In-situ methane enrichment in anaerobic digestion of food waste slurry by nano zero-valent iron: Long-term performance and microbial community succession.

In this work, nano zero-valent iron (nZVI) was added to a lab-scale continuous stirring tank reactor (CSTR) for food waste slurry treatment, and the effect of dosing rate and dosage of nZVI were attempted to be changed. The results showed that anaerobic digestion (AD) efficiency and biomethanation stability were optimum under the daily dosing and dosage of 0.48 g/gTCOD. The average daily methane (CH4) yield reached 495.38 mL/gTCOD, which was 43.65% higher than that at control stage, and the maximum CH4 content reached 95%. However, under single dosing rate conditions, high nZVI concentrations caused microbial cell rupture and loosely bound extracellular polymeric substances (LB-EPS) precipitation degradation. The daily dosing rate promoted the hydrogenotrophic methanogenesis pathway, and the activity of coenzyme F420 increased by 400.29%. The microbial analysis indicated that daily addition of nZVI could promote the growth of acid-producing bacteria (Firmicutes and Bacteroidetes) and methanogens (Methanothrix).

Cognitive performance was reduced by higher air temperature even when thermal comfort was maintained over the 24-28°C range.

This study managed to create thermal comfort conditions at three temperatures (24°C-T24, 26°C-T26, and 28°C-T28) by adjusting clothing and air velocity. Thirty-six subjects (18 males and 18 females) were exposed to each of the three conditions for 4.5 h in a design balanced for order of presentation of conditions. During each exposure, they rated the physical environment, their comfort, the intensity of acute subclinical health symptoms, and their mental load, and they performed a number of cognitive tasks. Their physiological reactions were monitored. The subjects rated T24 to be comfortably cool, T26 to be comfortably neutral, and T28 to be comfortably warm. Their self-estimated performance did not differ between conditions but 12 of 14 objective metrics of cognitive performance decreased significantly at the elevated temperatures: compared with T24, their average cognitive performance decreased by 10% at T26 and by 6% at T28. At the elevated temperatures, their parasympathetic nervous system activity (as indicated by PNN50) and their arterial blood oxygen saturation level (SpO2) were both lower, which would be expected to result in reduced cognitive performance. The subjects also rated their acute subclinical health symptoms as more intense and their workload as higher at the elevated temperatures. These results suggest that where cognitive performance is the priority, it is wise to ensure a comfortably cool environment. The present study also supports the use of fans or natural ventilation to reduce the need for mechanical cooling.

Enhanced biogas biological upgrading from kitchen wastewater by in-situ hydrogen supply through nano zero-valent iron corrosion.

The in-situ hydrogen supply by nano zero-valent iron (nZVI, nFe0) corrosion provided a feasible way to improve the efficiency of biogas biological upgrading. This work studied the effects of nZVI at different dosages (0, 2, 4, 6, 8 and 10 g/L) on anaerobic digestion of kitchen wastewater by two buffer systems 2-[4-(2-hydroxyethyl) piperazin-1-yl] ethanesulfonic acid (HEPES) and sodium hydrogen carbonate (NaHCO3). The addition of nZVI improved the content of methane (CH4) and stability of anaerobic digestion process. In HEPES buffer system, the CH4 was all increased and the maximum reached 90.51% with 10 g/L nZVI, higher than 32.25% compared to the control. The maximum hydrogen enrichment (HE) was 113 ppb after nZVI addition, indicating the mass transfer efficiency of hydrogen (H2) was improved. Microbial community analysis showed that the total relative abundance of Methanobacterium and Methanolinea at 10 g/L nZVI was 53.72%, which was 1.62 times of the control group. However, in the NaHCO3 buffer system with 10 g/L nZVI addition, the content of CH4 and the loosely bound extracellular polymeric substances (LB-EPS) was lower than the control. The results indicated that the addition of nZVI was feasible for biogas upgrading, and the bidirectional effect of nZVI on the promotion or inhibition of bio-methanation might be related to the buffer system of the anaerobic process.

Elevated airflow can maintain sleep quality and thermal comfort of the elderly in a hot environment.

The effect of elevated airflow on sleep quality was investigated with 18 elderly. Three airflow conditions were set: ceiling fan/30°C/max.0.8 m/s and mean 0.7 m/s, task fan/30°C/max.0.8 m/s and mean 0.6 m/s, and thermally neutral /27°C/0.2 m/s. Sleep quality was evaluated objectively by analysis of electroencephalogram signals that were continuously monitored during the sleeping period. Urinary cortisol concentrations were analyzed to measure the activity of sympathetic nervous system. No significant difference in sleep quality, thermal comfort, or cortisol concentration was found between the ceiling fan and the neutral condition. The duration of total sleep time decreased by 35 minutes, the duration of REM sleep decreased by 15 minutes, and the cortisol concentration in the morning increased by 50 ng/mL in the task fan than the other two conditions. Compared with ceiling fan, less heat load was removed in the task fan condition, possibly due to the lower air speed. This study shows that even small heat load led to reduced sleep quality and overactive sympathetic nervous system of the elderly. By supplying an airflow of 0.8 m/s evenly over the human body, the elderly could maintain sleep quality and thermal comfort at an air temperature that was 3 K higher than the neutral temperature.

Structural and Biochemical Characterization of SbnC as a Representative Type B Siderophore Synthetase.

Staphyloferrin B is a key siderophore secreted by Staphylococcus aureus to acquire ferric ions from a host during infection, and its biosynthetic pathway has been validated to develop efficient antibacterial agents. Herein, we report the crystal structure of AMP-bound SbnC from S. aureus (SaSbnC) as the first representative structure of type B synthetases in the biosynthesis of α-hydroxycarboxylate siderophores. While type B synthetases specifically use α-ketoglutarate (α-KG) as their carboxylic acid substrate, SaSbnC showed unique structural features in the substrate pocket compared with the type A and C synthetases. Screening of α-KG analogues suggested that the hydrogen-bonding interaction between the α-carbonyl group of α-KG and residue Lys552 is a key determinant for the substrate selectivity of type B synthetases. Interestingly, citrate, the product of the tricarboxylic acid cycle and the substrate of type A synthetases, was found to inhibit the activity of SaSbnC with an IC50 value of 83 µM by mimicking α-KG binding, suggesting a potential regulatory role of the tricarboxylic acid cycle, whose activity is under the control of the intracellular iron concentration, to SaSbnC and other type B synthetases. These results provide critical new information to understand the structure, function, and regulation of type B synthetases in the siderophore-based iron acquisition system employed by a large number of pathogenic microbes.

Structure-guided optimization and mechanistic study of a class of quinazolinone-threonine hybrids as antibacterial ThrRS inhibitors.

Aminoacyl-tRNA synthetases (aaRSs) are an attractive class of antibacterial drug targets due to their essential roles in protein translation. While most traditional aaRS inhibitors target the binding pockets of substrate amino acids and/or ATP, we recently developed a class of novel tRNA-amino acid dual-site inhibitors including inhibitor 3 ((2S,3R)-2-amino-N-((E)-4-(6,7-dichloro-4-oxoquinazolin-3(4H)-yl)but-2-en-1-yl)-3-hydroxybutanamide) against threonyl-tRNA synthetase (ThrRS). Here, the binding modes and structure-activity relationships (SARs) of these inhibitors were analyzed by the crystal structures of Salmonella enterica ThrRS (SeThrRS) in complex with three of them. Based on the cocrystal structures, twelve quinazolinone-threonine hybrids were designed and synthesized, and their affinities, enzymatic inhibitory activities, and cellular potencies were evaluated. The best derivative 8g achieved a Kd value of 0.40 µM, an IC50 value of 0.50 µM against SeThrRS and MIC values of 16-32 µg/mL against the tested bacterial strains. The cocrystal structure of the SeThrRS-8g complex revealed that 8g induced a bended conformation for Met332 by forming hydrophobic interactions, which better mimicked the binding of tRNAThr to ThrRS. Moreover, the inhibitory potency of 8g was less impaired than a reported ATP competitive inhibitor at high concentrations of ATP, supporting our hypothesis that tRNA site inhibitors are likely superior to ATP site inhibitors in vivo, where ATP typically reaches millimolar concentrations.

Discovery of Novel Antibiotics as Covalent Inhibitors of Fatty Acid Synthesis.

The steady increase in the prevalence of multidrug-resistant Staphylococcus aureus has made the search for novel antibiotics to combat this clinically important pathogen an urgent matter. In an effort to discover antibacterials with new chemical structures and mechanisms, we performed a growth inhibition screen of a synthetic library against S. aureus and discovered a promising scaffold with a 1,3,5-oxadiazin-2-one core. These compounds are potent against both methicillin-sensitive and methicillin-resistant S. aureus strains. Isolation of compound-resistant strains followed by whole genome sequencing revealed its cellular target as FabH, a key enzyme in bacterial fatty acid synthesis. Detailed mechanism of action studies suggested the compounds inhibit FabH activity by covalently modifying its active site cysteine residue with high selectivity. A crystal structure of FabH protein modified by a selected compound Oxa1 further confirmed covalency and suggested a possible mechanism for reaction. Moreover, the structural snapshot provided an explanation for compound selectivity. On the basis of the structure, we designed and synthesized Oxa1 derivatives and evaluated their antibacterial activity. The structure-activity relationship supports the hypothesis that noncovalent recognition between compounds and FabH is critical for the activity of these covalent inhibitors. We believe further optimization of the current scaffold could lead to an antibacterial with potential to treat drug-resistant bacteria in the clinic.

Structural Insights into Substrate Recognition and Activity Regulation of the Key Decarboxylase SbnH in Staphyloferrin B Biosynthesis.

Staphyloferrin B is a hydroxycarboxylate siderophore that is crucial for the invasion and virulence of Staphylococcus aureus in mammalian hosts where free iron ions are scarce. The assembly of staphyloferrin B involves four enzymatic steps, in which SbnH, a pyridoxal 5'-phosphate (PLP)-dependent decarboxylase, catalyzes the second step. Here, we report the X-ray crystal structures of S. aureus SbnH (SaSbnH) in complex with PLP, citrate, and the decarboxylation product citryl-diaminoethane (citryl-Dae). The overall structure of SaSbnH resembles those of the previously reported PLP-dependent amino acid decarboxylases, but the active site of SaSbnH showed unique structural features. Structural and mutagenesis analysis revealed that the citryl moiety of the substrate citryl-l-2,3-diaminopropionic acid (citryl-l-Dap) inserts into a narrow groove at the dimer interface of SaSbnH and forms hydrogen bonding interactions with both subunits. In the active site, a conserved lysine residue forms an aldimine linkage with the cofactor PLP, and a phenylalanine residue is essential for accommodating the l-configuration Dap of the substrate. Interestingly, the freestanding citrate molecule was found to bind to SaSbnH in a conformation inverse to that of the citryl group of citryl-Dae and efficiently inhibit SaSbnH. As an intermediate in the tricarboxylic acid (TCA) cycle, citrate is highly abundant in bacterial cells until iron depletion; thus, its inhibition of SaSbnH may serve as an iron-dependent regulatory mechanism in staphyloferrin B biosynthesis.