Lloyd et al examined 148 human pituitary adenomas for VEGF prote

Lloyd et al. examined 148 human pituitary adenomas for VEGF protein expression by immunohistochemistry, and showed positive staining in all groups with stronger staining in GH, ACTH, TSH, and gonadotroph adenomas and in pituitary carcinomas [27]. Our study detected 190 positive VEGF expression cases in 197 PAs and 58.9% of check details them are in high expression level, including 60.7% of PRL-secreting PAs, 78.4% FSH-secreting PAs, 51.9% ACTH-secreting PAs and 57.1% non-functioning

PAs. Niveiro et al. investigated VEGF expression in 60 human pituitary adenomas, and found that low expression of VEGF was seen predominantly in prolactin cell adenomas, and high in non-functioning adenomas, which is different from our data that 60.7% of prolactin cell adenomas verses 57.1% non-functioning adenomas [11]. Moreover, VEGF was considered also involved in conventional medical therapy for PAs. Octreotide was reported to down-regulate VEGF expression to achieve antiangiogenic effects on PAs see more [28]. Gagliano et al. demonstrated that cabergoline reduces cell viability in non-functioning pituitary adenomas by inhibiting VEGF secretion, of which the modulation might mediate the effects of DA agonists on cell proliferation in non-functioning adenoma [29]. Interestingly, in present study, we did GS-9973 spearman’s rank correlation analysis and found that D2R expression did not show a

correlation with VEGF expression. Although it is prospective to treat PAs by anti-VEGF, up to now, only one case of PA has been reported to be cured by bevacizumab [6]. The mechanisms of VEGF in PA genesis and progression are still unclear. More studies are needed to investigate the effects of anti-VEGF therapy on PA patients. To confirm the results, we also detected the expression of D2R, MGMT and VEGF by using western blot. The data supported the results of immunohistochemical staining. Two samples were selected for each PAs subtype. The positive expression of western blot indicated the immunohistochemical staining is available, and the thickness differences of the blot band revealed the expression level differences

in separate sample. Moreover, by spearman’s rank correlation analysis, we found that MGMT expression was positively associated with D2R and VEGF expression in PAs. As far Nintedanib (BIBF 1120) as we know, it is the first time to report the association of D2R and MGMT expression which is positive. Only one report by Moshkin et al. has ever mentioned the association of MGMT and VEGF expression in PA. They demonstrated a progressive regrowth and malignant transformation of a silent subtype 2 pituitary corticotroph adenoma, with significant VEGF and MGMT immunopositivity [30]. The association between VEGF and MGMT expression in PAs need further investigations, as well as D2R and MGMT expression. In addition, we analyzed the association of D2R, MGMT and VEGF expression with clinical features of PAs, but no association was found.

1:10 000) and were geo-statistically analysed using ArcGIS-ArcInf

1:10 000) and were geo-statistically analysed using ArcGIS-ArcInfo software, v. 9.2 (ESRI 2006–2009) and the program Fragstats 3.0 (McGarigal et al. 2002). Intersecting the two vector layers allowed demarcating areas where historically-old www.selleckchem.com/products/psi-7977-gs-7977.html meadows persisted, new meadows had been created, and historical meadows had been replaced by other Belnacasan price habitat types. Habitat fragmentation analysis examined the area covered by the target

meadow types in historical and recent times. For each study area and time period, individual grid maps (4 m × 4 m resolution) were produced illustrating the spatial distribution of (1) wet meadows, (2) species-rich mesic meadows, and (3) the combined area of the two meadow types. The grids were imported to Fragstats 3.0 and the following class-level landscape metrics were calculated: percentage

of the landscape (PLAND) covered by a given habitat type, number of patches (NP), patch density (PD), area-weighted mean of patch size (AM), total class area (CA) and effective mesh size (MESH) equalling the sum of patch area squared, summed across all patches of the corresponding patch type and divided by the total landscape area. For MESH, AM and total extent, Ipatasertib ic50 the significance of changes between the two time periods was tested by a Wilcoxon-test for pair-wise differences using R-software (R Development Core Team 2010). Results Changes in the extent of floodplain meadows In the six unprotected study areas, wet and species-rich mesic meadows declined enormously between the 1950/1960s and 2008 (differences significant at p ≤ 0.05; Fig. 2, Table 2). On average, wet meadows lost 85.2% of their former area, and species-rich mesic meadows decreased by 83.6%. Wet meadows were nearly completely lost at the Weser and the Luppe with <5 ha remaining, while species-rich

SSR128129E mesic meadows were reduced to about 8 ha. In the largest study area (Helme), a 83% loss led to a remaining wet meadow area of 100.3 ha, of which 77.5 ha were historically old and 22.8 ha were newly created after 1969. The Helme floodplain also harbours at present the largest area of species-rich mesic meadows (12.3 ha), of which 8.3 ha were newly created. The current extent of wet meadows in the Havel protected area was comparatively large (100.8 ha), but only about a third was historically old. While wet meadows at the Havel declined only slightly during the past decades (by 7.4%), the loss of species-rich mesic meadows was substantial (54.3%). Fig. 2 Areas of wet meadows (black) and species-rich mesic meadows (grey) in two of the seven study areas a Ems, b Havel, in the 1950/1960s and in 2008.

5) and frozen at -20°C for 15 min After thawing at room temperat

5) and frozen at -20°C for 15 min. After thawing at room temperature, the samples were centrifuged

at 10,000 × g. The supernatant containing the desired protein was applied onto affnity matrix of agarose coupled with p-aminobenzyl-1-thio-β-D-galactopyranoside (PABTG-agarose, Sigma) (10 ml column) equilibrated with four volumes of buffer A. The column was washed with 300 ml of the buffer A, and the recombinant β-D-galactosidase was eluted three times with 10 ml of 0.05 M sodium borate (pH 10.0) buffer at a flow rate of 0.5 ml/min. Active fractions containing the β-D-galactosidase were collected and dialyzed three times LY3039478 mouse against 3 L of buffer D (100 mM NH4HCO3). In case of the purification of the extracellular produced β-D-galactosidase in P. pastoris cultures, the yeast

cells were separated from the post-culture medium through centrifugation. Next, the ammonium sulphate was added to the post-culture medium to 60% w/w, at 4°C. The precipitated proteins were centrifugated at 20,000 × g, dissolved in buffer A and dialyzed overnight against the same buffer. For β-D-galactosidase purification the dissolved sample was applied further directly onto affnity matrix of agarose coupled with p-aminobenzyl-1-thio-β-D-galactopyranoside and purified as described above for bacterial system. The concentration of purified protein was determined by the Bradford method using bovine serum albumin (BSA) as a find more standard. β-D-galactosidase activity assays The activity of purified Arthrobacter sp. 32c β-D-galactosidase was determined by the use of chromogenic substrates as described elsewhere [4, 14]. The PRN1371 cost o-nitrophenol released from 10 mM of o-nitrophenyl-β-D-galactopyranoside (ONPG) by β-D-galactosidase at 0–70°C and pH range 4.5–9.5 (0.02 M citrate buffer for pH 4.5 and 5.5; 0.02 M K2HPO4-KH2PO4 for pH 6.5 and 7.0 and 0.02 M Tris-HCl for pH selleck products 8.5 and 9.5) was measured

at 405 nm. The reaction was stopped after 10 min with 1 M Na2CO3. One unit is defined as one micromolar of o-nitrophenol released per minute. Substrate specifiCity was estimated using 1 mM solution of chromogenic substrates: o-nitrophenyl-β-D-galactopyranoside (ONPG), p-nitrophenyl-β-D-galactopyranoside (PNPG), o-nitrophenyl-β-D-glucopyranoside (ONPGlu) and p-nitrophenyl-β-D-glucopyranoside (PNPGlu). Activity determination was carried out under standard conditions in 0.02 M K2HPO4-KH2PO4 (pH 6.5) buffer at 10, 20, 30, 40 or 50°C. The activity of the β-D-galactosidase towards lactose was monitored by HPLC analysis (column Bio-rad, Aminex HPX-87H) where 1% solutions of lactose, glucose, fructose and galactose were used as standards. In the combined enzyme assay glucose isomerase from Streptomyces murinus (Sigma G4166) was used in the amount of 0.01 g/ml of 5% w/v solution of lactose (0.02 M K2HPO4-KH2PO4, pH 6.5). The Arthrobacter sp. 32c β-D-galactosidase was used at concentration of 200 U/ml of the mixture.

However, this phenomenon has only been evaluated on a limited num

However, this phenomenon has only been evaluated on a limited number of strains [12–16]. Therefore, the objective of this study was to further explore the “seesaw effect” in 150 clinical strains with varying susceptibilities. Additionally, eight selleck kinase inhibitor strains were utilized in time–kill studies to determine if the response to CPT was affected by changing glyco- or lipopeptide NSC23766 in vivo susceptibilities in isogenic strain pairs. Materials and Methods Bacterial Strains A total of 150 clinical MRSA strains from the Anti-infective Research Laboratory (Detroit, MI,

USA) collected between 2008 to 2012 were chosen for evaluation of the “seesaw effect”. All strains were randomly chosen clinical blood isolates. Additionally, four isogenic strain pairs were selected for further evaluation of these antibiotics in time–kill curves to compare differences in kill between parent and reduced selleck chemicals llc susceptibility

to VAN mutant isolates. Antimicrobials Ceftaroline (Teflaro®) powder was provided by Forest Laboratories, Inc. (New York, NY, USA). DAP (Cubicin®) was purchased commercially from Cubist Pharmaceuticals (Lexington, MA, USA). VAN and TEI were purchased commercially from Sigma Chemical Co. (St. Louis, MO, USA). Media Due to the calcium-dependent mechanism of DAP, MHB was supplemented with 50 mg/L of calcium and 12.5 mg/L of magnesium for all experiments. Colony

counts were determined using tryptic soy agar (TSA) (Difco, heptaminol Detroit, MI, USA). Susceptibility Testing Minimum inhibitory concentrations (MIC) for all study antimicrobials were determined by Etest methods according to the manufacturer’s instructions. Additionally, broth microdilution MICs were performed in duplicate at 1 × 106 according to Clinical and Laboratory Standards Institute (CLSI) guidelines for isogenic strain pairs as a comparison/validation of MICs determined by Etest methodology [18]. All samples were incubated at 37 °C for 18–24 h. The following MIC data were determined for each tested antimicrobial: average MIC, MIC50, and MIC90. These MIC data were analyzed by linear regression to derive correlations coefficients between agents. In Vitro Time–Kills Four isogenic strain pairs were chosen as representative strains for evaluation in time–kill curves. Briefly, macro-dilution time–kill experiments were performed in duplicate using a starting inoculum of approximately 1 × 106 CFU/mL as previously described [17–19]. The 24-well culture plate was utilized with 100 μL of antibiotic stock solution, 200 μL of a 1:10 dilution of a 0.5 McFarland standard organism suspension, and sufficient volume of CAMHB for a total volume of 2 mL. Sample aliquots (0.1 mL) were removed over 0–24 h and serially diluted in cold 0.9% sodium chloride.

14(56): 10 (1985) [1984] ≡ Hygrocybe pratensis (Fr ) Murrill, Myc

14(56): 10 (1985) [1984] ≡ Hygrocybe pratensis (Fr.) Murrill, Mycologia 6(1): 2 (1914), ≡ Agaricus pratensis Fr., Observ. mycol. (Havniae) 2: 116 (1818), sanctioned by Fr., this website Syst. mycol. 1: 99 (1821).

Characters as in Cuphophyllus; basidiomes clitocyboid, pileus usually pigmented brown, orange, salmon, or buff, rarely cream; surface not or scarcely viscid; lamellae usually appearing opaque (chalky); pileipellis usually a cutis, not an ixocutis; basidiospores usually globose, subglobose or broadly ellipsoid, mean spore Q mostly 1.2–1.4, rarely up to 1.8. Phylogenetic support In our Supermatrix analysis (Fig. 2), sect. Cuphophyllus is a strongly supported (99 % MLBS) monophyletic group. Sect. Cuphophyllus is also highly supported in our LSU analysis (Fig. 3), but only species in the C. pratensis complex are included.

The ITS analysis by Dentinger et al. (unpublished) shows a strongly supported C. pratensis clade (100 % MLBS) comprising a terminal clade (100 % MLBS) and a subtending grade with very deep divergences, while C. pratensis var. pallida appears as a separate clade nearby (100 % MLBS). Species included Type species: Cuphophyllus pratensis. Molecular phylogenies indicate C. pratensis is a species complex. Cuphophyllus bicolor is included based on strong support in our Supermatrix analysis, morphology and pigments. Species included based on morphology alone are Camarophyllus panamensis Lodge & Ovrebo, Cuphophyllus neopratensis Courtec.

& Fiard, Camarophyllus subpratensis (Beeli) Heinem., Camarophyllus Fludarabine nmr subrufescens (Peck) Murrill, Selleck SN-38 Cuphophyllus umbrinus (Protein Tyrosine Kinase inhibitor Dennis) Courtec., Hygrocybe austropratensis A.M. Young, and Hygrocybe watagensis A.M. Young. Cuphophyllus pratensis var. pallidus (Cooke) Bon. is strongly supported in an ITS analysis by Dentinger et al. (unpublished data). Comments Sect. Cuphophyllus is strongly supported, but greater taxon sampling is needed as sequences are limited to the C. pratensis species complex. Support for inclusion of C. bicolor in sect. Cuphophyllus is strong in our Supermatrix analysis (99 % MLBS) and weak in our ITS-LSU analysis (55 % MLBS). Cuphophyllus bicolor, Cam. panamensis and Cuph. umbrinus differ from other species in sect. Cuphophyllus in having a central strand of nearly parallel hyphae bounded by lateral strata with interwoven hyphae in the lamellar context. Cuphophyllus sect. Virginei (Bataille) Kovalenko, in Nezdoiminogo, Opredelitel’ Gribov SSSR (Leningrad): 37 (1989) Type species: Cuphophyllus virgineus (Wulfen : Fr.) Kovalenko (1989) ≡ Hygrocybe virginea P.D. Orton & Watling, Notes R. bot. Gdn Edinb. 29(1): 132 (1969), ≡ Agaricus virgineus Wulfen, in Jacquin, Miscell. austriac. 2: 104 (1781), sanctioned by Fr., Syst. mycol. 1: 100 (1821).

However,

cerebral perfusion reduced significantly in the

However,

cerebral perfusion reduced significantly in the NF group compared to baseline and sham operated animals (Figure 4). Renal blood flow reduced significantly in both kidneys after hemorrhage compared to baseline levels, NF group reduced renal blood flow, in both kidneys, compared to all other groups (Figures 5A and 5B). Arterial blood flow to the liver was significantly reduced in the NF group compared to all other groups (Figure 6A). The portal blood flow to the liver was also significantly reduced in the NF group compared to baseline levels; there were no statistical differences amongst the other groups (Figure 6B). The NF group showed a significant reduction in the gastrointestinal blood flow compared to baseline and sham operated animals; there was no statistical Nutlin-3a molecular weight difference between NBP and PH groups (Figure 7). Blood flow to the spleen reduced significantly in the NF group compared to all other Wortmannin order groups (Figure 8). However, splenic blood flow in the NBP and PH groups were only statistically different compared to baseline (Figure 8). No statistical difference was noted in the blood flow to the myocardium (Figure 9A). Blood flow to the lungs reduced significantly

in all hemorrhage groups compared to baseline levels, regardless to the resuscitation regimen used (Figure 9B). Figure 4 Perfusion of the left cerebral hemisphere. * p < 0.05 NF vs. baseline and sham groups; no statistically significant difference between NBP vs. PH (p > 0.05). NF = No Fluid; NBP = Normal Blood AZD0156 order Pressure; PH = Permissive Hypotension. Figure 5 Perfusion of the kidneys. Right kidney (Figure 5A) and left kidney (Figure 5B), * p < 0.05 NBP and

5 FU PH vs. baseline; ** p < 0.05 NF vs. all other groups; no statistically significant difference between NBP vs. PH (p > 0.05). NF = No Fluid; NBP = Normal Blood Pressure; PH = Permissive Hypotension. Figure 6 Perfusion of the liver. Arterial perfusion to the liver (Figure 6A) and portal perfusion of the liver (Figure 6B). * p < 0.05 NF vs. all other groups; no statistically significant difference between NBP vs. PH (p > 0.05). NF = No Fluid; NBP = Normal Blood Pressure; PH = Permissive Hypotension. Figure 7 Gastrointestinal perfusion. * p < 0.05 NF vs. baseline and sham; no statistically significant difference between NBP vs. PH (p > 0.05). NF = No Fluid; NBP = Normal Blood Pressure; PH = Permissive Hypotension. Figure 8 Perfusion of the spleen. * p < 0.05 NBP and PH vs. baseline; ** p < 0.05 NF vs. all other groups, no statistically significant difference between NBP vs. PH (p > 0.05). NF = No Fluid; NBP = Normal Blood Pressure; PH = Permissive Hypotension. Figure 9 Perfusion of the myocardium and lung. Myocardial perfusion (Figure 9A) and lung perfusion (Figure 9B) after hemorrhage and resuscitation. * p < 0.05 NF, NBP, and PH vs. baseline and sham groups; no statistically significant difference between NBP vs. PH (p > 0.05).

The solid product was collected and washed repeatedly with THF un

The solid product was collected and washed repeatedly with THF until pH = 7 and dried under vacuum. The product was denoted as PAAGNPs. Reaction of PAA-GNPs and KH550

PAA-GNPs 100 mg, DCC 100 mg and THF 100 mg were mixed by sonication for 1 h. Then, the solution of KH550 was added dropwise into suspension at 60°C under nitrogen atmosphere. When completed, the reaction was kept at 60°C and vigorously stirred for 24 h. At last, the solid product was collected and washed MLN2238 mouse repeatedly with THF until pH = 7 and dried under vacuum. The KH550 functionalized GNPs were denoted as siloxane-GNPs. Preparation of SiO2/GNPs hybrid material Siloxane-GNPs (50 mg) were added into 10 ml deionized water and stirred for 24 h at room temperature to hydrolyze the alkoxysilane into Si-OH. Then, 0.6 g TEOS, 1.2 g ammonia solution, and 100 ml ethanol were added to the suspension and stirred for 8 h. Finally, the solid product was collected and washed repeatedly with THF until pH = 7 and dried under vacuum. In this process, the quantity of TEOS, the Hedgehog inhibitor quantity of ammonia, and the time of reaction can be different. Thus, we can control the size of SiO2 particles. Orthogonal array experimental design

In the present study, the experiment was based on an orthogonal array experimental design where the following three factors were analyzed: the quantity of TEOS, the quantity of ammonia and the reaction time. These variables were identified to have large effects on the growth of SiO2 particles.

So an orthogonal array of three factors and three levels was employed to assign the considered factors and levels as shown in Table  1. In principle, one column could be assigned to a factor. Here, the matrix denotes three factors, each with three levels (Table  2). Data analysis could be P-type ATPase carried out through the range analysis. Table 1 Levels of factor of orthogonal design Level   Factors     TEOS (g) NH3 · H2O (g) Time (h) 1 0.3 0.6 4 2 0.6 1.2 6 3 0.9 1.8 8 Table 2 Orthogonal AZD5153 supplier arrays for statistical experiment and results No. Experiment conditions Results   Ethanol (ml) Temperature (°C) TEOS (g) NH3 · H2O (g) Time (h) Average particle size (nm) 1 100 30 0.3 (1) 0.6 (1) 4 (1) 50 2 100 30 0.3 (1) 1.2 (2) 6 (2) 120 3 100 30 0.3 (1) 1.8 (3) 8 (3) 140 4 100 30 0.6 (2) 0.6 (1) 6 (2) 100 5 100 30 0.6 (2) 1.2 (2) 8 (3) 240 6 100 30 0.6 (2) 1.8 (3) 4 (1) 170 7 100 30 0.9 (3) 0.6 (1) 8 (3) 130 8 100 30 0.9 (3) 1.2 (2) 4 (1) 160 9 100 30 0.9 (3) 1.8 (3) 6 (2) 280 Characterizations Fourier transform infrared spectrometer (FTIR, Nexus 670, Valencia, CA, USA) was used to detect the functional groups on the surface of f-GNPs and f-GNPs/SiO2 hybrid materials, which was measured as pellets with KBr.

Meanwhile, the atomic percentage content of titanium in the tooth

Meanwhile, the atomic percentage content of titanium in the tooth shape particles is 12.14%; it is almost consistent with the experimental process in which the molar ratio of titanium and zinc is 1 to 10. It manifests that titanium is almost utterly doped in the ZnO. The crystalline characters of the samples are checked by selected area electron diffraction. Figure 5(a3) shows that samples synthesized from zinc acetate have certain crystalline state, and the crystalline grain size is slightly larger. The (101), (102), and (112) crystal #PXD101 ic50 randurls[1|1|,|CHEM1|]# faces are detected. This is consistent with the XRD. When the raw material is zinc sulfate, the diffraction pattern displays the ( 10)

lattice plane of Zn (SO4)2 · 3Zn (OH)2 and (101), (102), and (201) lattice

planes of ZnO (Figure 5(b2)). The result is consistent with the XRD. When the raw material is zinc nitrate, (101), (102), and (201) crystal planes of ZnO are detected, and the diffraction rings are obscure (Figure 5(c3)). It demonstrates that the samples are composed of amorphous and crystalline forms. The SAED pattern of the samples prepared from zinc chloride displays the (002), (101), (102), (110), (103), (200), and (201) crystal planes of ZnO (Figure 5(d3)). It indicates that the samples are hexagonal phase. Besides, there are some scattered bright spots in the diffraction pattern. It demonstrates that the grain size is slightly larger. Antibacterial properties of titanium-doped ZnO powders Tables 1 and 2 both show that the antibacterial activities of titanium-doped ZnO powders synthesized from selleck inhibitor different zinc salts is different. The antibacterial activities of the powders are optimal, which is prepared from zinc chloride, and their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) are lower than 0.25 g L−1. Moreover, the antibacterial properties of the powders synthesized from zinc nitrate are slightly

poorer than that of zinc chloride and are better than that of zinc acetate and zinc sulfate. Meanwhile, the antibacterial activities of the powders against E. coli are better than S. aureus. Table 1 Colony count of E. coli after antibacterial activities by titanium-doped ZnO powders Zinc salt Powder concentration (g/L) 0 0.25 0.5 0.75 1.0 1.5 2.0 2.5 Zn (Ac)2 1.25 × 108 2.1 × 107 1.95 × 107 1.75 × 107 1.2 × 107 3.85 × 106 Methane monooxygenase 2.9 × 103 1.65 × 103 ZnSO4 1.1 × 107 9.75 × 106 5.3 × 106 2.95 × 105 5.6 × 104 1.6 × 104 7.65 × 103 Zn (NO3)2 2.15 × 107 1.9 × 107 1.65 × 107 1.6 × 107 3.35 × 105 2.8 × 103 0 ZnCl2   3.05 × 104 6.55 × 103 3.9 × 103 2.5 × 103 2.3 × 103 2.0 × 103 0 The initial bacterial colony count is 8.75 × 105 CFU/mL. Table 2 Colony count of S. aureus after antibacterial activities by titanium-doped ZnO powders Zinc salt Powder concentration (g/L) 0 0.25 0.5 0.75 1.0 1.5 2.0 2.5 Zn (Ac)2 1.95 × 108 5.25 × 107 5.2 × 107 4.0 × 107 3.4 × 107 3.0 × 107 4.15 × 105 2.1 × 103 ZnSO4 8.85 × 107 8.

The supernatants were collected and subjected to Western blotting

The supernatants were collected and subjected to Western blotting with anti-WNV E protein monoclonal antibody. Discussion WNV NY strains have a highly virulent phenotype compared to the Eg strain which was isolated in Africa. Their enhanced replication click here in peripheral tissues may lead to long-lasting viremia resulting in increasing incidence of viral invasion

to CNS. The interaction of the virus with endothelial cells of blood capillaries could be involved in WNV invasion to target organs. In this study, we assessed the transport of WNV NY99 6-LP strain and Eg strain across human endothelial cells. Our data demonstrate that VLPs of the 6-LP strain were see more transported across human endothelial cells more than VLPs of the Eg strain. Microbial invasion across endothelial cells can occur through transcellular pathway mediated by vesicles, paracellular entry after buy MLN2238 disruption of the tight junctions,

or “”Trojan horse”" mechanism by transport within circulating phagocytic cells [35, 36]. Our data indicate that 6-LP VLPs are transported by a transcellular pathway, because the transport of VLPs was inhibited by the treatment with filipin, a modifier of lipid raft-associated membrane transport. Clathrin-dependent pathways seem to be less important because the treatment with chlorpromazine had no significant effect on the transport of VLPs. Paracellular entry is unlikely to be involved in transport of VLPs because the structure Etofibrate of ZO-1 and the permeability of Dx 70k were not altered during VLP transport. Our data partially support the results by Verma et al. [16] which suggested that WNV crosses HBMVE cells without altering the integrity

of tight junction. The authors concluded that WNV replicates in endothelial cells and the progeny viruses are transported from the apical to basolateral side. However, our data suggest that WNV can be transported across endothelial cells without viral replication. Cell type difference could be the most reasonable explanation, because several studies showed that there are differences between HBMVE cells and HUVEC in the production of growth factors, immunoregulatory factors and adhesion molecules [37–39]. HBMVE cells and HUVEC differentially respond to cytokine treatment resulting in the different cytokine production and leukocyte recruitment [40, 41]. Particularly, modulation of adhesion molecules can affect endocytosis [37]. Therefore, our data seem to reflect events that can occur in peripheral tissues having tight junction such as heart and muscles rather than in CNS. In WNV-infected mice, viral replication in peripheral tissues results in the inflammatory cytokine production such as TNF-α, IL-6 and macrophage migration inhibitory factor [42–45]. Although the role of these cytokines in infection still remains controversial, vascular permeability can be affected by the presence of these cytokines [45].

Circulation 2008;118:586–606 PubMedCrossRef 2 American

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