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Archaeal ribosomes have many domain-specific features; however, our understanding of these structures is limited. We present 10 cryo-electron microscopy (cryo-EM) structures of the archaeal ribosome from crenarchaeota Sulfolobus acidocaldarius (Sac) at 2.7-5.7 Å resolution. We observed unstable conformations of H68 and h44 of ribosomal RNA (rRNA) in the subunit structures, which may interfere with subunit association. These subunit structures provided models for 12 rRNA expansion segments and 3 novel r-proteins. Furthermore, the 50S-aRF1 complex structure showed the unique domain orientation of aRF1, possibly explaining P-site transfer RNA (tRNA) release after translation termination. Sac 70S complexes were captured in seven distinct steps of the tRNA translocation reaction, confirming conserved structural features during archaeal ribosome translocation. In aEF2-engaged 70S ribosome complexes, 3D classification of cryo-EM data based on 30S head domain identified two new translocation intermediates with 30S head domain tilted 5-6° enabling its disengagement from the translocated tRNA and its release post-translocation. Additionally, we observed conformational changes to aEF2 during ribosome binding and switching from three different states. Our structural and biochemical data provide new insights into archaeal translation and ribosome translocation.
Archaeal ribosomes display variations in their ribosomal proteins and ribosomal RNA (rRNA) expansion segments (ESs). Protein translation in archaea combines features in both bacterial and eukaryotic translation. In this study, we present 10 cryo-electron microscopy structures of the archaeal ribosome from crenarchaeota Sulfolobus acidocaldarius (Sac). The 50S and 30S subunit structures present 3 novel ribosomal proteins and 12 rRNA ESs. The 70S Sac ribosome structures were captured in seven distinct functional states, including pre-, intermediate- and post-translocation states. Specifically, we identified two novel translocation intermediates, in which the 30S subunit head domain tilts outward to release the translocated P-site transfer RNA. The structures of archaeal ribosomes provide insights into the archaeal translation and ribosome translocation.
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Ribossomos , Sulfolobus acidocaldarius , Microscopia Crioeletrônica , Proteínas Ribossômicas/metabolismo , Ribossomos/metabolismo , RNA Ribossômico/metabolismo , RNA de Transferência/metabolismo , Sulfolobus acidocaldarius/citologia , Sulfolobus acidocaldarius/metabolismoRESUMO
Ventricular septal defect (VSD) is the most common type of congenital heart disease. HAND1 gene plays a crucial role in the development of the heart, but the role of the variants in the HAND1 gene promoter region in patients with VSD has not been explored yet. From 588 participants (300 with isolated and sporadic VSD and 288 healthy controls), DNA was extracted from blood samples. Variants at the HAND1 gene promoter region were analyzed through Sanger sequencing. Subsequently, cell functional validation was conducted through cell experiments, including dual-luciferase reporter gene analysis, electrophoretic mobility shift analysis, and bioinformatics analysis was also conducted. The promoter region of HAND1 gene had a total of 9 identified variant sites. Among them, 4 variants were exclusively found in VSD patients, and 1 variant (g.3631A>C) was newly discovered. Cell functional experiments indicated that all four variants decreased the transcriptional activity of HAND1 gene promoter with three of them reached statistical significance (p < 0.05). Subsequent analysis using JASPAR (a transcription factor binding profile database) suggests that these variants may alter the binding sites of transcription factors, potentially contributing to the formation of VSD. Our study for the first time identified variants in the promoter region of HAND1 gene in Chinese patients with isolated and sporadic VSD. These variants significantly decreased the expression of HAND1 gene, impacting transcription factor binding sites, and thereby demonstrating pathogenicity. This study offers new insights into the role of HAND1 gene promoter region, contributing to a better understanding of the genetic basis of VSD formation.
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Patent ductus arteriosus (PDA) is a common congenital heart disease. CITED2 plays an important role in the development of the heart, and genetic variants in its coding region are significantly associated with cardiac malformations. However, the role of variants in the promoter region of CITED2 in the development of PDA remains unclear. We extracted the peripheral blood of 646 subjects (including 353 PDA patients and 293 unrelated healthy controls) for sequencing. We identified 13 promoter variants of the CITED2 gene (including 2 novel heterozygous variants). Of the 13 variants, 10 were found only in PDA patients. In mouse cardiomyocytes (HL-1) and rat cardiac myocytes (RCM), the transcriptional activity of the CITED2 gene promoter was significantly changed by the variants (p < 0.05). The results of the experiments of electrophoretic mobility indicated that these variants may affect the transcription of the CITED2 gene by influencing the binding ability of transcription factors. These results, combined with the JASPAR database analysis, showed that the destruction/production of transcription factor binding sites due to the variants in the promoter region of the CITED2 gene may directly or indirectly affect the binding ability of transcription factors. Our results suggest for the first time that variants at the CITED2 promoter region may cause low expression of CITED2 protein related to the formation of PDA.
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Permeabilidade do Canal Arterial , Cardiopatias Congênitas , Humanos , Animais , Camundongos , Ratos , Permeabilidade do Canal Arterial/genética , Permeabilidade do Canal Arterial/metabolismo , Cardiopatias Congênitas/genética , Fatores de Transcrição/genética , Miócitos Cardíacos/metabolismo , Regiões Promotoras Genéticas , Proteínas Repressoras/genética , Transativadores/genética , Transativadores/metabolismoRESUMO
Tetralogy of Fallot (TOF) is the most common cyanotic congenital heart disease. Highly penetrant copy number variants (CNVs) and genes related to the etiology of TOF likely exist with differences among populations. We aimed to identify CNV contributions to sporadic TOF cases in Han Chinese. Genomic DNA was extracted from peripheral blood in 605 subjects (303 sporadic TOF and 302 unaffected Han Chinese [Control] from cardiac centers in China) and analyzed by genome-wide association study (GWAS). The GWAS results were compared with existing Database of Genetic Variants. These CNVs were further validated by qPCR. Bioinformatics analyses were performed with protein-protein interaction (PPI) network and KEGG pathway enrichment. Across all chromosomes 119 novel "TOF-specific CNVs" were identified with prevalence of CNVs of 21.5% in chromosomes 1-20 and 37.0% including Chr21/22. In chromosomes 1-20, CNVs on 11q25 (encompasses genes ACAD8, B3GAT1, GLB1L2, GLB1L3, IGSF9B, JAM3, LOC100128239, LOC283177, MIR4697, MIR4697HG, NCAPD3, OPCML, SPATA19, THYN1, and VPS26B) and 14q32.33 (encompasses genes THYN1, OPCML, and NCAPD3) encompass genes most likely to be associated with TOF. Specific CNVs found on the chromosome 21 (6.3%) and 22(11.9%) were also identified in details. PPI network analysis identified the genes covering the specific CNVs related to TOF and the signaling pathways. This study for first time identified novel TOF-specific CNVs in the Han Chinese with higher frequency than in Caucasians and with 11q25 and 14q32.33 not reported in TOF of Caucasians. These novel CNVs identify new candidate genes for TOF and provide new insights into genetic basis of TOF.
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Variações do Número de Cópias de DNA , Tetralogia de Fallot , Povo Asiático/genética , Moléculas de Adesão Celular/genética , DNA , Variações do Número de Cópias de DNA/genética , Proteínas Ligadas por GPI/genética , Estudo de Associação Genômica Ampla , Humanos , Tetralogia de Fallot/genéticaRESUMO
2-Deoxycytidylate deaminase (dCD) is a member of the zinc-dependent cytidine deaminase family features in its allosterically regulated mechanism by dCTP and dTTP. The large double-stranded DNA-containing chlorovirus PBCV-1 encodes a dCD family enzyme PBCV1dCD that was reported to be able to deaminize both dCMP and dCTP, which makes PBCV1dCD unique in the dCD family proteins. In this study, we report the crystal structure of PBCV1dCD in complex with dCTP/dCMP and dTTP/dTMP, respectively. We further proved the ability of PBCV1dCD in the deamination of dCDP, which makes PBCV1dCD a multi-functional deaminase. The structural basis for the versatility of PBCV1dCD is analyzed and discussed, with the finding of a unique Trp121 residue key to the deamination and substrate binding ability. Our findings may broaden the understanding of dCD family proteins and provide novel insights into the multi-functional enzyme.
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DCMP Desaminase , Desoxicitidina Monofosfato , Cristalografia por Raios X , DCMP Desaminase/química , DCMP Desaminase/metabolismo , Especificidade por SubstratoRESUMO
Trace elements selenium (Se) and cobalt (Co) are essential in the human body, and a correlation between Se and cardiac surgery has been suggested. We investigated the plasma concentrations of Se and Co during and after coronary artery bypass grafting (CABG) surgery under cardiopulmonary bypass (CPB). From December 2019 to January 2020, preoperative plasma samples from isolated first-time CABG patients (n=20; 10 males, 10 females) were prospectively collected post-anesthesia and before CPB (T1), 45 min after CPB started (T2), 90 min after CPB started (T3), and postoperative days 1 (T4), and day 4 (T5). The plasma concentrations of Se and Co were measured. The Se concentration was significantly decreased at T2 (105.24±4.08 vs. 68.56±2.42 µg/L, p<0.001) and T3 (105.24±4.08 vs. 80.41±3.40 µg/L, p<0.001). The Co concentration was significantly decreased at T4 (0.35±0.19 vs. 0.26±0.13 µg/L, p<0.01) and T5 (0.35±0.19 vs. 0.23±0.11 µg/L, p<0.001). Five patients developed atrial fibrillation (AF); there was no other operative mortality or major morbidity. This is the first report of alterations of plasma Se and Co concentrations during and after CABG surgery. Our results may indicate that Se supplementation before or during CABG and Co supplementation after CABG may become necessary for patients undergoing CABG.
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Cobalto/sangue , Ponte de Artéria Coronária , Selênio/sangue , Oligoelementos/sangue , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
OBJECTIVE: To analyze the value of postoperative human leukocyte antigen-DR (HLA-DR) expression and high mobility group box 1 (HMGB1) level in predictive diagnosis of postoperative sepsis for patients with percutaneous nephrolithotomy (PCNL) surgery. METHODS: The present prospective observational study included 387 patients with renal calculus who received PCNL surgery from January 2017 to October 2020 in our hospital. After exclusion criteria, 33 patients with sepsis and 78 patients with no sepsis remained. All patients received PCNL surgery. Sepsis definition is according to the third international consensus definitions for sepsis and septic shock (Sepsis-3). The data of the HMGB1, c-reactive protein (CRP), interleukin-6 (IL-6), procalcitonin (PCT) and HLA-DR expression were collected within admission and 24âh and 72 h after surgery. Postoperative HMGB1 levels and HLA-DR expression at 24 h and 72 h were respectively compared between the two groups using t test. ROC cure was used to analyze the value of postoperative HLA-DR expression and HMGB1 level in predictive diagnosis of sepsis. RESULTS: The positive rate of urine culture and the time of hospitalization time in patients with sepsis were significantly higher than those in patients with no sepsis. Sepsis group had higher levels of HMGB1 at post-24 h ((93.07 ± 11.37) ng/mL vs (75.41 ± 4.85) ng/mL), p < 0.05) and 72 h ((96.58 ± 12.12) ng/mL vs (81.16 ± 8.86) ng/mL), p < 0.05) than nosepsis group. Meanwhile, sepsis group had lower expression of HLA-DR at post-24 h ((50.01 ± 7.42) % vs (69.32 ± 10.58) %), p < 0.05) and 72 h ((54.85 ± 9.45) % vs (69.98 ± 11.00) %), p < 0.05) than non-sepsis group. ROC analysis showed that the HLA-DR expression at postoperative 24 h had highest predictive value in the diagnosis of sepsis, the AUC of HLA-DR was 0.934, cutoff value 56.19%, with sensitivity 89.7%, specificity 81.8%. CONCLUSION: Postoperative HLA-DR and HMGB1 can both be used as a predictive diagnosis of sepsis for patients with renal calculus received PCNL surgery. HighlightsSepsis group had higher levels of high mobility group box 1 at post-24 h and 72 h than nosepsis group.Sepsis group had lower expression of HLA-DR at post-24 h and 72 h than nosepsis group.Postoperative HLA-DR and HMGB1 can both be used as a predictive diagnosis of sepsis for patients with renal calculus received PCNL surgery.
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Proteína HMGB1 , Cálculos Renais , Nefrolitotomia Percutânea , Sepse , Antígenos HLA-DR , Humanos , Cálculos Renais/diagnóstico , Cálculos Renais/cirurgia , Sepse/diagnósticoRESUMO
Owing to their numerous nutritional and bioactive functions, phospholipids (PLs), which are major components of biological membranes in all living organisms, have been widely applied as nutraceuticals, food supplements, and cosmetic ingredients. To date, PLs are extracted solely from soybean or egg yolk, despite the diverse market demands and high cost, owing to a tedious and inefficient manufacturing process. A microbial-based manufacturing process, specifically phospholipase D (PLD)-based biocatalysis and biotransformation process for PLs, has the potential to address several challenges associated with the soybean- or egg yolk-based supply chain. However, poor enzyme properties and inefficient microbial expression systems for PLD limit their wide industrial dissemination. Therefore, sourcing new enzyme variants with improved properties and developing advanced PLD expression systems are important. In the present review, we systematically summarize recent achievements and trends in the discovery, their structural properties, catalytic mechanisms, expression strategies for enhancing PLD production, and its multiple applications in the context of PLs. This review is expected to assist researchers to understand current advances in this field and provide insights for further molecular engineering efforts toward PLD-mediated bioprocessing.
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Fosfolipase D , Biocatálise , Catálise , Fosfolipase D/genética , Fosfolipase D/metabolismo , Fosfolipídeos , Glycine maxRESUMO
Neurological dysfunction commonly occurs after cardiac surgery with deep hypothermic circulatory arrest (DHCA). The mechanisms underlying DHCA-associated brain injury remain poorly understood. This study determined the changes in expression profiles of circular RNAs (circRNAs) in the hippocampus in rats that underwent DHCA, with an attempt to explore the potential role of circRNAs in the brain injury associated with DHCA. Adult male Sprague Dawley rats were subjected to cardiopulmonary bypass with DHCA. Brain injury was evaluated by neurological severity scores and histological as well as transmission electron microscope examinations. The expression profiles of circRNAs in the hippocampal tissues were screened by microarray. Quantitative real-time PCR (RT-qPCR) was used to validate the reliability of the microarray results. Bioinformatic algorithms were applied to construct a competing endogenous RNA (ceRNA) network, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were performed to explore the potential biological roles of the circRNAs. Out of 14 145 circRNAs screened, 56 were differentially expressed in the hippocampus between the DHCA and sham-operated rats, including 30 upregulated and 26 downregulated circRNAs. The expression changes of six selected circRNAs (upregulated: rno_circRNA_011190, rno_circRNA_012988, rno_circRNA_000544; downregulated: rno_circRNA_010393, rno_circRNA_012043, rno_circRNA_015149) were further confirmed by RT-qPCR. Bioinformatics analysis showed the enrichment of these confirmed circRNAs and their potential target mRNAs in several KEGG pathways including histidine metabolism, adipocytokine signaling, and cAMP signaling. By revealing the change expression profiles of circRNAs in the brain after DHCA, this study indicates possible involvements of these dysregulated circRNAs in brain injury and suggests a potential of targeting circRNAs for prevention and treatment of neurological dysfunction associated with DHCA.
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Parada Circulatória Induzida por Hipotermia Profunda , Hipocampo/metabolismo , RNA Circular/metabolismo , Algoritmos , Animais , Biologia Computacional/métodos , Modelos Animais de Doenças , Masculino , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo RealRESUMO
BACKGROUND: Although it is well known that aging impairs navigation performance, the underlying mechanisms remain largely unknown. Egocentric strategy requires navigators to remember a series of body-turns without relying on the relationship between environmental cues. Previous study suggested that the egocentric strategy, compared with non-egocentric strategy, was relatively unimpaired during aging. In this study, we aimed to examine strategy use during virtual navigation task and the underlying cognitive supporting mechanisms in older adults. METHODS: Thirty young adults and thirty-one older adults were recruited from the local community. This study adapted star maze paradigm using non-immersive virtual environment. Participants moved freely in a star maze with adequate landmarks, and were requested to find a fixed destination. After 9 learning trials, participants were probed in the same virtual star maze but with no salient landmarks. Participants were classified as egocentric or non-egocentric strategy group according to their response in the probe trial. RESULTS: The results revealed that older adults adopting egocentric strategy completed the navigation task as accurate as young adults, whereas older adults using non-egocentric strategy completed the navigation task with more detours and lower accuracy. The relatively well-maintained egocentric strategy in older adults was related to better visuo-spatial ability. CONCLUSIONS: Visuo-spatial ability might play an important role in navigation accuracy and navigation strategy of older adults. This study demonstrated the potential value of the virtual star maze in evaluating navigation strategy and visuo-spatial ability in older adults.
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Navegação Espacial , Idoso , Envelhecimento , Humanos , Aprendizagem em Labirinto , Rememoração MentalRESUMO
Congenital heart disease (CHD) associated with polydactyly involves various genes. We aimed to identify variations from genes related to complex CHD with polydactyly and to investigate the cellular functions related to the mutations. Blood was collected from a complex CHD case with polydactyly, and whole exome sequencing (WES) was performed. The CRISPR/Cas9 system was used to generate human pluripotent stem cell with mutations (hPSCs-Mut) that were differentiated into cardiomyocytes (hPSC-CMs-Mut) and analysed by transcriptomics on day 0, 9 and 13. Two heterozygous mutations, LTBP2 (c.2206G>A, p.Asp736Asn, RefSeq NM_000428.2) and TCTN3 (c.1268G>A, p.Gly423Glu, RefSeq NM_015631.5), were identified via WES but no TBX5 mutations were found. The stable cell lines of hPSCs-LTBP2mu /TCTN3mu were constructed and differentiated into hPSC-CMs-LTBP2mu /TCTN3mu . Compared to the wild type, LTBP2 mutation delayed the development of CMs. The TCTN3 mutation consistently presented lower rate and weaker force of the contraction of CMs. For gene expression pattern of persistent up-regulation, pathways in cardiac development and congenital heart disease were enriched in hPSCs-CM-LTBP2mu , compared with hPSCs-CM-WT. Thus, the heterozygous mutations in TCTN3 and LTBP2 affect contractility (rate and force) of cardiac myocytes and may affect the development of the heart. These findings provide new insights into the pathogenesis of complex CHD with polydactyly.
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Proteínas Adaptadoras de Transdução de Sinal/genética , Proteínas Reguladoras de Apoptose/genética , Predisposição Genética para Doença , Cardiopatias Congênitas/diagnóstico , Cardiopatias Congênitas/genética , Proteínas de Ligação a TGF-beta Latente/genética , Mutação , Polidactilia/genética , Alelos , Biomarcadores , Sistemas CRISPR-Cas , Biologia Computacional/métodos , Análise Mutacional de DNA , Edição de Genes , Perfilação da Expressão Gênica , Estudos de Associação Genética , Humanos , Miócitos Cardíacos/metabolismo , Fenótipo , Células-Tronco Pluripotentes/metabolismo , Radiografia , Ultrassonografia , Sequenciamento do ExomaRESUMO
The molecular mechanisms underlying pulmonary arterial hypertension (PAH) in congenital ventricular septal defects (VSD) are unclear. We aimed to reveal molecular pathways and potential biomarkers by multi-omics analysis in VSD-PAH. Plasma from 160 children, including 120 VSD patients with/without PAH and 40 healthy children was studied by integrated proteomics, metabolomics, and bioinformatics analyses. Proteomics identified 107 differential proteins (DPs) between patients with/without PAH including significantly increased adiponectin (ADIPO), dopamine ß-hydroxylase (DBH), alanyl membrane aminopeptidase (ANPEP), transferrin receptor 1, and glycoprotein Ib platelet α-subunit and decreased guanine nucleotide-binding protein Gs in VSD-PAH. Metabolomics discovered 191 differential metabolites between patients with/without PAH, including elevation of serotonin, taurine, creatine, sarcosine, and 2-oxobutanoate, and decrease of vanillylmandelic acid, 3,4-dihydroxymandelate, 15-keto-prostaglandin F2α, fructose 6-phosphate, l-glutamine, dehydroascorbate, hydroxypyruvate, threonine, l-cystine, and 1-aminocyclopropane-1-carboxylate. The DPs were validated in a new cohort of patients (n = 80). Integrated analyses identified key pathways, including cAMP, ECM receptor interaction, AMPK, hypoxia-inducible factor 1, PI3K-Akt signaling pathways, and amino acid metabolisms. Increased plasma protein levels of DBH, ADIPO, and ANPEP were found to be independently associated with the occurrence of PAH, with a new total risk score from these three proteins developed for clinical diagnosis. In this integrated multi-omics analysis in VSD-PAH patients, we have, for the first time, found that VSD-PAH patients present important differential proteins, metabolites, and key pathways. We have developed a total risk score (based on the plasma concentration of DBH, ANPEP, and ADIPO) as a predictor of development of PAH in CHD-VSD patients. Therefore, these proteins may be used as biomarkers, and the new total risk score has significant clinical implications in the diagnosis of PAH.
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Hipertensão Pulmonar Primária Familiar/metabolismo , Comunicação Interventricular/complicações , Hipertensão Pulmonar/metabolismo , Hipertensão Arterial Pulmonar/metabolismo , Biomarcadores/sangue , Criança , Pré-Escolar , Hipertensão Pulmonar Primária Familiar/fisiopatologia , Feminino , Genômica , Comunicação Interventricular/metabolismo , Humanos , Hipertensão Pulmonar/fisiopatologia , Masculino , Fosfatidilinositol 3-Quinases/metabolismo , Hipertensão Arterial Pulmonar/fisiopatologia , Fatores de RiscoRESUMO
AIM: Building on previous psychometric work, we aimed to further assess the minimally important difference (MID) of the Oral Health Impact Profile for Chronic Periodontitis (OHIP-CP). METHODS: In total, 240 consecutive patients with chronic periodontitis were recruited in the study. The OHIP-CP was completed at baseline and after six weeks. Methodology testing included the confirmatory factor analysis (CFA) and MID. Confirmatory factor analysis (CFA) was performed to assess the fit of the previously proposed three-factor model. The MID of this questionnaire was determined by applying anchor-based and distribution-based approaches. RESULTS: The CFA supported a three-factor model for the OHIP-CP with acceptable fit to the data. The fit indices were χ2 /df = 2.231, GFI = 0.935, TLI = 0.969 and CFI = 0.976, RMSEA = 0.076. The OHIP-CP scores showed significant improvements after treatment (p < .001). The anchor-based MIDs of OHIP-CP for "oral function restriction," "oral pain" and "psychological and social impact," and total score were 2, 1, 4 and 7 points, respectively. The effect sizes (ES) and standardized response mean (SRM) for the OHIP-CP were moderate to large. CONCLUSIONS: The MID of the OHIP-CP is recommended for interpreting clinically meaningful change in oral health-related quality of life (OHRQoL) over time.
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Periodontite Crônica , Humanos , Saúde Bucal , Dor , Qualidade de Vida , Inquéritos e QuestionáriosRESUMO
Patent ductus arteriosus is the third most common congenital heart disease and resulted from the persistence of ductal patency after birth. Ductus arteriosus closure involves functional and structural remodeling, controlled by many factors. The changes in plasma protein levels associated with PDA closure are not known. Here we for the first time demonstrate six key differential plasma proteins in human patent ductus arteriosus patients using proteomic technology and present a model to illustrate the constriction and closure of ductus arteriosus. Differentially expressed proteins were analyzed by using isobaric tags for relative and absolute quantification and validated by enzyme-linked immunosorbent assay in new samples. The proteomic data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXD008568. We found 74 upregulated and 98 downregulated proteins in the plasma of patients with PDA. Five decreased proteins (platelet factor 4, fibrinogen, von Willebrand factor, collagen, and mannose binding lectin-associated serine protease-2) and one increased protein (fibronectin) may increase the risk of patent ductus arteriosus. Those proteins are closely related to platelet activation and coagulation cascades, complement mannan-binding-lectin, and other systemic signaling pathways. Our findings for the first time indicate that the differential proteins involved in different pathways may play key roles in the nonclosure of the ductus arteriosus in humans and may be developed as biomarkers for diagnosis. All those findings may be served as the basis of understanding the etiology and pathogenesis of patent ductus arteriosus.
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Plaquetas/metabolismo , Proteínas Sanguíneas/metabolismo , Permeabilidade do Canal Arterial/metabolismo , Células Endoteliais/metabolismo , Biomarcadores/metabolismo , Pré-Escolar , Regulação para Baixo/fisiologia , Feminino , Humanos , Masculino , Proteômica/métodos , Transdução de Sinais/fisiologia , Regulação para Cima/fisiologiaRESUMO
Multi-functional metasurfaces have exhibited powerful abilities of manipulating electromagnetic (EM) wave in predetermined manners, largely improving their information capacities. However, most works are implemented with EM functions controlled by one of the intrinsic properties of EM wave, such as polarization, frequency, etc. Herein, we propose a coding scheme to design a broadband and high-efficient multi-functional metasurface independently controlled by both frequency and polarization. To achieve this goal, we design anisotropic coding particles to realize independent phase functions and polarization-selectivity in the microwave region. Meta-atoms are finally optimized to exhibit 2-bit phase responses insensitive to incident polarization in the X-band while showing a 1-bit phase shift sensitive to incident polarization in the Ku-band. As a proof of concept, a metasurface is configured as an isotropic lens in the X-band, whereas the metasurface is designed as an anisotropic beam deflector in the Ku-band with or without polarization-conversion functionality dependent on the input polarization. The measured results, which agree well with the simulated ones, show excellent performances in the designed dual bands. Such a multi-functional coding metasurface may provide a flexible and robust approach to manipulate EM wave of multiple frequencies, as well as to integrate diverse functionalities into a single flat device.
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BACKGROUND: Laminitis is considered as one of the most important causes of hoof lameness in dairy cows, which can lead to enormous economic losses. However, the etiology and pathogenesis of laminitis have not been clarified yet. Besides, it is of great significant to find alternative herbs for the prevention and treatment of dairy hooves to avoid the antibiotic abuse. In this study, the primary hoof dermal cells of dairy cows were isolated, the inflammatory model was induced by LPS, and treated with silymarin to find whether silymarin has protective effect on the inflammatory dermal cells. The viability of dermal cells, the levels of IL-1ß and TNF-α, the degree of p65 NF-κB and p38 MAPK phosphorylation, the expressions of CYP3A4 and CYP1A1 were measured. RESULTS: Hoof dermal cells of dairy cows were successfully isolated and cultured by tissue adherent culture method. Certain concentrations of LPS can increase the levels of IL-1ß and TNF-α, promote the phosphorylation of p65 NF-κB and p38 MAPK, and inhibit the mRNA expressions of CYP3A4 and CYP1A1. The optimal concentration for LPS to establish a hoof dermal cells inflammatory model was 10 µg/mL. Certain concentrations of silymarin can markedly decrease the secretions of IL-1ß and TNF-α, inhibit the phosphorylation of p65 NF-κB and p38 MAPK, and promote the mRNA expressions of CYP3A4 and CYP1A1 in LPS-induced dermal inflammatory model. CONCLUSIONS: LPS can be used for inducing the hoof dermal cells inflammatory model of dairy cows. Silymarin has protective effects on the LPS-induced inflammatory model.
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Sistema Enzimático do Citocromo P-450/metabolismo , Casco e Garras/citologia , Silimarina/farmacologia , Fator de Transcrição RelA/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Bovinos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP3A/genética , Citocromo P-450 CYP3A/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Derme/citologia , Derme/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Casco e Garras/efeitos dos fármacos , Inflamação/induzido quimicamente , Interleucina-1beta/metabolismo , Lipopolissacarídeos/toxicidade , Fosforilação , Fator de Transcrição RelA/genética , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/genéticaRESUMO
Phosphatidylserine is widely used in food, health, chemical and pharmaceutical industries. The phospholipase D-mediated green synthesis of phosphatidylserine has attracted substantial attention in recent years. In this study, the phospholipase D was heterologously expressed in Bacillus subtilis, Pichia pastoris, and Corynebacterium glutamicum, respectively. The highest activity of phospholipase D was observed in C. glutamicum, which was 0.25 U/mL higher than these in B. subtilis (0.14 U/mL) and P. pastoris (0.22 U/mL). System engineering of three potential factors, including (1) signal peptides, (2) ribosome binding site, and (3) promoters, was attempted to improve the expression level of phospholipase D in C. glutamicum. The maximum phospholipase D activity reached 1.9 U/mL, which was 7.6-fold higher than that of the initial level. The enzyme displayed favorable transphosphatidylation activity and it could efficiently catalyze the substrates L-serine and soybean lecithin for synthesis of phosphatidylserine after optimizing the conversion reactions in detail. Under the optimum conditions (trichloromethane/enzyme solution 4:2, 8 mg/mL soybean lecithin, 40 mg/mL L-serine, and 15 mM CaCl2, with shaking under 40 °C for 10 h), the reaction process showed 48.6% of conversion rate and 1.94 g/L of accumulated phosphatidylserine concentration. The results highlight the use of heterologous expression, system engineering, and process optimization strategies to adapt a promising phospholipase D for efficient phosphatidylserine production in synthetic application.
Assuntos
Biocatálise , Fosfatidilserinas/química , Fosfolipase D , Engenharia de Proteínas , Bacillus subtilis/enzimologia , Bacillus subtilis/genética , Corynebacterium glutamicum/enzimologia , Corynebacterium glutamicum/genética , Fosfolipase D/química , Fosfolipase D/genética , Pichia/enzimologia , Pichia/genética , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Especificidade por SubstratoRESUMO
Decline in successful conception decreases more rapidly after 38 years of age owing to follicular depletion and decreased oocyte quality. However, limited information is available regarding the underlying mechanism and the useful treatment. This study aimed to evaluate the effects of growth hormone supplementation on oocyte maturation in vivo in aged and young mice and to determine its effect on mitochondrial function. The influence of three different doses of recombinant human growth hormone (rhGH) (0.4, 0.8 and 1.6 mg/kg/day) for 8 weeks before ovarian stimulation was analyzed. Superovulated oocytes were released from the oviduct of 12-week-old and 40-week-old female C57BL/6J mice 14-16 h after administration of human chorionic gonadotropin. Ovarian follicle and morphological analysis and oocyte maturation parameters were then evaluated. This study is the first, to our knowledge, to report that medium- and high-dose rhGH significantly increases antral follicles in aged mice but anti-Müllerian hormone (AMH) levels. Furthermore, derived oocytes, MII-stage oocyte rate, ATP levels, mitochondrial membrane potential and frequencies of homogeneous mitochondrial distribution increased. In contrast, in both aged and young mice, the mtDNA copy numbers per oocyte were similar before rhGH administration, and upon saline administration, they did not differ significantly. We conclude that medium-dose rhGH supplementation before standard ovarian stimulation regimens improves oocyte quality in aged mice, probably by enhancing mitochondrial functionality.
Assuntos
Envelhecimento/fisiologia , Hormônio do Crescimento Humano/administração & dosagem , Mitocôndrias/fisiologia , Oócitos/fisiologia , Folículo Ovariano/fisiologia , Proteínas Recombinantes/administração & dosagem , Animais , Hormônio Antimülleriano/metabolismo , Feminino , Humanos , Técnicas de Maturação in Vitro de Oócitos , Potencial da Membrana Mitocondrial , Camundongos , Camundongos Endogâmicos C57BL , Mitocôndrias/efeitos dos fármacos , Oócitos/citologia , Oócitos/efeitos dos fármacos , Folículo Ovariano/citologia , Folículo Ovariano/efeitos dos fármacos , Indução da OvulaçãoRESUMO
Potent and selective chemical probes are valuable tools for discovery of novel treatments for human diseases. NF-κB-inducing kinase (NIK) is a key trigger in the development of liver injury and fibrosis. Whether inhibition of NIK activity by chemical probes ameliorates liver inflammation and injury is largely unknown. In this study, a small-molecule inhibitor of NIK, B022, was found to be a potent and selective chemical probe for liver inflammation and injury. B022 inhibited the NIK signaling pathway, including NIK-induced p100-to-p52 processing and inflammatory gene expression, both in vitro and in vivo Furthermore, in vivo administration of B022 protected against not only NIK but also CCl4-induced liver inflammation and injury. Our data suggest that inhibition of NIK is a novel strategy for treatment of liver inflammation, oxidative stress, and injury.-Ren, X., Li, X., Jia, L., Chen, D., Hou, H., Rui, L., Zhao, Y., Chen, Z. A small-molecule inhibitor of NF-κB-inducing kinase (NIK) protects liver from toxin-induced inflammation, oxidative stress, and injury.