Aromatic N-oxides are valuable due to their versatile chemical, pharmaceutical, and agricultural applications. Natural phenazine N-oxides possess potent biological activities and can be applied in many ways; however, few N-oxides have been identified. Herein, we developed a microbial system to synthesize phenazine N-oxides via an artificial pathway. First, the N-monooxygenase NaphzNO1 was predicted and screened in Nocardiopsis sp. 13-12-13 through a product comparison and gene sequencing. Subsequently, according to similarities in the chemical structures of substrates, an artificial pathway for the synthesis of a phenazine N-oxide in Pseudomonas chlororaphis HT66 was designed and established using three heterologous enzymes, a monooxygenase (PhzS) from P. aeruginosa PAO1, a monooxygenase (PhzO) from P. chlororaphis GP72, and the N-monooxygenase NaphzNO1. A novel phenazine derivative, 1-hydroxyphenazine N'10-oxide, was obtained in an engineered strain, P. chlororaphis HT66-SN. The phenazine N-monooxygenase NaphzNO1 was identified by metabolically engineering the phenazine-producing platform P. chlororaphis HT66. Moreover, the function of NaphzNO1, which can catalyze the conversion of 1-hydroxyphenazine but not that of 2-hydroxyphenazine, was confirmed in vitro. Additionally, 1-hydroxyphenazine N'10-oxide demonstrated substantial cytotoxic activity against two human cancer cell lines, MCF-7 and HT-29. Furthermore, the highest microbial production of 1-hydroxyphenazine N'10-oxide to date was achieved at 143.4 mg/L in the metabolically engineered strain P3-SN. These findings demonstrate that P. chlororaphis HT66 has the potential to be engineered as a platform for phenazine-modifying gene identification and derivative production. The present study also provides a promising alternative for the sustainable synthesis of aromatic N-oxides with unique chemical structures by N-monooxygenase.We study the bouncing dynamics of nanodroplets on superhydrophobic surfaces. We show that there are three velocity regimes with different scaling laws of the contact time, τ. Although τ remains constant over a wide velocity range, as seen for macroscale bouncing, we demonstrate that viscosity plays an essential role in nanodroplet bouncing even for low-viscosity fluids. We propose a new scaling τ ∼ (ρμR04/γ2)1/3 = (R0/v0)We2/3Re-1/3 to characterize the viscosity effect, which agrees well with the simulated results for water and argon nanodroplets with various radii and hydrophobicities. We also find pancake bouncing of nanodroplets, which is responsible for an abruptly reduced τ in a high-velocity regime.BACKGROUND An association between training load and changes in aerobic fitness has been established but the effect of training load on changes in strength/power remains controversial. METHODS Internal (Banister's TRIMP) and external (total distance, high-speed running and sprint distance) training load was collected from sixteen professional soccer players during and aerobic fitness and strength/power variables were measured before and after a 9-week pre-season. RESULTS Banister's TRIMP had a moderate correlation with changes in maximal oxygen uptake (r=0.46, 90% CI 0.04; 0.74). Total distance had a large and a moderate correlation with changes in velocity at 2M (r=0.60, 90% CI 0.23; 0.82) and changes in velocity at 4M (r=0.42, 90% CI -0.01; 0.72). High-speed running had moderate correlations with changes in maximal oxygen uptake (r=0.45, 90% CI 0.03; 0.74), velocity at 2M (r=0.45, 90% CI 0.03; 0.74) and velocity at 4M (r=0.39, 90% CI -0.00; 0.70). Sprint distance had a large and a moderate correlation with changes in maximal oxygen uptake (r=0.58, 90% CI 0.20; 0.81) and velocity at 4M (r=0.46, 90% CI 0.00; 0.74 respectively). High versus low total distance was associated with lower changes in squat jump and countermovement jump (ES=-0.90, 90% CI -1.57; -0.24 and ES=-1.06, 90% CI -1.89; -0.24) respectively. High versus low high-speed running was associated with higher changes in maximal oxygen uptake (ES=0.36, 90% CI 0.02; 0.70) but lower changes in squat jump (ES=-0.58, 90% CI -1.32; 0.15). CONCLUSIONS External rather internal training load had more pronounced correlations with changes in aerobic fitness. Higher compared with lower volumes of total distance and high-speed running were associated with lower gains in strength/power indices. Establishing a "dose-response" association between external/internal training load and endurance as well as strength adaptations, may maximize endurance gains with the least possible interference on strength/power gains, thus better informing soccer training practice.Dorsal root ganglion (DRG) neurons detect sensory inputs and are crucial for pain processing. They are often studied in vitro as dissociated cell cultures with the assumption that this reasonably represents in vivo conditions. However, to our knowledge, no study has directly compared genome-wide transcriptomes of DRG tissue in vivo versus in vitro, or between laboratories and culturing protocols. Mizoribine supplier Comparing RNA sequencing-based transcriptomes of native to cultured (4 days in vitro) human or mouse DRG, we found that the overall expression levels of many ion channels and GPCRs specifically expressed in neurons are markedly lower although still expressed in culture. This suggests that most pharmacological targets expressed in vivo are present under the condition of dissociated cell culture, but with changes in expression levels. The reduced relative expression for neuronal genes in human DRG cultures is likely accounted for by increased expression of genes in fibroblast-like and other proliferating cells, consistent with their mitotic status in these cultures. We found that the expression of a subset of genes typically expressed in neurons increased in human and mouse DRG cultures relative to the intact ganglion, including genes associated with nerve injury or inflammation in preclinical models such as BDNF, MMP9, GAL, and ATF3. We also found a striking upregulation of a number of inflammation-associated genes in DRG cultures, although many were different between mouse and human. Our findings suggest an injury-like phenotype in DRG cultures that has important implications for the use of this model system for pain drug discovery.