Systematic characterization of the chemical constituents in vitro and in vivo of Qianghuo by UPLC-Q-TOF-MS/MS.

The Chinese herb Qianghuo is an antiphlogistic herb with many effects and complex components. In this study, the chemical composition of Qianghuo and its components in rat plasma after oral administration were investigated using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS). The extracts, blank plasma, and plasma containing the drug were analyzed by mass spectrometry, and data collected in both positive and negative ion modes were analyzed using Masslynx software, and the structures were confirmed by combining the compound fragment ions and mass spectrometry cleavage pathways. A total of 62 in vitro chemical components were identified, including 27 coumarins, 18 organic acids, 5 amino acids, 5 glycosides, 2 flavonoids, 4 nucleotides, and 1 ester, which were summarized from the obtained compounds in terms of their possible cleavage patterns. Among the identified 31 compounds in rat plasma, 21 were prototypes, mostly coumarins, organic acids, and flavonoids, and 10 were metabolites, which were mainly generated via hydroxylation and methylation pathways. Based on these, this study provides a theoretical foundation for quality control and basic research on Qianghuo medicinal substances.

Searching for Afterglow: Light Dark Matter Boosted by Supernova Neutrinos.

A novel analysis is performed, incorporating time-of-flight (TOF) information to study the interactions of dark matter (DM) with standard model particles. After supernova (SN) explosions, DM with mass m_{χ}≲O(MeV) in the halo can be boosted by SN neutrinos (SNν) to relativistic speed. The SNν boosted DM (BDM) arrives on Earth with TOF which depends only on m_{χ} and is independent of the cross section. These BDMs can interact with detector targets in low-background experiments and manifest as afterglow events after the arrival of SNν. The characteristic TOF spectra of the BDM events can lead to large background suppression and unique determination of m_{χ}. New cross section constraints on sqrt[σ_{χe}σ_{χν}] are derived from SN1987a in the Large Magellanic Cloud with data from the Kamiokande and Super-Kamiokande experiments. Potential sensitivities for the next galactic SN with Hyper-Kamiokande are projected. This analysis extends the existing bounds on sqrt[σ_{χe}σ_{χν}] over a broad range of r_{χ}=σ_{χν}/σ_{χe}. In particular, the improvement is by 1-3 orders of magnitude for m_{χ}

Comparison of two methods for extracting exosomes from the nucleus accumbens in mice.

BACKGROUND: Exosomes bind to and are endocytosed by neurons of various brain regions. Methods for isolating and extracting exosomes from specific brain samples are critical. At present, the most important extractive methods for exosomes are Ultracentrifugation and exosome isolation kit extraction. Both of these extraction methods have applications in neuroscience. We compare these methods to reveal the differences. METHODS: We sectioned the nucleus accumbens of mice, and isolated exosomes. A culture medium containing exosomes was extracted using ultracentrifugation (UC) and a total exosome isolation kit (TEI). The exosomes were examined using transmission electron microscopy (TEM), measurement regarding the diameter of the exosomes was done, and the thermal allodynia and western blotting analysis were also conducted, respectively. RESULTS: Transmission electron microscopy observations showed that the ultracentrifugation samples had higher purity and fewer impurities than the kit samples. The results from the two methods were then compared with a number ratio regarding the percentage was not statistically significant. Marker protein tests showed that proteins were expressed under both methods. The thermal allodynia testing observed that the two extraction methods did not affect pain behavior regarding the detection. After the kit extraction method, there were substantial white subjects suspended by PBS. CONCLUSION: Our study compared the different protocols regarding exosome extraction from the nucleus accumbens and compared the quality of two principal methods for exosome extraction from a culture medium containing exosomes. It was found that the extraction quality of exosomes by ultracentrifugation was better, but the technical difficulty was greater.

Identification and characterization of a novel hydroxylamine oxidase, DnfA, that catalyzes the oxidation of hydroxylamine to N2.

Nitrogen (N2) gas in the atmosphere is partially replenished by microbial denitrification of ammonia. Recent study has shown that Alcaligenes ammonioxydans oxidizes ammonia to dinitrogen via a process featuring the intermediate hydroxylamine, termed "Dirammox" (direct ammonia oxidation). However, the unique biochemistry of this process remains unknown. Here, we report an enzyme involved in Dirammox that catalyzes the conversion of hydroxylamine to N2. We tested previously annotated proteins involved in redox reactions, DnfA, DnfB, and DnfC, to determine their ability to catalyze the oxidation of ammonia or hydroxylamine. Our results showed that none of these proteins bound to ammonia or catalyzed its oxidation; however, we did find DnfA bound to hydroxylamine. Further experiments demonstrated that, in the presence of NADH and FAD, DnfA catalyzed the conversion of 15N-labeled hydroxylamine to 15N2. This conversion did not happen under oxygen (O2)-free conditions. Thus, we concluded that DnfA encodes a hydroxylamine oxidase. We demonstrate that DnfA is not homologous to any known hydroxylamine oxidoreductases and contains a diiron center, which was shown to be involved in catalysis via electron paramagnetic resonance experiments. Furthermore, enzyme kinetics of DnfA were assayed, revealing a Km of 92.9 ± 3.0 µM for hydroxylamine and a kcat of 0.028 ± 0.001 s-1. Finally, we show that DnfA was localized in the cytoplasm and periplasm as well as in tubular membrane invaginations in HO-1 cells. To the best of our knowledge, we conclude that DnfA is the first enzyme discovered that catalyzes oxidation of hydroxylamine to N2.

Dirammox Is Widely Distributed and Dependently Evolved in Alcaligenes and Is Important to Nitrogen Cycle.

Nitrogen cycle is an essential process for environmental health. Dirammox (direct ammonia oxidation), encoded by the dnfT1RT2ABCD cluster, was a novel pathway for microbial N2 production defined in Alcaligenes ammonioxydans HO-1. Here, a copy of the cluster dnfT1RT2ABCD as a whole was proved to have existed and very conserved in all Alcaligenes genomes. Phylogenetic analyses based on 16S rRNA gene sequences and amino acid sequences of DnfAs, together with G + C content data, revealed that dnf cluster was evolved associated with the members of the genus Alcaligenes. Under 20% O2 conditions, 14 of 16 Alcaligenes strains showed Dirammox activity, which seemed likely taxon-related. However, the in vitro activities of DnfAs catalyzing the direct oxidation of hydroxylamine to N2 were not taxon-related but depended on the contents of Fe and Mn ions. The results indicated that DnfA is necessary but not sufficient for Dirammox activity. The fact that members of the genus Alcaligenes are widely distributed in various environments, including soil, water bodies (both freshwater and seawater), sediments, activated sludge, and animal-plant-associated environments, strongly suggests that Dirammox is important to the nitrogen cycle. In addition, Alcaligenes species are also commonly found in wastewater treatment plants, suggesting that they might be valuable resources for wastewater treatment.

Novel Alcaligenes ammonioxydans sp. nov. from wastewater treatment sludge oxidizes ammonia to N2 with a previously unknown pathway.

Heterotrophic nitrifiers are able to oxidize and remove ammonia from nitrogen-rich wastewaters but the genetic elements of heterotrophic ammonia oxidation are poorly understood. Here, we isolated and identified a novel heterotrophic nitrifier, Alcaligenes ammonioxydans sp. nov. strain HO-1, oxidizing ammonia to hydroxylamine and ending in the production of N2 gas. Genome analysis revealed that strain HO-1 encoded a complete denitrification pathway but lacks any genes coding for homologous to known ammonia monooxygenases or hydroxylamine oxidoreductases. Our results demonstrated strain HO-1 denitrified nitrite (not nitrate) to N2 and N2 O at anaerobic and aerobic conditions respectively. Further experiments demonstrated that inhibition of aerobic denitrification did not stop ammonia oxidation and N2 production. A gene cluster (dnfT1RT2ABCD) was cloned from strain HO-1 and enabled E. coli accumulated hydroxylamine. Sub-cloning showed that genetic cluster dnfAB or dnfABC already enabled E. coli cells to produce hydroxylamine and further to 15 N2 from (15 NH4 )2 SO4 . Transcriptome analysis revealed these three genes dnfA, dnfB and dnfC were significantly upregulated in response to ammonia stimulation. Taken together, we concluded that strain HO-1 has a novel dnf genetic cluster for ammonia oxidation and this dnf genetic cluster encoded a previously unknown pathway of direct ammonia oxidation (Dirammox) to N2 .

Fingerprints of Heavy-Element Nucleosynthesis in the Late-Time Lightcurves of Kilonovae.

The kilonova emission observed following the binary neutron star merger event GW170817 provided the first direct evidence for the synthesis of heavy nuclei through the rapid neutron capture process (r process). The late-time transition in the spectral energy distribution to near-infrared wavelengths was interpreted as indicating the production of lanthanide nuclei, with atomic mass number A≳140. However, compelling evidence for the presence of even heavier third-peak (A≈195) r-process elements (e.g., gold, platinum) or translead nuclei remains elusive. At early times (∼days) most of the r-process heating arises from a large statistical ensemble of ß decays, which thermalize efficiently while the ejecta is still dense, generating a heating rate that is reasonably approximated by a single power law. However, at later times of weeks to months, the decay energy input can also possibly be dominated by a discrete number of α decays, ^{223}Ra (half-life t_{1/2}=11.43 d), ^{225}Ac (t_{1/2}=10.0 d, following the ß decay of ^{225}Ra with t_{1/2}=14.9 d), and the fissioning isotope ^{254}Cf (t_{1/2}=60.5 d), which liberate more energy per decay and thermalize with greater efficiency than ß-decay products. Late-time nebular observations of kilonovae which constrain the radioactive power provide the potential to identify signatures of these individual isotopes, thus confirming the production of heavy nuclei. In order to constrain the bolometric light to the required accuracy, multiepoch and wideband observations are required with sensitive instruments like the James Webb Space Telescope. In addition, by comparing the nuclear heating rate obtained with an abundance distribution that follows the solar r abundance pattern, to the bolometric lightcurve of AT2017gfo, we find that the yet-uncertain r abundance of ^{72}Ge plays a decisive role in powering the lightcurve, if one assumes that GW170817 has produced a full range of the solar r abundances down to mass number A∼70.

Photobacterium sp. NNA4, an efficient hydroxylamine-transforming heterotrophic nitrifier/aerobic denitrifier.

An efficient heterotrophic nitrifying/aerobic denitrifying strain, Photobacterium sp. NNA4 was isolated from a recirculating aquaculture system (RAS). NNA4 was capable of utilizing ammonia, nitrate or nitrite as sole N-source with maximal removal rates of 12.5 mg/L/h for NH4+N, 16.4 mg/L/h for NO3--N, and 4.5 mg/L/h for NO2--N, respectively. Optimal nitrification conditions were: sodium succinate as C-source, 30-37°C, NaCl 1-4%, pH 7.0-8.0, dissolved oxygen 5.89 mg/L, C/N > 10. Gas chromatography/mass spectrometry and gas chromatography/isotope ratio mass spectrometry analyses showed that N2 and N2O were aerobic denitrification products of nitrite and nitrate. NNA4 could tolerate high concentration of hydroxylamine and displayed efficient hydroxylamine-transforming capability. Hydroxylamine oxidoreductase activity using potassium ferricyanide as electron acceptor was 0.042 U. Results revealed that strain NNA4 could oxidize NH2OH directly to N2O at aerobic conditions. In view of its high removal ability of inorganic nitrogen pollutants and broad salinity tolerance range, NNA4 has great potential in denitrification treatment of types of wastewater with either low salinity (e.g., municipal facilities) or high salinity (e.g., aquaculture, seafood processing).

Near-infrared photonic band structure in a semiconductor metamaterial photonic crystal.

In this study, we theoretically investigate the near-infrared (NIR) photonic band structure (PBS) in a one-dimensional semiconductor metamaterial (MM) photonic crystal (PC). The considered PC is (AB)N, where N is the stack number, A is a dielectric, and B is a semiconductor MM composed of Al-doped ZnO and ZnO. It is found that the photonic band gaps (PBGs) can be tunable by the variations in filling factor, and thicknesses of A and B. It is of particular interest to see that the PBG will vanish when the thicknesses of A and B satisfy a certain condition. The results provide fundamental information on a NIR PBS that could be of technical use in photonic applications using such a semiconductor MM. The band gap vanishing makes it possible to design a wider band pass filter at NIR based on the use of such a PC.