Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 32
Filter
1.
Inorg Chem ; 63(1): 902-908, 2024 Jan 08.
Article in English | MEDLINE | ID: mdl-38153042

ABSTRACT

Broadband near-infrared (NIR) phosphor-converted devices play a vital role in emerging applications of imaging, medicine, agriculture, etc. Herein, a series of economical broadband NIR-emitting Sr3Al10-xSiO20:xCr (SASO:xCr) phosphors with tunable bandwidths and emission peaks were realized by tailoring the Cr3+ doping concentration. The optimal Sr3Al9.8SiO20:0.2Cr (SASO:0.2Cr) phosphor exhibits a broadband emission with full width at half-maximum ∼ 140 nm, and the internal/external quantum efficiency and thermal stability of the SASO:0.2Cr were measured to be 68%/37% and 77%@380 K, respectively. An NIR phosphor-converted light-emitting diode (NIR pc-LED) device was fabricated by combining a blue LED chip with the SASO:0.2Cr phosphor. The applications of NIR pc-LED on plant growth promotion, night vision, and medical imaging were demonstrated. We reported an economical broadband NIR phosphor with multiple potential applications and highlighted the crystallographic site engineering strategy to explore broadband phosphors based on aluminates.

2.
Structure ; 31(10): 1220-1232.e5, 2023 Oct 05.
Article in English | MEDLINE | ID: mdl-37652001

ABSTRACT

Structural diverse natural products like ribosomally synthesized and posttranslationally modified peptides (RiPPs) display a wide range of biological activities. Currently, the mechanism of an uncommon reaction step during the biosynthesis of 3-thiaglutamate (3-thiaGlu) is poorly understood. The removal of the ß-carbon from the Cys in the TglA-Cys peptide catalyzed by the TglHI holoenzyme remains elusive. Here, we present three crystal structures of TglHI complexes with and without bound iron, which reveal that the catalytic pocket is formed by the interaction of TglH-TglI and that its activation is conformation dependent. Biochemical assays suggest a minimum of two iron ions in the active cluster, and we identify the position of a third iron site. Collectively, our study offers insights into the activation and catalysis mechanisms of the non-heme dioxygen-dependent holoenzyme TglHI. Additionally, it highlights the evolutionary and structural conservation in the DUF692 family of biosynthetic enzymes that produce diverse RiPPs.


Subject(s)
Iron , Peptides , Peptides/chemistry , Molecular Conformation , Holoenzymes/metabolism , Iron/metabolism , Protein Processing, Post-Translational
3.
Inorg Chem ; 62(25): 10021-10028, 2023 Jun 26.
Article in English | MEDLINE | ID: mdl-37312253

ABSTRACT

The discovery of violet-excitable blue-emitting phosphor is a significant breakthrough for the development of phosphor-converted full-spectrum white light-emitting diodes (WLEDs). However, the application of most known violet-excitable blue-emitting phosphors is limited by their low external quantum efficiency (EQE). In this work, we reported on how the EQE values of Eu2+-doped Ba(K)-ß-Al2O3 blue-emitting phosphor can be significantly improved through lattice site engineering. By partially substituting K+ for Ba2+, the Eu2+-occupied crystallographic site changes and the coordination polyhedron of Eu2+ shrinks, leading to the increase of crystal field splitting. Consequently, the excitation spectrum exhibits a continuous red shift to match the violet excitation, which enhances the PL intensity of solid solution phosphor (Ba0.4K1.6)0.84Al22O35-α:0.32Eu2+ ((B0.4K1.6)0.84AO:Eu) by 1.42 times compared to that of the end-member Ba1.68Al22O35-α:0.32Eu2+ (B1.68AO:Eu) phosphor. Correspondingly, under the 400 nm violet light excitation, the EQE of optimal blue-emitting (B0.4K1.6)0.84AO:Eu phosphor is up to 53%. Additionally, the phosphor also shows excellent resistance to luminescence thermal quenching (95% at 150 °C). Finally, the WLED fabricated based on (B0.4K1.6)0.84AO:Eu and commercial green and red phosphors exhibited an ultra-high color rending index with Ra = 95.5 and R1-R15 >90. This work offers guidance for tuning the spectral properties of phosphors through lattice site engineering.

4.
Opt Express ; 31(9): 14495-14508, 2023 Apr 24.
Article in English | MEDLINE | ID: mdl-37157313

ABSTRACT

Passive power generation has recently stimulated interest in thermoelectric generators (TEGs) using the radiative cooling mechanism. However, the limited and unstable temperature difference across the TEGs significantly degrades the output performance. In this study, an ultra-broadband solar absorber with a planar film structure is introduced as the hot side of the TEG to increase the temperature difference by utilizing solar heating. This device not only enhances the generation of electrical power but also realizes all-day uninterrupted electrical output due to the stable temperature difference between the cold and hot sides of the TEG. Outdoor experiments show the self-powered TEG obtains maximum temperature differences of 12.67 °C, 1.06 °C, and 5.08 °C during sunny daytime, clear nighttime, and cloudy daytime, respectively, and generates output voltages of 166.2 mV, 14.7 mV, and 95 mV, respectively. Simultaneously, the corresponding output powers of 879.25 mW/m2, 3.85 mW/m2, and 287.27 mW/m2 are produced, achieving 24-hour uninterrupted passive power generation. These findings propose a novel strategy to combine solar heating and outer space cooling by a selective absorber/emitter to generate all-day continuous electricity for unsupervised small devices.

5.
PLoS Pathog ; 19(1): e1011131, 2023 01.
Article in English | MEDLINE | ID: mdl-36701392

ABSTRACT

The rapid emergence of SARS-CoV-2 variants of concern, the complexity of infection, and the functional redundancy of host factors, underscore an urgent need for broad-spectrum antivirals against the continuous COVID-19 pandemic, with drug repurposing as a viable therapeutic strategy. Here we report the potential of RNA G-quadruplex (RG4)-targeting therapeutic strategy for SARS-CoV-2 entry. Combining bioinformatics, biochemical and biophysical approaches, we characterize the existence of RG4s in several SARS-CoV-2 host factors. In silico screening followed by experimental validation identify Topotecan (TPT) and Berbamine (BBM), two clinical approved drugs, as RG4-stabilizing agents with repurposing potential for COVID-19. Both TPT and BBM can reduce the protein level of RG4-containing host factors, including ACE2, AXL, FURIN, and TMPRSS2. Intriguingly, TPT and BBM block SARS-CoV-2 pseudovirus entry into target cells in vitro and murine tissues in vivo. These findings emphasize the significance of RG4 in SARS-CoV-2 pathogenesis and provide a potential broad-spectrum antiviral strategy for COVID-19 prevention and treatment.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Animals , Mice , SARS-CoV-2/metabolism , RNA , Pandemics , Antiviral Agents/metabolism , Virus Internalization , Spike Glycoprotein, Coronavirus
6.
Mol Biomed ; 3(1): 48, 2022 Dec 22.
Article in English | MEDLINE | ID: mdl-36547804

ABSTRACT

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is a tenacious pathogen that has latently infected one third of the world's population. However, conventional TB treatment regimens are no longer sufficient to tackle the growing threat of drug resistance, stimulating the development of innovative anti-tuberculosis agents, with special emphasis on new protein targets. The Mtb genome encodes ~4000 predicted proteins, among which many enzymes participate in various cellular metabolisms. For example, more than 200 proteins are involved in fatty acid biosynthesis, which assists in the construction of the cell envelope, and is closely related to the pathogenesis and resistance of mycobacteria. Here we review several essential enzymes responsible for fatty acid and nucleotide biosynthesis, cellular metabolism of lipids or amino acids, energy utilization, and metal uptake. These include InhA, MmpL3, MmaA4, PcaA, CmaA1, CmaA2, isocitrate lyases (ICLs), pantothenate synthase (PS), Lysine-ε amino transferase (LAT), LeuD, IdeR, KatG, Rv1098c, and PyrG. In addition, we summarize the role of the transcriptional regulator PhoP which may regulate the expression of more than 110 genes, and the essential biosynthesis enzyme glutamine synthetase (GlnA1). All these enzymes are either validated drug targets or promising target candidates, with drugs targeting ICLs and LAT expected to solve the problem of persistent TB infection. To better understand how anti-tuberculosis drugs act on these proteins, their structures and the structure-based drug/inhibitor designs are discussed. Overall, this investigation should provide guidance and support for current and future pharmaceutical development efforts against mycobacterial pathogenesis.

7.
Front Oncol ; 12: 929233, 2022.
Article in English | MEDLINE | ID: mdl-36033536

ABSTRACT

Glioma is the most common and fatal primary brain tumor in humans. A significant role for long non-coding RNA (lncRNA) in glioma is the regulation of gene expression and chromatin recombination, and immunotherapy is a promising cancer treatment. Therefore, it is necessary to identify necroptosis-related lncRNAs in glioma. In this study, we collected and evaluated the RNA-sequencing (RNA-seq) data from The Cancer Genome Atlas (TCGA, https://www.ncbi.nlm.nih.gov/, Data Release 32.0, March 29, 2022) glioma patients, and necroptosis-related lncRNAs were screened. Cox regression and least absolute shrinkage and selection operator (LASSO) analysis were performed to construct a risk score formula to explore the different overall survival between high- and low-risk groups in TCGA. Gene Ontology (GO) and pathway enrichment analysis (Kyoto Encyclopedia of Genes and Genomes (KEGG)) were performed to identify the function of screened genes. The immune correlation analysis showed that various immune cells and pathways positively associated with a patient's risk score. Furthermore, the analysis of the tumor microenvironment indicated many immune cells and stromal cells in the tumor microenvironment of glioma patients. Six necroptosis-related lncRNAs were concerned to be involved in survival and adopted to construct the risk score formula. The results showed that patients with high-risk scores held poor survival in TCGA. Compared with current clinical data, the area under the curve (AUC) of different years suggested that the formula had better predictive power. We verified that necroptosis-related lncRNAs play a significant role in the occurrence and development of glioma, and the constructed risk model can reasonably predict the prognosis of glioma. The results of these studies added some valuable guidance to understanding glioma pathogenesis and treatment, and these necroptosis-related lncRNAs may be used as biomarkers and therapeutic targets for glioma prevention.

8.
Chin Neurosurg J ; 8(1): 12, 2022 May 19.
Article in English | MEDLINE | ID: mdl-35585639

ABSTRACT

BACKGROUND: Low-grade gliomas (LGG) are WHO grade II tumors presenting as the most common primary malignant brain tumors in adults. Currently, LGG treatment involves either or a combination of surgery, radiation therapy, and chemotherapy. Despite the knowledge of constitutive genetic risk factors contributing to gliomas, the role of single genes as diagnostic and prognostic biomarkers is limited. The aim of the current study is to discover the predictive and prognostic genetic markers for LGG. METHODS: Transcriptome data and clinical data were obtained from The Cancer Genome Atlas (TCGA) database. We first performed the tumor microenvironment (TME) survival analysis using the Kaplan-Meier method. An analysis was undertaken to screen for differentially expressed genes. The function of these genes was studied by Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Following which a protein-protein interaction network (PPI) was constructed and visualized. Univariate and multivariate COX analyses were performed to obtain the probable prognostic genes. The key genes were selected by an intersection of core and prognostic genes. A clinical correlation analysis of single-gene expression was undertaken. GSEA enrichment analysis was performed to identify the function of key genes. Finally, a single gene-related correlation analysis was performed to identify the core immune cells involved in the development of LGG. RESULTS: A total of 529 transcriptome data and 515 clinical samples were obtained from the TCGA. Immune cells and stromal cells were found to be significantly increased in the LGG microenvironment. The top five core genes intersected with the top 38 prognostically relevant genes and two key genes were identified. Our analysis revealed that a high expression of HLA-DRA was associated with a poor prognosis of LGG. Correlation analysis of immune cells showed that HLA-DRA expression level was related to immune infiltration, positively related to macrophage M1 phenotype, and negatively related to activation of NK cells. CONCLUSIONS: HLA-DRA may be an independent prognostic indicator and an important biomarker for diagnosing and predicting survival in LGG patients. It may also be associated with the immune infiltration phenotype in LGG.

9.
MedComm (2020) ; 3(1): e99, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35281791

ABSTRACT

Listeria monocytogenes, a food-borne Gram-positive pathogen, often causes diseases such as gastroenteritis, bacterial sepsis, and meningitis. Newly discovered extracellular electron transfer (EET) from L. monocytogenes plays critical roles in the generation of redox molecules as electron carriers in bacteria. A Mg2+-dependent protein flavin mononucleotide (FMN) transferase (FmnB; UniProt: LMRG_02181) in EET is responsible for the transfer of electrons from intracellular to extracellular by hydrolyzing cofactor flavin adenine dinucleotide (FAD) and transferring FMN. FmnB homologs have been investigated in Gram-negative bacteria but have been less well studied in Gram-positive bacteria. In particular, the catalytic and inhibitory mechanisms of FmnB homologs remain elusive. Here, we report a series of crystal structures of apo-FmnB and FmnB complexed with substrate FAD, three inhibitors AMP, ADP, and ATP, revealing the unusual catalytic triad center (Asp301-Ser257-His273) of FmnB. The three inhibitors indeed inhibited the activity of FmnB in varying degrees by occupying the binding site of the FAD substrate. The key residue Arg262 of FmnB was profoundly affected by ADP but not AMP or ATP. Overall, our studies not only provide insights into the promiscuous ligand recognition behavior of FmnB but also shed light on its catalytic and inhibitory mechanisms.

10.
Cell Res ; 32(3): 302-314, 2022 03.
Article in English | MEDLINE | ID: mdl-35110668

ABSTRACT

Methanobactins (Mbns) are a family of copper-binding peptides involved in copper uptake by methanotrophs, and are potential therapeutic agents for treating diseases characterized by disordered copper accumulation. Mbns are produced via modification of MbnA precursor peptides at cysteine residues catalyzed by the core biosynthetic machinery containing MbnB, an iron-dependent enzyme, and MbnC. However, mechanistic details underlying the catalysis of the MbnBC holoenzyme remain unclear. Here, we present crystal structures of MbnABC complexes from two distinct species, revealing that the leader peptide of the substrate MbnA binds MbnC for recruitment of the MbnBC holoenzyme, while the core peptide of MbnA resides in the catalytic cavity created by the MbnB-MbnC interaction which harbors a unique tri-iron cluster. Ligation of the substrate sulfhydryl group to the tri-iron center achieves a dioxygen-dependent reaction for oxazolone-thioamide installation. Structural analysis of the MbnABC complexes together with functional investigation of MbnB variants identified a conserved catalytic aspartate residue as a general base required for MbnBC-mediated MbnA modification. Together, our study reveals the similar architecture and function of MbnBC complexes from different species, demonstrating an evolutionarily conserved catalytic mechanism of the MbnBC holoenzymes.


Subject(s)
Copper , Iron , Catalysis , Copper/metabolism , Holoenzymes/chemistry , Imidazoles , Oligopeptides
11.
Nurs Forum ; 57(1): 26-33, 2022 Jan.
Article in English | MEDLINE | ID: mdl-34473356

ABSTRACT

Psychological problems have become a significant public health problem. Appropriate mental health care is crucial in promoting patient care quality. This study aimed to test the feasibility of a Psychological Nursing Quality Evaluation Index in hospitalized patients. This is a pilot study with patients hospitalized with myocardial infarction from July to September 2020 in China. The researchers used an observational approach to examine nurses' psychological health care performance based on the Psychological Nursing Quality Evaluation Index. The results indicated high compliance rates of nurses' psychological care performance, which provides references for evaluating and monitoring inpatient psychological nursing care.


Subject(s)
Nursing Staff, Hospital , Quality of Health Care , China , Humans , Pilot Projects
12.
Appl Opt ; 60(32): 10176-10185, 2021 Nov 10.
Article in English | MEDLINE | ID: mdl-34807126

ABSTRACT

A compact square photonic crystal fiber polarization filter with high performance is proposed. Two larger holes filled with different metal and fluid are designed to break the strict structure symmetry and forming high birefringence. Four small pores are designed to provide a wider channel for the coupling between defect mode and core mode. Its filtering and coupling characteristics are analyzed by the full vector finite element method. In addition, the properties of PCF filled with different materials are compared and discussed. Different metal materials have different dielectric constants, different optical damping, interband transitions, molecular structure, and physicochemical properties, which make their transmission modes and coupling strength different. The results show that the performance of a PCF filter filled with gold and liquid is the best, which is very suitable for an adjustable PCF polarization filter. The calculation results show that the extinction ratio of the designed filter can reach 5383 dB for a device length of 3 mm; the unwanted fiber loss can reach 2073 dB/cm; and the applicable bandwidth of 2000 nm covers almost the whole communication band. The proposed polarization filter shows large unwanted loss, high extinction ratio, wide bandwidth and coordination, which make it a good candidate for excellent optical fiber filter devices.

13.
Opt Express ; 29(23): 37234-37244, 2021 Nov 08.
Article in English | MEDLINE | ID: mdl-34808800

ABSTRACT

Mid-infrared absorption spectroscopy is an effective method for detecting analyte fingerprints without labeling, but the inherent loss of metals in current methods is a main issue. Here, a sensing scheme was proposed that uses an all-dielectric grating metasurface and angular scanning of polarized light, and then it was verified by numerical simulation. The proposed fingerprint detection scheme could effectively couple a guided-mode resonance spectrum peak with the characteristic peak of the analyte's phonon-polariton in the mid-infrared region, significantly enhancing the interaction between light and the analyte. The novel scheme would realize broadband enhancement to detect a variety of substances, and facilitate mid-infrared sensing and analysis of trace substances.

14.
Article in English | MEDLINE | ID: mdl-31149354

ABSTRACT

S-adenosyl-1-methionine (SAM)-dependent enzymes regulate various disease-related behaviors in all organisms. Recently, the leporin biosynthesis enzyme LepI, a SAM-dependent enzyme, was reported to catalyze pericyclic reactions in leporin biosynthesis; however, the mechanisms underlying LepI activation and catalysis remain unclear. This study aimed to investigate the molecular mechanisms of LepI. Here, we reported crystal structures of LepI bound to SAM/5'-deoxy-5'-(methylthio) adenosine (MTA), S-adenosyl-homocysteine (SAH), and SAM/substrate states. Structural and biochemical analysis revealed that MTA or SAH inhibited the enzyme activities, whereas SAM activated the enzyme. The analysis of the substrate-bound structure of LepI demonstrated that this enzymatic retro-Claisen rearrangement was primarily driven by three critical polar residues His133, Arg197, Arg295 around the active site and assisted by SAM with unclear mechanism. The present studies indicate that the unique mechanisms underlying regulatory and catalysis of the unusual SAM-dependent enzyme LepI, not only strengthening current understanding of the fundamentally biochemical catalysis, but also providing novel insights into the design of SAM-dependent enzyme-specific small molecules.

15.
FEBS J ; 286(4): 750-764, 2019 02.
Article in English | MEDLINE | ID: mdl-30537137

ABSTRACT

Gram-negative bacteria defend against the toxicity of polymyxins by modifying their outer membrane lipopolysaccharide (LPS). This modification mainly occurs through the addition of cationic molecules such as phosphoethanolamine (PEA). EcEptC is a PEA transferase from Escherichia coli (E. coli). However, unlike its homologs CjEptC (Campylobacter jejuni) and MCR-1, EcEptC is unable to mediate polymyxin resistance when overexpressed in E. coli. Here, we report crystal structures of the C-terminal putative catalytic domain (EcEptCΔN, 205-577 aa) of EcEptC in apo and Zn2+ -bound states at 2.10 and 2.60 Å, respectively. EcEptCΔN is arranged into an α-ß-α fold and equipped with the zinc ion in a conserved mode. Coupled with isothermal titration calorimetry (ITC) data, we provide insights into the mechanism by which EcEptC recognizes Zn2+ . Furthermore, structure comparison analysis indicated that disulfide bonds, which play a key role in polymyxin resistance, were absent in EcEptCΔN. Supported by structural and biochemical evidence, we reveal mechanistic implications for disulfide bonds in PEA transferase-mediated polymyxin resistance. Significantly, because the structural effects exhibited by disulfide bonds are absent in EcEptC, it is impossible for this protein to participate in polymyxin resistance in E. coli. DATABASE: Structural data are available in the PDB under the accession numbers 6A82 and 6A83. ENZYME: EC 2.7.8.43.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Escherichia coli Proteins/chemistry , Escherichia coli Proteins/metabolism , Escherichia coli/enzymology , Membrane Proteins/chemistry , Membrane Proteins/metabolism , Polymyxins/pharmacology , Catalytic Domain , Crystallography, X-Ray , Disulfides/metabolism , Escherichia coli Proteins/genetics , Membrane Proteins/genetics , Mutagenesis, Site-Directed , Mutation , Protein Conformation , Zinc/metabolism
16.
iScience ; 3: 1-10, 2018 May 25.
Article in English | MEDLINE | ID: mdl-30428313

ABSTRACT

A clustered regularly interspaced short palindromic repeats (CRISPR)-like "mimivirus virophage resistance element" (MIMIVIRE) system, which contains specific cascade genes and a CRISPR array against virophages, was reported in mimiviruses. An essential component of the MIMIVIRE system is R354, encoding a nuclease and a likely functional homolog of Cas4. Here we show that R354 is a dual nuclease with both exonuclease and endonuclease activities. Structural analysis revealed that the catalytic core domain of R354 is similar to those of Cas4 and ? exonuclease despite their low sequence identity. R354 forms a homodimer that is important for its exonuclease but not endonuclease activity. Structural comparisons between the active and semi-active states of R354 demonstrated that an activation loop adjacent to the catalytic site is critical for enzymatic activity. Overall, the results suggest that R354 belongs to a novel MIMIVIRE system involved in innate virus immunity and provides a template for the identification of new CRISPR systems in other species.

17.
Nat Microbiol ; 3(11): 1285-1294, 2018 11.
Article in English | MEDLINE | ID: mdl-30323253

ABSTRACT

Communication is vital for all organisms including microorganisms, which is clearly demonstrated by the bacterial quorum-sensing system. However, the molecular mechanisms underlying communication among viruses (phages) via the quorum-sensing-like 'arbitrium' system remain unclear. Viral or host densities are known to be related to an increased prevalence of lysogeny; however, how the switch from the lytic to the lysogenic pathway occurs is unknown. Thus, we sought to reveal mechanisms of communication among viruses and determine the lysogenic dynamics involved. Structural and functional analyses of the phage-derived SAIRGA and GMPRGA peptides and their corresponding receptors, phAimR and spAimR, indicated that SAIRGA directs the lysis-lysogeny decision of phi3T by modulating conformational changes in phAimR, whereas GMPRGA regulates the lysis-lysogeny pathway by stabilizing spAimR in the dimeric state. Although temperate viruses are thought to share a similar lytic-lysogenic cycle switch model, our study suggests the existence of alternative strain-specific mechanisms that regulate the lysis-lysogeny decision. Collectively, these findings provide insights into the molecular mechanisms underlying communication among viruses, offering theoretical applications for the treatment of infectious viral diseases.


Subject(s)
Bacillus Phages/physiology , Bacteriolysis , Lysogeny , Viral Proteins/metabolism , Amino Acid Sequence , Bacillus Phages/drug effects , Bacillus subtilis/cytology , Bacillus subtilis/virology , Bacteriolysis/drug effects , Binding Sites , Crystallography, X-Ray , Lysogeny/drug effects , Models, Biological , Peptides/chemistry , Peptides/metabolism , Peptides/pharmacology , Species Specificity , Structure-Activity Relationship , Viral Proteins/chemistry
18.
PeerJ ; 6: e4728, 2018.
Article in English | MEDLINE | ID: mdl-29761048

ABSTRACT

Type I polyketide synthase 13 (Pks13) is involved in the final step of the biosynthesis of mycolic acid in Mycobacterium tuberculosis. Recent articles have reported that Pks13 is an essential enzyme in the mycolic acid biosynthesis pathway, and it has been deeply studied as a drug target in Tuberculosis. We report a high-resolution structure of the acyltransferase (AT) domain of Pks13 at 2.59 Å resolution. Structural comparison with the full-length AT domain (PDB code, 3TZW, and 3TZZ) reveals a different orientation of the C-terminal helix and rearrangement of some conserved residues.

19.
Clin Biochem ; 55: 28-35, 2018 May.
Article in English | MEDLINE | ID: mdl-29596792

ABSTRACT

BACKGROUND: Vitamin D is a lipid-soluble molecule that plays key physiological roles in the metabolism of calcium, phosphate and magnesium. Recent studies show that deficiency in vitamin D is linked to cardiovascular diseases, autoimmune diseases and cancer. As a result, regular monitoring of 25-OH vitamin D (the main circulating form of vitamin D) is becoming essential. Current 25-OH vitamin D testing methodologies are cumbersome (too many reagents, long incubation times, phase separation) and are not compatible with general clinical chemistry platforms. Here, we report on a novel method to detect 25-OH vitamin D that is fast (results in 10 min or less), simple (two reagents) and compatible with virtually all general clinical chemistry analyzers. METHODS: An immunoturbidimetric assay for 25-OH vitamin D (the Diazyme EZ Vitamin D Assay) has been developed using nanoparticles and vitamin D-specific antibodies. The performance of the assay kit, which consists of two reagents and five calibrators, was tested on the Beckman AU680 analyzer (AU680). RESULTS: The new assay was precise, sensitive (LOD = 7.2 nmol/L), linear (up to 390.1 nmol/L) and correlated strongly (R2 > 0.95) with major commercial 25-OH vitamin D assays. Additionally, the assay was found to be the fastest to date, with the first results obtained within 10 min. Throughput on the AU680 was estimated at over 300 tests per hour. CONCLUSIONS: The newly developed 25-OH vitamin D assay is fast, precise and accurate. It can be run on most general chemistry analyzers. This assay aims at providing vitamin D-testing capabilities to all clinical chemistry laboratories.


Subject(s)
Antibodies/chemistry , Immunoturbidimetry , Nanoparticles/chemistry , Vitamin D/analogs & derivatives , Female , Humans , Immunoturbidimetry/instrumentation , Immunoturbidimetry/methods , Male , Sensitivity and Specificity , Vitamin D/blood
20.
Sci Bull (Beijing) ; 63(11): 691-699, 2018 Jun 15.
Article in English | MEDLINE | ID: mdl-36658818

ABSTRACT

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR-associated (Cas) system is an adaptive immune system in bacteria and archaea that resists exogenous invasion through nucleic acid-mediated cleavage. In the type III-A system, the Csm complex contains five effectors and a CRISPR RNA, which edits both single stranded RNA and double stranded DNA. It has recently been demonstrated that cyclic oligoadenylates (cOAs), which are synthesized by the Csm complex, act as second messengers that bind and activate Csm6. Here, we report the crystal structures of Staphylococcus epidermidis Csm3 (SeCsm3) and an N-terminally truncated Csm6 (SeCsm6ΔN) at 2.26 and 2.0 Å, respectively. The structure of SeCsm3 highly resembled previously reported Csm3 structures from other species; however, it provided novel observations allowing further enzyme characterization. The homodimeric SeCsm6ΔN folds into a compact structure. The dimerization of the HEPN domain leads to the formation of the ribonuclease active site, which is consistent with the reported Csm6 structures. Altogether, our studies provide a structural view of the ribonuclease activity mediated by Csm3 and Csm6 of the type III-A CRISPR-Cas system.

SELECTION OF CITATIONS
SEARCH DETAIL