host immune system after expression in a human cell line, as shown in a recent study (Lad et al., 2007). Those authors considered these results as indicating a role for Tsp in a mechanism to evade the host immune system, which is obligate for intracellular pathogens as C. trachomatis.

The bacterial cells of a culture and the extracellular medium were separated in their different compartments as described in Materials and methods and CtpA was detected in the subcellular fractions by Western blot. CtpA could not be determined in fractionations prepared from the P. aeruginosa PAO1 wild-type strain after overnight culture in liquid media (data not shown). Variation of culture conditions such as different growth times and increased temperature also did not result in CtpA detection. This was probably due

to an extremely low redundancy of the protein, which may be only expressed in very low amounts or may only be present for a short time during the cell cycle in concentrations below the detection limit. Therefore,

an expression vector was constructed containing the coding sequence of the PA5134 MS-275 datasheet ORF, without the putative promoter region but maintaining the ribosome-binding site and the signal peptide. The coding region was under transcriptional control of the lac promoter which is constitutively transcribed in P. aeruginosa resulting in a moderate overexpression (Rosenau & Jaeger, 2004). When P. aeruginosa harbouring this plasmid was grown for 4 h and fractionated afterwards, Western blot analysis showed that see more CtpA was clearly detectable and resided exclusively in the periplasmic fraction (Fig. 2). The extracellular protein exotoxin A and the periplasmic protein DsbA detected with the respective specific antisera served as controls (Lory et al., 1983; Urban et al., 2001). Lane 1 of Fig. 2 represents the protein from whole cells and demonstrates that CtpA was expressed and could

be detected with the peptide-specific antibodies. CtpA showed the same distribution within the fractions as the known periplasmic protein DsbA (lane 3). Moreover, CtpA was not detected in the cytoplasmic fraction or in the membrane fraction, as lanes 4 and 5 indicate. The first localization study of Hara et al. (1991) in maxicells found that Prc was primarily present in the cytoplasmic membrane and periplasm. Silber et al. (1992) later identified Prc in the periplasm and membrane fractions. In our experiments, P. aeruginosa CtpA was present only in the periplasm, indicating that the presence of Prc found in the nonperiplasmic fraction of the former studies is likely due GPX6 to the artificial amounts of the protease present due to overexpression, as suggested by the authors. Lane 5 shows that no extracellular CtpA was present, whereas exotoxin A in lane 5 was solely detected in the extracellular fraction, as expected. A CTP from C. trachomatis, Tsp, was able to interfere with the NF-κB pathway by cleaving the p65 protein of the host immune system after expression in a human cell line, as shown in a recent study (Lad et al., 2007). Those authors considered these results as indicating a role for Tsp in a mechanism to evade the host immune system, which is obligate for intracellular pathogens as C. trachomatis.