Anti-heat shock protein-27 (Hsp-27) antibody levels in patients with chest pain: association with established cardiovascular risk factors

BACKGROUND Antibody titres to several heat shock proteins (Hsps) have been shown to be associated with risk factors for cardiovascular disease (CVD), but there are no data for Hsp-27. We developed an ELISA for total IgG antibody concentrations, applying this to individuals with and without acute coronary syndrome, and have assessed the relationship between antibody levels and individual coronary risk factors. METHODS Blood was collected from 63 healthy controls without a history of chest pain or CVD and 60 patients admitted to hospital with acute cardiac chest pain on admission and approximately 12 h after the acute event. RESULTS Patients with chest pain had significantly higher Hsp-27 antibody levels than controls [median 0.16 (range 0.01-0.51) vs. 0.10 (range 0.00-0.32); p312, p=0.01) respectively. CONCLUSIONS Raised antibody levels to Hsp-27 were associated only with age and hypertensio

Leishmania major: low infection dose causes short-lived hyperalgesia and cytokines upregulation in mice

Neural involvement was traditionally associated with leprosy. However, more recent studies have shown the presence of a persistent hyperalgesia in cutaneous leishmaniasis caused by the infection of BALB/c mice with a high dose of Leishmania major. In this study, we report the presence of hyperalgesia within the first two weeks of infection caused by a low dose of the parasite. Using BALB/c mice, we demonstrate the presence of hyperalgesia during the first 10 days of infection as assessed by thermal pain tests. After 10 days these decreased pain thresholds start to recover resulting in similar levels to those in uninfected controls during the third week of infection. This hyperalgesia is accompanied by a sustained upregulation of interleukin-1beta (IL-1beta) and an early upregulation of interleukin-6 (IL-6) which is restored to normal levels after five days of infection. In conclusion, this study shows that, during early infection, the low dose of L. major causes hyperalgesia accompanied by an upregulation of IL-1beta and IL-6 and that these effects are reversed within the first two weeks of infection

Wild carrot pentane-based fractions suppress proliferation of human HaCaT keratinocytes and protect against chemically-induced skin cancer

Abstract Background Previous studies in our laboratory showed that the Lebanese Daucus carota ssp. carota (wild carrot) oil extract possesses in vitro and in vivo anticancer activities. The present study aims to examine the cytotoxic effect of Daucus carota oil fractions on human epidermal keratinocytes and evaluate the chemopreventive activity of the pentane diethyl ether fraction on DMBA/TPA induced skin carcinogenesis in mice. Methods Wild carrot oil extract was chromatographed to yield four fractions (F1, 100% pentane; F2, 50:50 pentane:diethyl ether; F3, 100% diethyl ether; F4 93:7 chloroform:methanol). The cytotoxic effect of fractions (10, 25, 50 and 100\ua0\u3bcg/mL) was tested on human epidermal keratinocytes (non-tumorigenic HaCaT cells and tumorigenic HaCaT-ras variants) using WST a ssay. Cell cycle phase distribution of tumorigenic HaCaT-ras variants was determined by flow cytometry post-treatment with F2 fraction. Apoptosis related proteins were also assessed using western blot. The antitumor activity of F2 fraction was also evaluated using a DMBA/TPA induced skin carcinoma in Balb/c mice. Results All fractions exhibited significant cytotoxicity, with HaCaT cells being 2.4\u20133 times less sensitive than HaCaT-ras A5 (benign tumorigenic), and HaCaT-ras II4 (malignant) cells. GC-MS analysis revealed the presence of a major compound (around 60%) in the pentane/diethylether fraction (F2), identified as 2-himachalen-6-ol. Treatment of HaCaT-ras A5 and HaCaT-ras II4 cells with F2 fraction resulted in the accumulation of cells in the sub-G1 apoptotic phase and decreased the population of cells in the S and G2/M phases. Additionally, F2 fraction treatment caused an up-regulation of the expression of pro-apoptotic (Bax) and down-regulation of the expression of anti-apoptotic (Bcl2) proteins. A decrease in the phosphorylation of AKT and ERK was also observed. Intraperitoneal treatment with F2 fraction (50 or 200\ua0mg/kg) in the DMBA/TPA skin carcinogenesis mouse model showed a significant inhibition of papilloma incidence (mice with papilloma), yield (number of papilloma/mouse) and volume (tumor relative size) at weeks 15, 18 and 21. Conclusion The present data reveal that F2 fraction has a remarkable antitumor activity against DMBA/TPA-induced skin carcinogenesis, an effect that may be mediated through inhibition of the MAPK/ERK and PI3K/AKT pathways

Additional file 1: of Wild carrot pentane-based fractions suppress proliferation of human HaCaT keratinocytes and protect against chemically-induced skin cancer

Shebaby et al. Raw Data R4. (XLSX 56 kb

Characterization of polyfunctional cytokine expression by CSFV epitope-specific CD8 T cells.

<p>Cryopreserved PBMC from pigs AN5, AN7, AN11 and AN13 were stimulated with the identified antigenic peptides and the expression of IFN-γ, TNF-α and IL-2 by CD8 T cells was simultaneously assessed by flow cytometry. Panel A shows representative dot plots of the expression of TNF-α and IL-2 by peptide specific IFN-γ expressing CD8 T cells and Panel B the relative proportions of IFN-γ⁺ CD8 T cells expressing either of the two other cytokines. In panel C, the mean fluorescence intensity (MFI) of IFN-γ staining in each of the populations is presented. Values represent the mean values for triplicate cultures +/− SEM. Statistical analyses were performed using a one-way ANOVA followed by a Dunnett's multiple comparison test against CD8 T cells expressing only IFN-γ; ***p<0.001, **p<0.01, *p<0.05.</p

SLA class I low-resolution (Lr) haplotypes of C-strain vaccinated pigs and CD8 T cell reactivity against identified antigenic peptides.

#<p>C-strain vaccinated pigs were challenged with CSFV Brescia (AN), UK2000/7.1 (AD) and CBR/93 (AE).</p>∼<p>Medium-/high-resolution specificity not determined.</p><p>*16.0 mod.: SLA-1*11XX instead of SLA-1*04XX.</p

Recognition of the identified epitopes by CD8 T cells from C-strain vaccinated pigs challenged with divergent CSFV strains.

<p>PBMC from pigs vaccinated with C-strain CSFV and challenged with CSFV isolate UK2000/7.1 (A) or CBR/93 (B), collected 9 or 12 days post-challenge, were stimulated with antigenic peptides NS2 STVTGIFL and NS3 VEYSFIFLDEY. IFN-γ expression by CD8 T cells assessed by flow cytometry and graphs show the mean unstimulated-corrected % IFN-γ expressing CD8 T cells +/− SEM. Statistical analyses were performed using a one-way ANOVA followed by a Dunnett's multiple comparison test versus the un-stimulated control; **p<0.01, *p<0.05.</p

CD8 T cells from pigs vaccinated with C-strain and challenged with virulent CSFV display distinct profiles of antigen reactivity.

<p>Fourteen days after re-challenge with CSFV Brescia strain, PBMC from four selected pigs were stimulated with synthetic peptide pools representing the 12 CSFV proteins and IFN-γ expression was assessed by flow cytometry. Graphs show the mean unstimulated corrected % IFN-γ⁺ CD8 T cells for each animal in response to peptide pools +/− SEM. Statistical analyses were performed using a one-way ANOVA followed by a Dunnett's multiple comparison test versus the unstimulated cells; ***p<0.001, **p<0.01.</p