Reset of Inflammatory Priming of Joint Tissue and Reduction of the Severity of Arthritis Flares by Bromodomain Inhibition.

OBJECTIVE: We have recently shown that priming of synovial fibroblasts (SFs) drives arthritis flares. Pathogenic priming of SFs is essentially mediated by epigenetic reprogramming. Bromodomain and extraterminal motif (BET) proteins translate epigenetic changes into transcription. Here, we used a BET inhibitor (I-BET151) to target inflammatory tissue priming and to reduce flare severity in a murine experimental arthritis model. METHODS: BALB/c mice were treated by intraperitoneal injection or by local injection in the paw with I-BET151, which blocks the interaction of BET proteins with acetylated histones. We assessed the effects of I-BET151 on acute arthritis and/or inflammatory tissue priming in a model of repeated injections of monosodium urate crystals or zymosan into the mouse paw. I-BET151 was given before arthritis induction, at peak inflammation, or after healing of the first arthritis bout. We performed transcriptomic (RNA-Seq), epigenomic (ATAC-Seq), and functional (invasion, cytokine production, migration, senescence, metabolic flux) analyses of murine and human SFs treated with I-BET151 in vitro or in vivo. RESULTS: Systemic I-BET151 administration did not affect acute inflammation but abolished inflammatory tissue priming and diminished flare severity in both preventive and therapeutic treatment settings. I-BET151 was also effective when applied locally in the joint. BET inhibition also inhibited osteoclast differentiation, while macrophage activation in the joint was not affected. Flare reduction after BET inhibition was mediated, at least in part, by rolling back the primed transcriptional, metabolic, and pathogenic phenotype of SFs. CONCLUSION: Inflammatory tissue priming is dependent on transcriptional regulation by BET proteins, making them promising therapeutic targets for prevention of arthritis flares in previously affected joints.

The complement system drives local inflammatory tissue priming by metabolic reprogramming of synovial fibroblasts.

Arthritis typically involves recurrence and progressive worsening at specific predilection sites, but the checkpoints between remission and persistence remain unknown. Here, we defined the molecular and cellular mechanisms of this inflammation-mediated tissue priming. Re-exposure to inflammatory stimuli caused aggravated arthritis in rodent models. Tissue priming developed locally and independently of adaptive immunity. Repeatedly stimulated primed synovial fibroblasts (SFs) exhibited enhanced metabolic activity inducing functional changes with intensified migration, invasiveness and osteoclastogenesis. Meanwhile, human SF from patients with established arthritis displayed a similar primed phenotype. Transcriptomic and epigenomic analyses as well as genetic and pharmacological targeting demonstrated that inflammatory tissue priming relies on intracellular complement C3- and C3a receptor-activation and downstream mammalian target of rapamycin- and hypoxia-inducible factor 1α-mediated metabolic SF invigoration that prevents activation-induced senescence, enhances NLRP3 inflammasome activity, and in consequence sensitizes tissue for inflammation. Our study suggests possibilities for therapeutic intervention abrogating tissue priming without immunosuppression.

Pyrolysis of hydrochar from digestate: Effect of hydrothermal carbonization and pyrolysis temperatures on pyrochar formation.

Digestate from anaerobic digestion of biomass often contains more than 90% of water, which is economically unfavorable for pyrolysis. Hydrothermal carbonization (HTC) has potential to treat very wet biomass, however, the hydrochar may be acidic, contains polycyclic aromatic hydrocarbons (PAH) and toxic organic substances (e.g., phenolic compounds), and has very low Brunauer-Emmett-Teller (BET) surface area. In this study, pyrolysis of digestate derived hydrochar is performed at various pyrolysis and HTC temperatures. Solid chars were characterized for elemental analysis, pH, PAH, BET, pore size and volume, and phenolic substances, while HTC process liquids were characterized for pH, organic acids, furfural derivatives, and phenolic substances. Physicochemical characteristics of pyro-HTC char were compared with corresponding pyrochar and hydrochar. Pyro-HTC chars produced at higher HTC (i.e., 260°C) and pyrolysis temperatures (i.e., 800°C) showed highest BET surface area (63.5m(2)g(-1)), no PAH, relatively mild basic pH (9.34), and no phenolic compounds.

Influence of digestion temperature and organic loading rate on the continuous anaerobic treatment of process liquor from hydrothermal carbonization of sewage sludge.

This experimental study investigates the use of process liquor from hydrothermal carbonization (HTC) of sewage sludge as sole substrate for anaerobic digestion (AD). The process performance at 37°C (mesophilic) and 55°C (thermophilic) was determined based on two identical continuously-fed anaerobic filters (26 L each) operated for 20 weeks. During operation, the organic loading rate was stepwise increased from 1 to 5 gCOD L(-1)d(-)(1). This decreased the hydraulic retention time from 34 to 5 days. Significant differences in methane production were not observed as both reactors yielded up to 0.18 LCH4 gCOD(-)(1). Increased temperature had no effect on the steady-state COD removal efficiency with both reactors stabilized at 68-75%. Macro- and micronutrients were provided by HTC liquor in sufficient concentrations throughout the experiment. Methanogenesis was identified as the speed-limiting step in anaerobic digestion of HTC liquor. Hydrolysis was of an order of magnitude faster than methanogenesis.

Anaerobic degradation of increased phenol concentrations in batch assays.

Phenol is a wastewater contaminant depicting an environmental hazard. It can be found in effluents from various industrial processes and becomes even more common as a waste by-product of biomass-based bioenergy concepts. Because of its toxicity to anaerobic microorganisms, it can be recalcitrant during biogas production and anaerobic wastewater treatment. This study tested increased phenol loads (100 to 5000 mg L(-1)) as the sole carbon source in a semi-continuous mesophilic anaerobic adaption experiment using an unadapted microbial community from a standard biogas plant. Phenol was completely degraded at starting concentrations of up to 2000 mg L(-1). At 5000 mg L(-1), complete inhibition of the anaerobic community was observed. Lag times were reduced down to less than a day treating 2000 mg L(-1) after 16 weeks of adaption to gradually increased phenol concentrations. Specific degradation rates increased consecutively up to 7.02 mg gVS (-1) day(-1) at 2000 mg L(-1). This concentration was completely degraded within less than 12 days. The microbial community composition was assessed using 16S rRNA gene-based terminal restriction fragment length polymorphism (T-RFLP) analysis. In the bacterial community, no clear shift was visible. Clostridia were with the highest relative abundance of 27 %, the most prominent bacterial class. T-RFs representing Clostridia, Anaerolinaceae, Flavobacteria, and Bacteroidea appeared at similar relative abundance level throughout the experiment. The archaeal community, however, changed from a Methanosarcinales-dominated community (57%) to a community with a nearly even distribution of Methanobacteriales (21%) and Methanosarcinales (34%) with increasing starting phenol concentration.

Hydrothermal carbonization (HTC) of wheat straw: influence of feedwater pH prepared by acetic acid and potassium hydroxide.

In this study, influence of feedwater pH (2-12) was studied for hydrothermal carbonization (HTC) of wheat straw at 200 and 260°C. Acetic acid and KOH were used as acidic and basic medium, respectively. Hydrochars were characterized by elemental and fiber analyses, SEM, surface area, pore volume and size, and ATR-FTIR, while HTC process liquids were analyzed by HPLC and GC. Both hydrochar and HTC process liquid qualities vary with feedwater pH. At acidic pH, cellulose and elemental carbon increase in hydrochar, while hemicellulose and pseudo-lignin decrease. Hydrochars produced at pH 2 feedwater has 2.7 times larger surface area than that produced at pH 12. It also has the largest pore volume (1.1 × 10(-1) ml g(-1)) and pore size (20.2 nm). Organic acids were increasing, while sugars were decreasing in case of basic feedwater, however, phenolic compounds were present only at 260°C and their concentrations were increasing in basic feedwater.

Upflow anaerobic solid-state (UASS) digestion of horse manure: Thermophilic vs. mesophilic performance.

Energetic use of complex lignocellulosic wastes has gained global interest. Thermophilic digestion of horse manure based on straw was investigated using the upflow anaerobic solid-state (UASS) process. Increasing the organic loading rate from 2.5 to 5.5gvsL(-)(1)d(-)(1) enhanced the average methane production rate from 0.387 to 0.687LCH4L(-)(1)d(-)(1), whereas the yield decreased from 154.8 to 124.8LCH4kgvs(-)(1). A single-stage and two-stage process design showed almost the same performance. Compared to prior experiments at mesophilic conditions, thermophilic conditions showed a significantly higher efficiency with an increase of 59.8% in methane yield and 58.1% in methane production rate. Additional biochemical methane potential (BMP) tests with two types of horse manure and four different bedding materials showed that wheat straw obtained the highest BMP. The results show that the thermophilic UASS process can be the key to an efficient energy recovery from straw-based manures.

Behavior of selected hydrolyzed and dehydrated products during hydrothermal carbonization of biomass.

In this study, effects of reaction temperature and reaction time on both solid hydrochar and HTC process liquid products were studied for hydrothermal carbonization (HTC) of cellulose, wheat straw, and poplar. A novel slurry sampling system was designed and used with an 18.6L Parr reactor for 0-480 min in 200, 230, and 260 °C. Sugars (sucrose, glucose, and fructose), HMF, and furfural were found maximum in lower HTC temperature and time. However, they degrade following first order degradation kinetics. Activation energies of total sugars (glucose, fructose, sucrose, and xylose), furfural, and HMF for straw and poplar were 95-127, 130-135, and 74-90 kJ mol(-1), respectively and individuals were lower for HTC of cellulose than others. Organic acids (acetic acid, formic acid, and lactic acid) and phenolic compounds (phenol, catechol, and guaiacol) were increasing with higher HTC severity.

Anaerobic digestion of horse dung mixed with different bedding materials in an upflow solid-state (UASS) reactor at mesophilic conditions.

Aim of this study was to investigate the use of upflow anaerobic solid-state (UASS) digestion for treating horse manure. Biochemical methane potential (BMP) tests conducted for varying mixtures of dung (hay and silage feed) and bedding material (wheat straw, flax, hemp, wood chips) showed that straw mixed with hay horse dung has the highest potential of [Formula: see text] . Continuous mesophilic digestion was conducted for 238 days using a single-stage UASS reactor (27 L) and a two-stage UASS system with an anaerobic filter (AF, 21 L). Increasing the organic loading rate (OLR) from 2.5 to 4.5 g vs L(-1)d(-1) enhanced the methane rate of the single-stage reactor from 0.262 to 0.391 LL(-1)d(-1) while the methane yield declined from 104.8 to 86.9 L kg vs(-1). The two-stage system showed similar yields. Thus, for solid-state digestion of horse manure a single-stage UASS reactor appears sufficient.