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1.
Mol Neurobiol ; 61(8): 5882-5900, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38244148

ABSTRACT

Aurantii Fructus Immaturus total flavonoids (AFIF) is the main effective fraction extracted from AFI, which has a good effect on promoting gastrointestinal motility. This study aimed to investigate AFIF which regulates miR-5100 to improve constipation symptoms in mice by targeting Frizzled-2 (Fzd2) to alleviate interstitial cells of Cajal (ICCs) calcium ion balance and autophagy apoptosis. The constipated mouse model was induced by an antibiotic suspension, and then treated with AFIF. RNA-seq sequencing, luciferase assay, immunofluorescence staining, transmission electron microscopy, ELISA, flow cytometry, quantitative polymerase chain reaction (PCR), and Western blot were applied in this study. The results showed that AFIF improved constipation symptoms in antibiotic-induced constipated mice, and decreased the autophagy-related protein Beclin1 levels and the LC3-II/I ratio in ICCs. miR-5100 and its target gene Fzd2 were screened as key miRNAs and regulator associated with autophagy. Downregulation of miR-5100 caused increased expression of Fzd2, decreased proliferation activity of ICCs, increased apoptotic cells, and enhanced calcium ion release and autophagy signals. After AFIF treatment, miR-5100 expression was upregulated and Fzd2 was downregulated, while autophagy-related protein levels and calcium ion concentration decreased. Furthermore, AFIF increased the levels of SP, 5-HT, and VIP, and increased the expression of PGP9.5, Sy, and Cx43, which alleviated constipation by improving the integrity of the enteric nervous system network. In conclusion, AFIF could attenuate constipation symptoms by upregulating the expression of miR-5100 and targeting inhibition of Fzd2, alleviating calcium overload and autophagic death of ICCs, regulating the content of neurotransmitters, and enhancing the integrity of the enteric nervous system network.


Subject(s)
Autophagy , Calcium , Constipation , Flavonoids , Frizzled Receptors , Interstitial Cells of Cajal , MicroRNAs , Animals , Interstitial Cells of Cajal/metabolism , Interstitial Cells of Cajal/drug effects , Interstitial Cells of Cajal/pathology , Autophagy/drug effects , Frizzled Receptors/metabolism , Calcium/metabolism , Flavonoids/pharmacology , Mice , Constipation/drug therapy , Constipation/metabolism , MicroRNAs/genetics , MicroRNAs/metabolism , Male , Apoptosis/drug effects
2.
Mol Neurobiol ; 61(3): 1543-1561, 2024 Mar.
Article in English | MEDLINE | ID: mdl-37728849

ABSTRACT

Enteric glial cells (EGCs) are the major component of the enteric nervous system and affect the pathophysiological process of intestinal motility dysfunction. MicroRNAs (miRNAs) play an important role in regulating gastrointestinal homeostasis. However, the mechanism of miRNA-mediated regulation of EGCs in intestinal dysmotility remains unclear. In this study, we investigated the effect of EGC apoptosis on intestinal dysmotility, and the effect of miR-26b-3p on EGC proliferation and apoptosis in vivo and in vitro. A loperamide hydrochloride (Lop)-induced constipated mouse model and an in vitro culture system of rat EGCs were established. The transcriptome was used to predict the differentially expressed gene miR-26b-3p and the target gene Frizzled 10 (FZD10), and their targeting binding relationship was verified by luciferase. EGCs were transfected with miR-26b-3p mimic or antagomir, and the FZD10 expression was down-regulated by siRNA. Immunofluorescence and flow cytometry were used to detect EGC apoptosis. MiR-26b-3p and FZD10 expressions were examined using quantitative real-time PCR (qRT-PCR). The CCK-8 assay was used to detect EGC proliferation. The protein levels were detected by Western blotting and enzyme-linked immunosorbent assay (ELISA). The results showed that miR-26b-3p was up-regulated in the Lop group, whereas FZD10 was down-regulated, and EGC apoptosis was increased in the colon of intestinal dysmotility mice. FZD10 down-regulation and miR-26b-3p mimic significantly increased glycogen synthase kinase-3ß phosphorylation (p-GSK3ß) levels, decreased ß-catenin expression, and promoted EGC apoptosis. MiR-26b-3p antagomir alleviated intestinal dysmotility, promoted EGC increased activity of EGCs, and reduced EGC apoptosis in vivo. In conclusion, this study indicated that miR-26b-3p promotes intestinal motility disorders by targeting FZD10 to block GSK3ß/ß-catenin signaling and induces apoptosis in EGCs. Our results provide a new research target for the treatment and intervention of intestinal dysmotility.


Subject(s)
MicroRNAs , beta Catenin , Animals , Mice , Rats , Antagomirs , Apoptosis , beta Catenin/metabolism , Cell Proliferation , Glycogen Synthase Kinase 3 beta/metabolism , MicroRNAs/metabolism , Neuroglia/metabolism , Wnt Signaling Pathway/physiology
3.
BMC Biol ; 21(1): 195, 2023 09 19.
Article in English | MEDLINE | ID: mdl-37726763

ABSTRACT

BACKGROUND: Adelphocoris suturalis (Hemiptera: Miridae) is a notorious agricultural pest, which causes serious economic losses to a diverse range of agricultural crops around the world. The poor understanding of its genomic characteristics has seriously hindered the establishment of sustainable and environment-friendly agricultural pest management through biotechnology and biological insecticides. RESULTS: Here, we report a chromosome-level assembled genome of A. suturalis by integrating Illumina short reads, PacBio, 10x Chromium, and Hi-C mapping technologies. The resulting 1.29 Gb assembly contains twelve chromosomal pseudomolecules with an N50 of 1.4 and 120.6 Mb for the contigs and scaffolds, respectively, and carries 20,010 protein-coding genes. The considerable size of the A. suturalis genome is predominantly attributed to a high amount of retrotransposons, especially long interspersed nuclear elements (LINEs). Transcriptomic and phylogenetic analyses suggest that A. suturalis-specific candidate effectors, and expansion and expression of gene families associated with omnivory, insecticide resistance and reproductive characteristics, such as digestion, detoxification, chemosensory receptors and long-distance migration likely contribute to its strong environmental adaptability and ability to damage crops. Additionally, 19 highly credible effector candidates were identified and transiently overexpressed in Nicotiana benthamiana for functional assays and potential targeting for insect resistance genetic engineering. CONCLUSIONS: The high-quality genome of A. suturalis provides an important genomic landscape for further investigations into the mechanisms of omnivory, insecticide resistance and survival adaptation, and for the development of integrated management strategies.


Subject(s)
Genomics , Insecticide Resistance , Insecticide Resistance/genetics , Phylogeny , Agriculture , Crops, Agricultural , Chromosomes
4.
Polymers (Basel) ; 15(12)2023 Jun 08.
Article in English | MEDLINE | ID: mdl-37376251

ABSTRACT

Poly(2-hydroxyethyl methacrylate) (polyHEMA) hydrogels are commonly used in biomaterials such as contact lenses. However, water evaporation from these hydrogels can cause discomfort to wearers, and the bulk polymerization method used to synthesize them often results in heterogeneous microstructures, reducing their optical properties and elasticity. In this study, we synthesized polyHEMA gels using a deep eutectic solvent (DES) instead of water and compared their properties to traditional hydrogels. Fourier-transform infrared spectroscopy (FTIR) showed that HEMA conversion in DES was faster than in water. DES gels also demonstrated higher transparency, toughness, and conductivity, along with lower dehydration, than hydrogels. The compressive and tensile modulus values of DES gels increased with HEMA concentration. A DES gel with 45% HEMA showed excellent compression-relaxation cycles and had the highest strain at break value in the tensile test. Our findings suggest that DES is a promising alternative to water for synthesizing contact lenses with improved optical and mechanical properties. Furthermore, DES gels' conduction properties may enable their application in biosensors. This study presents an innovative approach to synthesizing polyHEMA gels and provides insights into their potential applications in the biomaterials field.

5.
Behav Sci (Basel) ; 13(5)2023 May 03.
Article in English | MEDLINE | ID: mdl-37232612

ABSTRACT

In this study, we examined influencer marketing and consumption of non-alcoholic beer by adolescents to determine how these factors could affect the intentions of adolescents to purchase and drink alcohol. A total of 3121 high-school students recruited from 36 schools in Taiwan completed a self-administered questionnaire during the COVID-19 pandemic in 2022. The results indicate that 19% of these adolescents consumed non-alcoholic beer and 28% consumed alcohol in the past year. Multivariate analysis positively associated adolescents' exposure to influencer marketing with their purchase and consumption of non-alcoholic beer. Adolescents' exposure to influencer marketing of non-alcoholic beer combined with lower levels of parental restrictive mediation was associated with increased odds of the purchase and consumption of alcohol. For individuals who did not purchase alcohol in the past year, both the exposure to influencer marketing and the consumption of non-alcoholic beer were associated with intending to purchase alcohol in the future. Similarly, individuals who previously abstained from the consumption of alcohol, both the exposure to influencer marketing and the consumption of non-alcoholic beer were associated with intending to consume alcohol. In conclusion, when adolescents were exposed to influencer marketing of non-alcoholic beer they were more likely to consume it, which resulted in an increased likelihood that they would then purchase and consume alcohol.

6.
Chem Sci ; 14(17): 4620-4626, 2023 May 03.
Article in English | MEDLINE | ID: mdl-37152263

ABSTRACT

Single-atom Cu supported on CeO x nanorod catalysts (Cu1/CeO x ) have been synthesized through the anchoring of copper by terminal hydroxyl groups on the CeO x surface. The oxygen defect characteristics of the CeO x nanorods promote electron transfer between Cu and CeO x through a Ce-O-Cu interface, which realizes flexible electronic regulation of the Cu sites. Single-atom Cu species with an oxidation state of between +1 and +2 were formed, which was confirmed by X-ray photoelectron spectroscopy, X-ray fine structure spectroscopy, and electron paramagnetic resonance spectroscopy. Cu1/CeO x emerged as a catalyst with advanced catalytic performance for elemental sulfur in S-arylation using aryl iodides, achieving 97.1% iodobenzene conversion and 94.8% selectivity toward diphenyl disulfide. The substituted iodobenzene with different electronic or steric groups successfully realized S-arylation and produced the corresponding diaryl disulfides with high selectivity. The fully exposed single-atom Cu with flexible electronic characteristics successively realized oxidative addition or coordination of multiple substrates, making it possible to obtain diaryl disulfide with high selectivity.

7.
Nat Commun ; 14(1): 2118, 2023 04 25.
Article in English | MEDLINE | ID: mdl-37185249

ABSTRACT

The growth in world population, climate change, and resource scarcity necessitate a sustainable increase in crop productivity. Photosynthesis in major crops is limited by the inefficiency of the key CO2-fixing enzyme Rubisco, owing to its low carboxylation rate and poor ability to discriminate between CO2 and O2. In cyanobacteria and proteobacteria, carboxysomes function as the central CO2-fixing organelles that elevate CO2 levels around encapsulated Rubisco to enhance carboxylation. There is growing interest in engineering carboxysomes into crop chloroplasts as a potential route for improving photosynthesis and crop yields. Here, we generate morphologically correct carboxysomes in tobacco chloroplasts by transforming nine carboxysome genetic components derived from a proteobacterium. The chloroplast-expressed carboxysomes display a structural and functional integrity comparable to native carboxysomes and support autotrophic growth and photosynthesis of the transplastomic plants at elevated CO2. Our study provides proof-of-concept for a route to engineering fully functional CO2-fixing modules and entire CO2-concentrating mechanisms into chloroplasts to improve crop photosynthesis and productivity.


Subject(s)
Carbon Dioxide , Ribulose-Bisphosphate Carboxylase , Ribulose-Bisphosphate Carboxylase/metabolism , Carbon Dioxide/metabolism , Chloroplasts/metabolism , Organelles/metabolism , Photosynthesis
8.
Biomolecules ; 13(5)2023 04 25.
Article in English | MEDLINE | ID: mdl-37238608

ABSTRACT

Bacterial RNA polymerases (RNAP) form distinct holoenzymes with different σ factors to initiate diverse gene expression programs. In this study, we report a cryo-EM structure at 2.49 Å of RNA polymerase transcription complex containing a temperature-sensitive bacterial σ factor, σ32 (σ32-RPo). The structure of σ32-RPo reveals key interactions essential for the assembly of E. coli σ32-RNAP holoenzyme and for promoter recognition and unwinding by σ32. Specifically, a weak interaction between σ32 and -35/-10 spacer is mediated by T128 and K130 in σ32. A histidine in σ32, rather than a tryptophan in σ70, acts as a wedge to separate the base pair at the upstream junction of the transcription bubble, highlighting the differential promoter-melting capability of different residue combinations. Structure superimposition revealed relatively different orientations between ßFTH and σ4 from other σ-engaged RNAPs and biochemical data suggest that a biased σ4-ßFTH configuration may be adopted to modulate binding affinity to promoter so as to orchestrate the recognition and regulation of different promoters. Collectively, these unique structural features advance our understanding of the mechanism of transcription initiation mediated by different σ factors.


Subject(s)
Escherichia coli Proteins , Escherichia coli , Escherichia coli/metabolism , DNA-Directed RNA Polymerases/metabolism , Transcription, Genetic , Escherichia coli Proteins/metabolism , Promoter Regions, Genetic , Sigma Factor/metabolism , Bacterial Proteins/metabolism , DNA, Bacterial/genetics
9.
Structure ; 31(6): 677-688.e4, 2023 06 01.
Article in English | MEDLINE | ID: mdl-37015227

ABSTRACT

Carboxysomes are proteinaceous bacterial microcompartments that sequester the key enzymes for carbon fixation in cyanobacteria and some proteobacteria. They consist of a virus-like icosahedral shell, encapsulating several enzymes, including ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO), responsible for the first step of the Calvin-Benson-Bassham cycle. Despite their significance in carbon fixation and great bioengineering potentials, the structural understanding of native carboxysomes is currently limited to low-resolution studies. Here, we report the characterization of a native α-carboxysome from a marine cyanobacterium by single-particle cryoelectron microscopy (cryo-EM). We have determined the structure of its RuBisCO enzyme, and obtained low-resolution maps of its icosahedral shell, and of its concentric interior organization. Using integrative modeling approaches, we have proposed a complete atomic model of an intact carboxysome, providing insight into its organization and assembly. This is critical for a better understanding of the carbon fixation mechanism and toward repurposing carboxysomes in synthetic biology for biotechnological applications.


Subject(s)
Cyanobacteria , Ribulose-Bisphosphate Carboxylase , Cryoelectron Microscopy , Ribulose-Bisphosphate Carboxylase/chemistry , Ribulose-Bisphosphate Carboxylase/metabolism , Organelles/metabolism , Photosynthesis , Bacterial Proteins/metabolism
10.
Plant Cell ; 35(2): 795-807, 2023 02 20.
Article in English | MEDLINE | ID: mdl-36471570

ABSTRACT

Ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) performs most of the carbon fixation on Earth. However, plant Rubisco is an intrinsically inefficient enzyme given its low carboxylation rate, representing a major limitation to photosynthesis. Replacing endogenous plant Rubisco with a faster Rubisco is anticipated to enhance crop photosynthesis and productivity. However, the requirement of chaperones for Rubisco expression and assembly has obstructed the efficient production of functional foreign Rubisco in chloroplasts. Here, we report the engineering of a Form 1A Rubisco from the proteobacterium Halothiobacillus neapolitanus in Escherichia coli and tobacco (Nicotiana tabacum) chloroplasts without any cognate chaperones. The native tobacco gene encoding Rubisco large subunit was genetically replaced with H. neapolitanus Rubisco (HnRubisco) large and small subunit genes. We show that HnRubisco subunits can form functional L8S8 hexadecamers in tobacco chloroplasts at high efficiency, accounting for ∼40% of the wild-type tobacco Rubisco content. The chloroplast-expressed HnRubisco displayed a ∼2-fold greater carboxylation rate and supported a similar autotrophic growth rate of transgenic plants to that of wild-type in air supplemented with 1% CO2. This study represents a step toward the engineering of a fast and highly active Rubisco in chloroplasts to improve crop photosynthesis and growth.


Subject(s)
Nicotiana , Ribulose-Bisphosphate Carboxylase , Nicotiana/metabolism , Ribulose-Bisphosphate Carboxylase/genetics , Ribulose-Bisphosphate Carboxylase/metabolism , Photosynthesis/genetics , Chloroplasts/metabolism , Plants, Genetically Modified/metabolism , Carbon Dioxide/metabolism
11.
Dalton Trans ; 51(48): 18473-18479, 2022 Dec 13.
Article in English | MEDLINE | ID: mdl-36421021

ABSTRACT

The development and transformation of biomass-derived platform compounds is a sustainable way to deal with the fossil fuel crisis. 5-Hydroxymethylfurfural (HMF) can be reduced or oxidized to produce many high-value compounds; however, it is challenging to effectively produce 2,5-diformylfuran (DFF) due to overoxidation. In this work, a carbon-doped V2O5 (C-V2O5) material was obtained through pyrolysis of MIL-47(V) nanorods, a typical metal-organic framework material. The X-ray diffraction patterns and X-ray photoelectron spectra showed that the graphitized carbon species were incorporated in C-V2O5. High-efficiency HMF oxidation, high specific selectivity for DFF and excellent recycling could be achieved with the C-V2O5 catalyst. Fourier-transform infrared spectroscopy combined with density functional theory (DFT) calculation revealed that graphitized carbon weakens the VO bond and promotes the formation of oxygen vacancies in C-V2O5, thus improving the catalytic activity in the oxidation of furfuryl alcohols. The V4+ induced by oxygen vacancies will be oxidized by O2 to form V5+, so that the cycle can be realized. It exhibits remarkable selectivity in the oxidation of different alcohols produced from biomass based on the relatively constant active sites in C-V2O5.


Subject(s)
Carbon , Oxides , Furaldehyde/chemistry , Vanadium , Oxygen
12.
Inorg Chem ; 61(28): 11011-11021, 2022 Jul 18.
Article in English | MEDLINE | ID: mdl-35795917

ABSTRACT

Ruthenium, which is relatively cheap in precious metals, has become a popular alternative for a hydrogen evolution reaction (HER) catalyst because of its corrosion resistance and appropriate metal-H bond strength. Convenient synthesis and active site regulation are conducive to stimulating the excellent catalytic performance of Ru as much as possible. Herein, using the mature mesoporous nitrogen-doped carbon material as the support, the catalytic materials containing both single atom Ru and Ru nanoparticles were synthesized by impregnation using the solid-phase reduction method. The effect of reduction temperature on the dispersion state and electronic structure of Ru species has been fully studied using electronic and spectroscopic characterizations. The sample reduced at 300 °C has excellent HER activity with overpotentials of 10.8 and 53.8 mV to deliver 10 mA/cm2 in alkaline and acidic media, respectively, which is among the best activities in the reported results. Electrochemical impedance analysis shows that the reduction temperature has a great influence on the number of active sites and charge transfer impedance of the catalyst.

13.
Curr Issues Mol Biol ; 44(5): 2107-2121, 2022 May 09.
Article in English | MEDLINE | ID: mdl-35678671

ABSTRACT

Taraxacum officinale (dandelion) is often used in traditional Chinese medicine for the treatment of cancer; however, the downstream regulatory genes and signaling pathways mediating its effects on breast cancer remain unclear. The present study aimed to explore the effects of luteolin, the main biologically active compound of T. officinale, on gene expression profiles in MDA-MB-231 and MCF-7 breast cancer cells. The results revealed that luteolin effectively inhibited the proliferation and motility of the MDA-MB-231 and MCF-7 cells. The mRNA expression profiles were determined using gene expression array analysis and analyzed using a bioinformatics approach. A total of 41 differentially expressed genes (DEGs) were found in the luteolin-treated MDA-MB-231 and MCF-7 cells. A Gene Ontology analysis revealed that the DEGs, including AP2B1, APP, GPNMB and DLST, mainly functioned as oncogenes. The human protein atlas database also found that AP2B1, APP, GPNMB and DLST were highly expressed in breast cancer and that AP2B1 (cut-off value, 75%) was significantly associated with survival rate (p = 0.044). In addition, a Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the DEGs were involved in T-cell leukemia virus 1 infection and differentiation. On the whole, the findings of the present study provide a scientific basis that may be used to evaluate the potential benefits of luteolin in human breast cancer. Further studies are required, however, to fully elucidate the role of the related molecular pathways.

14.
Clin Transl Med ; 12(5): e876, 2022 05.
Article in English | MEDLINE | ID: mdl-35605028

ABSTRACT

OBJECTIVE: Apurinic endonuclease 1 (APE1) has been suggested as an oncogene of lung tumours and our bioinformatics analysis identified the association between Erlotinib resistance and interleukin-6 (IL-6). Thus, we performed this work to delineate the mechanistic actions of APE1/IL-6 signalling in Erlotinib resistance of non-small cell lung cancer (NSCLC). METHODS: We selected human NSCLC cell lines HCC827 and PC9 to establish Erlotinib-resistant HCC827R and PC9R cells. Cancer stem cells (CSCs) were isolated from Erlotinib-sensitive HCC827P and PC9P cells (PCSCs) and from HCC827R and PC9R cells (RCSCs). Further, extracellular vesicles (EVs) were separated from PCSCs (PCSC-EVs) and RCSCs (RCSC-EVs) and co-cultured with RCSCs with or without short hairpin RNA (shRNA)-targeting APE1 (APE1 shRNA) transduction. In addition, functional assays were conducted to determine the effect of APE1 shRNA on malignant phenotypes of cancer cells in vitro and in vivo and the activation of IL-6/STAT3 signalling. RESULTS: It was found that NSCLC cells could internalize both RCSC-EVs and PCSC-EVs. RCSC-EVs augmented the resistance of NSCLC cells to Erlotinib. The overexpression of APE1 occurred in NSCLC tissues, and IL-6 was enriched in serum samples of patients with NSCLC. APE1 shRNA was demonstrated to restrict the Erlotinib resistance of NSCLC cells by inactivating the IL-6/STAT3 signalling. Additionally, shAPE1-loaded RCSC-EVs suppressed the Erlotinib resistance of NSCLC via the IL-6/STAT3 axis both in vitro and in vivo, as reflected by impeded malignant phenotypes and xenograft tumour formation. CONCLUSIONS: Collectively, these data indicate that APE1 confers Erlotinib resistance by activating the IL-6/STAT3 signalling, suggesting targeting APE1 as a possible therapeutic target in Erlotinib-resistant NSCLC.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Extracellular Vesicles , Lung Neoplasms , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Cell Line, Tumor , DNA-(Apurinic or Apyrimidinic Site) Lyase/genetics , DNA-(Apurinic or Apyrimidinic Site) Lyase/metabolism , DNA-(Apurinic or Apyrimidinic Site) Lyase/therapeutic use , Erlotinib Hydrochloride/metabolism , Erlotinib Hydrochloride/pharmacology , Erlotinib Hydrochloride/therapeutic use , Extracellular Vesicles/metabolism , Extracellular Vesicles/pathology , Humans , Interleukin-6/metabolism , Interleukin-6/therapeutic use , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , RNA, Small Interfering/metabolism , RNA, Small Interfering/therapeutic use , STAT3 Transcription Factor/genetics , STAT3 Transcription Factor/metabolism , STAT3 Transcription Factor/therapeutic use
15.
mBio ; 13(2): e0362921, 2022 04 26.
Article in English | MEDLINE | ID: mdl-35343789

ABSTRACT

Carboxysomes are anabolic bacterial microcompartments that play an essential role in carbon fixation in cyanobacteria and some chemoautotrophs. This self-assembling organelle encapsulates the key CO2-fixing enzymes, Rubisco, and carbonic anhydrase using a polyhedral protein shell that is constructed by hundreds of shell protein paralogs. The α-carboxysome from the chemoautotroph Halothiobacillus neapolitanus serves as a model system in fundamental studies and synthetic engineering of carboxysomes. In this study, we adopted a QconCAT-based quantitative mass spectrometry approach to determine the stoichiometric composition of native α-carboxysomes from H. neapolitanus. We further performed an in-depth comparison of the protein stoichiometry of native α-carboxysomes and their recombinant counterparts heterologously generated in Escherichia coli to evaluate the structural variability and remodeling of α-carboxysomes. Our results provide insight into the molecular principles that mediate carboxysome assembly, which may aid in rational design and reprogramming of carboxysomes in new contexts for biotechnological applications. IMPORTANCE A wide range of bacteria use special protein-based organelles, termed bacterial microcompartments, to encase enzymes and reactions to increase the efficiency of biological processes. As a model bacterial microcompartment, the carboxysome contains a protein shell filled with the primary carbon fixation enzyme Rubisco. The self-assembling organelle is generated by hundreds of proteins and plays important roles in converting carbon dioxide to sugar, a process known as carbon fixation. In this study, we uncovered the exact stoichiometry of all building components and the structural plasticity of the functional α-carboxysome, using newly developed quantitative mass spectrometry together with biochemistry, electron microscopy, and enzymatic assay. The study advances our understanding of the architecture and modularity of natural carboxysomes. The knowledge learned from natural carboxysomes will suggest feasible ways to produce functional carboxysomes in other hosts, such as crop plants, with the overwhelming goal of boosting cell metabolism and crop yields.


Subject(s)
Carbonic Anhydrases , Halothiobacillus , Carbon Cycle , Carbonic Anhydrases/metabolism , Escherichia coli/genetics , Escherichia coli/metabolism , Halothiobacillus/genetics , Halothiobacillus/metabolism , Organelles , Ribulose-Bisphosphate Carboxylase/genetics , Ribulose-Bisphosphate Carboxylase/metabolism
16.
Cancer Manag Res ; 14: 103-106, 2022.
Article in English | MEDLINE | ID: mdl-35023973

ABSTRACT

Pulmonary carcinosarcoma (PC) is a rare and highly malignant type of non-small cell lung cancer (NSCLC) that is insensitive to radiotherapy and chemotherapy and has a poor prognosis. Here, we report a case of an 88-year-old patient with inoperable PC and a history of cerebral infarction who was treated with first-line anlotinib combined with stereotactic body radiation therapy (SBRT). The therapeutic response has sustained for 10 months. Our work suggests that SBRT combined with anlotinib may be a safe and effective treatment strategy for octogenarians with PC.

17.
Med Oncol ; 39(3): 36, 2022 Jan 20.
Article in English | MEDLINE | ID: mdl-35059870

ABSTRACT

SOX2 is related to drug resistance in many types of cancer, including lung cancer. Herein, we investigated the role of SOX2 and its regulatory signaling in cisplatin-treated non-small-cell lung cancer (NSCLC). The effects of SOX2 on cell viability, proliferation, and apoptosis were evaluated in vitro. Western blotting, real-time quantitative PCR, immunohistochemistry, and luciferase reporter assays were used to investigate the underlying mechanism. Kaplan-Meier survival analysis and the log-rank test were used to assess the relationship between SOX2 expression and patient survival. A549/CDDP cells had marked resistance to cisplatin and stronger colony formation ability than A549 cells. The expression of SOX2 protein or mRNA in A549/CDDP was higher than that in A549. Knockdown of SOX2 in A549/CDDP-induced apoptosis by inhibiting colony formation and decreasing viability, but overexpression of SOX2 reversed these effects. Interestingly, Genomatix software predicted that the APE1 promoter has some SOX2 binding sites, while the SOX2 promoter has no APE1 binding sites. Furthermore, luciferase reporter assays proved that SOX2 could bind the promoter of APE1 in 293T cells. We further verified that SOX2 expression was not affected by shAPE1 in A549/CDDP. As expected, colony formation was obviously inhibited and apoptosis was strongly enhanced in A549/CDDP treated with SOX2 siSOX2 alone or combined with CDDP compared with control cells. Meaningfully, patients with low expression of SOX2, and even including its regulating APE1, survived longer than those with high expression of SOX2, and APE1. siSOX2 overcomes cisplatin resistance by regulating APE1 signaling, providing a new target for overcoming cisplatin resistance in NSCLC.


Subject(s)
Carcinoma, Non-Small-Cell Lung/drug therapy , Cisplatin/therapeutic use , Drug Resistance, Neoplasm/drug effects , SOXB1 Transcription Factors/metabolism , A549 Cells/drug effects , Antineoplastic Agents/therapeutic use , DNA Repair , DNA-(Apurinic or Apyrimidinic Site) Lyase , Humans , RNA, Small Interfering , Signal Transduction
18.
ACS Synth Biol ; 11(1): 154-161, 2022 01 21.
Article in English | MEDLINE | ID: mdl-34664944

ABSTRACT

The carboxysome is a versatile paradigm of prokaryotic organelles and is a proteinaceous self-assembling microcompartment that plays essential roles in carbon fixation in all cyanobacteria and some chemoautotrophs. The carboxysome encapsulates the central CO2-fixing enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), using a polyhedral protein shell that is selectively permeable to specific metabolites in favor of Rubisco carboxylation. There is tremendous interest in repurposing carboxysomes to boost carbon fixation in heterologous organisms. Here, we develop the design and engineering of α-carboxysomes by coexpressing the Rubisco activase components CbbQ and CbbO with α-carboxysomes in Escherichia coli. Our results show that CbbQ and CbbO could assemble into the reconstituted α-carboxysome as intrinsic components. Incorporation of both CbbQ and CbbO within the carboxysome promotes activation of Rubisco and enhances the CO2-fixation activities of recombinant carboxysomes. We also show that the structural composition of these carboxysomes could be modified in different expression systems, representing the plasticity of the carboxysome architecture. In translational terms, our study informs strategies for engineering and modulating carboxysomes in diverse biotechnological applications.


Subject(s)
Ribulose-Bisphosphate Carboxylase , Tissue Plasminogen Activator , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Carbon Cycle , Carbon Dioxide/metabolism , Organelles/metabolism , Ribulose-Bisphosphate Carboxylase/genetics , Tissue Plasminogen Activator/metabolism
19.
Integr Zool ; 17(1): 93-104, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34216516

ABSTRACT

Habitat structure has been considered as an important factor affecting the acoustic evolution of birds, and bird songs are increasingly affected by artificial environmental variation. Invasive plants sometimes can dramatically alter native habitats, but the song variation of native songbirds migrating into invaded habitats has received little attention. The invasion of smooth cordgrass Spartina alterniflora in the coastal wetlands of eastern China has drastically altered the vegetation structure and some small passerines have begun to use invaded habitats to breed. In this study, we compared the song type prevalence and the song characteristics of male plain prinia Prinia inornata to identify differences in vocal behavior between native and invaded habitats. We also tested for differences in vocal behavior in relation to singing perch and wind speed variation between different habitats. The results indicated that males of plain prinia in invaded habitats sang shorter songs than those in native habitats and had a lower song diversity. The homogeneous vegetation structure and higher wind speed in invaded habitats likely leads to males changing the traditional perched singing style. The song variation may be related to the founder effect, the alteration of vegetation structure and microclimate in invaded habitats. This finding highlights the need for better understanding the behavioral evolution of native species in the process of adapting to the invaded habitat. In the future, experimental manipulation is needed to ascertain how the invasive plant drove these vocal behavior changes of native songbirds.


Subject(s)
Poaceae , Songbirds , Vocalization, Animal , Animals , China , Introduced Species , Wetlands
20.
Biodivers Data J ; 10: e90724, 2022.
Article in English | MEDLINE | ID: mdl-36761590

ABSTRACT

Background: Tiaozini, the core area of the Yellow (Bohai) Sea Migratory Bird Habitat in Dongtai, Jiangsu Province and a World Heritage Site, has provided an ideal habitat for migratory birds. As an important hub on the East Asian-Australasian Flyway (EAAF), Tiaozini Wetland provides pivotal stopover and wintering sites for tens of thousands of migratory waterbirds, including some global critically endangered species, such as Spoon-billed Sandpiper (Calidrispygmaea) and Spotted Greenshank (Tringaguttifer). Although many researchers have conducted a lot of studies on waterbirds in Tiaozini Wetland, there is still a lack of a dataset on waterbird species composition and individual quantity in Tiaozini Wetland throughout the year. Here, we conducted a one-year waterbird survey in the Tiaozini Wetland during 2020-2021 and provided an occurrence dataset with detailed species and geographic information. New information: This occurrence dataset is the first public record of species and number of waterbirds in Tiaozini Wetland for a whole year, which includes the taxonomic information, location information, number, investigation date and endangered level for each species. All data have been published on GBIF.

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