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Biocatalysis has become the main approach to produce cytidine diphosphate choline (CDP-choline), which has been applied for treatment of acute craniocerebral injury and consciousness after brain surgery. However, salt accumulates with the production and inhibits enzyme activity, and eventually reduces yield and product accumulation rate. Our work provided a possible solution to this problem by applying a computational designed halophilic choline kinase. The halotolerant CKI (choline kinase) was designed following a unique strategy considering the most variable residue positions on the protein surface among target enzymes from different sources. The basic and neutral surface residues were replaced with acidic ones. This approach was enlightened by features of natural halophilic enzymes. Mutants in the work represented higher catalytic activities and IC50 (inhibit activity by 50%) at high salt concentrations (over 1200 mM). Furthermore, when the mutant was used in fed-batch production, the CDP-choline accumulation rate doubled comparing with process using wild-type CKI at acetate concentration of over 700 mM. The maximum titer was 151 ± 3.2 mM, the productivity was 5.8 ± 0.1 mM·L−1 h−1, and molar yield to CMP and utilization efficiency of energy were 85.3 and 63.5%. The idea of computational design in our work can also be applied to modify other enzymes in industry, and sheds light on alleviating effect of salt accumulation during industrial manufacturing process.

AP25 is an anti-tumor peptide with a high affinity for integrins. It exerts its anti-tumor activity by inhibiting angiogenesis and by directly inhibiting the growth of tumor cells. Its half-life time in vivo is only about 50 minutes, which limits its clinical application. In order to prolong the half-life time of AP25 while preserving its anti-tumor activity, several fusion proteins of AP25 and IgG4 Fc were designed and expressed; their anti-tumor activity and pharmacokinetics properties were evaluated. Firstly, four AP25-Fc fusion protein sequences were designed, and the corresponding proteins were expressed and purified. Based on the results of HUVEC migration inhibition assay, HUVEC and tumor cell proliferation inhibition assay and yields of expression by HEK293 cells, the fusion protein designated PSG4R was selected for further evaluation. The anti-tumor effect of PSG4R was then evaluated in vivo on HCT-116 nude mice xenograft model. And the pharmacokinetics properties of PSG4R were investigated in rats. The results showed that PSG4R could inhibit the growth of xenografts of human colon cancer cell line HCT-116 in nude mice by intravenous administration of 40 mg/kg once every two days. The half-life time of PSG4R was 56.270 ± 15.398 h. This study showed that the construction of AP25-Fc fusion protein could significantly prolong the half-life of AP25 while retaining its anti-tumor activity, which provides a new direction for new drug development of AP25.

We studied Phellinus lonicerinus to determine the cytotoxic effect and the dual estrogenic activities of methyl-hispolon and their relation to estrogen signals in vivo and in vitro. The Glide scores of methyl-hispolon-estrogen receptor α (ERα) and methyl-hispolon-ERβ docked complexes were -7.29 kcal/mol and -6.68 kcal/mol in docking simulations. Methyl-hispolon had a significant antiproliferative effect for estrogen-sensitive ER(+) MCF-7 cells in the absence of estrogen, and it exhibited dual estrogen activities. Methyl-hispolon increased the serum E2 in rats with premature ovarian failure and fulfilled the estrogenic function in the uterus and ovary. Methyl-hispolon significantly inhibited the expression of Ras, API, ERα, C-myc, and cyclinDl, as well as their gene transcription in RL95-2 cells. The phosphorylation of ERK1/2 was inhibited by methyl-hispolon. Thus, methyl-hispolon has potential use in treating estrogen deficiency-related diseases, with good antitumor effects and estrogenic activity.

Long-term and excessive herbicide use has led to some environmental concerns and especially, herbicide resistance evolution in weeds. Here, we confirmed acetolactate synthase (ALS) inhibiting herbicide penoxsulam resistance and cross resistance to acetyl-coenzyme carboxylase (ACCase) inhibiting herbicides (cyhalofop-butyl and metamifop) in a global weed Echinochloa crus-galli population resistant to these herbicides (R). Penoxsulam metabolism study indicated that degradation rate was significantly higher in R than susceptible E. crus-galli population (S). RNA-sequencing revealed that a cytochrome P450 (P450) gene, CYP81A68, expressed higher in R versus S. Rice seedlings overexpressing this CYP81A68 gene are resistant to penoxsulam, cyhalofop-butyl and metamifop, and penoxsulam resistance is due to enhanced metabolism via O-demethylation. Deletion analysis of the CYP81A68 gene promoter identified an efficient region, in which differential methylation of CpG islands occurred between R and S. Collectively, these results demonstrate that upregulation of E. crus-galli CYP81A68 gene endows generalist metabolic resistance to commonly used ALS- and ACCase-inhibiting herbicides in rice fields and epigenetic regulation may play a role in the resistance evolution. This research could contribute to strategies reducing herbicide environmental impacts by judicious selection of alternative herbicide and non-chemical control tactics.

The aim of this study was to elucidate the gastroprotective activity and possible mechanism of involvement of araloside A (ARA) against ethanol- and aspirin-induced gastric ulcer in mice. The experimental mice were randomly divided into control, model, omeprazole (20 mg/kg, orally) and ARA (10, 20 and 40 mg/kg, orally). Gastric ulcer in mice was induced by intragastric administration of 80% ethanol (10 mL/kg) containing 15 mg/mL aspirin 4 h after drug administration on day 7. The results indicated that ARA could significantly raise gastric juice volume and acidity; ameliorate gastric mucosal blood flow, gastric binding mucus volume, ulcer index and ulcer inhibition rate; suppress H+/K+-ATPase activity, which was confirmed by computer-aided docking simulations; inhibit the release of mitochondrial cytochrome c into the cytoplasm; inhibit caspase-9 and caspase-3 activities and down-regulate mRNA expression levels; down-regulate the mRNA and protein expressions of apoptosis protease-activating factor-1 and protein expression of cleaved poly(ADP ribose) polymerase-1; and up-regulate Bcl-2 mRNA and protein expressions and down-regulate Bax mRNA and protein expressions, thus elevating the Bcl-2/Bax ratio in a dose-dependent manner. Histopathological observations further provided supportive evidence for the aforementioned results. The results demonstrated that ARA exerted beneficial gastroprotective effects on alcohol- and aspirin-induced gastric ulcer in mice, which was related to suppressing H+/K+-ATPase activity as well as pro-apoptotic protein expression, and promoting anti-apoptotic protein expression, thus alleviating gastric mucosal injury and cell death.

Accurate determination of biological activity is essential in quality control of recombinant human brain natriuretic peptide (rhBNP). In previous study, we successfully developed a genetically modified cell line 293GCAC3-based ELISA assay for rhBNP. But ELISA procedure is still tedious, so this study was aimed to develop a rapid and simple bioassay for rhBNP using GloSensor technology, which provides a platform of flexible luciferase-based biosensors for real-time detection of signaling events in live cells, including cGMP production. A reporter cell line 293GCAGlo-G1 was constructed by transfecting pGloSensor™ 40 F plasmid into 293GCAC3. The reporter assay based on 293GCAGlo-G1 showed high precision with intra-assay CV being 8.3% and inter-assay CV being 14.1%; high accuracy with 80%, 100% and 120% recovery rate being 99.2%, 102.4% and 99.0% respectively; and great linearity with R2 of linear fitting equation being 0.99. Besides, no significant difference was found in test results of reporter assay and 293GCAC3-based ELISA assay (paired t test, p = 0.630). All these results suggested that the reporter assay was a viable assay for biological determination of rhBNP. Keywords: RhBNP, cGMP, GloSensor technology, Reporter assay

Calcitonin gene-related peptide (CGRP) is critical for the pathophysiology of migraine, and four therapeutic antibodies targeting CGRP and its corresponding receptors have been approved by the Food and Drug Administration (FDA), while many others are in the different stages of clinical trials. Bioactivity determination is essential for the quality control and clinical application of therapeutic monoclonal antibodies (mAbs). However, no bioassay has been reported to date. In this study, we developed a reporter gene assay (RGA) based on SK-N-MC cells stably expressing firefly luciferase driven by cAMP response element (CRE). The key assay parameters were optimized according to signal-to-noise (SNR), the response value, and the fitted dose-response curve. Validation of the RGA in accordance with ICH-Q2 guidelines showed that the method had good specificity, accuracy, linearity, and precision. The established RGA can be utilized as a reference method for release testing and stability studies of relevant antibodies.

Measurements of first and second harmonic radiation from nonlinear Thom- son scattered light as a function of laser polarization ellipticity are reported. Polarization-resolved patterns were observed and connected to the underlying electron motion, confirming half-century old theoretical predictions.

Background Exploring herbicide resistance mechanisms in weeds is regarded as an important part of designing resistance management strategies and rationalizing the use of herbicides. Beckmannia syzigachne is one of the most important agricultural weeds in China. Long-term use of acetyl-CoA carboxylase (ACCase)-inhibiting herbicides has led to evolution of herbicide resistance in B. syzigachne. ACCase-inhibiting herbicides consist of three chemical families, including the aryloxyphenoxypropionates (APPs), cyclohexanediones (CHDs) and phenylpyraxoline (DENs). Results Based on whole-plant dose-response experiments, a B. syzigachne population (BS-R) was confirmed to be 20- and 12-fold resistant to the APP herbicides clodinafop-propargyl and quizalofop-P-ethyl; 2.2-, 2.8-, and 2.8-fold resistant to fenoxaprop-P-ethyl, the CHD herbicide sethoxydim and PPZ herbicide pinoxaden, respectively, compared to the susceptible counterpart (BS-S). Resistance to clodinafop-propargyl in the BS-R population could not be reversed by the known cytochrome P450 (CYP450) inhibitor malathion and glutathione S-transferase (GST) inhibitor 4-chloro-7-nitrobenzoxadiazole. In addition, no difference in CYP450 and GST activity was confirmed in BS-R and BS-S population. ACCase gene sequencing revealed an Ile-2041-Val mutation in the BS-R population. A derived cleaved amplified polymorphic sequence (dCAPS) marker was developed for rapid detection of the specific Ile-2041-Val mutation. Correlation quantification of resistance in homo- and hetro-resistant versus WT plants showed that the resistance to clodinafop-propargyl in this population is conferred by the Ile-2041-Val mutation. Conclusion Unlike the previous reports on the unique cross-resistance pattern by the 2041 mutation, this study demonstrates that the BS-R population confers resistance to certain ACCase-inhibiting APP, CHD and PPZ herbicides in B. syzigachne. This article is protected by copyright. All rights reserved.

DR21 south filament (DR21SF) is a unique component of the giant network of filamentary molecular clouds in the north region of Cygnus X complex. Unlike the highly fragmented and star-forming active environment it resides, DR21SF exhibits a coherent profile in the column density map with very few star formation signposts, even though the previously reported linear density of the filament is an order of magnitude higher than the thermal stable threshold. We derive the size (3.6~pc by 0.13~pc), temperature (10 to 15~K), and mass (1048~\textit{M$_\odot$}) of DR21SF from Shanghai 65 m TianMa Radio Telescope (TMRT) observations of NH$_3$ (1, 1) and (2, 2) inversion lines in conjunction with the column density map from our previous work. Star-forming sites are identified along the filament where gas temperature excesses. We find clear gradients in radial velocity and intrinsic line-width along the spine of the filament. The gradients can be well interpreted with a scenario of an accretion flow feeding DR 21 at a mass transfer rate of $1.1 \times 10^{-3}$~\textit{M$_\odot$} yr$^{-1}$. Based on the analysis of its kinematic temperature, intrinsic line-width and mass distribution, we conclude that DR21SF is in an overall trans-critical status, which indicates an early evolutionary stage., 15 pages, 4 figures, to be published in Astrophysical Journal

The tumor necrosis factor alpha (TNF-α)/nuclear factor-kappa B (NF-κB) signaling pathway plays a crucial role in the pathogenesis of inflammatory diseases. Several therapeutic monoclonal antibodies (mAbs) and biosimilars against TNF-α have been developed to treat patients who suffer from inflammatory diseases caused by disordered expression of TNF-α. Hence, quality control of biopharmaceuticals is crucial during research and development. The high-order structure of these complex molecules cannot be entirely identified by physiochemical attributes; however, they can be inferred by observing biological activities. Thus, we developed a U937-based bioassay to determine the biological activities of mAbs and biosimilars against TNF-α using a low-basal NF-κB-inducible lentiviral reporter gene. The reporter gene assay (RGA) can be induced with a high signal-to-noise ratio (SNR) in a short time by TNF-α. Validation of the RGA showed accuracy (% relative standard deviation [RSD] = 4.64%), linearity (r2 = 0.9856), and precision (Interday RSD = 4.6%, between analysts RSD = 3.51%) as well as reasonable specificity and robustness. The measured potency values of a biosimilar to adalimumab were between 90% and 110%. Results showed our RGA is suitable for mAb quality control and lot release, and for evaluation of the biological activity similarity of the biosimilar.

OX40 plays a prominent role in the onset and development of solid tumors, and OX40-targeted monoclonal antibodies (mAbs) have entered clinical trials for various tumors. Bioactivity determination of therapeutic mAbs is of great significance in product quality, however, mechanism of action-based bioassays to determine the bioactivity of anti-OX40 mAbs is still lacking. Here, we established a reporter gene assay system based on two cell lines, namely Jurkat-OX40-NFκB-Luc which stably expresses NFκB-controlled luciferase, and Raji cells which inherently express FcγRs. In the model, FcγRs on Raji cells could crosslink the Fc of anti-OX40 mAbs, which leads to the further crosslinking between Fab of anti-OX40 mAbs and OX40 on Jurkat-OX40-NFκB-Luc cells. OX40 crosslinking could activate Jurkat-OX40-NFκB-Luc cells, and induce the expression of NFκB-controlled luciferase, the extent of which could reflect the bioactivity of anti-OX40 mAbs in a dose-dependent manner. After the optimization of various assay conditions, the validation of the cell-based bioassay showed good assay performance characteristics, including specificity, accuracy, precision, linearity, and stability. This innovative assay that is based on the OX40-NFκB pathway can be a powerful pool to measure the bioactivity of OX40-targeted mAbs.

We present a study of outflows at extremely early stages of high-mass star formation obtained from the ALMA Survey of 70 $\mu \rm m$ dark High-mass clumps in Early Stages (ASHES). Twelve massive 3.6$-$70 $\mu \rm m$ dark prestellar clump candidates were observed with the Atacama Large Millimeter/submillimeter Array (ALMA) in Band 6. Forty-three outflows are identified toward 41 out of 301 dense cores using the CO and SiO emission lines, yielding a detection rate of 14%. We discover 6 episodic molecular outflows associated with low- to high-mass cores, indicating that episodic outflows (and therefore episodic accretion) begin at extremely early stages of protostellar evolution for a range of core masses. The time span between consecutive ejection events is much smaller than those found in more evolved stages, which indicates that the ejection episodicity timescale is likely not constant over time. The estimated outflow dynamical timescale appears to increase with core masses, which likely indicates that more massive cores have longer accretion timescales than less massive cores. The lower accretion rates in these 70 $\mu \rm m$ dark objects compared to the more evolved protostars indicate that the accretion rates increase with time. The total outflow energy rate is smaller than the turbulent energy dissipation rate, which suggests that outflow induced turbulence cannot sustain the internal clump turbulence at the current epoch. We often detect thermal SiO emission within these 70 $\mu \rm m$ dark clumps that is unrelated to CO outflows. This SiO emission could be produced by collisions, intersection flows, undetected protostars, or other motions., Comment: 32 pages, 9 figures, 4 tables, accepted for publication in ApJ

Fermentation and chemical methods for industrial cytidine diphosphate choline (CDP-choline) catalytic production both suffer from several disadvantages such as relatively low efficiency and productivity. To overcome these problems, we applied the concept of synthetic biology to develop a new one-pot multi-enzyme system to produce CDP-choline from orotic acid. Enzymes from different sources were selected and optimized as building blocks of the system, and parameters such as oxygen supply were also optimized. This system shows a titer of 37.6 ± 1.1 mM and a reaction rate of 1.6 mM L

Objective This work designs an in vitro multi-enzyme system to produce CDP-choline form D-ribose, and develop an optimization procedure for one-pot multi-enzyme catalytic system. Results The entire process integrated ten enzymes, and an efficient acetate kinase/acetyl phosphate-based ATP regeneration module was applied. Then some optimizations to this system were made including selecting optimum enzyme building blocks and improving expression parameters. The process improved the final yield of CDP-choline form 0.2g/L to 6g/L (CDP-choline titer 12.2 mM). Conclusions This new one pot CDP-choline producing system has a potential for industrial use, and the optimization procedure shed light on improving other one pot multi-enzyme system for industrial production of energy rich compounds. This article is protected by copyright. All rights reserved.

Middle East Respiratory Syndrome coronavirus (MERS-CoV) is a highly virulent pathogen that causes Middle East Respiratory Syndrome (MERS). Anti-MERS-CoV antibodies play an integral role in the prevention and treatment against MERS-CoV infections. Bioactivity is a key quality attribute of therapeutic antibodies, and high accuracy and precision are required. The major methods for evaluating the antiviral effect of antiviral antibodies include neutralization assays using live viruses or pseudoviruses are highly variable. Recent studies have demonstrated that the antibody-dependent cellular cytotoxicity (ADCC) activity of antiviral antibodies is more consistent with the virus clearance effect in vivo than neutralization activity. However, no reports evaluating the ADCC activity of anti-MERS antibodies have been published to date. Here, we describe the development of a robust and reliable cell-based reporter gene assay for the determination of ADCC activity of anti-MERS antibodies using 293T/MERS cells stably expressing the spike protein of MERS-CoV (MERS-S) as target cells and the engineered Jurkat/NFAT-luc/FcγRIIIa stably expressing FcγRIIIA and NFAT reporter gene as effector cells. According to the ICH-Q2 analytical method guidelines, we carefully optimized the experimental conditions and assessed the performance of our assay. In addition, we found that the ADCC activity of afucosylated anti-MERS antibodies is higher than their fucosylated counterparts. The establishment of this ADCC determination system provides a novel method for evaluating the bioactivity of anti-MERS antibodies and improving ADCC activity through modification of N-glycosylation of the Fc segment.

Using the Five-hundred-meter Aperture Spherical radio Telescope (FAST), we search for H I narrow-line self-absorption (HINSA) features in twelve Planck Galactic cold clumps (PGCCs), the starless core L1521B, and four star forming sources. Eight of the 12 PGCCs have detected emission of J = 2–1 of cyanoacetylene (HC3N). With an improved HINSA extraction method more robust for weaker and blended features with high velocity resolution, the detection rates of HINSA in PGCCs are high, at 92% overall (11/12) and 87% (7/8) among sources with HC3N J = 2–1 emissions. Combining the data of molecular spectra and Planck continuum maps, we studied the morphologies, dynamics, abundances and excitations of H I, CO and HC3N in PGCCs. The spatial distribution of HINSA is similar to that of CO, implying that HINSA features are confined to regions within and around CO emission kernels. HINSA tends to be not detected in regions associated with warm dust and background ionizing radiation, as well as regions associated with stellar objects. The L-band continnum and average background H I emission may be non-ignorable for the excitation of HINSA. The abundances of cold H I in PGCCs are approximately 3 × 10−4, and vary within a factor of ~3. The non-thermal velocity dispersions traced by C18O J = 1–0 and HINSA are consistent with each other (0.1–0.4 km s−1), larger than the typical value of HC3N (~0.1 km s−1). Carbon chain molecule (CCM) abundant PGCCs provide a good sample to study HINSA.

Type 2 inflammatory cytokines, including IL-4, IL-5 and IL-13, contribute considerably to the pathogenesis of asthma. Anti-IL-4R monoclonal antibody (mAb) has been approved for the therapeutic treatment of asthma, and many mAbs with the same target are in the different stages of R&D and clinical trials. Bioactivity determination is required to ensure the quality control of mAbs. However, current ELISA and SPR assays or cell-based anti-proliferation assays for IL-4R mAbs are either not mechanism-of-action (MOA) representative or tedious and time consuming. Therefore, we developed a reporter gene assay (RGA) based on the HEK-293 cell line that stably expressed signal transducer and activator of transcription 6 (STAT6) and the luciferase reporter controlled by STAT6 binding elements. Anti-4R mAb could bind to IL-4R, and block the interaction between IL-4 and IL-4R, resulting in the reduction of IL-4 induced STAT6 controlled luciferase expression. After careful optimization of the experiment parameters, the RGA method demonstrated optimal dose-response curve between anti-IL-4R mAb concentration and luciferase expression level. Validation according ICH-Q2 proved the excellent assay performance characteristics of the established RGA, including specificity, accuracy, precision, linearity and range. The established transgenic cell line was stable for the bioactivity determination of anti-IL-4R mAb up to 46 generations, and the RGA was also suitable for the bioactivity determination of anti-IL-4 mAbs, and potentially of anti-IL-13 mAbs. The established RGA could be adopted to determine the bioactivity during the development, characterization, lot release, stability, and comparability studies of anti-IL-4R mAbs.

We report studies of the relationships between the total bolometric luminosity ($L_{\rm bol}$ or $L_{\rm TIR}$) and the molecular line luminosities of $J=1-0$ transitions of H$^{13}$CN, H$^{13}$CO$^+$, HCN, and HCO$^+$ with data obtained from ACA observations in the "ATOMS" survey of 146 active Galactic star forming regions. The correlations between $L_{\rm bol}$ and molecular line luminosities $L'_{\rm mol}$ of the four transitions all appear to be approximately linear. Line emission of isotopologues shows as large scatters in $L_{\rm bol}$-$L'_{\rm mol}$ relations as their main line emission. The log($L_{\rm bol}$/$L'_{\rm mol}$) for different molecular line tracers have similar distributions. The $L_{\rm bol}$-to-$L'_{\rm mol}$ ratios do not change with galactocentric distances ($R_{\rm GC}$) and clump masses ($M_{\rm clump}$). The molecular line luminosity ratios (HCN-to-HCO$^+$, H$^{13}$CN-to-H$^{13}$CO$^+$, HCN-to-H$^{13}$CN and HCO$^+$-to-H$^{13}$CO$^+$) all appear constant against $L_{\rm bol}$, dust temperature ($T_{\rm d}$), $M_{\rm clump}$ and $R_{\rm GC}$. Our studies suggest that both the main lines and isotopologue lines are good tracers of the total masses of dense gas in Galactic molecular clumps. The large optical depths of main lines do not affect the interpretation of the slopes in star formation relations. We find that the mean star formation efficiency (SFE) of massive Galactic clumps in the "ATOMS" survey is reasonably consistent with other measures of the SFE for dense gas, even those using very different tracers or examining very different spatial scales., Comment: Published on MNRAS. The full tables are included in Tables.pdf or Tables.tex files, which can be downloaded from source files

In most galaxies like the Milky Way, stars form in clouds of molecular gas. Unlike the CO emission that traces the bulk of molecular gas, the rotational transitions of HCN and CS molecules mainly probe the dense phase of molecular gas, which has a tight and almost linear relation with the far-infrared luminosity and star formation rate. However, it is unclear if dense molecular gas exists at very low metallicity, and if exists, how it is related to star formation. In this work, we report ALMA observations of the CS $J$=5$\rightarrow$4 emission line of DDO~70, a nearby gas-rich dwarf galaxy with $\sim7\%$ solar metallicity. We did not detect CS emission from all regions with strong CO emission. After stacking all CS spectra from CO-bright clumps, we find no more than a marginal detection of CS $J$=5$\rightarrow$4 transition, at a signal-to-noise ratio of $\sim 3.3$. This 3-$\sigma$ upper limit deviates from the $L^\prime_{\rm CS}$-$L_{\rm IR}$ and $L^\prime_{\rm CS}$-SFR relationships found in local star forming galaxies and dense clumps in the Milky Way, implying weaker CS emission at given IR luminosity and SFR. We discuss the possible mechanisms that suppress CS emission at low metallicity., Comment: 5 pages, 4 figures. Accepted for publication in MNRAS Letters

We present observations of HCN 3-2 emissions towards 37 local galaxies using 10-m Submillimeter Telescope (SMT). HCN 3-2 emission is detected in 23 galaxies. The correlation of infrared luminosity (LIR) and the luminosity of HCN 3-2 line emission measured in our sample is fitted with a slope of 1.11 and correlation coefficient of 0.91, which follows the linear correlation found in other dense gas tracers in the literatures. Although molecular gas above a certain volume density threshold (i.e., $n_{\rm H_2}\geq$ 10$^4$ cm$^{-3}$) statistically gave similar relation with infrared luminosity, the large scatter of HCN 3-2/HCN1-0 ratios for galaxies with different LIR indicates that dense gas masses estimated from the line luminosities of only one transition of dense gas tracers should be treated with caution for individual galaxies., 15 pages, 5 figures, accepted for publication in PASJ

BACKGROUND Asia minor bluegrass (Polypogon fugax) is one of the main weeds invading Chinese canola fields. The P. fugax resistant population SC-R, which survived quizalofop-p-ethyl at the field-recommended rate (67.5 g ha-1 ), was collected from a canola field in Qingsheng County in China. The present study aimed to (1) characterize the SC-R resistance pattern to acetyl-CoA carboxylase (ACCase)-inhibiting herbicides, and (2) investigate the mechanism of quizalofop-p-ethyl resistance in this population. RESULTS Dose-response studies showed that resistance to quizalofop-p-ethyl and haloxyfop occurred in the SC-R population. Four transcripts/genes encoding the plastidic ACCase carboxyl-transferase domain were isolated from the P. fugax plants. No mutations in the four ACCase genes were detected in the SC-R population compared to the SC-S population. Pre-treatment with the known glutathione S-transferase (GST) inhibitor 4-chloro-7-nitrobenzoxadiazole (NBDCl), reversed resistance to quizalofop-p-ethyl and partially reversed resistance to haloxyfop-R-methyl in the resistant population (SC-R). However, the cytochrome P450 inhibitor malathion did not reverse the resistance. There was no difference in basal GST activity (using CDNB as a substrate), but there was higher inducible GST activity in SC-R relative to SC-S. Two GST genes, GST2c and GSTL3, were constitutively overexpressed in the resistant SC-R population. CONCLUSION This study confirmed that resistance to quizalofop-p-ethyl in the resistant P. fugax population is likely nontarget-site based involving GST, and this resistance mechanism also partially confers haloxyfop-R-methyl resistance. © 2020 Society of Chemical Industry.

Environmental disturbances may result in dysregulation of interleukin-23 (IL-23), which is a crucial modulator of immunity. Several therapeutic monoclonal antibodies (mAbs) have been developed for treating IL-23-related autoimmune inflammation, such as ustekinumab, guselkumab, tildrakizumab, and risankizumab. Accurate bioactivity determination of therapeutic mAbs is essential for their quality control and clinical application. However, the current methods are tedious and complicated. In the present study, we employed low-background lentivirus carrying sis-inducible element (SIE)-driven firefly luciferase to generate a stable DB-SIE-Luc cell line that expresses endogenous IL-23 receptors and developed a sensitive and straightforward reporter gene assay (RGA) based on DB-SIE-Luc cells. After the optimization of various assay parameters, we set up a bioassay with the best fit of a four-parameter model and an appropriate signal-to-noise ratio (SNR) for bioactivity determination of guselkumab. We further verified the excellent assay performance characteristics of our RGA, including specificity, linearity, accuracy, precision, and stability, according to ICH-Q2. Taken together, we established a reliable and robust cell-based RGA, which potentially serves as a valubale alternative bioactivity determination assay for the release control and stability study of anti-IL-23 mAbs.

We report the detection of an emission feature at the 12 sigma level with FWHM line width of about 450 km/s toward the nearest quasi-stellar object, QSO Mrk 231. Based on observations with the IRAM 30 m telescope and the NOEMA Interferometer, the 11-10 transition of molecular oxygen is the likely origin of line with rest frequency close to 118.75 GHz. The velocity of the O2 emission in Mrk 231 coincides with the red wing seen in CO emission, suggesting that it is associated with the outflowing molecular gas, located mainly at about ten kpc away from the central AGN. This first detection of extragalactic molecular oxygen provides an ideal tool to study AGN-driven molecular outflows on dynamic time scales of tens of Myr. O2 may be a significant coolant for molecular gas in such regions affected by AGN-driven outflows. New astrochemical models are needed to explain the implied high molecular oxygen abundance in such regions several kpc away from the center of galaxies., 11 pages, 5 figures, published in ApJ

Lyman-α blobs (LABs) are spatially extended nebulae of emission in the Lyman-α (Lyα) line of hydrogen, seen at high redshifts1,2, and most commonly found in the dense environment of star-forming galaxies3,4. A recent study showed that nearly 100% of the sky is covered by Lyα emission around high-redshift galaxies5–7. The origin of Lyα emission in the LABs is still unclear and under debate8. It may be powered by photoionization involving galactic superwinds/outflows, resonant scattering of Lyα photons from starbursts or active galactic nuclei9–16, or by cooling radiation from cold streams of gas accreting onto galaxies17,18, as demonstrated by recent simulations19. Here we analyse the gas kinematics within a LAB, providing rare observational evidence for infalling gas. This is consistent with the release of gravitational accretion energy as cold streams radiate Lyα photons. It also provides direct evidence for possible cold streams feeding the central galaxies. The mass of the infalling gas is not important in comparison to the gas mass consumed by star formation, and is also not the major powering source of Lyα emission, but it hints at another mechanism to explain the origin of the extended Lyα emission around young galaxies. The gas kinematics within a Lyman-α blob show clear evidence of infall, consistent with the release of gravitational accretion energy as cold streams radiate Lyman-α photons. These cold streams are apparently responsible for feeding central galaxies.

In this study, the estrogenic effect of GL-1, a component of the Ganoderma lucidum, was studied, and the possible mechanism was discussed preliminarily. The binding ability of GL-1 to estrogen receptor was calculated by computer aided simulation. The effects of GL-1 on the proliferation of estrogen sensitive estrogen receptor (ER) (+) MCF-7 cells and estrogen insensitive ER (−) MDA-MB-231 cells were detected by MTT method. The effects of GL-1 on the proliferation of estrogen-induced MCF-7 cells, and the effects of the estrogen receptor inhibitor ICI182780 on the proliferation of GL-1-induced MCF-7 cells were detected by MTT assay. The expression of ERα and ERβ monoclonal antibody were detected by Western blot. The results showed that GL-1 has a good binding ability to estrogen receptor β, and has estrogen-like effect, which might be related to secretion of estrogen and expression of ERβ by binding to ERs.

In order to search for shocks in the very early stage of star formation, we performed single-point surveys of SiO J=1-0, 2-1 and 3-2 lines and the H$_2$CO $2_{12}-1_{11}$ line toward a sample of 100 high-mass starless clump candidates (SCCs) by using the Korean VLBI Network (KVN) 21-m radio telescopes. The detection rates of the SiO J=1-0, 2-1, 3-2 lines and the H$_2$CO line are $31.0\%$, $31.0\%$, $19.5\%$ and $93.0\%$, respectively. Shocks seem to be common in this stage of massive star formation. The widths of the observed SiO lines (full width at zero power (FWZP)) range from 3.4 to 55.1 km s$^{-1}$. A significant fraction ($\sim29\%$) of the detected SiO spectra have broad line widths (FWZP $>20~km~s^{-1}$), which are very likely associated with fast shocks driven by protostellar outflows. This result suggests that about one third of the SiO-detected SCCs are not really starless but protostellar. On the other hand, about 40$\%$ of the detected SiO spectra show narrow line widths (FWZP, 19 pages, 13 figures

We demonstrate a novel approach to the generation of femtosecond electron bunch trains via laser-driven wakefield acceleration. We use two independent high-intensity laser pulses, a drive, and injector, each creating their own plasma wakes. The interaction of the laser pulses and their wakes results in a periodic injection of free electrons in the drive plasma wake via several mechanisms, including ponderomotive drift, wake-wake interference, and pre-acceleration of electrons directly by strong laser fields. Electron trains were generated with up to 4 quasi-monoenergetic bunches, each separated in time by a plasma period. The time profile of the generated trains is deduced from an analysis of beam loading and confirmed using 2D Particle-in-Cell simulations., Comment: 11 pages, 5 figures, accepted by Physics of Plasmas

The "ATOMS," standing for {\it ALMA Three-millimeter Observations of Massive Star-forming regions}, survey has observed 146 active star forming regions with ALMA Band 3, aiming to systematically investigate the spatial distribution of various dense gas tracers in a large sample of Galactic massive clumps, to study the roles of stellar feedback in star formation, and to characterize filamentary structures inside massive clumps. In this work, the observations, data analysis, and example science of the "ATOMS" survey are presented, using a case study for the G9.62+0.19 complex. Toward this source, some transitions, commonly assumed to trace dense gas, including CS $J = 2-1$, HCO$^+$ $J = 1-0$ and HCN $J = 1-0$, are found to show extended gas emission in low density regions within the clump; less than 25\% of their emission is from dense cores. SO, CH$_3$OH, H$^{13}$CN and HC$_3$N show similar morphologies in their spatial distributions and reveal well the dense cores. Widespread narrow SiO emission is present (over $\sim$1 pc), which may be caused by slow shocks from large--scale colliding flows or H{\sc ii} regions. Stellar feedback from an expanding H{\sc ii} region has greatly reshaped the natal clump, significantly changed the spatial distribution of gas, and may also account for the sequential high-mass star formation in the G9.62+0.19 complex. The ATOMS survey data can be jointly analyzed with other survey data, e.g., "MALT90", "Orion B", "EMPIRE", "ALMA\_IMF", and "ALMAGAL", to deepen our understandings of "dense gas" star formation scaling relations and massive proto-cluster formation., Comment: published on MNRAS

We have introduced a new method of estimating the electron temperature and density of H II regions by using single dish observations. In this method, multiple hydrogen radio recombination lines of different bands are computed under the assumption of low optical depth. We use evolutionary hydrodynamical models of H II regions to model hydrogen recombination line emission from a variety of H II regions and assess the reliability of the method. According to the simulated results, the error of the estimated temperature is commonly < 13%, and that of the estimated density is < 25% for a < 1% uncertainty of the observed line fluxes. A reasonable estimated value of electron density can be achieved if the uncertainty of the line fluxes are lower than 3%. In addition, the estimated values are more representative of the properties in the relatively high-density region if the gas density gradient is present in the H II region. Our method can be independent of the radio continuum observations. But the accuracy will be improved if a line-to-continuum ratio at millimeter wavelengths is added to the estimation. Our method provides a way to measure the temperature and density in ionized regions without interferometers., 17 pages, 12 figures

We present Submillimeter Array (SMA) observations of seven massive molecular clumps which are dark in the far-infrared for wavelengths up to 70 $\mu$m. Our 1.3 mm continuum images reveal 44 dense cores, with gas masses ranging from 1.4 to 77.1 M$_{\odot}$. Twenty-nine dense cores have masses greater than 8 M$_{\odot}$ and the other fifteen dense cores have masses between 1.4 and 7.5 M$_{\odot}$. Assuming the core density follows a power-law in radius $\rho \propto r^{-b}$, the index $b$ is found to be between 0.6 and 2.1 with a mean value of 1.3. The virial analysis reveals that the dense cores are not in virial equilibrium. CO outflow emission was detected toward 6 out of 7 molecular clumps and associated with 17 dense cores. For five of these cores, CO emissions appear to have line-wings at velocities of greater than 30 km s$^{-1}$ with respect to the source systemic velocity, which indicates that most of the clumps harbor protostars and thus are not quiescent in star formation. The estimated outflow timescale increase with core mass, which likely indicates that massive cores have longer accretion timescale than that of the less massive ones. The fragmentation analysis shows that the mass of low-mass and massive cores are roughly consistent with thermal and turbulent Jeans masses, respectively., Comment: 24 pages, 7 figures. Accepted for publication in ApJ

To overcome the lack of availability of fresh human whole blood for pyrogen detection, we explored the feasibility of utilizing cryopreserved pooled human blood to detect the responses of the pro-inflammatory cytokines IL-6 and IL-1β to LPS. Whole blood was obtained from five donors and incubated with LPS. The quantities of pro-inflammatory cytokines were measured using ELISA, and the results were compared among the samples. After the blood was cryopreserved with Dimethyl sulfoxide (DMSO) (10% v/v) and stored for 4 mo at –196℃, the detection limits of the IL-6/IL-1β responses to LPS were 0.2/0.4 endotoxin units (EU)/ml, respectively, and IL-6/IL-1β release increased in response to LPS in a dose-dependent manner. When these experiments were performed in three separate laboratories, the within-laboratory reproducibility of the IL-6/IL-1β responses was 100%/86.7%, 93.3%/100%, and 86.7%/80%, and the inter-laboratory reproducibility was 92.9%/85.7%, 64.3%/63.6%, and 57.1%/66.7%, respectively. The sensitivity (the probability of correctly classifying positive samples) and specificity (the probability of correctly classifying negative samples) of the IL-6/IL-1β tests were 81.7%/82.5% and 100%/100%, respectively. The results of this study suggest that cryopreserved pooled blood is a convenient and viable alternative for evaluating in vitro pyrogenicity. Additionally, maintaining cryopreserved pooled blood promotes safety for the user because it is released only after pretesting for infection parameters and has lower variation than fresh donations from a variety of donors.

The management of hand-foot-and-mouth disease(HFMD) epidemic is difficult due to the frequent emergence of non-EV71 and non-CVA16 enteroviruses and some cases testing negative for HFMD-associated causative agents. To clarify the virus spectrum of mild and severe HFMD, a comprehensive virome analysis of 238 samples was performed using next-generation sequencing (NGS). The data revealed total thirteen mammalian- and plant- virus families and diverse viral populations including enteroviruses, common respiratory viruses, diarrhea-related viruses, plant viruses and anelloviruses. A total of 18 viruses from 7 virus families were identified in severe cases, versus 37 viruses from 12 virus families in mild cases. Moreover, complicated mixed-infections of enteroviruses with common respiratory viruses were mainly found in severe cases(P = 0.013), while diarrhea-related viruses were mainly found in mild cases(P 0.001). This study provides the preliminary understanding of viromes both in mild and severe cases, which may benefit the detection of etiologic agents and prevention of HFMD.

In this work, novel cyclic-brush polymers with high grafting densities were successfully constructed via the grafting onto approach by combining a light-induced Diels–Alder ring-closure approach and activated ester chemistry as a post-modification method. The cyclic activated ester containing polymer precursor [poly(pentafluorophenyl 4-vinylbenzoate), c-PPF4VB4.0k] was synthesized by combining reversible addition–fragmentation chain transfer (RAFT) polymerization and the light-induced Diels–Alder click reaction, which was fully characterized by NMR, SEC, MALDI TOF mass and UV-Vis spectroscopy. Using activated ester chemistry, this cyclic polymer (c-PPF4VB4.0k), bearing a pentafluorophenyl group, was post-modified with PEG-NH2 to obtain a cyclic-brush polymer (c-PPF4VB4.0k-g-PEG) with hydrophilic grafting polymer chains and a hydrophobic cyclic polymer backbone. Meanwhile, c-PPF4VB4.0k was post-functionalized using propargyl amine to obtain a clickable cyclic polymer (c-P1). The cyclic polymer (c-P1) was further coupled with an azide-containing polymeric chain (l-PS-N3) to produce the cyclic-brush polymer (c-P1-g-PS). This work illustrated a concept for constructing cyclic-brush polymers by combining the ring-closure strategy and activated ester chemistry. Notably, these two cyclic-brush polymers exhibited high grafting densities (almost quantitatively) and narrow molecular weight distribution (Mw/Mn < 1.10).

Eosinophilic asthma is characterized by the eosinophilic inflammation with the allergen independent activation of Th2 lymphocytes. Since IL-5 plays an important role in the maturation, survival and migration of eosinophils, hence the pathogenesis of eosinophilic asthma, biotherapeutics targeting IL-5/IL-5Rα have been developed and/or marketed, including Mepolizumab, Reslizumab, and Benralizumab. Accurate determination of bioactivity is crucial for the safety and efficacy of therapeutic antibodies. The current mode of action (MOA) based method used in the quality control and stability tests for anti-IL-5 mAbs is anti-proliferation assay, which is tedious with long duration and high variation. We describe here the development and validation of a reporter gene assay (RGA), based on an IL-5-dependent TF-1 cell line variant we established that stably expresses the luciferase reporter under the control of STAT5 response elements. After careful optimization, we demonstrate the excellent specificity, precision, accuracy and linearity of the established RGA. Our study also proves that the assay is superior on precision, sensitivity and assay simplicity to the anti-proliferation assay. The established RGA is also applicable to another anti-IL-5Rα mAb. These results show for the first time that this novel RGA, based on the IL-5-IL-5R-STAT5 pathway, can be a valuable supplement to the anti-proliferation assay and employed in the bioactivity determination of anti-IL-5/anti-IL-5Rα biotherapeutics.

Arthrobacter sp. CGMCC 3584 is able to produce high yields of extracellular cyclic adenosine monophosphate (cAMP), which plays a vital role in the field of treatment of disease and animal food, during aerobic fermentation. However, the molecular basis of cAMP production in Arthrobacter species is rarely explored. Here, for the first time, we report the comparative transcriptomic and proteomic study of Arthrobacter cells to elucidate the higher productivity of cAMP under high oxygen supply. We finally obtained 14.1% and 19.3% of the Arthrobacter genome genes which were up-regulated and down-regulated notably, respectively, with high oxygen supply, and identified 54 differently expressed proteins. Our results revealed that high oxygen supply had two major effects on metabolism: inhibition of glycolysis, pyruvate metabolism, nitrogen metabolism, and amino acid metabolism (histidine, branched-chain amino acids and glutamate metabolism); enhancement of the tricarboxylic acid cycle and purine metabolism. We also found that regulation of adenylate cyclase and phosphodiesterase was not significant under high oxygen supply, suggesting efficient cAMP export might be important in cAMP production. These findings may contribute to further understanding of capacities of Arthrobacter species and would be highly useful in genetic regulation for desirable production.

This study focused on analysis of the chemical characteristics of mine waters. The aim of this study is to correlate the degree of different ionic components in mine water and the influence of their convergence using a combination of the three-scale AHP and fuzzy evaluation methods for the comprehensive evaluation of water quality. Ion chromatography (ICS 1100) has been used to analyze the content of the water sample while portable pH/EC/TDS/Tem- perature meters (SX 811 and SX 813) were used to test physical-chemical parameters. The results of this study show that chemistry of in No.11 gushing mine is dominated by HCO3-Na and HCO3-Ca, and had a pH between 7.1 and 8.00, belonging to neutral or slightly alkaline water. In addition, water were found to have the hardness between 18 mg/L and 542.5 mg/L. Results also show that the TDS of the roof sandstone and goaves water are higher than Cambrian limestone water, while the turbidity of the mixed water is 20 NTU in the sump, again higher than in other samples such as Cambrian limestone water. Total dissolved solids and the total hardness of Cambrian limestone groundwater mainly depend on the content of K+ + Na+, Ca2+, B={b1,b2,…,bj} and SO2-4. Thus, chemical composition changes remarkably after mine water mixing. Results showed that the coal roof sandstone water is class V while that in the sump is class III, and the Cambrian limestone groundwater is class I. In gushing, the quality of water can vary greatly; thus, water from the coal face roof sandstone and the Cambrian limestone should be stored and treated separately before being utilized.

More than 100 monoclonal antibodies (mAbs) have been approved by FDA. The mechanism of action (MoA) involves in neutralization of a specific target via the Fab region and Fc effector functions through Fc region, while the latter include complement-dependent cytotoxicity (CDC), antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP). ADCP has been recognized one of the most important MoAs, especially for anti-cancer mAbs in recent years. However, traditional bioassays measuring ADCP always introduced primary macrophages and flow cytometry, which are difficult to handle and highly variable. In this study, we engineered a monoclonal Jurkat/NFAT/CD32a-FceRIγ effector cell line that stably expresses CD32a-FceRIγ chimeric receptor and NFAT-controlled luciferase. The corresponding mAb could bind with the membrane antigens on the target cells with its Fab fragment and CD32a-FceRIγ on the effector cells with its Fc fragment, leading to the crosslinking of CD32a-FceRIγ and the resultant expression of subsequent NFAT-controlled luciferase, which represents the bioactivity of ADCP based on the MoA of the mAb. With rituximab as the model mAb, Raji cells as the target cells, and Jurkat/NFAT/CD32a-FceRIγ cells as the effector cells, we adopted the strategy of Design of Experiment (DoE) to optimize the bioassay. Then we fully validated the established bioassay according to ICH-Q2(R1), which proved the good assay performance characteristics of the bioassay, including specificity, accuracy, precision, linearity, stability and robustness. This RGA can be applied to evaluate the -ADCP bioactivity for anti-CD20 mAbs in lot release, stability testing as well as biosimilar comparability. The engineered cells may also potentially be used to evaluate the ADCP bioactivity of mAbs with other targets.

Local metal-poor galaxies are ideal analogues of primordial galaxies with the interstellar medium (ISM) barely being enriched with metals. However, it is unclear whether carbon monoxide remains a good tracer and coolant of molecular gas at low metallicity. Based on the observation with the upgraded Northern Extended Millimeter Array (NOEMA), we report a marginal detection of CO $J$=2-1 emission in IZw18, pushing the detection limit down to $L^\prime_{\rm CO(2-1)}$=3.99$\times$10$^3$ K km$^{-1}$pc$^{-2}$, which is at least 40 times lower than previous studies. As one of the most metal-poor galaxies, IZw18 shows extremely low CO content despite its vigorous star formation activity. Such low CO content relative to its infrared luminosity, star formation rate, and [CII] luminosity, compared with other galaxies, indicates a significant change in the ISM properties at a few percent of the Solar metallicity. In particular, the high [CII] luminosity relative to CO implies a larger molecular reservoir than the CO emitter in IZw18. We also obtain an upper limit of the 1.3mm continuum, which excludes a sub-millimetre excess in IZw18., Comment: 7 pages, 5 figures, 2 tables, accepted by A&A Letter

Peptide bonds, as the molecular bridges that connect amino acids, are crucial to the formation of proteins. Searches and studies of molecules with embedded peptide-like bonds are thus important for the understanding of protein formation in space. Here we report the first tentative detection of propionamide (C2H5CONH2), the largest peptide-like molecule detected in space toward Sagittarius B2(N1) at a position called N1E that is slightly offset from the continuum peak. A new laboratory measurements of the propionamide spectrum were carried out in the 9-461 GHz, which provide good opportunity to check directly for the transition frequencies of detected interstellar lines of propionamide. Our observing result indicates that propionamide emission comes from the warm, compact cores in Sagittarius B2, in which massive protostellars are forming. The column density of propionamide toward Sgr B2(N1E) was derived to be 1.5\times 10^{16} cm^-2, which is three fifths of that of acetamide, and one nineteenth of that of formamide. This detection suggests that large peptide-like molecules can form and survive during star-forming process and may form more complex molecules in the interstellar medium. The detection of propionamide bodes well for the presence of polypeptides, as well as other complex prebiotic molecules in the interstellar medium., 49 pages, 13 figures, accepted by ApJ

Seasonal inactivated quadrivalent influenza vaccines are currently formulated to include antigens from two strains of influenza A and a strain from each of the two circulating influenza B virus lineages. However, the applicability of the potency assay currently required for the release of vaccines has been hindered due to cross-reactivity between the two B strains. In this study, a reversed-phase high-performance liquid chromatography method previously developed for the separation and quantitative determination of the hemagglutinin content in trivalent influenza vaccine preparations was further extended and found to be adaptable for the assessment of all four hemagglutinin antigens present in quadrivalent influenza vaccines. Vaccines prepared from monovalent bulks and commercial quadrivalent products from the past three vaccination seasons in the Northern Hemisphere were tested with the new method. The results showed excellent resolution of all four hemagglutinins from frequently interfering formulation agents such as surfactants. This method provides a simple approach for fast evaluation of quality and hemagglutinin strain identification in influenza vaccines. It is also the only physicochemical method capable of distinguishing the B strains in quadrivalent influenza vaccines.

The interaction mechanisms of a series of bisabolonalone hydrazone carboxamides as H+,K+-ATPase reversible inhibitors were studied by molecular docking, QM/MM calculation, and MM/GBSA binding free energy analysis methods. The correlation coefficient, R, between calculated ΔG bind (the binding free energy with H+,K+-ATPase homology model) and experimental pIC50 (negative logarithms of 50% inhibition concentration) of ligands is −0.9386. A suitable molecular volume of ligand is needed to dock into the whole binding pocket, which includes Gln127–Leu141 in the loop of TM1–TM2; Arg328–Ala335 in the loop of TM4; Tyr799–Thr815 in the loop of TM5–TM6; Trp899–Phe917 in the loop of TM7–TM8; Leu921–Tyr928 in the loop of TM8; and Asn986–Asn989 in the loop of TM9–TM10. By the decomposed energy comparisons of residues in binding sites, the naphthyl group substituting benzene ring in bisabolonalone hydrazone carboxamides could increase the hydrophobic interaction with Cys813, which is a very important binding site for enhancing the inhibition activity. In addition, Asp137 is another key residue binding with ligand through the hydrogen bond and electrostatic interactions. The results of this analysis could help in further rational design of novel H+,K+-ATPase reversible inhibitors.

Context. We present our findings for the HCN/H13CN 1-0 line ratio in the molecular outflow of Arp 220 west based on high-resolution ALMA data. Aims. Molecular gas masses in the outflowing gas of galaxies driven by active galactic nuclei (AGNs) or starbursts are important parameters for understanding the feedback of these latter two phenomena and star-formation quenching. The conversion factor of line luminosities to masses is related to the optical depth of the molecular lines. Methods. Using H13CN 1-0, the isotopic line of HCN 1-0, to obtain the line ratio of HCN/H13CN 1-0 is an ideal way to derive the optical depth of HCN 1-0 in outflowing gas. Results. With the nondetection of H13CN 1-0 in the outflowing gas, a 3{\sigma} lower limit of HCN/H13CN 1-0 line ratio is obtained, which is at least three times higher than that found in the whole of the whole system of Arp 220. The high HCN/H13CN 1-0 line ratio indicates low opacity of HCN 1-0 in the outflowing gas, even though the upper limit of HCN 1-0 opacity obtained here is still not good enough to draw any robust conclusions if HCN 1-0 is optically thin. A lower conversion factor of HCN 1-0 luminosity to dense gas mass in the outflowing gas should be used than that used for the host galaxy of Arp 220., Comment: 5 pages, 3 figures, A&A in press; Corresponding authors: Junzhi Wang (jzwang@shao.ac.cn) and Shu Liu (liushu@nao.cas.cn)

The cusp-core problem is one of the main challenges of the cold dark matter paradigm on small scales: the density of a dark matter halo is predicted to rise rapidly toward the center as rho ~ r^alpha with alpha between -1 and -1.5, while such a cuspy profile has not been clearly observed. We have carried out the spatially-resolved mapping of gas dynamics toward a nearby ultra-diffuse galaxy (UDG), AGC 242019. The derived rotation curve of dark matter is well fitted by the cuspy profile as described by the Navarro-Frenk-White model, while the cored profiles including both the pseudo-isothermal and Burkert models are excluded. The halo has alpha=-(0.90+-0.08) at the innermost radius of 0.67 kpc, Mhalo=(3.5+-1.2)E10 Msun and a small concentration of 2.0+-0.36. AGC 242019 challenges alternatives of cold dark matter by constraining the particle mass of fuzzy dark matter to be < 0.11E-22 eV or > 3.3E-22 eV , the cross section of self-interacting dark matter to be < 1.63 cm2/g, and the particle mass of warm dark matter to be > 0.23 keV, all of which are in tension with other constraints. The modified Newtonian dynamics is also inconsistent with a shallow radial acceleration relationship of AGC 242019. For the feedback scenario that transforms a cusp to a core, AGC 242019 disagrees with the stellar-to-halo-mass-ratio dependent model, but agrees with the star-formation-threshold dependent model. As a UDG, AGC 242019 is in a dwarf-size halo with weak stellar feedback, late formation time, a normal baryonic spin and low star formation efficiency (SFR/gas)., ApJ accepted, 25 pages, 17 figures

To develop a new one-pot polyphosphate kinase (PPK) system with low cost and high efficiency for ATP regeneration in industrial CTP production. We developed a new one-pot PPK system by applying a three-enzyme cascade (CMK, NDK and PPK) with an in vitro polyP-based ATP regeneration system. The PPK was selected from twenty sources, and was made solvable by fusion expressing with soluble protein and constructing polycistronic plasmids, or co-expressing with molecular chaperones GroES/EL. Activities of other enzymes were optimized by employing fusion expression, tac-pBAD system, Rosetta host and codon optimization. After 24 h, the concentration of CDP and CTP reached 3.8 ± 0.2 and 6.9 ± 0.3 mM l−1 respectively with a yield of approximately 79%. The molar conversion rate of CTP was 51%, and its yield and conversion rate increased 100% from the traditional system. A new one-pot ATP regeneration system applying polyphosphate kinase for CTP production was developed.

Excessive activation of macrophages has been implicated in various types of inflammatory injury. Suppression of macrophage activation would have therapeutic benefits, leading to the alleviation of the progression of inflammatory diseases. Eburicoic acid (EA) is one of main bioactive components isolated from Laetiporus sulphureus (Bull.:Fr.) Murrill. In our previous study, we found that EA possessed anti-inflammatory activities. However, the cellular and molecular mechanisms underlying its anti-inflammatory activities remain to be poorly understood. The present study aimed to further evaluate its effect on lipopolysaccharide (LPS)-induced inflammatory responses in RAW264.7 macrophage cells. We investigated the anti-inflammatory effect by modulating LPS-induced activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR)/nuclear transcription factor-κB (NF-κB) pathway in RAW264.7 cells. The results showed that EA caused no obvious cytotoxicity, and its suitable concentrations on RAW264.7 cells were in the range from 0.02 to 0.08 μM. EA significantly inhibited the releases of inflammatory mediators, nitrite oxide (NO) and prostaglandin E2 (PGE2); suppressed mRNA and protein expression levels of inducible nitrite oxide synthase (iNOS) and cyclooxygenase-2 COX-2 and pro-inflammatory cytokine TNF-α, IL-6, and IL-1β; and reduced levels of phosphorylated PI3K, Akt, mTOR, and NF-κBp65 in LPS-induced RAW264.7 cells in dose- and time-dependent manners. These aforementioned results indicated that EA executed anti-inflammatory effect on LPS-induced RAW264.7 cells, and this effect might be achieved via suppressing the PI3K/Akt/mTOR/NF-κB signaling pathway and inhibiting the LPS-induced productions of inflammatory mediators and pro-inflammatory cytokines.