Onion (Allium cepa L.) Flavonoid Extract Ameliorates Osteoporosis in Rats Facilitating Osteoblast Proliferation and Differentiation in MG-63 Cells and Inhibiting RANKL-Induced Osteoclastogenesis in RAW 264.7 Cells.

Osteoporosis, a prevalent chronic health issue among the elderly, is a global bone metabolic disease. Flavonoids, natural active compounds widely present in vegetables, fruits, beans, and cereals, have been reported for their anti-osteoporotic properties. Onion is a commonly consumed vegetable rich in flavonoids with diverse pharmacological activities. In this study, the trabecular structure was enhanced and bone mineral density (BMD) exhibited a twofold increase following oral administration of onion flavonoid extract (OFE). The levels of estradiol (E2), calcium (Ca), and phosphorus (P) in serum were significantly increased in ovariectomized (OVX) rats, with effects equal to alendronate sodium (ALN). Alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) levels in rat serum were reduced by 35.7% and 36.9%, respectively, compared to the OVX group. In addition, the effects of OFE on bone health were assessed using human osteoblast-like cells MG-63 and osteoclast precursor RAW 264.7 cells in vitro as well. Proliferation and mineralization of MG-63 cells were promoted by OFE treatment, along with increased ALP activity and mRNA expression of osteoprotegerin (OPG)/receptor activator of nuclear factor-kappaB ligand (RANKL). Additionally, RANKL-induced osteoclastogenesis and osteoclast activity were inhibited by OFE treatment through decreased TRAP activity and down-regulation of mRNA expression-related enzymes in RAW 264.7 cells. Overall findings suggest that OFE holds promise as a natural functional component for alleviating osteoporosis.

Pore architecture of TRIC channels and insights into their gating mechanism.

Intracellular Ca2+ signalling processes are fundamental to muscle contraction, neurotransmitter release, cell growth and apoptosis. Release of Ca2+ from the intracellular stores is supported by a series of ion channels in sarcoplasmic or endoplasmic reticulum (SR/ER). Among them, two isoforms of the trimeric intracellular cation (TRIC) channel family, named TRIC-A and TRIC-B, modulate the release of Ca2+ through the ryanodine receptor or inositol triphosphate receptor, and maintain the homeostasis of ions within SR/ER lumen. Genetic ablations or mutations of TRIC channels are associated with hypertension, heart disease, respiratory defects and brittle bone disease. Despite the pivotal function of TRIC channels in Ca2+ signalling, their pore architectures and gating mechanisms remain unknown. Here we present the structures of TRIC-B1 and TRIC-B2 channels from Caenorhabditis elegans in complex with endogenous phosphatidylinositol-4,5-biphosphate (PtdIns(4,5)P2, also known as PIP2) lipid molecules. The TRIC-B1/B2 proteins and PIP2 assemble into a symmetrical homotrimeric complex. Each monomer contains an hourglass-shaped hydrophilic pore contained within a seven-transmembrane-helix domain. Structural and functional analyses unravel the central role of PIP2 in stabilizing the cytoplasmic gate of the ion permeation pathway and reveal a marked Ca2+-induced conformational change in a cytoplasmic loop above the gate. A mechanistic model has been proposed to account for the complex gating mechanism of TRIC channels.

Integration of DSF and Temperature Signals for RpfC/RpfG Two-Component System Modulating Protease Production in Stenotrophomonas maltophilia FF11.

Two-component signal system (TCS) is the predominant bacterial sense-and-response machinery. RpfC/RpfG TCS involved in quorum sensing molecule Diffuse Signal Factor (DSF) signal perception and transduction was well studied in many bacteria. However, whether other environmental factors participating in the signal perception and transduction of RpfC/RpfG was still unclear. Here, we showed that RpfC/RpfG could integrate temperature and DSF signal partially controlling the production of the temperature-dependent protease (SmtP) in S. maltophilia FF11, a strain isolated from frozen Antarctic krill, exhibited spoilage potential due to secret more protease at low temperatures involving in protein degradation. qRT-PCR analysis revealed rpf system mediating approximately 60% transcription activity of Clp, a critical transcription factor linking with LotS/LotR, consisting a signal network controlling completely the SmtP production in previous study. Protease production was partially reduced in rpfF (coding DSF synthetase) mutant strains at 15 °C or 25 °C, not be increased through addition DSF or overexpression RpfF in WT at 37 °C, indicating that DSF was effective for protease production only at low temperatures in S. maltophilia. Additionally, biochemical analysis revealed the enzymatic activity of RpfG from strain FF11 cultured at 37 °C or DSF-deficient strains grown at 25 °C was significantly reduced compared to that of RpfG from strain FF11 cultured at 25 °C. These findings outline an interplay mechanism that allows S. maltophilia to integrate quorum sensing and temperature cues controlling protease production, and imply a potential relationship between two distinct systems of RpfC/RpfG and LotS/LotR.

Imaging and Inhibiting: A Dual Function Molecular Flare for Cancer Cells.

The Wnt pathway is dysregulated and activated in many human malignancies. More than 90% of colon cancers have variations in the Wnt pathway. Sulindac, a drug that targets protein Dvl of the Wnt/Dvl/ß-catenin pathway, which regulates cancer gene expression, has been reported to significantly reduce the incidence and the risk of death from colorectal cancer and other types of cancer. Herein, a dual functional compound (SLN) containing Sulindac and a linked fluorophore is first reported, combining the functions of lighting up colon cancer cells as a flare and inhibiting colon tumors as a drug. SLN can not only mark the Dvl protein in the Wnt pathway to recognize tumors layer by layer but also achieve effective inhibition of colon cancer, providing a promising reagent for chemotherapy and a fluorescent indicator for surgery during the removal the colon tumors in situ.

Functional daidzein enhances the anticancer effect of topotecan and reverses BCRP-mediated drug resistance in breast cancer.

Topotecan (TPT), a semisynthetic derivative of camptothecin, has been used in cancer chemotherapy, but side effects and drug resistance limit its clinical application. Daidzein (DAI), a natural isoflavone and bioactive food component widely existing in fruits, nuts, soybeans and soy-based products, is a type of phytoestrogen. Combination treatment with DAI and TPT showed a strong synergistic effect on tumor cells, with a 0.10˜0.66 combined index, by increasing TPT inhibition on Topo Ⅰ, resulting in more cells arresting at the G2/M phase and inducing more cells to undergo apoptosis. In addition, the resistance of MCF7/ADR cells to TPT was reversed (the resistance index decreased from 7.17 to 0.77) by inhibiting the expression of ERα and BCRP to increase TPT accumulation intracellularly. Moreover, the combination of DAI and TPT showed a stronger inhibitory effect (P < 0.01) on tumor growth in both MCF7 and MCF7/ADR xenograft models than the 9 mg/kg TPT monotherapy group. Our results may provide a reasonable, new approach to develop safe and efficient nutrition components from foods for breast cancer combination treatment.

Mechanical coupling of the multiple structural elements of the large-conductance mechanosensitive channel during expansion.

The prokaryotic mechanosensitive channel of large conductance (MscL) is a pressure-relief valve protecting the cell from lysing during acute osmotic downshock. When the membrane is stretched, MscL responds to the increase of membrane tension and opens a nonselective pore to about 30 Å wide, exhibiting a large unitary conductance of ∼ 3 nS. A fundamental step toward understanding the gating mechanism of MscL is to decipher the molecular details of the conformational changes accompanying channel opening. By applying fusion-protein strategy and controlling detergent composition, we have solved the structures of an archaeal MscL homolog from Methanosarcina acetivorans trapped in the closed and expanded intermediate states. The comparative analysis of these two new structures reveals significant conformational rearrangements in the different domains of MscL. The large changes observed in the tilt angles of the two transmembrane helices (TM1 and TM2) fit well with the helix-pivoting model derived from the earlier geometric analyses based on the previous structures. Meanwhile, the periplasmic loop region transforms from a folded structure, containing an ω-shaped loop and a short ß-hairpin, to an extended and partly disordered conformation during channel expansion. Moreover, a significant rotating and sliding of the N-terminal helix (N-helix) is coupled to the tilting movements of TM1 and TM2. The dynamic relationships between the N-helix and TM1/TM2 suggest that the N-helix serves as a membrane-anchored stopper that limits the tilts of TM1 and TM2 in the gating process. These results provide direct mechanistic insights into the highly coordinated movement of the different domains of the MscL channel when it expands.

Ion- and water-binding sites inside an occluded hourglass pore of a trimeric intracellular cation (TRIC) channel.

BACKGROUND: Trimeric intracellular cation (TRIC) channels are crucial for Ca2+ handling in eukaryotes and are involved in K+ uptake in prokaryotes. Recent studies on the representative members of eukaryotic and prokaryotic TRIC channels demonstrated that they form homotrimeric units with the ion-conducting pores contained within each individual monomer. RESULTS: Here we report detailed insights into the ion- and water-binding sites inside the pore of a TRIC channel from Sulfolobus solfataricus (SsTRIC). Like the mammalian TRIC channels, SsTRIC is permeable to both K+ and Na+ with a slight preference for K+, and is nearly impermeable to Ca2+, Mg2+, or Cl-. In the 2.2-Å resolution K+-bound structure of SsTRIC, ion/water densities have been well resolved inside the pore. At the central region, a filter-like structure is shaped by the kinks on the second and fifth transmembrane helices and two nearby phenylalanine residues. Below the filter, the cytoplasmic vestibule is occluded by a plug-like motif attached to an array of pore-lining charged residues. CONCLUSIONS: The asymmetric filter-like structure at the pore center of SsTRIC might serve as the basis for the channel to bind and select monovalent cations. A Velcro-like plug-pore interacting model has been proposed and suggests a unified framework accounting for the gating mechanisms of prokaryotic and eukaryotic TRIC channels.

Bifurcation and chaos in the simple passive dynamic walking model with upper body.

We present some rich new complex gaits in the simple walking model with upper body by Wisse et al. in [Robotica 22, 681 (2004)]. We first show that the stable gait found by Wisse et al. may become chaotic via period-doubling bifurcations. Such period-doubling routes to chaos exist for all parameters, such as foot mass, upper body mass, body length, hip spring stiffness, and slope angle. Then, we report three new gaits with period 3, 4, and 6; for each gait, there is also a period-doubling route to chaos. Finally, we show a practical method for finding a topological horseshoe in 3D Poincaré map, and present a rigorous verification of chaos from these gaits.

Combined fabrication of zeolitic imidazolate framework-8 and lanthanide towards coordination polymers: A dual-signal fluorescent probe for sensing Cu2+ based on synergistic effect of aggregation-induced emission and antenna effect.

Copper ion (Cu2+) detection remains an important task for monitoring water quality because of its specific toxicity. Herein, a new dual-signal fluorescent probe was developed by combining zeolitic imidazolate framework-8 (ZIF-8) and lanthanide for the detection of Cu2+ for the first time. The lanthanide coordination polymer (guanosine monophosphate and Eu3+, GMP/Eu) was initially incorporated into ZIF-8 to yield ZIF-8/GMP/Eu nanomaterials with extremely weak single emission fluorescence at 618 nm. It was found that the resulted nanomaterials could display a dual emission fluorescence at 515 nm and 618 nm after the introduction of tetracycline (TC) due to the synergistic effect of aggregation-induced emission effect (AIE, TC induced by ZIF-8) and antenna effect (AE, between TC and GMP/Eu). Interestingly, in the presence of Cu2+, the AIE of TC was destroyed because of the interaction of Cu2+ with ZIF-8 and TC. The AE between TC and GMP/Eu disappeared due to the formation of complex between TC and Cu2+. A dual-signal fluorescent probe of ZIF-8/GMP/Eu/TC was thereby established for sensing Cu2+ in the range of 0.5-100 µM. Such a dual-signal response strategy that intelligently utilized the "ON"/"OFF" of AIE and AE can not only eliminate the background interference, but also ensure the improved selectivity of Cu2+ sensing. Subsequently, the dual-signal fluorimetric strategy was applied for the detection of Cu2+ in environmental water samples, indicating the potential feasibility of applications for water quality monitoring.

Selenite Ameliorates Cadmium-induced Cytotoxicity Through Downregulation of ROS Levels and Upregulation of Selenoprotein Thioredoxin Reductase 1 in SH-SY5Y Cells.

Cadmium (Cd) as a ubiquitous toxic heavy metal in the environment, causes severe hazards to human health, such as cellular stress and organ injury. Selenium (Se) was reported to reduce Cd toxicity and the mechanisms have been intensively studied so far. However, it is not yet crystal clear whether the protective effect of Se against Cd-induced cytotoxicity is related to selenoproteins in nerve cells or not. In this study, we found that Cd inhibited selenoprotein thioredoxin reductase 1 (TrxR1; TXNRD1) and decreased the expression level of TrxR1, resulting in cellular oxidative stress, and Se supplements ameliorated Cd-induced cytotoxicity in SH-SY5Y cells. Mechanistically, the detoxification of Se against Cd is attributed to the increase of the cellular TrxR activity and upregulated TrxR1 protein level, culminating in strengthened antioxidant capacity. Results showed that Se supplements attenuated the ROS production and apoptosis in SH-SY5Y cells, and significantly mitigated Cd-induced SH-SY5Y cell death. This study may be a valuable reference for shedding light on the mechanism of Cd-induced cytotoxicity and the role of TrxR1 in Se-mitigated cytotoxicity of Cd in neuroblast cells, which may be helpful for understanding the therapeutic potential of Cd and Se in treating or preventing neurodegenerative diseases, like Alzheimer's disease (AD) and Parkinson's disease (PD).

Thioredoxin reductase 1 inhibitor shikonin promotes cell necroptosis via SecTRAPs generation and oxygen-coupled redox cycling.

Shikonin, a naturally occurring naphthoquinone with potent anti-tumor activity, has been reported to induce cancer cell death via targeting selenoenzyme thioredoxin reductase 1 (TrxR1; TXNRD1). However, the interaction between shikonin and TrxR1 remains unclear, and the roles of the cellular antioxidant system in shikonin induced cell death are obscure. Here, we found that shikonin modified the Sec498 residue of TrxR1 to fully inhibit its antioxidant activity, however, the shikonin-modified TrxR1 still remained intrinsic NADPH oxidase activity, which promotes superoxide anions production. Besides, TrxR1 efficiently reduced shikonin in both selenocysteine dependent and selenocysteine independent manners, and the oxygen-coupled redox cycling of shikonin also generates excessive superoxide anions. The inhibitory effects and the redox cycling of shikonin towards TrxR1 caused cancer cell ROS-dependent necroptosis. Interestingly, as we evaluated, some cancer cell lines were insensitive to shikonin, especially kelch-like ECH associated protein 1 (KEAP1)-mutant non-small cell lung cancer (NSCLC) cells, which harbor constitutive activation of the nuclear factor-erythroid 2-related factor 2 (NRF2). NADPH bankruptcy caused by glucose starvation or glucose limitation (inhibiting glucose transporter 1 by BAY-876) could efficiently overcome the resistance of KEAP1-mutant NSCLC cells to shikonin. Glucose-6-phosphate dehydrogenase (G6PD), was known as a rate-limiting enzyme in the pentose phosphate pathway, however, the pharmacological inhibition of G6PD by 6-aminonicotinamide (6-AN), enhanced the shikonin-induced cytotoxicity but has no selectivity on KEAP1-mutant NSCLC cells. This study will be helpful in applying shikonin for potential chemotherapy, and in combinational treatment of KEAP1-mutant NSCLC.

Nursing Countermeasures of Continuous Renal Replacement Treatment in End-Stage Renal Disease with Refractory Hypotension in the Context of Smart Health.

This work is aimed at exploring the nursing strategies and effects of continuous renal replacement therapy (CRRT) for end-stage renal disease (ESRD) with refractory hypotension under the background of smart health. 40 ESRD patients with refractory hypotension who received CRRT treatment were enrolled as the research objects and were randomly rolled into the intervention group and the control group, with 20 cases in each group. Patients in the control group received routine nursing, and those in the intervention group received individualized nursing. The incidence of hypotension, dry body weight, serous cavity effusion, renal function indicators (blood urea nitrogen (BUN) and creatinine (Cre)), and patient satisfaction were compared between the two groups. The results showed that the probability of hypotension in the intervention group was 9.38%, which was lower than that in the control group (34.38%). The probability of early termination of dialysis in the intervention group was 0%, which was lower than that in the control group (18.75%), and the difference was statistically significant (P < 0.05). The decreases of BUN and Cre in the intervention group were significantly greater than those in the control group, and the differences were statistically significant (P < 0.05). The proportion of water growth less than 10% during dialysis in the intervention group was 98.44%, which was greater than that in the control group (93.45%), and the difference was statistically significant (P < 0.05). The ultrafiltration volume after dialysis in the intervention group was 2850 ± 400 mL, which was greater than that in the control group 2350 ± 350 mL. After intervention, the proportion of patients with pleural effusion in the intervention group was 10% less than that in the control group (20%), and the difference was statistically significant (P < 0.05). The satisfaction rate of the intervention group was 97.66%, which was higher than that of the control group (65.63%). In conclusion, individualized nursing was more helpful to the recovery of ESRD patients with refractory hypotension treated with CRRT than routine nursing.

Plumbagin reduction by thioredoxin reductase 1 possesses synergy effects with GLUT1 inhibitor on KEAP1-mutant NSCLC cells.

Thioredoxin reductase 1 (TrxR1 or TXNRD1) is a major enzyme in cellular redox regulation and is considered as a drug target for cancer therapy. Previous studies have reported that plumbagin caused reactive oxygen species (ROS)-dependent apoptosis via inhibiting TrxR1 activity or being reduced by TrxR1, leading to selectively cancer cell death. However, the mechanism of TrxR1-mediated redox cycling of plumbagin is obscure and the evidence for plumbagin targeting TrxR1 is still lacking. Herein, we demonstrated that TrxR1 catalyzed plumbagin reduction in both selenocysteine (Sec)-dependent and independent manners, and its activity relied on the intact N-terminal motif of TrxR1, but a high-efficiency reduction was supported by the C-terminal thiols. During the redox cycling of plumbagin, excessive ROS production was observed coupled with oxygen. Using LC-MS and TrxR1 mutants, we found that the Sec residue of TrxR1 was modified by plumbagin, which converted the enzyme from antioxidant to pro-oxidant. Furthermore, we evaluated the therapeutic potential of plumbagin in non-small cell lung cancer (NSCLC), and found that Kelch-like ECH-associated protein 1 (KEAP1)-mutant NSCLC cells, which possess constitutive nuclear factor erythroid 2-related factor 2 (NRF2) activity, were insensitive to plumbagin; however, inhibition of glucose transporter 1 (GLUT1) by small-molecule BAY-876 or inhibiting glucose-6-phosphate dehydrogenase (G6PD) by 6-aminonicotinamide (6-AN) overcame the plumbagin-resistance of KEAP1-mutant NSCLC cells. Taken together, this study elucidated the pharmacological mechanism of plumbagin by targeting TrxR1 and revealed the synergy effect of plumbagin and BAY-876, which may be helpful for applying naphthoquinone compounds to chemotherapy, particularly for treating KEAP1-mutant NSCLC cells.

Proteomic analysis of functional dyspepsia in stressed rats treated with traditional Chinese medicine "Wei Kangning".

BACKGROUND AND AIM: Chinese traditional medical science is generally used as a therapeutic method against functional dyspepsia (FD) in China. Although great effort is made to understand the pharmaceutical mechanisms of Chinese traditional medicine, such as typical traditional Chinese medicine, Wei Kangning, there are still many mysteries to be uncovered. METHODS: The model of FD was established by stimulating rats via tail damping and the rats were treated with traditional Chinese medicine, Wei Kangning. The proteins of the rat gastrointestinal tissues were extracted and run by 2-DE, then the differential proteins were identified using matrix-assisted laser desorption ionisation time-of-flight mass spectrometry and validated with Western blotting or fluorescent quantitation polymerase chain reaction. RESULTS: A total of 228 unique proteins in FD model rats were detected with significant changes in their expression levels corresponding with traditional Chinese medicine, Wei Kangning, administration. Twenty-eight of these proteins were identified, which are involved in many biological functions, such as organism antioxidant enzymes, energy metabolism, glutathione S-transferase, pi2, superoxide dismutase 2 and alpha-enolase and so on. CONCLUSIONS: These proteomic results presented therefore provide additional support to the hypothesis that glutathione S-transferase, pi2, superoxide dismutase 2, α-enolase and voltage-dependent anion channel are the targets of FD treated with traditional Chinese medicine, Wei Kangning.

Formaldehyde suppresses neuronal apoptosis via inhibition of outward K(+) currents in rat hippocampus.

Formaldehyde (FA) is widely present in the environment, and is also a mammalian metabolite. However, its biological role has not been well understood. Here, we show that FA plays an anti-apoptotic role in cultured hippocampal neurons: FA suppressed staurosporine-induced neuronal apoptosis and inhibited the activity of apoptosis-associated caspase-3/7 proteases in a concentration-dependent manner. Moreover, FA suppressed outward K(+) currents and attenuated the enhanced IK currents that are associated with neuronal apoptosis. As an increase in outward K(+) currents is critical for cell apoptosis, our results suggest that FA exerts its anti-apoptotic effects on neuronal cells, probably through its inhibitory effect on the outward K(+) currents.

The effect of psychological nursing on the short- and long-term negative emotions and quality of life of cervical cancer patients undergoing postoperative chemotherapy.

OBJECTIVE: The purpose of this study was to analyze the effect of psychological nursing intervention on the short- and long-term negative emotions and changes in the quality of life in patients with cervical cancer who underwent postoperative chemotherapy. METHODS: 141 patients with cervical cancer who received postoperative chemotherapy in our hospital were recruited as the study cohort. They were divided into the study group (80 cases) and the control group (61 cases) according to the different nursing methods each underwent. The patients in the control group underwent routine nursing, and the study group also underwent psychological nursing. The changes in the quality of life and the negative emotions of the patients in the two groups before and after the intervention were compared, and the correlation between the quality of life and the negative emotions were explored. RESULTS: The patients' Quality of Life Questionnaire (EROTC-QLQ-C30) and Self-rating Anxiety Scale (SAS) scores in the two groups before the intervention were not significantly different (P > 0.05). A re-evaluation at the end of the 90 day-intervention showed that the EROTC-QLQ-C30 scores in the study group were significantly higher than they were in the control group (P < 0.05). A dynamic evaluation showed that the proportion of patients with mild anxiety in the study group was higher than it was in the control group at 30, 60, and 90 days of intervention (P < 0.05). A Spearman correlation analysis showed that the SAS scale and EROTC-QLQ-C30 scores were negatively correlated (r=-0.4438, P < 0.05). CONCLUSION: The implementation of psychological intervention can help alleviate the short- and long-term negative emotions of cervical cancer patients who underwent postoperative chemotherapy, and it is feasible and conducive to the patients' quality of life. We recommend carrying out the clinical promotion and application of this psychological intervention.

Enhancing the understanding of hydrological responses induced by ecological water replenishment using improved machine learning models: A case study in Yongding River.

The ecological water replenishment (EWR) of Yongding River has been an important project implemented in response to the Development of an Ecological Civilization policy in China since 2016. A reasonable amount of EWR requires a systematic understanding of the relationship among the surface water, groundwater, ecology and economy. However, studying surface water-groundwater interactions still remains an important issue. Thus, a coupled model integrating a Muskingum method-based open channel flow model and machine learning-based groundwater model is developed to describe the dynamic changes in streamflow and groundwater level in response to the EWR of Yongding River. The model is calibrated using observed streamflow data as well as groundwater level data on a daily scale for the spring EWR in 2020. The simulated results match well with the observed data and suggest that significant groundwater level increases occur only around the main channel of Yongding River. Fifteen scenarios under different EWR schemes are set to obtain reasonable streamflow during EWR, and then the responses of streamflow and groundwater level changes are simulated. Reasonable streamflow at the Guanting Reservoir need to be above 65 m3/s to ensure the streamflow can pass through Beijing and significant groundwater level recoveries of 170 million m3 through EWR. The developed models can improve the understanding of the interaction between surface water and groundwater and provide a quick assessment of the factors influencing the different EWR schemes and thus aid in effective EWR project management.

Chlorophyllin Inhibits Mammalian Thioredoxin Reductase 1 and Triggers Cancer Cell Death.

Food colorants are widely used by humans in food production and preparation; however, their potential toxicity requires an in-depth analysis. In this study, five out of 15 commercial food colorants, namely, lutein, betanin, caramel, crocin and chlorophyll, significantly inhibited wild type selenoprotein thioredoxin reductase 1 (TrxR1, TXNRD1) in vitro. The hyperactive Sec498 residue of TrxR1 was targeted by those five colorants, which was confirmed by the site-directed mutagenesis of TrxR1. Furthermore, two colorants, chlorophyll and betanin, triggered the oligomerization of TrxR1. A chlorophyll-derived compound, chlorophyllin, irreversibly inhibited the 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) reducing activity of TrxR1 with Kinact = 6.96 × 10-3 ± 0.49 × 10-3 µM-1 min-1. Moreover, chlorophyllin reduced the cellular TrxR activity, leading to reactive oxygen species (ROS) accumulation and, subsequently, promoting cancer cell death. In conclusion, this study might contribute to understand the food safety of commercial colorants and provide chemotherapeutic compounds by targeting TrxR1.

TMEM120A contains a specific coenzyme A-binding site and might not mediate poking- or stretch-induced channel activities in cells.

TMEM120A, a member of the transmembrane protein 120 (TMEM120) family, has a pivotal function in adipocyte differentiation and metabolism, and may also contribute to sensing mechanical pain by functioning as an ion channel named TACAN. Here we report that expression of TMEM120A is not sufficient in mediating poking- or stretch-induced currents in cells and have solved cryo-electron microscopy (cryo-EM) structures of human TMEM120A (HsTMEM120A) in complex with an endogenous metabolic cofactor (coenzyme A, CoASH) and in the apo form. HsTMEM120A forms a symmetrical homodimer with each monomer containing an amino-terminal coiled-coil motif followed by a transmembrane domain with six membrane-spanning helices. Within the transmembrane domain, a CoASH molecule is hosted in a deep cavity and forms specific interactions with nearby amino acid residues. Mutation of a central tryptophan residue involved in binding CoASH dramatically reduced the binding affinity of HsTMEM120A with CoASH. HsTMEM120A exhibits distinct conformations at the states with or without CoASH bound. Our results suggest that TMEM120A may have alternative functional roles potentially involved in CoASH transport, sensing, or metabolism.

Cryo-electron microscopy structure of CLHM1 ion channel from Caenorhabditis elegans.

Calcium homeostasis modulators (CALHMs/CLHMs) comprise a family of pore-forming protein complexes assembling into voltage-gated, Ca2+ -sensitive, nonselective channels. These complexes contain an ion-conduction pore sufficiently wide to permit the passing of ATP molecules serving as neurotransmitters. While their function and structure information is accumulating, the precise mechanisms of these channel complexes remain to be full understood. Here, we present the structure of the Caenorhabditis elegans CLHM1 channel in its open state solved through single-particle cryo-electron microscopy at 3.7-Å resolution. The transmembrane region of the channel structure of the dominant class shows an assembly of 10-fold rotational symmetry in one layer, and its cytoplasmic region is involved in additional twofold symmetrical packing in a tail-to-tail manner. Furthermore, we identified a series of amino acid residues critical for the regulation of CeCLHM1 channel using functional assays, electrophysiological analyses as well as structural-based analysis. Our structure and function analyses provide new insights into the mechanisms of CALHM channels.