Controls on the Geometry and Evolution of Deep-water Fold Thrust Belt of the NW Borneo.

The key driving mechanisms for establishing deep-water fold-thrust belt are either lithospheric stress or gravity-driven associated with margin instability or a combination of both. Despite long academic interest, we still lack of detailed understanding of the interaction between the deformation mechanisms (gravity- and tectonic-driven). The results of an evaluation of the interaction between the deformation mechanisms, with focused attention upon the NW Borneo deep-water fold-thrust belt, are reported. A methodology integrating a detailed structural analysis of the deep-water fold-thrust belt from the available subsurface data and equivalent onshore outcrop is utilized in this study. Detailed structural analysis of 2D seismic profiles is used to present a basin-scale seismic-stratigraphic framework and detailed description of the general appearance of the deformational style along the deltaic system. Sub-seismic scale investigation of well-exposed outcrops onshore NW Sabah is used to extract information on onshore tectonic deformation, making it possible to evaluate the differences of structural architecture related to different deformation mechanisms. The result has led to an improved understanding of the regional-scale structural geometry along the NW Borneo margin. Regional scale cross-sections are used to demonstrate a regional-scale analysis of the NW Borneo margin that includes structural restoration. The results allow an assessment of the relative timing of deformation, the domain interaction and the possible processes and parameters that control deformation. This has led to an improved insight relating to the kinematic nature of the allochthon and the interaction between the deformation mechanisms. Structural restorations are also used to evaluate of areas of compressionally and extensionally dominated systems, in order to verify the main proses responsible for the margin evolution. This study illustrates outcrop-scale to seismic-scale analysis and quantitative measurements combined with seismic interpretations, with the aim to identify the interaction between gravity-and tectonic-driven deformation, and their controls on the geometry and evolution of deep-water fold-thrust systems. Additionally, the margin evolution and the implications on NW Borneo are evaluated

Survival of wild type and <i>ralp3</i> expression strains in human whole blood and human serum.

<p>The data of the survival assays represent the mean values ± standard deviations from four independent biological experiments. The number of surviving CFU was determined by plating serial dilutions and subsequent counting. The <i>y</i> axis shows the resulting multiplication factor for each strain calculated from the percentage of surviving CFU related to the inoculum. The level of significance was calculated by U-test.</p

Anaerobic Co-Culture of Mesenchymal Stem Cells and Anaerobic Pathogens - A New <i>In Vitro</i> Model System

<div><p>Background</p><p>Human mesenchymal stem cells (hMSCs) are multipotent by nature and are originally isolated from bone marrow. In light of a future application of hMSCs in the oral cavity, a body compartment with varying oxygen partial pressures and an omnipresence of different bacterial species i.e. periodontitis pathogens, we performed this study to gain information about the behavior of hMSC in an anaerobic system and the response in interaction with oral bacterial pathogens.</p><p>Methodology/Principal Findings</p><p>We established a model system with oral pathogenic bacterial species and eukaryotic cells cultured in anaerobic conditions. The facultative anaerobe bacteria <i>Fusobacterium nucleatum</i>, <i>Porphyromonas gingivalis</i> and <i>Aggregatibacter actinomycetemcomitans</i> were studied. Their effects on hMSCs and primary as well as permanent gingival epithelial cells (Ca9-22, HGPEC) were comparatively analyzed. We show that hMSCs cope with anoxic conditions, since 40% vital cells remain after 72 h of anaerobic culture. The Ca9-22 and HGPEC cells are significantly more sensitive to lack of oxygen. All bacterial species reveal a comparatively low adherence to and internalization into hMSCs (0.2% and 0.01% of the initial inoculum, respectively). In comparison, the Ca9-22 and HGPEC cells present better targets for bacterial adherence and internalization. The production of the pro-inflammatory chemokine IL-8 is higher in both gingival epithelial cell lines compared to hMSCs and <i>Fusobacterium nucleatum</i> induce a time-dependent cytokine secretion in both cell lines. <i>Porphyromonas gingivalis</i> is less effective in stimulating secretion of IL-8 in the co-cultivation experiments.</p><p>Conclusions/significance</p><p>HMSCs are suitable for use in anoxic regions of the oral cavity. The interaction with local pathogenic bacteria does not result in massive pro-inflammatory cytokine responses. The test system established in this study allowed further investigation of parameters prior to set up of oral hMSC <i>in vivo</i> studies.</p></div

Enzyme-linked immunosorbent assay of IL-8.

<p>The hMSCs and Ca9-22 cells were challenged with a MOI of 1∶100 for 1, 2, 4 and 24 h with the microorganism (A) <i>Fusobacterium nucleatum</i> ATTCC 23726 (B)<i>Fusobacterium nucleatum</i> ATTCC 25586 (C) <i>Porphyromonas gingivalis</i> W50 or (D) <i>Porphyromonas gingivalis</i> W83. The Enzyme-linked immunosorbent assay was performed with supernatant from MSC (black bars) and Ca9-22 (white bars) after the indicated time points.</p

Direct interaction of oral bacteria on hMSC surface.

<p>(A) Scanning electron microscope and (B) immunofluorescence microscope pictures. The immunofluorescence microscope samples were stained with Live/Dead dyes. Live cells are stained in green, dead cells light up in red.</p

Bacterial growth in cell culture medium.

<p>Growth rates of bacteria in DMEM cell culture medium under anaerobic conditions (10% CO₂, 10% H₂, 80% N₂), 37°C. Bacteria were grown in PYG medium supplemented with 5 µg/ml hemin and 1% vitamin K to the stationary phase. Subsequently, they were centrifuged, washed in PBS, and each bacterial suspension was diluted in DMEM 1∶10. The OD₆₀₀ was measured at 600 nm from timepoint 0 every hour to timepoint 12 h with a final measurement after 24 h.</p

Cell survival under anaerobic conditions.

<p>Survival of hDFSC, hBMSC, hGiF and Ca9-22 cells under anaerobic compared with aerobic conditions. Anaerobic atmosphere: (10% CO₂, 10% H₂, 80% N₂) and 37°C. Viable cells were counted at timepoints of 24, 48 and 72 h in a Neubauer hemacytometer by exclusion of trypan blue. Values are expressed as means ± SD (standard deviation), *p<0.05 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22.</p

IL-8 and IL-10 secretion.

<p>The assays were carried out as described in the text. IL-8 and IL-10 levels were assayed by ELISA. Absorbance was read at 450 nm. Values represent the means ± SD, *p<0.05, **p = 0.01 (T-test), n = 4. Asterisks indicate statistically significant differences between hDFSC and hBMSC, hGiF, and Ca9-22. Cytokine secretion by hDFSC was compared with secretion by hBMSC, hGiF, and Ca9-22 cells at the same timepoint. <b>A</b>) IL-8 measured in the supernatant of the <i>F.nucleatum</i> ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. <b>B</b>) IL-8 measured in the supernatant of the <i>P. gingivalis</i> W50 and W83 co-culture after 1, 2, 4, and 24 h. <b>C</b>) IL-10 measured in the supernatant of the <i>F.nucleatum</i> ATCC 23727 and 25586 co-culture after 1, 2, 4, and 24 h. <b>D</b>) IL-10 measured in the supernatant of the <i>P. gingivalis</i> W50 and W83 co-culture after 1, 2, 4, and 24 h.</p

Survival of hMSC and Ca9-22 under aerobic and anaerobic conditions.

<p>(A) Percentage of living cells in comparison of aerobic to anaerobic cultivation conditions. The significance was calculated via Mann-Whitney-U test. P≤0.05 was considered as significant. (B) Immunofluorescence microscope images of hMSC and Ca9-22 stained with Live/Dead dyes after 24 h, 48 h and 72 h after anaerobic conditions.</p