As reported from several other studies, both within Norway [17] a

As reported from several other studies, both within Norway [17] and from other countries like UK [34] and the US [35], there was a significant seasonal variation in the occurrence of hip fractures in our study. In a study comparing and observing seasonal variation selleck inhibitor of hip fractures in Scotland, Hong Kong and New Zealand [36] as well as in Taiwan [37], it was claimed evidence against a major influence of conditions underfoot causing extra falls and increased risk of fracture

during winter [36]. In our study, we had information about place of injury in 90% of all cases; 64% occurred indoors with no significant seasonal variation. For the fractures happening outdoors, there was a significant seasonal variation, which can be connected to falls on ice or slippery surfaces. Unfortunately, the data from the Harstad Injury Registry do not provide enough information for exact studies of the mechanisms leading to falls and fracture indoors. The mean age at hip fracture in persons above 50 years in Harstad, were not different from the mean age at hip fracture in Oslo, which

was 82.1 years in women and 76.6 years in men [8]. A lower mean age at fracture in men, compared to women, are also reported by others [26]. With 73% of the hip fractures occurring in women, the gender distribution of hip fractures in Harstad did not differ in comparison with Oslo (78%) or other comparable studies [12, 14]. Increased mortality risk up to 10 years PF-6463922 has been reported for hip fractures

[38], although mortality is highest in the first year [3, 38]. A sex difference in mortality after hip fracture has also been indicated, with higher rates in men compared with women [2, 3, 38, 39]. In our study, mortality Aprepitant was higher in men than in women 3 months after fracture and persisted at 6 and 12 months after adjustment for age of hip fracture. This is in accordance with other Norwegian data showing higher mortality in men throughout the first year after hip fracture [40], and with a recent meta-analyses showing that, although the sex difference in mortality persists, the difference is greatest in the first 3 months after hip fracture, with reported relative all-cause mortality hazard of 5.75 (95% CI, 4.94–6.67) in women and 7.95 (CI, 6.13–10.30) in men [41]. One of the strengths of this study is the possibility to study the incidence of hip fractures in a well-defined municipality over a long time period and the accessibility of a well-established injury registry, which also provides the opportunity for quality assessment of the hip fracture registration. Furthermore, the injury registry provided valuable information on date and place of fracture and through the medical records we got access to mortality data. There are, however, several limitations in our dataset.

1999; Hopkinson et al 2000) The third gap is located between di

1999; Hopkinson et al. 2000). The third gap is located between different disciplines of science, thus it is a disciplinary gap. One particularly booming field selleck products of biodiversity research deals with the analysis of potential consequences of biodiversity loss for ecosystem processes such as seed dispersal or element cycling (e.g. Hooper et al. 2005). While in this functional biodiversity research

species loss serves as the starting point, the questions addressed are usually generic, e.g. related to investigate whether complex ecosystems generally function differently from more simple ones. To answer such questions, researchers often apply strictly controlled experiments, either in the field or in contained laboratory microcosms, e.g. by artificially creating (plant) communities with different levels of diversity and/or structural complexity (e.g. Schmid and Hector 2004). Biodiversity Batimastat ic50 experiments provide innovative research platforms that may generate hundreds of papers, such as in the case of the Jena Experiment (Roscher et al. 2004). A second recent approach in biodiversity research is that of comparative studies in real landscapes, with plots that are managed differently. Land use is a main driver of biodiversity loss

and comparing the effects of land use on biodiversity and ecosystem processes, such as in the Biodiversity Exploratories (Fischer et al. 2010), again provides a platform for interdisciplinary research that potentially yields outcomes relevant for conservation. However, there appears to be a disciplinary gap between fundamental biodiversity science and conservation science that does not just include differences in the Astemizole topics being addressed, but apparently there are also different subsets of scientists addressing the different topics. While scientists conducting functional biodiversity research often argue that their work is relevant

to conservation (Hector et al. 2001), this is regularly questioned (Srivastava and Vellend 2005). As a consequence, the importance of functional biodiversity research for conservation is often reduced to providing a general argument for why conservation is necessary for humankind, such as in the Millennium Ecosystem Assessment that classified the ecosystem services that are potentially adversely affected by a loss in biodiversity (Millennium Ecosystem Assessment 2005a, b). Another click here example is given by population genetics where fundamental research often focuses on the genetics of natural indigenous grazers, while applied conservation research focuses, for example, on the mechanistic effect of grazing by domestic animals on plant recovery in nature reserves. A link between these types of research is often lacking.

In image e, B bacteriovorus HD100 (blue) are shown attached at o

In image e, B. bacteriovorus HD100 (blue) are shown attached at one pole to P. tolaasii 2192T (yellow), a crucial first step in the predatory process. Images d and e both show rounded P. tolaasii 2192T cells, characteristic of the bdelloplast structures formed after Bdellovibrio invades the host cell and begins replication. 1 μm scale bar shown. Where B. bacteriovorus HD100 was added to the mushroom surface both before (Figure 3e) and after P. tolaasii 2192T (Figure 3d), B. bacteriovorus

HD100 attachment CP-690550 to P. tolaasii 2192T cells was observed: a crucial first step in the predatory process. In addition, bdelloplasts, the rounded, dead P. tolaasii structures in which Bdellovibrio establish, grow and replicate after attachment and invasion, were also observed where B. bacteriovorus HD100 was added before or after P. tolaasii 2192T. Although a valid statistical survey is not possible in these SEM samples, AZD0156 price bdelloplasts were most clearly visible on the mushroom surface where B. bacteriovorus HD100 was added before

P. tolaasii 2192T (Figure 3d). This correlates with the greater reduction in lesion intensity measurements on mushrooms where B. bacteriovorus HD100 was added before P. tolaasii 2192T (Figure 2): Bdellovibrio attachment to prey and subsequent bdelloplast formation may be easier, and occur more rapidly, where P. tolaasii cells have not had time to accumulate, adapt and adhere to the mushroom surface, preventing P. tolaasii from producing as much tolaasin, and thus reducing the extent of the characteristic brown blotch symptoms. A King’s Medium B control addition to LY2835219 the pileus resulted in the growth of different types of bacterial cells, with different morphologies that were distinct from that of P. tolaasii 2192T & B. bacteriovorus HD100 (Figure 3f); however, typically, no bacterial cells were observed on untreated mushroom tissue (Figure 3a). This indicates that the supermarket mushrooms carry a small, indigenous bacterial microflora that replicates about readily in added growth medium, which may impact upon P. tolaasii CFU numbers recovered from experimentally inoculated tissue, as described

below. Application of Bdellovibriobefore inoculation with P. tolaasiireduced the number of P. tolaasiiin infected mushroom tissue To determine whether the reduction in lesion intensity after treatment with B. bacteriovorus HD100 correlated with a reduction in P. tolaasii 2192T cell numbers, CFU were recovered and enumerated from mushroom tissue that had been inoculated with P. tolaasii 2192T and pre-treated with B. bacteriovorus HD100, compared with a P. tolaasii 2192T inoculated, non-B. bacteriovorus HD100 treated control (Figure 4). A mean number of 4.5 × 107 and 3.9 × 107 CFU were recovered from mushrooms pre-treated with 2.9 × 106 or 1.4 × 107 PFU live B. bacteriovorus HD100 respectively, which were both significantly lower than the mean 1.9 × 108 CFU recovered from mushrooms inoculated with P.

Coculture of breast stromal fibroblasts with primary mammosphere

Coculture of breast stromal fibroblasts with primary mammosphere cells Coculture of primary mammosphere cells (1 × 105 cells/dish) with breast stromal fibroblasts

(1 × 105 cells/dish) were performed by using a transwell (BD) cell culture system, which allows free diffusion see more of substances without contact between cancer cells and stromal fibroblasts. Stromal fibroblasts in the insert layer were subcultured on a transwell cell culture membrane (7.5 cm in diameter; pore size: 0.4 μm), and mammosphere cells in the bottom layer were maintained in a 10-cm Petri dish. Stromal fibroblasts were precultured in DMEM/F12 with 10% FBS for 48 h before the start of coculture. Stromal fibroblasts were maintained in fresh serum-free DMEM/F12 medium, and mammosphere cells were cultured in Stem Cells inhibitor suspension for six days. Coinoculation of mammosphere cells with different stromal fibroblasts in vivo Mammospheres and fibroblasts were collected, enzymatically dissociated, washed in PBS, and kept at 4°C. Mice were

maintained in laminar flow rooms under constant temperature and humidity and received an estradiol supplementation (0.6 mg/kg, s.i., mTOR inhibitor review Sigma) every 7 days for 28 days before cell injection. The mammosphere cells (1 × 105) admixed with either CAFs (1 × 105) or NFs (1 × 105) were suspended in 0.1 ml of DMEM/F12 and then inoculated into the mammary fat pad of 5-week-old female NOD/SCID mice (Shanghai Experimental Animal Center, Chinese Academy of Sciences, Shanghai, China). Mice were examined by palpation for tumor formation for up to 12 weeks, and then were sacrificed Telomerase by cervical dislocation. The histologic features of the xenografts were examined by hematoxylin and eosin staining. All experimentation performed with NOD/SCID mice, as well as routine care of the animals, was carried out in accordance with the institutional guide of animal care & use committee. Measurement of SDF-1 The baseline level of SDF-1 production was determined by coculture of mammosphere cells with stromal fibroblasts

for six days at a density of 1 × 105/bottle. The concentration of SDF-1 in the supernatant was measured by using a human SDF-1 antibody and enzyme immunoassay kit (R&D Systems, Minneapolis, MN), according to the manufacturer’s instructions. Statistical analysis Statistical analysis was performed by using GraphPad Prism 4.0 software© (San Diego, CA). Student’s t-test (for comparison between two groups) or ANOVA with Tukey post test (for comparison between more than two groups) were used to determine whether there exists statistically significance. Fisher exact probability test was used to analyze tumorigenicity in NOD/SCID mice. Data is presented as the mean ± SEM. P values of ≤ 0.05 were regarded as being statistically significant.

Significantly, there is an increased risk for HIV seroconversion

Significantly, there is an increased risk for HIV seroconversion in both women

and men following infection with T. vaginalis [12–17]. On the other hand, T. tenax is a commensal of the human oral cavity found under conditions of poor oral hygiene and advanced periodontal disease. Its prevalence in the mouth ranges from 4% to GSI-IX 53% [18]. Interestingly, both T. vaginalis and T. tenax have SN-38 recently been reported to be associated with broncho-pulmonary infections in patients with Pneumocystis carinii or with underlying cancers or other lung diseases [18–24]. Although speculative to date, the organisms of both species are believed to enter the respiratory tract by aspiration from the oropharynx. While lung infection by the oral trichomonads can be envisioned, the mechanisms by which the urogenital parasites establish residence in the oral cavity for subsequent oropharyngeal and respiratory infections is unclear. Furthermore and importantly, these reports question the extent of the genetic interrelatedness and host site tropisms between these two species. The phylogenetic analyses based on the rRNA and selleck compound class II fumerase gene sequences have shown that Trichomonas species formed a closely related clade, including isolates of Trichomonas gallinae, T. tenax, and T. vaginalis [25, 26]. Given the common host specificity of T. vaginalis

and T. tenax, and the relatedness with respect to rRNA sequences, we felt it important to attempt to determine the extent of genetic identity between the two species. One strategy by us was to identify uniquely-expressed genes of T. vaginalis that may represent determinants that contribute to urogenital virulence and pathogenesis. We, therefore, used two approaches. The first involved the subtraction cDNA library approach and the second involved screening a cDNA expression library with pooled patient sera adsorbed with 3-mercaptopyruvate sulfurtransferase T. tenax antigens. We hypothesized that T. vaginalis and T. tenax would be significantly

genetically unrelated to permit isolation of many uniquely-expressed genes of T. vaginalis. However, to our surprise, while a few T. vaginalis genes were identified, the genes were found to be identical with those of T. tenax. We determined that the isolated T. vaginalis genes had increased amounts of mRNAs, indicating elevated expression at the transcriptional level. While functional analyses of these up-regulated genes may provide insight about the role of these proteins in trichomonal virulence, our data suggest that both T. vaginalis and T. tenax have remarkable genetic identity but different rates of gene expression. Results PCR-based cDNA subtractive hybridization We have successfully used the PCR-based cDNA subtraction method to isolate differentially expressed cDNAs among two different cDNA populations called tester (T. vaginalis) and driver (T. tenax) [27].

J Electro Mater 2009, 38:586–595 CrossRef 37

J Electro Mater 2009, 38:586–595.CrossRef 37. AZD5582 datasheet Li S, Bi H, Cui B, Zhang F, Du Y, Jiang X, Yang C, Yu Q, Zhu Y: Anomalous magnetic properties in Co 3 O 4 nanoparticles covered with polymer decomposition residues. J Appl Phys 2004, 95:7420–7422.CrossRef 38. Zhang S, Pelligra CI, Keskar G, Majewski PW, Ren F, Pfefferle LD, Osuji CO: Liquid crystalline

order and magnetocrystalline anisotropy in magnetically doped semiconducting ZnO nanowires. ACS Nano 2011, 5:8357–8364.CrossRef 39. Pelligra CI, Majewski PW, Osuji CO: Large area vertical alignment of ZnO nanowires in semiconducting polymer thin films directed by magnetic fields. Nanoscale 2013, 5:10511–10517.CrossRef 40. Singhal RK, Dhawan MS, Gaur SK, Dolia SN, Kumar S, Shripathi T, Deshpande UP, Xing YT, Saitovitch E, Garg KB: PI3K Inhibitor Library cell line Room-temperature

ferromagnetism in Mn-doped dilute ZnO semiconductor: an electronic structure study using X-ray photoemission. J Alloys Compd 2009, 477:379–385.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions BSK and SL designed and planned the experiments. BSK performed powder and nanowire synthesis and measurements. BSK, SL, and SYJ performed data analysis and interpretation. WKK, JHP, and YCC assisted with sample characterization and contributed to measurement discussions. JK, CRC, and SYJ wrote the manuscript with help from the co-authors. All authors discussed the results and reviewed the manuscript. All authors

read and approved the final manuscript.”
“Background Nowadays, the rapid development of microfluidic/nanofluidic systems has been seen in many applications such as fluid mixing [1, 2], drug delivery [3], ion transporters [4], and DNA selleck kinase inhibitor translocators [5]. The micro/nanochannels are the key components in the microfluidic/nanofluidic systems. Recently, more complex nanochannels (e.g., with some Gemcitabine datasheet nanostructures at the bottom) are designed to study the influences on the flowing characteristic of fluid in the nano/microchannels [2]. The successful fabrication of these micro/nanochannels urgently needs to be solved. At present, the nanochannel fabrication methods mainly include focused ion beam milling [5], nanoimprint lithography [6], electron beam drilling [7], and wet chemical etching [8]. However, the complexity and/or cost of these methods greatly restrict the nanochannel fabrication, especially for the nanochannel with complex nanostructures at the bottom. Since atomic force microscopy (AFM) was invented, the AFM tip-based nanomachining method had emerged as one of the essential technologies for nanostructure fabrication [9]. A lot of works have already been carried out to fabricate nanochannels on the surfaces of different kinds of materials using this method [10–15]. For example, Zhang et al. [13] presented an AFM-based high-rate tunable nanolithography technique to scratch nanochannels on PMMA surfaces. Kawasegi et al.

Endogenous peroxidase activity was blocked by immersion in 0 3% m

Endogenous peroxidase activity was blocked by immersion in 0.3% methanolic peroxide for 30 minutes. Next, sections were microwaved in citrate buffer for antigen retrieval. Rabbit polyclonal anti-HBO1 (1:100 dilutions) was used as primary antibodies. Staining was completed with Polink-2-plus kit, in accordance with the manufacturer’s instructions. Color reaction product was developed with diaminobenzidine. All PFT�� mw sections were counterstained with hematoxylin. Two

pathologists separately blinded to the clinical outcome of the patients evaluated all samples. The immunoreactivity intensity was evaluated as 0 (absent); 1 (weak nuclear staining); 2 (moderate nuclear staining of intermediate level); 3 (more coarse nuclear staining). Positive cells were quantified Blasticidin S cost as a percentage of the total number of tumor cells with observation of 1000 cells in 5 high power field (×400), and assigned to one of five categories: 0: < 5%, 1: 5-25%, 2: 26-50%, 3: 51-75% and 4: > 75%. HBO1 expression was scored semi-quantitatively using the Remmele-score (immunoreactive score (IS) = score of percentage of positive

tumor cells × score of staining intensity). Then the slide of IS > 3 was classified as a positive case [11]. Reproducibility of the scoring method used by both observers was greater than 95%. Total RNA extraction and real-time PCR Total mRNA samples of T47 D and MCF-7 breast cancer cells were extracted using trizol reagent according to the manufacturer’s instructions. One microgram of total RNA extracted from the cells was subjected to reverse transcription (RT). The RT and real-time

PCR were performed by using TaKaRa RNA PCR Kit (AMV) Ver.3.0 and SYBR Methocarbamol Premix Ex Taq II according to the manual respectively. Primers used for real-time PCR were as follows: HBO1-F 5′-GATGCCCACTGTATCATAACC-3′ and HBO1-R 5′-TTCTTCCTGAGTTCAGCCACT-3′; GAPDH-F 5′-GGCTGAGAACGGGAAGCTTGTCAT-3′ and GAPDH-R 5′-CAGCCTTCTCCATGGTGGTGAAGA-3′. Real-time PCR was performed using 7500 multicolor real-time PCR detection system (ABI) with the following cycling conditions: (i) 30s at 95°C and (ii) 40 cycles, with 1 cycle consisting of 5s at 95°C, 34s at 60°C. GAPDH was employed as an internal control under the same experimental conditions. Data were analyzed by using 7500 software (ABI). The values were obtained through normalizing to GAPDH copies. Statistical see more analysis Statistical data analyses were performed using SPSS 11.5 statistical software package. The relationship between protein levels and different clinical and pathological features were explored using cross tabulation and Pearson’s x2. P values less than 0.05 were selected. Results HBO1 protein level correlates positively with ERα In order to explore the biological role of HBO1 in human breast cancer in vivo, we examined the expression of HBO1 in tumor samples of primary breast cancer (n = 112) by IHC analysis.

The P1 fragments were expressed in E coli system and all these f

The P1 fragments were expressed in E. coli system and all these fragments were expressed in inclusion bodies. A protocol was developed to purify these protein fragments to near homogeneity and to obtain

these proteins in large amount. The testing of P1 protein fragments with anti-M. pneumoniae sera and sera of M. pneumoniae infected patients revealed that all these protein fragments were recognized by these sera, thereby suggesting that the immunodominant regions are distributed across PND-1186 the entire length of the protein. These results are in agreement with a number of previous reports that showed the presence of immunodominant regions either in the N-, middle and C-terminal segments of P1 protein [21, 23, 25, 27]. A number of previous reports have shown the presence of immunodominant epitopes usually in the C-terminal of M. pneumoniae P1 protein [21, 23, 36], but few reports also showed immunodominant regions in the middle and extreme N-terminal [25, 27]. A comparative summary of these results is presented in additional figure file 4 [see Additional file 4]. However, our’s is the first

study that systematically scanned the full P1 protein for their immunodominant and cytadherence. Sotrastaurin chemical structure Since P1 protein is considered to be the major ligand mediating attachment, we next tested the ability of the antibodies raised against the four P1 fragments for adhesion detection, surface exposure and adhesion inhibition assays to identify the cytadherence regions. Previously, a number of studies have identified a few M. pneumoniae P1 regions involved in cytadherence. Trypsinization of

M. pneumoniae medroxyprogesterone P1 protein and ability of various fragments or peptides so generated to block cytadherence buy TSA HDAC provided first evidence for the role of P1 protein in cytadherence [4]. Baseman et al., later showed that the treatment of M. pneumoniae with protease blocked its adherence to tracheal explants which was restored when P1 was re-generated [32]. Role of M. pneumoniae P1 protein in cytadherence was further substantiated by a study where pre-treatment of M. pneumoniae with antiserum directed against the P1 protein blocked its cytadherence to hamster tracheal ring up to 80% [37]. Gerstenecker and Jacobs [11] and Opitz and Jacobs [24], showed the involvement of N-terminal, middle and C-terminal segment of M. pneumoniae (P1) as well as M. genitalium (MgPa) in cytadherence. Although a number of above mentioned studies have highlighted the role of M. pneumoniae P1 protein in cytadherence, however a systematic study spanning the entire length of P1 protein is missing. We performed a systematic analysis of surface exposure and cytadherence region(s) for M. pneumoniae P1 protein fragments spanning the entire length.

The theoretical value of A** can be calculated using A** = 4πm*qk

The theoretical value of A** can be calculated using A** = 4πm*qk 2/h 3, where h is Planck’s constant. For BAY 11-7082 in vivo n-type GaN, m* = 0.22m o is the effective electron mass for GaN and the value of A** is determined to be 26.4 A/(cm2K2). Zhou et al. [21] also reported

that the value of A** determined by a modified Richardson plot in the GaN material is close to the theoretical value. The values were calculated using both values of σ so obtained GW3965 in vitro for the temperature ranges of 100 to 220 and 220 to 340 K. Thus, in Figure 6, the circles represent the plot calculated with σ so = 90 mV (straight line 1) in the temperature range of 100 to 200 K, and the squares represent the plot calculated with σ so = 176 mV (straight line 2) in the temperature range of 200 to 380 QNZ molecular weight K. The best linear fits to the modified experimental data are depicted by solid lines in Figure 6 which represent the true activation energy plots in respective temperature ranges. The calculations have yielded zero-bias mean SBH ϕ bo of 0.92 eV (in the range of 100 to 220 K) and 1.82 eV (in the range of 220 to 340 K). In Figure 6, the intercepts at the ordinate give the Richardson constant A** as 72.4 A/(cm2K2) (in the range of 100 to 220 K) and 32.2A/(cm2K2) (in the range of 220 to 340 K) without using the temperature coefficient

of the SBHs. This value of the Richardson coefficient at room temperature is close to the theoretical value 26.4A/(cm2K2) [14, 16–20, 23]. It can be pointed out that although a barrier inhomogeneity is visible in Pt/GaN diodes, But highlighting

feature of these diodes, is high Schottky barrier height observed. The quality of the metal–semiconductor interface is affected by the process steps and deposition vacuum since contamination and oxide layer growth at the interface may result in SBH reduction and high leakage current by inducing local nanoscopic patches of low barrier heights. Studies by Iucolano et al. revealed that this kind of inhomogeneous behavior is observed in all semiconductors and results in overall decreased barrier heights [10]. The contamination level and oxide layer can be minimized by following fabrication steps in a clean room and depositing 2-hydroxyphytanoyl-CoA lyase Schottky metals in UHV. By selecting high work function metal Pt, a high gate potential can be achieved. These kinds of high barrier heights are suitable for many high-power and switching applications. The reverse characteristics of these devices are also quite good as compared to those of other Schottky metal combinations. Very low reverse leakage current and high breakdown voltages are good for high-power applications where losses should be low. A high rectifying ratio is desired for switching applications. These diodes are better in terms of observed Schottky barrier height and reverse characteristics. Figure 6 Modified Richardson plot, [ln( I 0 / T 2 ) -  q 2 σ so 2 /2 k 2 T 2 ] versus 1/ T , for the Pt/n-GaN Schottky diode.

Since PC required 6-fold more PhlA than lecithin for induction of

Since PC required 6-fold more PhlA than lecithin for induction of 50% hemolysis (Fig. 4A), the egg yolk lecithin used in this study might have contained enough LPL for hemolysis. However, no hemolysis was induced by lecithin without PhlA treatment (Fig. 4D). Taken together, these results indicated that PhlA phospholipase activity hydrolyzed PL and produced LPL. Since LPL is known to be a surfactant [33], it may have been the final effector leading to destabilization of the RBC membrane and hemolysis.

Cytotoxicity of PhlA in the Bafilomycin A1 solubility dmso presence of phospholipid We examined buy GSK872 the cytotoxicity of PhlA using HeLa and 5637 cells. PhlA had cytotoxic activity against both HeLa and 5637 cells in the presence of lecithin (Fig. 4E). To investigate the cytolytic activity of late log phase S. marcescens culture

supernatants, S. marcescens was grown at 37°C for 6 h in LB containing PL. Up to 48-fold dilutions of the S. marcescens culture supernatant induced cell death of both HeLa and 5637 cells, while supernatant of S. marcescens ΔphlAB cultured under the same conditions had no effect on HeLa or 5637 cells, indicating that PhlA was an extracellular secretion product (data not shown). Discussion A wide range of pathogenic bacteria produce phospholipases, selleck screening library and the putative role of PLA in virulence has been studied in some of these pathogens. Outer membrane-associated PLAs (OMPLAs) were first identified in E. coli [34] and orthologs were subsequently reported in numerous gram-negative bacteria, including H. pylori (PldA) [9]. The OMPLAs have been well-characterized and are thought to enhance bacterial growth, colonization, and survival. In addition to modulation of the bacterial membrane, some OMPLAs were shown to have contact-dependent hemolytic/cytolytic activities [35]. Another group of PLAs (e.g., YplA [12], ExoU [36], PlaA [10], and SlaA [37]) is secreted from bacterial cells. Purified ExoU and SlaA [38, 39] recombinant proteins do not show cytotoxic activity when added exogenously, and there is little information next on the cytotoxicity of other secretory

PLAs. To our knowledge, ShlA is the only previously reported hemolysin from S. marcescens. Although, a ΔshlAB mutant showed hemolytic activity on blood agar plates, it did not exhibit contact-dependent hemolytic activity (Fig. 1C). Therefore, we performed functional cloning, which identified PhlA as an S. marcescens candidate hemolytic factor (Fig. 2A). In the experiments reported here, we described the hemolytic and cytotoxic activities of S. marcescens PhlA. PhlA itself did not directly induce the destabilization of target cell membranes, but the LPL produced from PL by PhlA phospholipase activity showed hemolytic and cytolytic activities. Therefore, PhlA and ShlA have different hemolytic mechanisms. In addition, ShlA was expressed at lower temperature, but its expression decreased at 37°C [17].