Compared with other screening techniques such as transcriptomics

Compared with other screening techniques such as transcriptomics or proteomics, SEREX offers a crucial advantage that subtle changes in the protein expressions can be detected through immunological reactions [32, 33]. Several authors have already applied SEREX to glioma, and some Selinexor datasheet antigens, including glioma-expressed antigen 2 (GLEA2) [7], PHD finger protein 3 (PHF3) [7, 34], and SRY-box 6 (SOX6) [8] have been identified. It should be noted that we found autologous antibodies against SH3GL1 to be a low-grade glioma-specific marker with similar experimental systems to others. Our

unique approach was the quantitative comparison of the levels of serum antibodies using the ELISA, while the approach of others Akt inhibitor was qualitative analysis. The application of ELISA in the validation step could lead to the discovery of a low-grade glioma-specific high titer of the Entospletinib in vitro autoantibody and the decrease in high-grade gliomas. Although some candidates of glioma biomarkers have been identified by various screening methods [6–8, 34–37], no serum marker for early diagnosis has been found yet. Therefore,

it is quite valuable to find a novel serum biomarker for its early diagnosis, prediction of the prognosis in each patient, and development of a new molecular target. Indeed, The results of an overlap peptide array and ELISA using deletion mutants of SH3GL1 showed that 12 amino acids in the C-terminal portion, FPLSYVEVLVPL, were indicated as a major epitope site. By using a synthetic peptide corresponding to the epitope as an antigen, a more accurate screening for the patients

with low-grade gliomas and a specific peptide vaccine therapy would be achieved in the future. Author details 1Departments of Neurological Surgery, Chiba University, Graduate School of Medicine, 1-8-1, Inohana, Chuo-ku, Chiba 260-8670, Japan. 2Genetics and Biochemistry, Chiba University, Graduate School of Medicine, 1-8-1, Inohana, Chuo-ku, Chiba 260-8670, Japan. 3Diagnostic Pathology, Chiba University, Graduate School of Medicine, 1-8-1, Rho Inohana, Chuo-ku, Chiba 260-8670, Japan. 4Department of Biochemistry, Graduate School of Life Science, Nagoya Women’s University, 3-40, Shioji-cho, Mizuho-ku, Nagoya 467-8610, Japan. References 1. Ohgaki H, Kleihues P: Epidemiology and etiology of gliomas. Acta Neuropathol 2005, 109:93–108.PubMedCrossRef 2. Anderson E, Grant R, Lewis SC, Whittle IR: Randomized Phase III controlled trials of therapy in malignant glioma: where are we after 40 years? Br J Neurosurg 2008, 22:339–349.PubMedCrossRef 3. van den Bent MJ, Afra D, de Witte O, Ben Hassel M, Schraub S, Hoang-Xuan K, Malmstrom PO, Collette L, Pierart M, Mirimanoff R, Karim AB: Long-term efficacy of early versus delayed radiotherapy for low-grade astrocytoma and oligodendroglioma in adults: the EORTC 22845 randomised trial. Lancet 2005, 366:985–990.PubMedCrossRef 4. Sanai N, Berger MS: Glioma extent of resection and its impact on patient outcome.

In contrast, most atypical antipsychotics like clozapine, olanzap

In contrast, most atypical antipsychotics like clozapine, olanzapine, quetiapine, and low-dose risperidone have a higher affinity for the 5-hydroxytryptamine-2A (5-HT2A) receptor than for dopamine D2 receptors [4]. Blocking of the 5-HT2A receptor has been associated with lowered prolactin levels. In contrary, the stimulating

of 5-HT2A receptors has been linked to increased prolactin levels [7]. The latter is the case when using a selective serotonin reuptake inhibitor (SSRI). Elevated serum prolactin may reduce bone mineral density (BMD) in the long-term [6, 8, 9]. O’Keane and Meaney [10] found that the BMD of patients using prolactin-raising antipsychotics was significantly lower than that of users of antipsychotics without prolactin-raising properties. In line with Sapitinib concentration these results are the findings that patients using SSRIs also experience a lower BMD [11] and have an increased risk of fracture [12]. Several epidemiological studies have reported an increased risk of

hip or femur fracture among users of antipsychotics [13–19]. One study found a relationship between dose and use of antipsychotics, regardless of timing of exposure, although this was not reported for current users [17]. Liperoti et al. found no difference in fracture risk between conventional and atypical antipsychotics [15], whereas Howard et al. found an increased risk for individuals using prolactin-raising SC79 purchase antipsychotics [13]. In addition, there is some evidence to suggest that men using antipsychotics have a greater risk of fracture than women [13]. The aims of this study were to evaluate the association between the use of antipsychotics and the risk of fracture of the hip or femur for men and women, to derive risk estimates separately for conventional and atypical antipsychotics, and to investigate the risk associated with dose and pharmacological properties. Methods Setting

and study design We conducted a case–control study within the Dutch PHARMO Record Linkage System (RLS) (www.​pharmo.​nl). The database includes the demographic Epigenetics inhibitor details and complete medication histories for about one million community-dwelling residents in the Netherlands representing isothipendyl some 7% of the general population. Data are available from 1986 onwards and are linked to hospital discharge records as well as several other health registries, including pathology, clinical laboratory findings, and general practitioner data. Almost every individual in the Netherlands is registered with a single community pharmacy, independent of prescriber and irrespective of his or her health insurance or socioeconomic status. Pharmacy records have a high degree of completeness with regard to dispensed drugs [20]. Pharmacy data include information about the drug dispensed, the date of dispensing, the prescriber, the amount dispensed, the prescribed dosage regimen, and the estimated duration of use.

Studies on the hearing of

Studies on the hearing of musicians in symphony orchestras have indicated that their pure-tone hearing thresholds do not really deviate from that of

a HM781-36B solubility dmso non-exposed population (e.g. Kähäri et al. 2001a, b; Eaton and Gillis 2002; Obeling and Poulsen 1999). It has been hypothesized that specific “musician characteristics” are responsible for this result: wanted sounds such as music could be less harmful than unwanted sounds such as industrial noise (Karlsson et al. 1983), or musicians perform relatively good on pure-tone audiometry because of a strong motivation and familiarity with detecting pure tones (Dowling and Harwood 1986). The musicians participated on a voluntary basis. We are aware that this could have produced HMPL-504 clinical trial a selection bias, probably towards the better hearing musicians, as musicians with hearing complaints may have been reluctant of having their hearing tested. Most musicians judged their hearing as good, though slightly worse than before (5 or 10 years ago). As far as we could check, the self BYL719 in vivo reports on medical history did not show deviations from the general population. When categorizing

the musicians’ pure-tone audiograms in absolute terms, almost half of the tested musicians’ ears can be categorized as normal. Among the larger groups (i.e. HS, LS, BW and WW), age seems to be more predictive for audiogram category than the instrument played: the percentage of brass-wind players, who had the lowest average age, was smallest in the sloping-loss category in contrast to the low-string players who had a relatively higher average age and were better represented in this category. Audiograms corrected for age and gender resulted in better threshold levels for low-string players, as compared to high-string Progesterone and wood-wind players. This could suggest an effect of exposure as low-string players are usually the least exposed group (Boasson 2002). It was unexpected that the more heavily exposed group (i.e. brass-wind players) did not show a larger increase in the thresholds than the other groups, except for the already

mentioned low-string players. All the instrument categories show an evenly profound notch in the hearing-thresholds at 6 kHz, a frequency that is known to be very sensitive for noise-induced hearing loss. When the relative audiometric group results were compared to that of the ISO 7029 (2000) population, musicians showed better hearing thresholds on all tested frequencies, except on 6 kHz. This supports the observation that professional musicians perform relatively good on pure-tone audiometry despite intense exposure. It is possible that this effect is able to mask early signs of NIHL and in that case screening techniques other than the pure-tone hearing thresholds could be more adequate for the detection of early stages of NIHL in professional musicians (e.g. Kähäri 2001b).

In contrast, in an in vivo

study of bacteriocins employin

In contrast, in an in vivo

study of bacteriocins employing the same mouse model as described here, did not detect an increased persistence of colicinogenic enteric bacteria [24]. However, in that study persistence was monitored for only 15 days. Our data suggest that over a longer period of time, 112 days in the present study, the benefit of colicinogenicity becomes more apparent (Figure 1), with see more colicin producers maintaining significantly higher densities than their non-colicin producing counterparts. The colicin-based advantage observed in the present in vivo study reflects a similar advantage to colicin Adriamycin production as has been detected in prior in silico and in vitro studies [20]. Our results are even more promising with respect to the advantage gained from colicin production when the sampling method employed here is considered, as fecal-based sampling will generally underestimate

the actual density of the strain in the GI tract [25, 26]. There is one further colicin-based in selleck inhibitor vitro study, which employed the same mouse model described here, but which differed significantly in experimental design. In this latter study the focus was on the interaction (or competition) between colicinogenic and non-colicinogenic strains, while the current study focuses on the ability of colicinogenicity to enhance strain maintenance [12]. This prior colicin competition study revealed that colicin production enhances strain persistence when mice equilibrated with colicin producing strains are co-caged with mice equilibrated with colicin sensitive strains [12]. Thus, although the intent of the

two studies is quite different, both reveal that colicinogenicity has a significant and positive effect on the ability of a strain to be maintained in the GI tract of a streptomycin-treated mouse. Many studies in humans and livestock have shown that probiotic bacteria have the ability to re-establish an indigenous microflora after perturbations of the normal intestinal flora [27–31]. Probiotic bacteria provide this health benefit in Tolmetin many ways and the production of toxins, in particular bacteriocins, was proposed as a leading candidate in this process [21]. E. coli strain Nissle 1917, a producer of microcins H47 and M [32], is a well characterized probiont in humans and livestock [3, 5, 33, 8]. This strain was found to be effective in treating chronic inflammatory bowel disease [33] and in inhibiting the adhesion of enteric pathogens to the GI epithelial cells of infants [5]. E. coli strain H22 inhibits the invasion of the enetric pathogen Shigella flexneri in germ-free mice, probably due to the production of microcin C7 [34], colicins E1 and Ib, as well as aerobin and an unidentified phage [4]. In order for a probiotic strain to exert its beneficial effect in the GI tract, it is essential for the cells to become established.

Effect of EGFR knockdown on LRIG1-induced cell

Effect of EGFR knockdown on LRIG1-induced cell proliferation and signal pathway regulation To determine whether EGFR expression is critical for the effect of LRIG1 on bladder cancer cells in vitro, we next used specific genetic inhibition of EGFR to assess the consequences of its inhibition on LRIG1 mediated cell proliferation and signal pathway regulation. First, we confirmed that the EGFR siRNA effectively reduced the EGFR

protein level in T24 and 5637 cells (Figure 6A). Then we found EGFR knockdown significantly decreased the effect of LRIG1 cDNA on cell proliferation compared with control-siRNA-transfected cells (Figure 6B). Luminespib clinical trial And EGFR siRNA significantly weakened the effect of LRIG1 cDNA on the EGFR signaling pathway regulation in both cell lines compared with cells transfected with control siRNA

(Figure 6C). Figure 6 Effect of EGFR knockdown on LRIG1-induced cell proliferation and signal pathway regulation. A: Genetic suppression of EGFR by EGFR-siRNA transfection. B: Proliferation of cells treated with LRIG1 cDNA after selleck compound transfection with EGFR siRNA or control siRNA. *P < 0.05 vs cells transfected with control siRNA. C: Effects of silencing EGFR on the LRIG1-induced regulation of the expression of AKT, MAPK, and their phosphorylated forms. Discussion Kekkon proteins negatively regulate the epidermal growth factor receptor (EGFR) during oogenesis in Drosophila. Their structural relative in mammals, LRIG1, is a transmembrane protein, could restrict growth factor signaling by enhancing receptor ubiquitylation Selleckchem Rucaparib and degradation [13]. The feasibility and efficacy of

the inhibitory effects of LRIG1 on tumor through inhibiting EGFR signaling activity have been studied in renal cancer, glioma, squamous cell carcinoma of skin, colorectal cancer and prostate cancer [19–23]. In this study, we attempted to evaluate the inhibitory effects of LRIG1 on aggressive bladder cancer cells. EGFR is a well-studied, versatile signal transducer that is overexpressed in many types of tumour cells, including lung, colon and prostatic carcinoma, and up-regulation of EGFR is associated with poor clinical prognosis [24, 25]. EGFR is a 170 kDa tyrosine kinase receptor consisting of an extracellular ligand-binding domain, a transmembrane lipophilic domain, and an intracellular tyrosine kinase domain and the C-terminus region with Alvocidib supplier multiple tyrosine residues [26]. EGFR mediates signals that stimulate proliferation, migration, and metastasis in many tumour types [25, 27], and its signal transduction is regulated by stimulatory and inhibitory inputs.

These pulses lead to a superposition of excitonic states, an exci

These pulses lead to a superposition of excitonic states, an excitonic wavepacket, with the target to populate just a single chromophore at a given time. The theoretical framework is given by

the multi-exciton density matrix, and although the dissipation is damping the wavepacket buy Doramapimod at low temperatures, the target can be reached quite well. In a follow-up article, the additional effects of inhomogeneous broadening and orientational averaging were included (Brüggemann et al. 2006). Again, the target could be reached although to a lesser extend. The introduction of a laser field, shaped in both polarization directions, led to a larger target state population, partially working against MK-8931 cell line the energetic and oriental averaging. Under conditions encountered by the FMO complex in vivo it is very likely that multiple excitations occur within one complex. These double-excited states are more complicated than its single counterpart and are less well studied. Often 2D spectra are obscured by overlapping contributions of single and double exciton resonances. By looking at a smart representation of the 2D spectra using a particular set of pulses, the correlated dynamics of the double excited states can be probed (Abramavicius et al. 2008a). Strong peaks are observed for double exciton states 1, 7, and 18 that also happen to be the most delocalized states in the system. In addition, weaker signals

of exciton states 9, 16, and 17 are observed. Instead of calculating the wavefunctions of the different exciton states, an alternative method can be used to describe the behavior of

excitons in aggregates. In the quasiparticle approach, all the properties of the system are described in terms of scattering and double exciton energies are simply given by a sum of single exciton energies. Comparing the spectra resulting from the full calculation with that of the quasiparticle approach shows that the energies at which the peaks appear in the spectra agree, while the fine structure in the spectra of the quasiparticle this website approach is distorted. In order to approximate the spectra, the quasiparticle approach can be used, however, because the exciton coupling is strong, which is neglected in this approach, and the nonbosonic nature of the excitons a full calculation of the spectra is necessary for detailed analysis. New types of 2D techniques can be developed by introducing pulse polarizations as variables into standard 2D schemes, as described in the previous section. This, amongst others enables the dissection of the congested 2D spectra into incoherent and coherent contributions and provides interesting perspective for new control strategies (Abramavicius et al. 2008b; Voronine et al. 2008). Current consensus and future directions Slowly the choice of parameters used to simulate the results obtained from various optical techniques is converging.

Nucleic Acids Res 2004, 32:W665–667 PubMedCrossRef 75 Schuttelko

Nucleic Acids Res 2004, 32:W665–667.PubMedCrossRef 75. Schuttelkopf AW, van Aalten DM:

PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallogr D Biol Crystallogr 2004, 60:1355–1363.PubMedCrossRef 76. Inagaki K, Tanizawa K, Badet B, Walsh CT, Tanaka H, Soda K: Thermostable alanine racemase from Bacillus stearothermophilus : molecular cloning of the gene, enzyme purification, and characterization. Biochemistry 1986, 25:3268–3274.PubMedCrossRef 77. Noda M, Matoba Y, Kumagai T, ON-01910 concentration Sugiyama M: A novel assay method for an amino acid racemase reaction based on circular dichroism. Biochem J 2005, 389:491–496.PubMedCrossRef 78. Badet B, Walsh C: Purification of an alanine racemase from Streptococcus faecalis and analysis of its inactivation by (1-aminoethyl)phosphonic acid enantiomers. Biochemistry 1985, 24:1333–1341.PubMedCrossRef click here Authors’ contributions HI performed research, helped draft the manuscript, analyzed results and prepared figures. MS helped to refine the structure and draft the manuscript, analyzed results and prepared figures. US and MD performed research and critically appraised the manuscript. KK designed research, supervised the work, organized financial support, and critically appraised the manuscript. All authors read and approved the final manuscript.”
“Background Staphylococci are common commensal bacteria of the skin [1], as well as important pathogens

in foreign-body infections [2]. The gram-positive Staphylococcus (S.) aureus is a major human pathogen. Inflammation inhibitor It is the cause of many nosocomial infections,

including life-threatening diseases such as toxic shock syndrome, sepsis and endocarditis [3]. S. aureus infections (-)-p-Bromotetramisole Oxalate account for approximately 19,000 deaths per year in the United States [4]. The emergence of multi-drug resistant strains of S. aureus, such as methicillin-resistant S. aureus (MRSA), has intensified the need for new treatments [5]. The danger of untreatable staphylococcal infections highlights the importance of new anti-microbial drug discovery. It has been discovered that chronic, infected wounds are often infected with strong biofilm forming bacteria, such as S. aureus [6], and it is now thought that the presence of biofilm actively prevents the healing of these wounds [7]. Chronic wounds can arise as a result of pressure sores, venous leg ulcers, diabetic foot ulcers or combat wounds, for example. While physical debridement can assist the healing of these wounds, biofilm-focused therapeutic approaches can promote more rapid healing in a large percent of patients [7]. This biofilm-centric philosophy may represent a modern strategy to treat chronic, infected wounds in which reducing the ability of the bacteria to form biofilm is itself the critical goal. In this strategy, subsequent healing by the body or treatment with antibiotics is then more effective.

The dark-acclimated membrane without qE is shown on the left Exc

The dark-acclimated membrane without qE is shown on the left. Excitation energy can be absorbed at any nodes and transferred on the picosecond (10−12s) timescale along the lattice grid lines until it reaches a RC (gray nodes) (van Amerongen et al. 2000). Once it reaches a RC, the excitation energy GDC-0973 nmr can be “photochemically” quenched and converted into chemical

energy. The \(\Updelta\hboxpH\) triggers a series of changes in the membrane (Fig. 6, right) that change the energy transfer network on a timescale of tens of seconds to minutes. Some antennae (Havaux et al. 2007) (white nodes) gain a photophysical pathway or mechanism with a rate of relaxation to the ground state that is fast relative to fluorescence and ISC. Efficient quenching of chlorophyll excitations could prevent the excitation from reaching a RC that is susceptible to damage. To alter the PI3K targets properties of the pigments such that they become quenching sites may require a rearrangement of the proteins in the membrane, which is indicated by the changes in the connectivity of the network. Fig. 6 A schematic of a possible CHIR-99021 purchase configuration of

chlorophyll connectivity of a portion of the grana membrane when qE is off (left) and when qE is on (right). The black circles represent non-quenching chlorophyll, such as those in LHCII. The gray circles represent PSII reaction centers, and the white circles represent qE quenching sites. At both reaction centers and qE sites, there is a rate for removing excitation from the grid. The grid lines display the connectivity for energy transfer between different groups of chlorophyll While this general picture of quenching is agreed on, nearly all of the details remain controversial. The energetic connectivity

of pigments in the membrane is determined HSP90 by their orientation, separation from other pigments, and their local protein environments. However, it is not possible at present to acquire the nearly atomic level resolution necessary for obtaining that information. Instead, a few approaches are used to study intact photosynthetic organisms. We categorize these approaches into four groups: spectroscopic measurements of pigment–pigment interactions, imaging and microscopy, fluorescence lifetimes, and transient absorption (TA) spectroscopy. Combined with modeling, these techniques can provide insight on aspects of both the membrane changes and on the site and mechanism of qE (Fig. 1). Spectroscopic measurements of pigment–pigment interactions To switch a pigment from participating in light harvesting (black node in Fig. 6) to quenching (white node) requires an alteration of its physical properties by changing its protein environment or by interactions with other pigments. Pigment–pigment interactions can be tuned by small changes in the protein conformation (van Oort et al. 2011) or by changes in the structure of a neighboring pigment, as when zeaxanthin replaces violaxanthin in high light (Crimi et al. 2001).

Experiments to investigate the expression and function of these g

Experiments to investigate the expression and function of these genes in vivo are in progress. Methods Bacterial strains and culture conditions Selleck CHIR98014 Bacteroides fragilis strains used in this study are presented in Table 7. All strains were purchased from the United Kingdom National Culture Collection (UKNCC) except

638R which was a kind gift from Dr Sheila Patrick, Queen’s University, Belfast. Both B. fragilis strains and B. thetaiotaomicron VPI-5482 [42] were grown in an anaerobic chamber at 37°C. Cultures were grown without shaking in Brain Heart Infusion (BHI) broth supplemented with 50 μg/ml hemin and 0.5 μg/ml menadione. Media for plating was made from Brain Heart Infusion agar supplemented with 5% defibrinated sheep blood, 50 μg/ml

hemin and 0.5 μg/ml menadione. Bioinformatics and sequence analysis Members of the C10 protease family in B. fragilis were detected by BLAST analysis [43]. Sequences were aligned by CLUSTAL W [44] or T-Coffee [45]. Protein secondary structure was predicted using GorIV [46] and JPred [47]. Protein export signals were identified using the AZD2281 concentration algorithms using LipPred [23], LipoP [48], SignalP [25] and PSORTb [26]. Phylogenetic and molecular evolutionary analyses were conducted using genetic-distance-based neighbour-joining algorithms [49] within MEGA Version 4.0 http://​www.​megasoftware.​net/​. Bootstrap analysis for 1000 replicates was performed to estimate the confidence of tree topology [50]. MegaBLAST [51] was used to search all NCBI genomes for Bfgi1 and Bfgi2. Molecular techniques Standard techniques were employed for molecular analysis [52]. Bacteroides genomic PARP inhibitor DNA was prepared as described by [53]. Total microbial DNA was extracted from human faeces, collected under an ethically approved protocol, by a glass beads-Qiagen Stool kit method previously described [54]. PCR reactions were carried using 10-30 ng of genomic DNA from B. fragilis 638R as template and using Phusion Polymerase (New England Biolabs).

The primers Bfp3_F and Bfgi2_Int_F (Table 4) were used for detecting the attP sites for Bfgi2. Bfgi2_attB_F and Bfgi2_attB_R (Table 4) were used for determining the attB attachment sites for Bfgi2 integration. The primers TraQ_F and Int_F were used in testing for the presence of the circular intermediate for Bfgi1. Primers to detect the circular intermediate for both Bfgi1 and Bfgi2 were designed, pointing outwards, flanking the ends of each predicted element. Primers to detect the attB site in Bfgi2 were designed, pointing inwards, flanking the proposed excision point for the Bfgi2 prophage DNA. Total RNA isolation for Reverse Transcription analysis B. fragilis 638R and B. thetaiotaomicron VPI-5482 were cultured under anaerobic conditions until early logarithmic phase and the cultures were then immediately centrifuged for 15 minutes at 4000 × g. Total RNA extraction from B. fragilis 638R and B.

HIF1A, IRF1, and STAT1, were expressed to a greater extent in DBA

HIF1A, IRF1, and STAT1, were expressed to a greater extent in DBA/2 compared to selleck chemical C57BL/6 mice, and YY1 to a lesser extent.

STAT1 is the largest hub representing the transcription factor regulating the most differentially expressed genes and it was previously selected as a target for RT-qPCR confirmation from the top 100 modulated genes (P005091 mouse Figure 2). YY1 is a transcription factor whose “yin-yang” designation reflects its ability to both activate and repress transcription through interactions with histone acetylases and deacetylases, respectively [17]. A novel finding from the protein network analysis was the hub HIF-1α, which is a transcription factor that plays a central role in the cellular and systemic responses to hypoxia. HIF-1α is regulated at the post-translational level, which CAL-101 in vivo results in increases in protein half-life, and also at the transcriptional level

by NF-κB [18, 19]. HIF1A was selected for gene expression confirmation by RT-qPCR, as was interleukin 6 (IL6), since it is a transcriptional target of both HIF-1α and Stat1 [20, 21]. Figure 6 Direct protein interaction network constructed from the genes differentially expressed with a fold change ≥ 2 or ≤ -2 (log 2 fold change ≥ 1 or ≤ -1, respectively) between DBA/2 and C57BL/6 mice at day 14 following C. immitis infection (N = 416). MetaCore was used to identify protein-protein and protein-DNA interactions L-NAME HCl between the protein products of differentially expressed genes and Cytoscape was used to visualize the network. Log2 fold changes were superimposed on this protein network such that red indicates greater expression in DBA/2 versus C57BL/6 mice, and blue

lesser expression, as indicated by the scale bar. Each node represents a gene and the size of a node is indicative of the number of interactions the product of each gene makes at the protein level. The largest nodes are labeled HIF1A, IRF1, STAT1 and YY1, and represent hubs that correspond to transcription factors. Stat1 and Irf1 are both transcription factors that upregulate the expression of ISGs and thus corroborate the presence of ISGs in the top 100 modulated genes (Figure 2), as well as the identification of chemokine related pathways (Figure 4). The well-characterized ISGs selected for RT-qPCR analysis, IRGM1, ISG20 and PSMB9[22, 23], were targets of Stat1 regulation in protein network analysis (Figure 6). In contrast, Ubd (also known as Fat10) and Cxcl9, were not identified as regulatory targets of STAT1 in protein network analysis. However, they were both retained for RT-qPCR analysis since these genes are clearly regulated by IFN-γ as previously demonstrated using promoter/reporter gene constructs in the case of Ubd[24, 25] and gene expression studies in the case of Cxcl9[26].