Macrophage Subset Sensitivity to Endotoxin Tolerisation by Porphyromonas gingivalis

Macrophages (MΦs) determine oral mucosal responses; mediating tolerance to commensal microbes and food whilst maintaining the capacity to activate immune defences to pathogens. MΦ responses are determined by both differentiation and activation stimuli, giving rise to two distinct subsets; pro-inflammatory M1- and anti-inflammatory/regulatory M2- MΦs. M2-like subsets predominate tolerance induction whereas M1 MΦs predominate in inflammatory pathologies, mediating destructive inflammatory mechanisms, such as those in chronic P.gingivalis (PG) periodontal infection. MΦ responses can be suppressed to benefit either the host or the pathogen. Chronic stimulation by bacterial pathogen associated molecular patterns (PAMPs), such as LPS, is well established to induce tolerance. The aim of this study was to investigate the susceptibility of MΦ subsets to suppression by P. gingivalis. CD14hi and CD14lo M1- and M2-like MΦs were generated in vitro from the THP-1 monocyte cell line by differentiation with PMA and vitamin D3, respectively. MΦ subsets were pre-treated with heat-killed PG (HKPG) and PG-LPS prior to stimulation by bacterial PAMPs. Modulation of inflammation was measured by TNFα, IL-1β, IL-6, IL-10 ELISA and NFκB activation by reporter gene assay. HKPG and PG-LPS differentially suppress PAMP-induced TNFα, IL-6 and IL-10 but fail to suppress IL-1β expression in M1 and M2 MΦs. In addition, P.gingivalis suppressed NFκB activation in CD14lo and CD14hi M2 regulatory MΦs and CD14lo M1 MΦs whereas CD14hi M1 pro-inflammatory MΦs were refractory to suppression. In conclusion, P.gingivalis selectively tolerises regulatory M2 MΦs with little effect on pro-inflammatory CD14hi M1 MΦs; differential suppression facilitating immunopathology at the expense of immunity

Neuroticism Modifies Psychophysiological Responses to Fearful Films

Background: Neuroticism is a personality component frequently found in anxious and depressive psychiatric disorders. The influence of neuroticism on negative emotions could be due to its action on stimuli related to fear and sadness, but this remains debated. Our goal was thus to better understand the impact of neuroticism through verbal and physiological assessment in response to stimuli inducing fear and sadness as compared to another negative emotion (disgust).¦Methods: Fifteen low neurotic and 18 high neurotic subjects were assessed on an emotional attending task by using film excerpts inducing fear, disgust, and sadness. We recorded skin conductance response (SCR) and corrugator muscle activity (frowning) as indices of emotional expression.¦Results: SCR was larger in high neurotic subjects than in low neurotics for fear relative to sadness and disgust. Moreover, corrugator activity and SCR were larger in high than in low neurotic subjects when fear was induced.¦Conclusion: After decades of evidence that individuals higher in neuroticism experience more intense emotional reactions to even minor stressors, our results indicate that they show greater SCR and expressive reactivity specifically to stimuli evoking fear rather than to those inducing sadness or disgust. Fear processing seems mainly under the influence of neuroticism. This modulation of autonomic activity by neurotics in response to threat/fear may explain their increased vulnerability to anxious psychopathologies such as PTSD (post traumatic stress disorder)

Ionizing radiation modulates human macrophages towards a pro-inflammatory phenotype preserving their pro-invasive and pro-angiogenic capacities

In order to improve the efficacy of conventional radiotherapy, attention has been paid to immune cells, which not only modulate cancer cell response to therapy but are also highly recruited to tumours after irradiation. Particularly, the effect of ionizing radiation on macrophages, using therapeutically relevant doses, is not well understood. To evaluate how radiotherapy affects macrophage behaviour and macrophage-mediated cancer cell activity, human monocyte derived-macrophages were subjected, for a week, to cumulative ionizing radiation doses, as used during cancer treatment (2Gy/fraction/day). Irradiated macrophages remained viable and metabolically active, despite DNA damage. NF-kappaB transcription activation and increased Bcl-xL expression evidenced the promotion of pro-survival activity. A significant increase of pro-inflammatory macrophage markers CD80, CD86 and HLA-DR, but not CCR7, TNF and IL1B was observed after 10Gy cumulative doses, while anti-inflammatory markers CD163, MRC1, VCAN and IL-10 expression decreased, suggesting the modulation towards a more proinflammatory phenotype. Moreover, ionizing radiation induced macrophage morphological alterations and increased their phagocytic rate, without affecting matrix metalloproteases (MMP)2 and MMP9 activity. Importantly, irradiated macrophages promoted cancer cell-invasion and cancer cell-induced angiogenesis. Our work highlights macrophage ability to sustain cancer cell activities as a major concern that needs to be addressed to improve radiotherapy efficacy

Influence of Caloric Restriction on Constitutive Expression of NF-κB in an Experimental Mouse Astrocytoma

Many of the current standard therapies employed for the management of primary malignant brain cancers are largely viewed as palliative, ultimately because these conventional strategies have been shown, in many instances, to decrease patient quality of life while only offering a modest increase in the length of survival. We propose that caloric restriction (CR) is an alternative metabolic therapy for brain cancer management that will not only improve survival but also reduce the morbidity associated with disease. Although we have shown that CR manages tumor growth and improves survival through multiple molecular and biochemical mechanisms, little information is known about the role that CR plays in modulating inflammation in brain tumor tissue.Phosphorylation and activation of nuclear factor κB (NF-κB) results in the transactivation of many genes including those encoding cycloxygenase-2 (COX-2) and allograft inflammatory factor-1 (AIF-1), both of which are proteins that are primarily expressed by inflammatory and malignant cancer cells. COX-2 has been shown to enhance inflammation and promote tumor cell survival in both in vitro and in vivo studies. In the current report, we demonstrate that the p65 subunit of NF-κB was expressed constitutively in the CT-2A tumor compared with contra-lateral normal brain tissue, and we also show that CR reduces (i) the phosphorylation and degree of transcriptional activation of the NF-κB-dependent genes COX-2 and AIF-1 in tumor tissue, as well as (ii) the expression of proinflammatory markers lying downstream of NF-κB in the CT-2A malignant mouse astrocytoma, [e.g. macrophage inflammatory protein-2 (MIP-2)]. On the whole, our date indicate that the NF-κB inflammatory pathway is constitutively activated in the CT-2A astrocytoma and that CR targets this pathway and inflammation.CR could be effective in reducing malignant brain tumor growth in part by inhibiting inflammation in the primary brain tumor

Structural control, hydrothermal alteration zonation, and fluid chemistry of the concealed, high-grade 4EE iron orebody at the paraburdoo 4E deposit, Hamersley Province, Western Australia

High-grade iron ore of the 4EE orebody of the 4E deposit (>200 Mt at 63.5 wt % Fe) occurs as a southerly dipping sheet within banded iron formation (BIF) of the Paleoproterozoic Dales Gorge and Joffre members of the Brockman Iron Formation. Structural reconstruction of the 4E deposit shows that reactivation of the 18E fault and development of the NW-striking, steeply SW dipping 4E and 4EE normal faults resulted in preservation of the 4EE orebody below the 4E deposit, and 400 m below the modern topographic surface. Three hypogene alteration zones between low-grade BIF and high-grade iron ore are observed: (1) distal magnetite-quartz-dolomite-stilpnomelane-hematite ± pyrite, (2) intermediate magnetite-dolomite-hematitechlorite-quartz- stilpnomelane, and (3) proximal hematite-dolomite-chlorite ± pyrite ± magnetite. Hydrothermal alteration is temporally and spatially constrained by NW-trending dolerite dikes that intruded the 4E and 4EE faults prior to hypogene alteration. Six vein types (V1-V6) are recognized at the 4E deposit. The veins both cut and parallel the primary BIF layers and were emplaced contemporaneously with the hydrothermal alteration zones that record the transformation of low-grade BIF to high-grade iron ore. Our integrated structural-hydrothermal alteration and fluid flow model proposes that during early stage 1a, hypogene fluid flow in the 4E orebody occurred during a period of continental extension and enhanced heat flow within sedimentary basins to the south of the Paraburdoo Range. Heated basinal brines were focused by the NW-striking, steeply SW dipping 4E and 4EE normal faults and reacted with BIF of the Dales Gorge and Joffre members. The warm to hot (160° -255°C), Ca-rich (26.6-31.9 equiv wt % CaCl2) basinal brine interacted with magnetite-chert layers, transforming them into magnetite-quartz-dolomite- stilpnomelane-hematite-pyrite BIF. The iron-rich brine (up to 2.8 wt % Fe) likely originated from evaporated seawater that had lost Mg and Na and gained Li and Ca through fluid-rock reactions with volcaniclastic rocks and carbonate successions within the Wittenoom Formation. The first incursion of deeply circulating, low-salinity (5.8-9.5 wt % NaCl equiv), heated (106°-201°C) modified meteoric water is recorded in late stage 1a minerals. This modified meteoric water had lost some of its Na through wall rock interaction with plagioclase, possibly by interaction with dolerite of the Weeli Wooli Formation that directly overlies the Joffre and Dales Gorge members. Stage 1b involved continuing reactions between the hydrothermal fluids and the magnetite-quartz-dolomite-stilpnomelane-hematite-pyrite BIF, and produced both the intermediate magnetite-dolomite-hematite-chlorite-pyrite and the proximal hematite-dolomite-magnetite-stilpnomelane alteration assemblages. Microplaty (10-80 μm), platy (100-250 μm), and anhedral hematite increasingly replace magnetite in the intermediate alteration zone, forming the proximal alteration zones that consist of microplaty, platy, anhedral hematite and magnetite. The intermediate and proximal alteration zones represent the mixing of a hot (250°-400°C), high-salinity, Ca-rich (30-40 wt % CaCl2 equiv), Sr-rich basinal brine with low-temperature and low-salinity (~5 wt % NaCl equiv) modified meteoric water that was heated (~100°-200°C) during its descent into the upper crust. Heterogeneous mixing of the two end-member fluids resulted in the trapping of primary fluid inclusion assemblages containing a wide range of trapping temperatures (up to 200°C) and salinities (up to 25 wt % NaCl equiv). Stage 1c of the hypogene hydrothermal fluid is characterized by low-temperature (<110°C), low-salinity (~5 wt % NaCl) meteoric water that interacted with the proximal hematite-dolomite-magnetite-stilpnomelane- altered BIF, leaving a porous, hematite-apatite high-grade ore. Supergene alteration affected the orebody since the Cretaceous and produced a hematite-goethite alteration assemblage, resulting in destruction of the hypogene alteration zones that are only preserved below the depth of modern weathering. Discovery of the concealed 4EE orebody of the 4E deposit demonstrates that structural geology plays a critical role in the exploration for high-grade iron orebodies. Structural reconstruction should be considered a critical exploration activity in structurally complex terranes where concealed orebodies may exist