tuberculosis infections. This TLR-2-dependent negative regulation of the IFN-I response during M. tuberculosis infections is likely to be beneficial to the host by limiting the harmful effects of IFN-I. This inhibitory mechanism may also play a positive role during other bacterial infections as TLR-2 recognizes a wide range of bacterial pathogens. What is interesting is that TLR-2 signalling impairs TLR-7-,

TLR-9- but not TLR-3-induced IFN-I synthesis [42, 43]. This in turn explains why influenza virus co-infections in M. tuberculosis-infected mice KU-60019 impairs bacterial control in an IFN-I-dependent manner [44]. Influenza virus generates multiple ligands of pattern recognition receptors during Decitabine chemical structure the viral replication cycle, which includes dsRNA (TLR-3 agonist) and

ssRNA (TLR-7 agonist). Thus, influenza virus infections can override TLR-2-dependent inhibition of IFN-I responses in M. tuberculosis-infected mice through TLR-3 signalling and induce IFN-I responses that ultimately result in outgrowth of M. tuberculosis. These findings provide answers as to why the risk of influenza death was higher among patients with tuberculosis than non-tuberculosis patients during an influenza pandemic [37]. Recent studies have focused on the mechanism of how primary viral infections render the host vulnerable to a sequel of bacterial infections. Severe forms of viral–bacterial co-infections are rare and only seen when the virus itself is highly virulent such as the 1918 Spanish influenza virus [23]. In fact, according to the Centre for Disease Control and Prevention, only 29% of fatal cases of patients with H1N1 influenza had bacterial co-infection [45]. When the primary viral infection is highly pathogenic, it is difficult to ascertain whether the increased susceptibility Cytidine deaminase is due to suppression of antibacterial immunity or the consequence of viral pathology

itself. We hypothesize that severe forms of viral–bacterial co-infection are an exception to the rule and that in most cases, that is, with less virulent viruses, primary infections do not lead to severe secondary bacterial pathology. Thus, there have to exist immune mechanisms that limit secondary co-infections. Our current understanding of the biology of IFN-I is that it is beneficial and essential to recover from most if not all acute viral infections, but may be detrimental to the host when fighting off bacterial pathogens. We also know from our previous studies [16] and reports from others [21] that IFN-I deficiency as a consequence of exhaustion occurs after primary viral infections and the host is rendered more susceptible to secondary unrelated viral infections during this transient period of IFN-I exhaustion.

While its expression is observed only in the convoluted proximal tubules of the normal Tg mouse, de novo expression of hL-FABP is also found in the straight portion of the proximal tubules during renal injury in a nephropathy model using the Tg mouse. In the setting of kidney disease, the distribution of hL-FABP expression is similar between human kidney and Tg mouse kidney. However, whether the different distribution this website of hL-FABP expression in human kidney and the Tg mouse kidney under normal conditions affects the mechanisms

by which urinary excretion of hL-FABP from the proximal tubules increases in kidney disease has not been evaluated yet, thus, further studies are needed to clarify this point. Urinary protein is widely known to be an aggravating factor for tubulointerstitial damage. Therefore, elucidation of the mechanisms by which urinary protein induces tubulointerstitial damage is needed in order to inhibit the progression of kidney disease or to check details develop new strategies against kidney disease. In the experimental model of protein overload nephropathy, a massive amount of bovine serum albumin (BSA), approximately 250 mg per sample, is intraperitoneally-injected into mice. The injected BSA is absorbed in the peritoneum, circulated via the systemic vasculature, filtered through glomeruli by overflow and reabsorbed

into proximal tubules, ultimately provoking tubulointerstitial damage without glomerular

injury. This model is suitable for clarification of the relationship between urinary protein and tubulointerstitial damage and is used to evaluate the pathophysiology of tubulointerstitial damage in nephrotic syndrome, which develops to end stage renal failure. The establishment of this model in the above-mentioned hL-FABP Tg mice background shows that the administration of abundant BSA causes severe tubulointerstitial damage, upregulation of hL-FABP gene expression, and from increases urinary excretion of hL-FABP.13 From these results, urinary excretion of hL-FABP reflects stress of urinary protein overload on the proximal tubules, which causes tubulointerstitial damage. Furthermore, in the protein overload nephropathy model, hL-FABP expression in the proximal tubules reduced macrophage infiltration and mildly inhibited the development of tubulointerstitial damage. We consider that hL-FABP may reduce accumulation of overload FFAs in the proximal tubules, inhibit production of inflammatory factors, attenuate macrophage infiltration and mildly inhibit the progression of the tubulointerstitial damage. Intraperitoneally injected streptozotocin (STZ) damages the endocrine part of the pancreas and induces type1 diabetes, thus, STZ-induced diabetic mice are widely used as a type 1 diabetes model.

Methods: Four groups of Japanese white rabbits underwent either

PBOO by mild ligation of the urethra (2- and 4-week PBOO) or no obstruction (2- and 4-week sham). Histopathological examination was performed by Elastica van Gieson staining, scanning electron microscopy, transmission electron microscopy, and ultra-high voltage electron microscopy. The number of pixels representing elastin fibers in computerized images was analyzed using Adobe Photoshop Version 2.0. Results: Bladder weight significantly increased after PBOO. Increase in the thickness of the bladder wall was observed after obstruction on histopathological examination. On scanning electron microscopy, elastin was very thick and CFTR activator was found in large configurations. 3-D analysis using electron microscopic tomography revealed that elastic fibers in the bladder had a coil-like appearance in the muscle layer, with each fiber composed of several fibrils. Such structures may be closely related to the physiological function 3MA of the bladder. Conclusion:

Elastin in the bladder assumes the form of a coil during micturition. We examined that the increase in elastin makes it difficult for elastin to stretch linearly resulting in reduced elasticity. This change may be one of the factors involved in the decrease in compliance mediated by PBOO. “
“Most pelvic organ prolapse (POP) patients have lower urinary tract symptoms (LUTS) before and after POP surgery. LUTS of POP patients consist of various storage and voiding symptoms from anatomical causes. Videourodynamic examination for POP patients provides accurate information about morphological findings of the bladder and urethra, and lower urinary tract (LUT) function. The leak point pressure (LPP) measurement at cough maneuver in the standing position is important to detect urodynamic stress urinary incontinences (UDS SUI). Prolapse reduction procedure is not perfect for the detection of SUI. Most pelvic organ prolapse (POP) patients have lower urinary tract symptoms (LUTS) Succinyl-CoA before and after POP surgery. LUTS of POP patients consist of various storage

and voiding symptoms due to anatomical causes.1 Evaluation of lower urinary tract (LUT) function is very important; however, there are few reports2,3 of urodynamic studies of patients with POP surgery. Tension-free vaginal mesh (TVM) procedure4 is choice for POP surgery. In the present paper we report video urodynamic examination of preoperative POP patients with TVM procedure and/or combined TVM and transobturator tape (TOT) procedure.5 Seventy-nine patients with POP-Q Stage 2 or higher underwent POP repairs conducted at Shinshu University Hospital between July 2008 and December 2010 using polypropylene mesh (GyneMesh PSTM, Ethicon, Somerville, NJ, USA) cut by the surgeon according to the TVM procedure.

The absence of correlation between trough IgG levels and annual dose of IgG in relation to body size (height, weight or body mass index) [45] suggests that dosing based on ideal body weight maybe equally effective, but this hypothesis remains to be proved. While the questions physicians face are challenging and continually keep pace with progress itself, the future for patients in need of immunoglobulin therapies appears

brighter than ever before. Through understanding the needs, specifics and clinical outcomes of patients in need of immune replacement therapy or immunomodulation, the application of IgG therapy can be improved by focusing upon the metrics derived from the patients themselves. The administration route, regimen and diversity of applications for IgG preparations are continually being optimized. A deeper understanding of immunoglobulin molecules, gene check details variability and its impact on the susceptibility of patients with certain gene patterns to IgG therapy may allow pharmacogenetic prediction of individual IgG dose requirements for patients and redefining IgG therapy. The authors are grateful for the help of Mary Lucas for data collation, Helen Chapel, Jennifer Lortan and Smita Patel Alvelestat for inclusion of data on their patients, and nursing colleagues Janet Burton, Nicola Salome-Bentley and Carol Ross

are gratefully acknowledged. Dr Misbah has received honoraria for advisory board membership on immunoglobulin therapy from CSL Behring, Baxter and Biotest;

Nintedanib (BIBF 1120) Dr Kuijpers, honoraria for advisory activities of Sanquin; Dr van der Heijden, support from Sanquin for his scientific work as PhD student; Dr Grimbacher is a member of the IgPro20 Steering Committee and Advisory Boards, honoraria for presentations from Baxter and Grifols; Dr Orange, consultant fees from CSL Behring, Baxter Biosciences, IBT Reference Laboratories, and Octapharma research grants review committee. No other potential conflicts of interest were reported. “
“Cyclooxygenase (Cox) inhibitors are among the most widely used and commonly prescribed medications. Relatively little is understood about their influence on human immune responses. Herein, we discovered a novel and important mechanism whereby non-steroidal anti-inflammatory drugs (NSAIDs) blunt human B-cell antibody production. We demonstrate that the Cox-2 selective small molecule inhibitors SC-58125 and NS-398 attenuate the production of human antibody isotypes including immunoglobulin M (IgM), IgG1, IgG2, IgG3 and IgG4. In addition, inhibition of Cox-2 significantly reduced the generation of CD38+ IgM+ and CD38+ IgG+ antibody-secreting cells. Interestingly, we discovered that inhibition of Cox-2 activity in normal human B cells severely reduced the messenger RNA and protein levels of the essential plasma cell transcription factor, Blimp-1.

Although the treatment for leishmaniasis was introduced in the early 20th century, parenteral administration of pentavalent antimony compounds (meglumine antimoniate and

sodium stibogluconate) remains the first-choice treatment for all forms of leishmaniasis [7]. In the case of antimonial resistance, the second-choice treatment includes amphotericin B (deoxycholate or liposomal formulation) [7]. However, each of these therapies has important limitations, such as long-term check details parenteral administration, toxic side effects, high cost in endemic countries and an increase in number of resistance cases [8]. A major breakthrough in chemotherapy of VL was the discovery of miltefosine, an analogue of phosphatidylcholine initially developed as an anticancer agent [9]. It is not recommended during pregnancy as teratogenicity has been observed in one species during preclinical development. Moreover,

its cost is another limiting factor [10]. Till date, no ideal drugs are available that fulfil the major requirements for efficient antileishmanial therapy, including high efficacy, low toxicity, easy administration, low costs and avoiding occurrence of drug-resistant parasites [11]. Cisplatin (cis-diamminedichloroplatinum II; CDDP) is a platinum-based anticancerous drug, which mediates its action by forming cross-link of DNA ultimately triggering apoptosis, or programmed cell death [12], and is also known to enhance the cytotoxic immunity [13]. An in vivo antileishmanial study with cisplatin at low dose also resulted in decreased parasite burden, increased find protocol delayed-type hypersensitivity (DTH) response, initial transient and reversible increase in various liver and kidney function tests [14]. It is well known that nephrotoxicity is a dose-limiting factor of cisplatin, so later on, Sharma et al. [15] investigated the protective efficacy of high dose of cisplatin in combination with antioxidants (Silibinin, vitamin C and

vitamin E) which effectively reversed the toxic side effects caused by the drug. So an auxiliary therapeutic measure that might enhance the efficacy of these antileishmanials or reduce the resulting toxicity would be valuable. Immunochemotherapy Thiamet G has been used with various combinations of drugs and vaccines mostly in case of cutaneous leishmaniasis. Some of them are sodium stibogluconate with poly ICLC (Polyinosinic-po lycytidilic acid) plus arginine [16], antimony with interferon–gamma [17], N-methyl meglumine antimoniate with recombinant Leish-110f plus MPL-SE vaccine [18], killed Leishmania promastigotes with antimonials [19] and alum precipitated autoclaved Leishmania promastigote (ALUM/ALM) plus BCG with sodium stibogluconate [20]. Chemotherapy of leishmaniasis is often compromised due to suppression of immune function during the course of infection.

By contrast, on Ag-experienced CD8+ T cells we found that whereas IFN-α enhances the effector functions, it decreases fold cell expansion. No differences were found between IFN-α2b and IFN-α5 subtypes, suggesting redundancy in the system. The magnitude of the stimuli and

the inputs from different stimulatory/inhibitory receptors are critical parameters for the outcome of the T-cell response. Thus, the need of choosing a fixed dose of stimuli, a single costimulatory signal and few time points for the array analyses provides a limited and static picture of the transcriptional changes induced on human T cells. Despite this limitation, our array data provided a baseline definition of the IFN-α transcriptional effects on human CD8+ Ixazomib in vivo T cells and will form the basis for further and more detailed studies. The results of the transcriptional analysis of human CD8+ T cells stimulated with IFN-α alone agree with previous studies of IFN-α stimulation of unfractionated PBL 18, 19. The overall similarity suggests that IFN-α imprints a common transcriptional

signature on the peripheral blood immune cell populations. Despite induction of relevant genes for effector functions, human CD8+ T cells treated only with IFN-α experienced no sign of activation. However IFN-α-derived signals synergize with signals elicited by CD3/CD28-triggering and promote the acquisition of effector functions on human CD8+ T cells. The biological meaning of the regulation of all these genes relevant for CD8+ T-cell functions by IFN-α itself

is still unknown. One possibility is that pre-exposure to IFN-α induces mRNA others that facilitate T-cell activation Caspase activity upon an eventual Ag encounter. Transcriptional analyses performed in human CD8+CD45RO− cells stimulated with Beads and either IFN-α2b and/or IFN-α5 show that, as a signal-3 cytokine, IFN-α regulates outstanding genes involved in the overall activation of T cells. Among these genes we found IL2. IL-2 is an important cytokine for survival, clonal expansion and differentiation of T cells 20. The fact that IFN-α also promotes the surface expression of CD25 strengthens the idea that IFN-α may promote the CD8+ T-cell response, at least in part, by inducing additional cytokines that could further stimulate CD8+ T cells in an autocrine manner. Importantly, the chief transcriptional signature of IFN-α, as a third signal, encompasses the up-regulation of transcripts involved in effector functions (IFNG, GZB and TRAIL) as well as production of chemokines (CXCL10 and CXCL11). A similar transcriptional signature has been found in OT1 cells stimulated in vitro with artificial DC and IFN-α 14, suggesting that IFN-α may promote the conversion of CD8+ T cells not only into highly effector cells but also into efficient chemotactic attractants of additional effector cells. This transcriptional effect was substantiated at the protein level and verified by functional assays.

(Carlsbad, CA). Human peripheral blood mononuclear cells (PBMC) were isolated and purified MDV3100 cost from blood (Red Cross Blood Bank) by density gradient centrifugation and adherence as described by us previously (Liao et al., 1994). PBMC were then cultured in serum-free macrophage media (37 °C, 5% CO2) overnight with lipopolysaccharide (Escherichia coli, 100 ng mL−1) or vehicle alone. Doxycycline was added at final concentrations ranging from 0.1 to 20 μM. Conditioned media were analyzed for the cytokines [tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β)] and MMP-9 by enzyme-linked immunosorbent

assay (ELISA). In separate assays, PBMC at 5 × 105 cells mL−1 were cultured with macrophage medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) and containing 100 U mL−1 penicillin and 100 μg mL−1 streptomycin in Teflon beakers for 7 days with different concentrations of doxycycline. At the end of the 7-day incubation, conditioned media were analyzed by gelatin zymography as

described by us previously (Golub et al., 1995). Western blot, gelatinase and collagenase activity assays were carried out as described below. The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)/fluorography of [3H]-labeled type I collagen was scanned using a laser densitometer to quantify the effect of doxycycline on the collagenase activity, the latter assessed by the production of [3H]-labeled collagen degradation Abiraterone fragments as described by us previously (Yu et al., 1993). R22 rat heart smooth muscle Demeclocycline cells were cultured in minimum essential medium supplemented with FBS, tryptose phosphate broth and cefotaxime (Gu et al., 2005). The R22 cells were plated onto multiwell tissue culture

plates at an initial density of 2.5 × 104 cells cm−2 and were maintained at 37 °C in 5% CO2. At confluence, the medium was supplemented with [3H]-fucose, which were incorporated into a complete interstitial ECM elaborated by the cells. Every 4 days, 50 μg mL−1 ascorbic acid was added to ensure maximal formation of an insoluble collagen-rich ECM. After culturing for at least 1 week in radiolabeled medium, cells were lysed by brief exposure to 25 mM NH4OH without disrupting the ECM. The wells were washed three times with sterile H2O and once in phosphate-buffered saline (PBS) containing 0.02% NaN3. Excess PBS was then removed and plates were stored at 4 °C until use. Before use, the ECM was rehydrated by rinsing three times with sterile buffer. PBMC in serum-free media were applied to R22 ECM-coated wells of microplates at a density of 5 × 105 cells mL−1 and incubated for 2 days at 37 °C in 5% CO2 in the presence or absence of doxycycline. After the 2-day incubation, the supernatants were collected and the remaining undegraded ECM in each well was solubilized by overnight incubation with 2 M NaOH. The radioactivity in the supernatants and in the NaOH was determined in an LKB liquid scintillation counter (Gu et al., 2005).

5% in 2000 to 70% in 2010. No differences were found between C. albicans and C. non-albicans episodes in terms of demographics, risk factors or mortality. The highest resistance rates (overall 7.6%) were observed for fluconazole (4.3% in C. albicans, 7.1% in C. parapsilosis

and 13.8% in other Candida species). Resistance Selleckchem MK 2206 to amphotericin B (2.5%) was limited to non-albicans isolates. The dynamic changes in species distribution and increasing resistance of fungal pathogens confirm the importance of epidemiological surveillance. “
“We report for the first time the environmental isolation of Cryptococcus neoformans from decaying wood and bark debris of living trees in Guindy National Park, Chennai, South India. Of the 40 trees screened, four isolates of Cryptococcus species were recovered of which two were Cryptococcus gattii, one was C. neoformans and one was untypable. The isolation of C. neoformans from Eucalyptus globulus and C. gattii from Cassia marginata HTS assay in this study constitutes the first record of the natural occurrence of C. neoformans varieties in these tree species anywhere in the world. The isolation of C. gattii from Syzygium cumini represents the first isolation from South India. “
“Typically, the onset of candidiasis is characterised by the appearance of a

biofilm of Candida albicans, which is associated with several diseases including oral candidiasis in young and elderly people. The objective of this work was to investigate the in vitro fungicidal activity as well as the antibiofilm activity of ambroxol (AMB) against C. albicans

growth. In the present investigation, the fungicidal activity of AMB was established using the cell viability 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Isotretinoin Also the minimum inhibitory concentration (MIC) of AMB required to inhibit the fungal growth was determined. Simultaneously, the antibiofilm activity of AMB was evaluated using fluorescence microscopy. The study revealed that 2 mg ml−1 of AMB exhibited higher fungicidal activity than 3.3 mg ml−1 of terbinafine, one of most common commercial antifungals. A MIC of 1 mg ml−1 was determined for AMB to interfere with C. albicans growth. Furthermore, AMB was found to be effective in inhibiting the biofilm formation of C. albicans and exerted its fungicidal activity against the fungal cells interspersed in the preformed biofilm. The study suggests a potential role of the mucolytic agent, AMB, as an interesting therapeutic alternative in the treatment of oral candidiasis. “
“Peptidorhamnomannans (PRMs), rhamnomannans and α-glucans are especially relevant for the architecture of the Scedosporium/Pseudallescheria boydii cell wall, but many of them are immunologically active, with great potential as regulators of pathogenesis and the immune response of the host.

Data in bar graphs are given Gefitinib supplier as the mean ± standard deviation (s.d.).

A value of P < 0·05 was considered significant. Monocytes were isolated and cultured with GM-CSF and IL-4; the resulting iDCs were exposed to hypoxia on day 5 for 48 h or to LPS for 24 h to induce cell maturation. Figure 1a shows the analysis of different cellular subpopulations during the differentiation and maturation of DCs. At day 0 we had a high percentage of monocytes (CD14+) and the presence of several lymphocyte subtypes (CD3+, CD20+ and CD56+). During differentiation, the CD14+ population expressed DCs markers (HLA-DR+ and CD11c+) and the lymphocyte percentage diminished after removing the medium and replacing it with fresh culture medium. At the end of the differentiation (at day 7) the purity of DCs was greater than 90% (Fig. 1b). DC population was gathered in two subpopulations, depending on the degree of maturation according to the forward-/side-scatter LY2157299 cost profile and specific phenotypic markers established in our previous study [8]. We also performed

a follow-up of DC differentiation at different time-points. We observed that after hypoxia or LPS stimulus, cells changed their morphology, acquiring a stellate form characteristic of the mDCs shifting to the upper window. LPS stimulus induced a more homogeneous and stronger maturation response, while hypoxia stimulus showed a different magnitude of response (Fig. 1b). To evaluate

further the changing phenotype after stimuli cAMP of the DC population, FACS analysis was performed at days 1, 5 and 7. CD40 mean fluorescence revealed that mDCs appeared at day 5 of decreasing monocytes and iDCs populations. After LPS and hypoxia stimuli at day 7, DCs were well differentiated from non-stimulated cells. To characterize mDCs we used DC-LAMP, a type I transmembrane glycoprotein restricted to mDCs and expressed in the endosomal/lysosomal compartment. DCs exposed to LPS or hypoxia showed a clear DC LAMP-positive up-regulation, confirming the mature phenotype. Dual staining with the Pgp (JSB1) or MRP1 (4124) antibodies also showed an over-expression of Pgp and MRP1 in those DC-LAMP-positive DCs, differing from non-stimulated cells (P < 0·05) (Fig. 2a,b, respectively). This may indicate that in DC maturation there is an increase in Pgp and MRP1 in the cell membrane. Furthermore, this effect was more evident after LPS stimuli than after hypoxia. To evaluate the ABC transporters involvement in DC maturation, PSC833, MK571 or PBN were added to inhibit MDR1, MRP1 and MRP2, respectively. After hypoxia stimulation the percentage of mature DCs was evaluated by the forward-/side-scatter profile. Hypoxia resulted in an induction of 67·8% of mDCs versus 32·2% of iDCs (Fig. 3), lower compared to LPS, which induced 80·8% of mDCs and 19·2% of iDCs (P < 0·05).

Prior to use, S1P was dissolved in 4 mg/mL fatty acid-free BSA solution. Protease inhibitors Complete and Pefabloc SC were obtained from Roche Applied Science (Mannheim, Germany), while phosphatase inhibitor okadaic Poziotinib research buy acid was from Alexis Biochemicals (Grünberg, Germany). Mononuclear cells were routinely isolated from citrated blood of healthy single donor by pancoll (PanBiotech, Aidenbach, Germany) density centrifugation

and counter flow elutriation as described previously 40. The resulting monocyte fraction consisted of more than 97% pure monocytes. Cells were cultured in RPMI 1640 supplemented with 5% FBS and 2 mM L-glutamine (all from Biochrom (Berlin, Germany)) and treated with stimuli and inhibitors essential as published earlier 3. Approval for these studies was obtained from the Institutional Review board at the University of Lübeck (Lübeck, Germany), and informed consent was provided according to the Declaration of Helsinki. Generation of ROS was determined in a microplate luminometer (LB 96V; Berthold (Wildbach, Germany)) by measurement of chemiluminescence in the presence of 60 μg/mL luminol Ceritinib datasheet (5-amino-2,3-dihydro-1,4-phthalazindione; Roche Applied Science) essentially as described

elsewhere 2, 3. In brief, monocytes were stimulated with 4 μM CXCL4 or increasing concentrations of S1P and chemiluminescence was recorded for 60 min. Individual assay backgrounds were determined in samples of unstimulated cells

in the presence or absence of inhibitors run in parallel and were substracted. Data were expressed as relative light units and quantified by integration over the time periods indicated. Determination of apoptotic and necrotic cells was done by double labeling with annexin V-FITC and PI, according to manufacturer’s recommendations (Bender MedSystems, Heidelberg, Germany) 3. The populations of apoptotic and necrotic cells were defined by their characteristic binding patterns annexin Vhigh/PIlow, and annexin Vhigh/PIhigh, respectively. Phosphorylated Erk MAPK was detected in cell lysates by western blot analysis with antibodies specific for the phosphorylated (activated) kinases essential Fossariinae as described earlier 3. Proteins derived from cell lysates (40 or 80 μg/lane) were separated by SDS-PAGE 41 using 12% polyacrylamide gels and blotted onto polyvinylidene fluoride membranes (Roth). Immunodetection was performed as described in detail elsewhere 3, 42. Band intensities on blot membranes were quantified using Odyssey software 2.1 and presented either as integrated intensities or fold induction from unstimulated control. Total RNA was purified using NucleoSpin RNA II kit (Macherey-Nagel, Düren, Germany) according to manufacturer’s recommendations followed by reverse transcription into cDNA using First Strand cDNA Synthesis Kit (Fermentas, St. Leon-Rot, Germany).