The absolute

CD4 cell count before vaccination, find more the magnitude of the CD4 increase, or whether or not CD4 increased to ≥200 cells/μL in the respective study year was not associated with persistence of significant antibody responses to any of the three serotypes from years 3 to 5 after vaccination, which may be attributable to the smaller sample size in the later years of follow-up. In this cohort study, the analysis showed that HIV-infected patients with CD4 counts <100 cells/μL at vaccination had significantly lower antibody responses to the three serotypes studied and faster loss of antibody responses than patients with CD4 counts of ≥100 cells/μL. During follow-up for 5 years, CD4 <100 cells/μL at vaccination and failure to achieve HIV suppression were the two independent negative predictors for maintaining significant antibody responses to 23-valent PPV despite continued increases in CD4 cell counts following HAART among the vaccine recipients. Studies investigating short-term serological responses to 23-valent PPV in HIV-infected patients have not produced consistent results [14–22,24–27,30–38], and only one study assessed the rate of antibody decline for five consecutive years after vaccination in 16 HIV-infected patients with short-term exposure to HAART

and declining CD4 lymphocyte counts [23]. The discrepancy may result from enrolment of subjects with different degrees

of immunosuppression, use of different vaccination schedules or vaccines (polysaccharide vs. conjugated vaccine) [22,24,37,38], receipt of different types Ixazomib mw of antiretroviral therapy (mono or dual antiretroviral therapy vs. HAART) [23,25–27,36,38], different immunological or virological responses to HAART, and different durations of observation. In this study we used a single dose of 23-valent PPV and the overall response rate was estimated to be 50% for those patients with CD4 counts of ≥100 cells/μL at vaccination and 25% for those with CD4 counts of <100 cells/μL at vaccination. Reverse transcriptase The lower overall response rate is likely to be related to our enrolment of patients with moderate to severe immunosuppression, as indicated by low nadir CD4 cell counts. Furthermore, we did not find statistically significant differences between patients with CD4 counts of <200 cells/μL and those with CD4 counts of ≥200 cells/μL in terms of serological responses throughout the 5-year study period. For example, at year 1, 28 of 70 patients (40.0%) with CD4 counts <200 cells/μL developed twofold or greater increases in antibody titres to serotype 14 compared with 45 of 98 (45.9%) with CD4 counts of ≥200 cells/μL (risk ratio 0.871; 95% confidence interval 0.609, 1.247). This finding may be explained by the small sample size of our study population.

, 2007). A range of compounds structurally similar to the quorum-sensing molecules produced by P. aeruginosa NVP-LDE225 nmr were tested for their inhibitory properties. These were decanol, decanoic acid, octanoic acid, tetradecanol and dodecanol (Sigma-Aldrich). These were solubilized in ethyl acetate containing 0.01% (v/v) glacial acetic acid (Fisher Scientific, UK) to a 1 M stock concentration. All solutions were stored at −20 °C for a maximum of 1 month. Each compound was diluted to 100 mM in MOPS-buffered RPMI and, using the CLSI broth microdilution M28-A assay

(CLSI, 2008), their effect on conidia, biofilm formation and the resultant biomass was evaluated (Mowat et al., 2007). Eight replicates were tested for each compound concentration on three separate occasions with all A. fumigatus strains. For biomass data, an angular transformation

was performed and the transformed data were analysed using one-way anova with Bonferroni’s multiple comparisons post-test. P<0.05 was considered significant. The analyses were performed using graphpad prism version 4.0 for Windows (GraphPad Software, CA). When A. fumigatus conidia were exposed to live P. aeruginosa cells overnight, the resultant fungal biomass was significantly reduced to 14.5% (P<0.001) of the untreated controls. Methanol-treated this website P. aeruginosa cells also showed this effect (Fig. CHIR-99021 order 1a). Exposure to the P. aeruginosa supernatant resulted in the inhibition of hyphal growth, restricting the biomass to 19.1%. The heat-treated supernatant did not significantly reduce this effect, restricting the biomass to 23.0%. When mature A. fumigatus biofilms were exposed to live P. aeruginosa cells, the fungal biomass was minimally affected (84.8%). SEM analysis revealed individual P. aeruginosa (PA01) cells and microcolonies distributed throughout the intertwined filamentous

networks of the mature A. fumigatus biofilms (Fig. 1b). All nine P. aeruginosa isolates examined showed similar effects. Aspergillus fumigatus conidia were exposed to live cells from two P. aeruginosa quorum-sensing knockout strains: PAO1:ΔLasI (unable to synthesize HSL) and PAO1:ΔLasR (synthesizes HSL, but cannot respond) (Fig. 2). Aspergillus fumigatus growth was significantly greater (P<0.001) during direct coculture with PAO1:ΔLasI (58.3%) and PAO1:ΔLasR (52.6%) in comparison with the wild-type PAO1 (22.9%). When the Transwell® system was used to determine an indirect effect on A. fumigatus biofilm development, the biomass was restricted to 30.1% of the unchallenged control by the wild-type PAO1. In comparison, the levels of inhibition were significantly less than the wild type (P<0.001) during an indirect coculture with PAO1:ΔLasI (58.8%) and PAO1:ΔLasR (56.8%). All the compounds tested reduced the cellular viability of A.

, 2007). A range of compounds structurally similar to the quorum-sensing molecules produced by P. aeruginosa Selleck HSP inhibitor were tested for their inhibitory properties. These were decanol, decanoic acid, octanoic acid, tetradecanol and dodecanol (Sigma-Aldrich). These were solubilized in ethyl acetate containing 0.01% (v/v) glacial acetic acid (Fisher Scientific, UK) to a 1 M stock concentration. All solutions were stored at −20 °C for a maximum of 1 month. Each compound was diluted to 100 mM in MOPS-buffered RPMI and, using the CLSI broth microdilution M28-A assay

(CLSI, 2008), their effect on conidia, biofilm formation and the resultant biomass was evaluated (Mowat et al., 2007). Eight replicates were tested for each compound concentration on three separate occasions with all A. fumigatus strains. For biomass data, an angular transformation

was performed and the transformed data were analysed using one-way anova with Bonferroni’s multiple comparisons post-test. P<0.05 was considered significant. The analyses were performed using graphpad prism version 4.0 for Windows (GraphPad Software, CA). When A. fumigatus conidia were exposed to live P. aeruginosa cells overnight, the resultant fungal biomass was significantly reduced to 14.5% (P<0.001) of the untreated controls. Methanol-treated BIBW2992 P. aeruginosa cells also showed this effect (Fig. Farnesyltransferase 1a). Exposure to the P. aeruginosa supernatant resulted in the inhibition of hyphal growth, restricting the biomass to 19.1%. The heat-treated supernatant did not significantly reduce this effect, restricting the biomass to 23.0%. When mature A. fumigatus biofilms were exposed to live P. aeruginosa cells, the fungal biomass was minimally affected (84.8%). SEM analysis revealed individual P. aeruginosa (PA01) cells and microcolonies distributed throughout the intertwined filamentous

networks of the mature A. fumigatus biofilms (Fig. 1b). All nine P. aeruginosa isolates examined showed similar effects. Aspergillus fumigatus conidia were exposed to live cells from two P. aeruginosa quorum-sensing knockout strains: PAO1:ΔLasI (unable to synthesize HSL) and PAO1:ΔLasR (synthesizes HSL, but cannot respond) (Fig. 2). Aspergillus fumigatus growth was significantly greater (P<0.001) during direct coculture with PAO1:ΔLasI (58.3%) and PAO1:ΔLasR (52.6%) in comparison with the wild-type PAO1 (22.9%). When the Transwell® system was used to determine an indirect effect on A. fumigatus biofilm development, the biomass was restricted to 30.1% of the unchallenged control by the wild-type PAO1. In comparison, the levels of inhibition were significantly less than the wild type (P<0.001) during an indirect coculture with PAO1:ΔLasI (58.8%) and PAO1:ΔLasR (56.8%). All the compounds tested reduced the cellular viability of A.

Mice with mOFC Talazoparib lesions acquired the reversal but failed to inhibit responding on the previously reinforced aperture, while mice with prelimbic prefrontal cortex lesions were unaffected. When tested on a progressive ratio schedule of reinforcement, mice with prelimbic cortical lesions were unable to maintain responding, resulting in declining response levels. Mice with

mOFC lesions, by contrast, escalated responding. Neither lesion affected sensitivity to satiety-specific outcome devaluation or non-reinforcement (i.e. extinction), and neither had effects when placed after animals were trained on a progressive ratio response schedule. Lesions of the ventral hippocampus, which projects to the mOFC, resulted in similar response patterns, while lateral OFC and dorsal hippocampus lesions resulted in response acquisition, though not inhibition, deficits in an instrumental reversal. Our findings thus selectively implicate the rodent mOFC in braking reinforced goal-directed action when reinforcement requires the acquisition of novel response contingencies. “
“Repetitive transcranial magnetic stimulation (rTMS) is an effective tool for inducing functional plastic changes in the brain. rTMS can also potentiate the effects of other interventions such as tactile coactivation, a form of repetitive stimulation, Dabrafenib purchase when both

are applied simultaneously. In this study, we investigated the interaction of these techniques in

affecting tactile acuity and cortical excitability, measured with somatosensory evoked potentials after paired median nerve stimulation. We first applied a session of 5-Hz rTMS, followed by a session of tactile repetitive stimulation, consisting of intermittent high-frequency tactile stimulation (iHFS) to a group of 15 healthy volunteers Terminal deoxynucleotidyl transferase (“rTMS + iHFS” group). In a second group (“rTMS w/o iHFS”), rTMS was applied without iHFS, with a third assessment performed after a similar wait period. In the rTMS w/o iHFS group, the 5-Hz rTMS induced an increase in cortical excitability that continued to build for at least 25 min after stimulation, with the effect on excitability after the wait period being inversely correlated to the baseline state. In the rTMS + iHFS group, the second intervention prevented the continued increase in excitability after rTMS. In contrast to the effect on cortical excitability, rTMS produced an improvement in tactile acuity that remained stable until the last assessment, independent of the presence or absence of iHFS. Our results show that these methods can interact homeostatically when used consecutively, and suggest that different measures of cortical plasticity are differentially susceptible to homeostatic interactions.

To address the question of whether pORF102 specifically recognizes telomeric DNA, we aimed to produce recombinant learn more protein in E. coli for use in EMSA. All attempts to prepare hexahistidine-tagged pORF102 or fusions of pORF102 with a chitin-binding domain failed, because all proteins precipitated with the insoluble fraction of cell extracts (data not shown). An N-terminal fusion with MBP yielded soluble protein, which could be purified to near electrophoretic homogeneity (Fig. 3a). Because cleavage of MBP-pORF102 with factor Xa protease and the subsequent attempt

to remove MBP by affinity chromatography again resulted in loss of soluble protein, EMSAs were performed with the fusion protein. Migration of ssDNA was retarded by MBP-pORF102 Vincristine mouse (Fig. 3b), whereas the mobility of double-stranded DNA was not affected by an up

to 1000-fold molar excess of protein (not shown). However, the shift in retardation with increasing protein concentrations suggests nonstoichiometric binding of pORF102 to the ssDNA, and interaction of the fusion protein with ssDNA representing an internal coding sequence of pAL1 indicated that the MBP-pORF102 protein was not able to specifically recognize telomeric DNA sequences (Fig. 3b). However, it cannot be excluded that recognition fails because the conformation of ssDNA under the experimental conditions differs from the native in vivo conformation of telomeric 3′-overhangs of pAL1 or because the MBP fusion (which, as shown above, did not prevent Arthrobacter from using MBP-pORF102 for in vivo replication of pAL1) impedes specific in vitro DNA binding. In this context, it is noteworthy that binding of the terminal protein TpgL of Streptomyces lividans to ssDNA corresponding to the 3′-overhang of plasmid pSLA2 telomeres also showed little specificity (Bao & Cohen, 2003). Axenfeld syndrome Similar to what was

observed in the Streptomyces system, recruitment of pORF102 to the termini of pAL1 might require additional proteins. To investigate whether pORF102 can act as a replication priming protein, we used an in vitro deoxynucleotidylation assay, which contained an ssDNA template representing the 3′-terminal 70 nucleotides of the ‘left’ end of pAL1, purified MBP-pORF102 protein, a crude extract of A. nitroguajacolicus Rü61a, MBP-pORF101 fusion protein that exhibits DNA polymerase activity (unpublished data), ATP, and different [α-32P]dNTPs in a Mg2+-containing buffer. As shown in Fig. 4, dCMP was specifically incorporated into the 64.1-kDa MBP-pORF102 protein. The deoxynucleotidylation was not detected in the absence of pORF102 or pORF101 (Fig. 4), or in the absence of crude extract, ATP, or Mg2+ (data not shown). When the single-stranded ‘left70’ DNA was omitted from the reaction, dNMP incorporation into pORF102-MBP likewise was not observed (not shown), indicating that the reaction requires a DNA template. Specific dCMP incorporation, complementary to the 3′-end of the S.

To address the question of whether pORF102 specifically recognizes telomeric DNA, we aimed to produce recombinant ERK inhibitor clinical trial protein in E. coli for use in EMSA. All attempts to prepare hexahistidine-tagged pORF102 or fusions of pORF102 with a chitin-binding domain failed, because all proteins precipitated with the insoluble fraction of cell extracts (data not shown). An N-terminal fusion with MBP yielded soluble protein, which could be purified to near electrophoretic homogeneity (Fig. 3a). Because cleavage of MBP-pORF102 with factor Xa protease and the subsequent attempt

to remove MBP by affinity chromatography again resulted in loss of soluble protein, EMSAs were performed with the fusion protein. Migration of ssDNA was retarded by MBP-pORF102 buy Epacadostat (Fig. 3b), whereas the mobility of double-stranded DNA was not affected by an up

to 1000-fold molar excess of protein (not shown). However, the shift in retardation with increasing protein concentrations suggests nonstoichiometric binding of pORF102 to the ssDNA, and interaction of the fusion protein with ssDNA representing an internal coding sequence of pAL1 indicated that the MBP-pORF102 protein was not able to specifically recognize telomeric DNA sequences (Fig. 3b). However, it cannot be excluded that recognition fails because the conformation of ssDNA under the experimental conditions differs from the native in vivo conformation of telomeric 3′-overhangs of pAL1 or because the MBP fusion (which, as shown above, did not prevent Arthrobacter from using MBP-pORF102 for in vivo replication of pAL1) impedes specific in vitro DNA binding. In this context, it is noteworthy that binding of the terminal protein TpgL of Streptomyces lividans to ssDNA corresponding to the 3′-overhang of plasmid pSLA2 telomeres also showed little specificity (Bao & Cohen, 2003). Casein kinase 1 Similar to what was

observed in the Streptomyces system, recruitment of pORF102 to the termini of pAL1 might require additional proteins. To investigate whether pORF102 can act as a replication priming protein, we used an in vitro deoxynucleotidylation assay, which contained an ssDNA template representing the 3′-terminal 70 nucleotides of the ‘left’ end of pAL1, purified MBP-pORF102 protein, a crude extract of A. nitroguajacolicus Rü61a, MBP-pORF101 fusion protein that exhibits DNA polymerase activity (unpublished data), ATP, and different [α-32P]dNTPs in a Mg2+-containing buffer. As shown in Fig. 4, dCMP was specifically incorporated into the 64.1-kDa MBP-pORF102 protein. The deoxynucleotidylation was not detected in the absence of pORF102 or pORF101 (Fig. 4), or in the absence of crude extract, ATP, or Mg2+ (data not shown). When the single-stranded ‘left70’ DNA was omitted from the reaction, dNMP incorporation into pORF102-MBP likewise was not observed (not shown), indicating that the reaction requires a DNA template. Specific dCMP incorporation, complementary to the 3′-end of the S.

Although the dd-CPases are usually the most abundant PBPs in the cell, they are not essential for bacterial survival (Denome et al., 1999) and the in vivo purposes of these seemingly nonessential and redundant enzymes are mostly unknown. The

exception to the above statement is the E. coli protein PBP 5, which helps maintain the normal morphology of this organism even in the absence of seven other PBPs (Nelson & Young, 2001). In the absence of PBP 5 by itself, the cells exhibit small morphological aberrations, but as more PBPs are deleted, the cells become considerably misshapen (Nelson & Young, 2000, 2001). PBP 5 consists of two major domains, I and II, oriented almost at right angles to one another (Davies et al., 2001; Nicholas et al., 2003). The dd-CPase active selleck kinase inhibitor site Staurosporine price is located in domain I and is responsible for maintaining normal cell shape (Nelson et al., 2002; Ghosh & Young, 2003). Domain II is composed mostly of β-sheets and may lift the enzymatic domain away from the inner membrane and into the periplasm toward the peptidoglycan

substrate (Nelson et al., 2002; Ghosh & Young, 2003). At its extreme carboxyl terminus, at the base of domain II, PBP 5 has a short 18-amino acid (Jackson & Pratt, 1987) amphipathic helix that tethers the protein to the outer face of the inner membrane (Nelson et al., 2002; Ghosh & Young, 2003). The closest homologue to PBP 5 from any organism is PBP 6, from E. coli itself. Interestingly, PBP 6, although ∼65% identical to PBP 5, cannot restore normal shape to aberrant cells, as can PBP 5 (Ghosh & Young, 2003). Domain swap and mutagenesis experiments indicate that the relevant differences

between the two enzymes localize to domain I, and, in fact, to a small stretch of 20 amino acids that surrounds the canonical KTG motif of the active site (Nelson et al., 2002; Ghosh & Young, 2003). For convenience, we will refer to this 20-amino acid segment as the ‘morphology maintenance domain’ (MMD) (Ghosh & Young, 2003) (Fig. 1). When PBP 6 is engineered so that its MMD is replaced by that from PBP 5, the mosaic protein (PBP 656) complements the shape defects of the E. coli mutants as well as wild-type PBP 5 (Ghosh Carnitine palmitoyltransferase II & Young, 2003). Conversely, replacing the MMD of PBP 5 with that from PBP 6 (generating PBP 565) eliminates the ability to complement. PBPs 5 and 6 differ by seven residues in this peptide fragment, but only two, Asp 218 and Lys 219, seem to be necessary to confer on PBP 656 the complementation attributes of PBP 5 (Ghosh & Young, 2003) (Fig. 1). However, mutation of these two amino acids does not eliminate the ability of PBP 5 to complement shape defects, suggesting that other structural features blunt the effects of these changes in the wild-type protein.

A auto Ruxolitinib perceção do estado de saúde e da qualidade de vida na amostra em estudo podem estar sobrestimados em relação ao que se passará à média dos doentes celíacos portugueses. As características de idade

e escolaridade levam a levantar a hipótese de que se está perante uma amostra de doentes celíacos muito pró-ativos na procura de soluções que minimizem as limitações impostas pela doença, nomeadamente pela procura de informação e de novos produtos alimentares, bem como soluções para a sua preparação. Estas competências permitir-lhes-ão conviver melhor com a doença, fazendo com que afete menos a sua qualidade de vida. O facto de a grande maioria dos participantes terem referido que, após o diagnóstico, a relação social com os familiares, amigos, colegas de find more trabalho não tinha sofrido alterações; que a alimentação se tinha tornado mais saudável e ainda que se sentiam satisfeitos por terem sido diagnosticados, mesmo atendendo a todas as mudanças que tiveram que efetuar, são razões que podem ajudar a explicar os resultados obtidos. É razoável supor-se que o mal-estar associado aos sintomas prévios ao diagnóstico, que podem demorar anos, e a ansiedade associada ao desconhecimento do mesmo fazem com que, após o diagnóstico, os

doentes consigam controlar melhor a doença e manifestarem melhor qualidade de vida. Os autores declaram que para esta investigação não se realizaram experiências em seres humanos e/ou animais. Os autores declaram que não aparecem dados de pacientes neste artigo. Os autores declaram que não aparecem dados de pacientes neste artigo. Os autores declaram não haver conflito de interesses.

“
“A doença celíaca (DC), com uma prevalência de 0,5‐1%, é uma doença autoimune caracterizada por inflamação da mucosa do intestino delgado RAS p21 protein activator 1 com hiperplasia das criptas e atrofia das vilosidades. Pode ser classificada de diferentes formas: 1) clássica com sintomas e sinais sugestivos de má absorção de nutrientes, associada a atrofia das vilosidades e com resoluções clínica e histológica após dieta isenta de glúten entre poucas semanas a alguns meses; 2) atípica em que prevalecem as queixas extraintestinais, embora a maioria dos pacientes apresente lesão grave da mucosa do intestino; 3) assintomática ou silenciosa clinicamente, embora existam alterações da mucosa intestinal; 4) latente com sintomas minor ou assintomática e sem alterações intestinais mesmo com dieta com glúten. Não se conhece a sua etiologia, sabe‐se, no entanto, que é mediada por fatores ambientais, genéticos e imunológicos. Em relação aos primeiros existe uma associação evidente entre a DC e a gliadina, componente do glúten presente no trigo, na cevada, no centeio e, em pequenas quantidades, na aveia.

As revealed here via cytotoxicity assays, both PAMAM-coated and citrate-coated AuNps induced cytotoxicity in HepG2 cells or PBMC. A decrease

in cell viability upon incubation with AuNps-citrate and AuNps-PAMAM for both HepG2 and PBMC has been observed using the MTT assay. A change in morphology of HepG2 cells upon AuNps treatment also indicated the toxicity effects (data not shown). The genotoxicity assays employed here, as shown in Table 2 (HepG2 cells) and Table 3 (PBMC), can be related to the nanometric dimensions of AuNps, which may undergo cell uptake (Lewinski et al., 2008). It was evidenced that AuNps-PAMAM and AuNps-citrate induced DNA damage, as an indicative of genotoxicity. This effect is related to the cellular toxicity of gold nanoparticles, Alectinib clinical trial which in our case is also related to the small size of the particles that easily undergo cell uptake through diffusion, in agreement with Pernodet et al. (2006). Li et al. (2008) demonstrated that serum coated 20 nm AuNps were also able to induce genotoxicity in the form of single-strand this website lesions in DNA in human lung fibroblasts. Our analyses provided convincing evidence of the toxic effect of AuNps, indicating that surface charge or size may be a major determinant of how AuNps impact cellular processes. Furthermore, the DNA damage index for AuNps-PAMAM

and AuNps-Citrate was statistically significant analyzed for HepG2 cells, except at 1.0 μM AuNps-Citrate. The genotoxicity for PBMC was statistically significant only upon incubation with AuNps-PAMAM at 50.0 μM. The tendency of the AuNps to accumulate in the cells nuclei was associated with their small size, which allows the nanoparticles to freely diffuse through pore complex (Zhao

and Nalwa, 2007). Since the comet assay evaluates the reversible DNA damage, our genotoxicity results also suggest that PBMC, a primary cell culture, were less sensitive to DNA damage to a certain extent the nanoparticles than HepG2 cancer cells. The purpose was to analyze the repair system in comet assay evidencing the DNA repair. The use of SSC parameter obtained via flow cytometry has been DCLK1 proposed as an efficient way to investigate cell uptake (Suzuki et al., 2007). In our analyses, the uptake of both types of AuNps was monitored by SSC (Table 4), revealing that for HepG2 cells, the relative SSC values were significantly increased (p < 0.05) only for cells incubated with AuNps-PAMAM at 50.0 μM. In contrast, the PBMC exhibited an increase in the SSC values for cells incubated with both types of nanoparticles at 50.0 μM. Furthermore, a significantly increase in SSC was also observed for PBMC upon incubation with AuNps-PAMAM at the lower concentration investigated (1.0 μM).

Studies of esophageal precancers revealed that the degree of clonal diversity was found to increase the probability of progression from esophageal precancer

to adenocarcinoma [22]. Minor subpopulations of primary tumors were shown to be responsible for relapse after drug administration [34]. Intratumor heterogeneity of PTEN protein expression corresponded with loss of heterozygosity and shorter OS in glioblastoma [35]. Tumor heterogeneity of Ki-67 protein in prostate cancer correlated with more aggressive tumor characteristics [5]. In this study, we have demonstrated that heterogeneity of check details individual proteins, namely PIK3CA, MYC, TOP2A, ESR1, PGR, RUNX1, RAD21, and CDKN2A, correlates with more aggressive tumor behavior and, in case of MYC, TOP2A, ESR1, and RAD21, also confers poor prognosis. Interestingly, prognostic significance of the studied proteins depends on whether the heterogeneity or the expression level is being analyzed. Apart from ESR1, PGR, and TOP2A, which were significantly correlated with prognosis in terms of both the heterogeneity and the expression level, there

were also proteins that were either informative in the context of tumor heterogeneity (PIK3CA, MYC, CDKN2A, RAD21, and RUNX1) or protein expression level (ERBB2, ERBB3, and TP53). Thus, protein heterogeneity and staining intensity might be two distinct phenomena, differently reflecting the course of the disease. Correlations between protein heterogeneity of ESR1 and PGR, ESR1 and RAD21, and ERBB1 and pAKT1 were especially strong. ESR1 and PGR1 expression was found to correlate

strongly in EC [36]. Investigation of ERBB1 and GSI-IX pAKT1 expression revealed strong correlation between those two proteins in head and neck squamous cell carcinoma [37]. Similarly, we have found statistically significant correlations between ESR1 and PGR, ESR1 and RAD21, and ERBB1 and pAKT1 (data not shown). Mentioned proteins are functionally related. Perhaps if their expression is co-dependent, so could be the heterogeneity. Cumulative tumor heterogeneity of selected proteins’ heterogeneity proved to be an independent predictor most of survival and showed the strongest correlations with clinicopathologic data. Apparently, simultaneous analysis of a large number of protein markers gives more thorough image of clonal diversity present in the tumor. Therefore, we conclude that the larger the extent of intratumor heterogeneity in EC, the more aggressive the tumor behavior is and thus the worse the prognosis is. One of the limitations of the study was relatively short follow-up period. Furthermore, due to variable quality and sometimes small amount of collected material, reliable analysis of all four cores per patient not always could have been achieved. This issue was even greater in case of global protein heterogeneity determination. However, despite TMA limitations, there is an increasing number of publications based on tumor microarrays due to their convenience.