Hydrodynamic coupling in microbially mediated fracture mineralization : formation of self-organized groundwater flow channels

Evidence of fossilized microorganisms embedded within mineral veins and mineral-filled fractures has been observed in a wide range of geological environments. Microorganisms can act as sites for mineral nucleation and also contribute to mineral precipitation by inducing local geochemical changes. In this study, we explore fundamental controls on microbially induced mineralization in rock fractures. Specifically, we systematically investigate the influence of hydrodynamics (velocity, flow rate, aperture) on microbially mediated calcite precipitation. Our experimental results demonstrate that a feedback mechanism exists between the gradual reduction in fracture aperture due to precipitation, and its effect on the local fluid velocity. This feedback results in mineral fill distributions that focus flow into a small number of self-organizing channels that remain open, ultimately controlling the final aperture profile that governs flow within the fracture. This hydrodynamic coupling can explain field observations of discrete groundwater flow channeling within fracture-fill mineral geometries where strong evidence of microbial activity is reported

Rock fracture grouting with microbially induced carbonate precipitation

Microbially induced carbonate precipitation has been proposed for soil stabilization, soil strengthening and permeability reduction as an alternative to traditional cement and chemical grouts. In this paper we evaluate the grouting of fine aperture rock fractures with calcium carbonate, precipitated through urea hydrolysis, by the bacteria Sporosarcina pasteurii. Calcium carbonate was precipitated within a small-scale and a near field-scale (3.1 m2) artificial fracture consisting of a rough rock lower surfaces and clear polycarbonate upper surfaces. The spatial distribution of the calcium carbonate precipitation was imaged using time-lapse photography and the influence on flow pathways revealed from tracer transport imaging. In the large-scale experiment, hydraulic aperture was reduced from 276 μm to 22 μm, corresponding to a transmissivity reduction of 1.71x10-5 m2/s to 8.75x10-9 m2/s, over a period of 12 days under constantly flowing conditions. With a modified injection strategy a similar three orders of magnitude reduction in transmissivity was achieved over a period of three days. Calcium carbonate precipitated over the entire artificial fracture with strong adhesion to both upper and lower surfaces and precipitation was controlled to prevent clogging of the injection well by manipulating the injection fluid velocity. These experiments demonstrate that microbially induced carbonate precipitation can successfully be used to grout a fracture under constantly flowing conditions and may be a viable alternative to cement based grouts when a high level of hydraulic sealing is required and chemical grouts when a more durable grout is required

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

The comparative effects of high fat diet or disturbed blood flow on glycocalyx integrity and vascular inflammation

Abstract Background and aims Endothelial surface glycocalyx shedding plays a role in endothelial dysfunction and increases vessel wall permeability, which can lead to inflammation and atherogenesis. We sought to elucidate whether a high fat diet (HFD) or disturbed blood flow conditions, both of which are atherogenic risk factors, would contribute more detrimentally to pre-atherosclerotic loss of endothelial glycocalyx integrity and vascular inflammation. Methods Six to seven week-old C57BL/6-background apolipoprotein-E-knockout (ApoE-KO) male mice were either fed a chow diet, fed a modified Western HFD, and/or subjected to a partial left carotid artery (LCA) ligation procedure to induce disturbed blood flow patterns in the LCA. Mice were sacrificed after 1 week of experimental conditions. Both LCA and right carotid artery (RCA) vessels were dissected and preserved to compare glycocalyx coverage and thickness as well as macrophage accumulation in carotid arterial walls amongst and between cohorts. Results Glycocalyx coverage of the endothelium was significantly reduced in the LCAs of HFD fed mice when compared to the control. More significant reduction in glycocalyx coverage occurred in the LCAs of mice exposed to disturbed flow by partial LCA ligation when compared to the control. No differences were found in glycocalyx coverage of RCAs from all cohorts. Regarding inflammation, no difference in macrophage accumulation in carotid arterial walls was observed when comparing the LCAs and RCAs of control and HFD fed mice. However, macrophage infiltration in vessel walls showed a 20-fold increase in the LCAs exposed to disturbed flow following ligation, when compared to control LCAs, while no such statistical difference was observed between the RCAs of the group. Conclusions In our mouse model, endothelial glycocalyx integrity was compromised more by disturbed blood flow patterns than by exposure of the carotid vessel to HFD conditions. The pathophysiological implications include endothelial dysfunction, which correlates to macrophage infiltration in vessel walls and promotes atherogenesis

Glucose consumption and lactate production increase under hypoxic conditions.

<p>A and B, Cal33 and OSC19 cells consume more glucose after 48 hours of hypoxic culturing than cells grown in normoxic conditions. C and D, a concomitant rise in lactate production was detected in media from Cal33 and OSC19 cells grown in hypoxic conditions for 48 hours. OSC19 cells are more glycolytic than Cal33 cells. Each dot represents one of four replicate dishes used for each time point and culturing condition. The experiment was performed twice with two replicate dishes. P values indicate a significant difference in metabolite concentrations and were calculated using a two-way ANOVA.</p