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1.
PLoS One ; 18(7): e0278814, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37428738

RESUMEN

For the optimal design of the sustainable supply chain network, considering the comprehensiveness of the problem factors, considering the three aspects of economy, environment and society, the goal is to minimize the establishment cost, minimize the emission of environ-mental pollution and maximize the number of labor. A mixed integer programming model is established to maximize the efficiency of the supply chain network. The innovation of this paper, first, is to consider the impact of economic, environmental and social benefits in a continuous supply chain, where the environmental benefits not only consider carbon emissions but also include the emissions of plant wastewater, waste and solid waste as influencing factors. Second, a multi-objective fuzzy affiliation function is constructed to measure the quality of the model solution in terms of the overall satisfaction value. Finally, the chaotic particle ant colony algorithm is proposed, and the problem of premature convergence in the operation of the particle swarm algorithm is solved. Experimental results show that the PSCACO algorithm proposed in this paper is compared with MOPSO, CACO and NSGA-II algorithms, and the convergence effect of the algorithm is concluded to be more effective to verify the effectiveness and feasibility of chaotic particle ant colony algorithm for solving multi-objective functions, which proposes a new feasible solution for the supply chain management.


Asunto(s)
Residuos Sólidos , Aguas Residuales , Algoritmos , Carbono
2.
Adv Sci (Weinh) ; 10(26): e2302443, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37409423

RESUMEN

The accomplishment of condition monitoring and intelligent maintenance for cantilever structure-based energy harvesting devices remains a challenge. Here, to tackle the problems, a novel cantilever-structure freestanding triboelectric nanogenerator (CSF-TENG) is proposed, which can capture ambient energy or transmit sensory information. First, with and without a crack in cantilevers, the simulations are carried out. According to simulation results, the maximum change ratios of natural frequency and amplitude are 1.1% and 2.2%, causing difficulties in identifying defects by these variations. Thus, based on Gramian angular field and convolutional neural network, a defect detection model is established to achieve the condition monitoring of the CSF-TENG, and the experimental result manifests that the accuracy of the model is 99.2%. Besides, the relation between the deflection of cantilevers and the output voltages of the CSF-TENG is first built, and then the defect identification digital twin system is successfully created. Consequently, the system is capable of duplicating the operation of the CSF-TENG in a real environment, and displaying defect recognition results, so the intelligent maintenance of the CSF-TENG can be realized.

3.
EMBO Mol Med ; 15(5): e16775, 2023 05 08.
Artículo en Inglés | MEDLINE | ID: mdl-37013609

RESUMEN

Topoisomerase 3α (TOP3A) is an enzyme that removes torsional strain and interlinks between DNA molecules. TOP3A localises to both the nucleus and mitochondria, with the two isoforms playing specialised roles in DNA recombination and replication respectively. Pathogenic variants in TOP3A can cause a disorder similar to Bloom syndrome, which results from bi-allelic pathogenic variants in BLM, encoding a nuclear-binding partner of TOP3A. In this work, we describe 11 individuals from 9 families with an adult-onset mitochondrial disease resulting from bi-allelic TOP3A gene variants. The majority of patients have a consistent clinical phenotype characterised by bilateral ptosis, ophthalmoplegia, myopathy and axonal sensory-motor neuropathy. We present a comprehensive characterisation of the effect of TOP3A variants, from individuals with mitochondrial disease and Bloom-like syndrome, upon mtDNA maintenance and different aspects of enzyme function. Based on these results, we suggest a model whereby the overall severity of the TOP3A catalytic defect determines the clinical outcome, with milder variants causing adult-onset mitochondrial disease and more severe variants causing a Bloom-like syndrome with mitochondrial dysfunction in childhood.


Asunto(s)
Enfermedades Mitocondriales , Enfermedades Musculares , Humanos , Mitocondrias/genética , ADN Mitocondrial/genética , Enfermedades Mitocondriales/genética , Síndrome , Inestabilidad Genómica
4.
Methods Mol Biol ; 2615: 89-98, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-36807786

RESUMEN

Proper mitochondrial DNA (mtDNA) levels are critical for many cellular biological functions and are associated with aging and many mitochondria disorders. Defects in core subunits of the mtDNA replication machinery lead to decreased mtDNA levels. Other indirect mitochondrial contexts including ATP concentration, lipid composition, and nucleotide composition also contribute to mtDNA maintenance. Furthermore, mtDNA molecules are distributed evenly throughout the mitochondrial network. This uniform distribution pattern is required for oxidative phosphorylation and ATP production and has been linked to many diseases when perturbed. Thus, it is important to visualize mtDNA in the cellular context. Here we provide detailed protocols for cellular visualization of mtDNA using fluorescence in situ hybridization (FISH). The fluorescent signals are targeted to the mtDNA sequence directly, ensuring both sensitivity and specificity. This mtDNA FISH method can be combined with immunostaining and used for visualizing mtDNA-protein interactions and dynamics.


Asunto(s)
ADN Mitocondrial , Mitocondrias , ADN Mitocondrial/genética , Hibridación Fluorescente in Situ , Mitocondrias/genética , Replicación del ADN , Adenosina Trifosfato
5.
International Eye Science ; (12): 996-1000, 2023.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-973793

RESUMEN

AIM: To analyze the clinical effect of 577nm subthreshold micropulse laser(SML)photocoagulation combined with intravitreal injection of Conbercept in the treatment of diabetic macular edema(DME)after vitrectomy in patients with proliferative diabetic retinopathy(PDR).METHODS:A retrospective analysis was performed on 29 cases(30 eyes)of PDR patients who had DME after vitrectomy in our hospital from January 2019 to June 2021. They were divided into two groups according to different treatment methods: 14 cases(14 eyes)in the single injection group received intravitreal injection of Conbercept, and 15 cases(16 eyes)in the combined treatment group received 577nm SML photocoagulation in the macular area combined with intravitreal injection of Conbercept. The changes in best corrected visual acuity(BCVA)and central macular thickness(CMT)before and at 6 and 12mo after treatment, as well as the changes of multifocal electroretinogram(mfERG)before and at 12mo after treatment were compared between the two groups.RESULTS: The BCVA(LogMAR)of patients in both groups improved and CMT decreased after treatment for 6 and 12mo(all P<0.001). There were no significant differences in BCVA(LogMAR)and CMT before treatment and 6mo, 12mo after treatment between single injection group and combined treatment group(all P>0.05). Compared with the combined treatment group, the amplitude was slightly lower(23.02±3.13 vs. 26.50±3.33 μV/deg2)and the latency time was prolonged(38.75±1.62 vs. 34.21±3.06ms)in single injection group at 12mo(all P≤0.001). The average injection times in single injection group was 8.14±1.46, and 5.05±1.51 in combined treatment group at 12mo after treatment(P<0.05).CONCLUSION: 577nm SML photocoagulation combined with intravitreal injection of conbercept can effectively relieve macular edema, improve BCVA and visual function of macular area and reduce the injection times of conbercept for DME patients.

6.
Mol Cell ; 82(19): 3646-3660.e9, 2022 10 06.
Artículo en Inglés | MEDLINE | ID: mdl-36044900

RESUMEN

The human mitochondrial genome must be replicated and expressed in a timely manner to maintain energy metabolism and supply cells with adequate levels of adenosine triphosphate. Central to this process is the idea that replication primers and gene products both arise via transcription from a single light strand promoter (LSP) such that primer formation can influence gene expression, with no consensus as to how this is regulated. Here, we report the discovery of a second light strand promoter (LSP2) in humans, with features characteristic of a bona fide mitochondrial promoter. We propose that the position of LSP2 on the mitochondrial genome allows replication and gene expression to be orchestrated from two distinct sites, which expands our long-held understanding of mitochondrial gene expression in humans.


Asunto(s)
Genoma Mitocondrial , Adenosina Trifosfato/metabolismo , ADN Mitocondrial/metabolismo , Humanos , Mitocondrias/genética , Mitocondrias/metabolismo , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Transcripción Genética
7.
Nucleic Acids Res ; 50(15): 8749-8766, 2022 08 26.
Artículo en Inglés | MEDLINE | ID: mdl-35947649

RESUMEN

The in vivo role for RNase H1 in mammalian mitochondria has been much debated. Loss of RNase H1 is embryonic lethal and to further study its role in mtDNA expression we characterized a conditional knockout of Rnaseh1 in mouse heart. We report that RNase H1 is essential for processing of RNA primers to allow site-specific initiation of mtDNA replication. Without RNase H1, the RNA:DNA hybrids at the replication origins are not processed and mtDNA replication is initiated at non-canonical sites and becomes impaired. Importantly, RNase H1 is also needed for replication completion and in its absence linear deleted mtDNA molecules extending between the two origins of mtDNA replication are formed accompanied by mtDNA depletion. The steady-state levels of mitochondrial transcripts follow the levels of mtDNA, and RNA processing is not altered in the absence of RNase H1. Finally, we report the first patient with a homozygous pathogenic mutation in the hybrid-binding domain of RNase H1 causing impaired mtDNA replication. In contrast to catalytically inactive variants of RNase H1, this mutant version has enhanced enzyme activity but shows impaired primer formation. This finding shows that the RNase H1 activity must be strictly controlled to allow proper regulation of mtDNA replication.


Asunto(s)
ADN Mitocondrial , Ribonucleasa H , Ratones , Animales , ADN Mitocondrial/química , Ribonucleasa H/genética , Ribonucleasa H/metabolismo , ARN/química , Replicación del ADN/genética , Mitocondrias/genética , Mamíferos/genética
8.
Cell ; 185(13): 2309-2323.e24, 2022 06 23.
Artículo en Inglés | MEDLINE | ID: mdl-35662414

RESUMEN

The mitochondrial genome encodes 13 components of the oxidative phosphorylation system, and altered mitochondrial transcription drives various human pathologies. A polyadenylated, non-coding RNA molecule known as 7S RNA is transcribed from a region immediately downstream of the light strand promoter in mammalian cells, and its levels change rapidly in response to physiological conditions. Here, we report that 7S RNA has a regulatory function, as it controls levels of mitochondrial transcription both in vitro and in cultured human cells. Using cryo-EM, we show that POLRMT dimerization is induced by interactions with 7S RNA. The resulting POLRMT dimer interface sequesters domains necessary for promoter recognition and unwinding, thereby preventing transcription initiation. We propose that the non-coding 7S RNA molecule is a component of a negative feedback loop that regulates mitochondrial transcription in mammalian cells.


Asunto(s)
ADN Mitocondrial , Proteínas Mitocondriales , Animales , ADN Mitocondrial/genética , ARN Polimerasas Dirigidas por ADN/metabolismo , Dimerización , Humanos , Mamíferos/metabolismo , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , ARN/metabolismo , ARN Mitocondrial , ARN Citoplasmático Pequeño , Partícula de Reconocimiento de Señal , Transcripción Genética
9.
Zhongguo Zhong Yao Za Zhi ; 47(4): 1064-1072, 2022 Feb.
Artículo en Chino | MEDLINE | ID: mdl-35285207

RESUMEN

This study aims to establish a rapid and sensitive UPLC-MS/MS method for simultaneously determining the content of strychnine and paeoniflorin in plasma and brain tissue of rats, and compare the pharmacokinetic behavior and brain tissue distribution of paeoniflorin combined with normal and toxic doses of strychnine in rats after percutaneous administration. Compared with those in the toxic-dose strychnine group, the AUC_(0-t), AUC_(0-∞), and C_(max) of strychnine decreased by 51.51%, 45.68%, and 46.03%, respectively(P<0.01), and the corresponding values of paeoniflorin increased by 91.41%, 102.31%, and 169.32%, respectively(P<0.01), in the compatibility group. Compared with the normal-dose strychnine group, the compatibility group showed insignificantly decreased C_(max), AUC_(0-t), and AUC_(0-∞) of strychnine, increased C_(max) and T_(max) of paeoniflorin(P<0.01), 66.88% increase in AUC_(0-t), and 70.55% increase in AUC_(0-∞) of paeoniflorin. In addition, the brain tissue concentration of strychnine decreased and that of paeoniflorin increased after compatibility. The combination of paeoniflorin with normal dose and toxic dose of strychnine can inhibit the percutaneous absorption of strychnine, and greatly promote the percutaneous penetration of paeoniflorin, whereas the interaction mechanism remains to be explored. The UPLC-MS/MS method established in this study is easy to operate and has good precision. It is suitable for in vivo study of pharmacokinetic behavior and brain tissue distribution of paeoniflorin and strychnine after percutaneous administration in rats, which provides reference for the safe and rational clinical use of strychnine and the combined use of drugs, and lays a solid foundation for the development of external preparations containing Strychni Semen.


Asunto(s)
Estricnina , Espectrometría de Masas en Tándem , Administración Cutánea , Animales , Encéfalo , Hidrocarburos Aromáticos con Puentes/farmacología , Cromatografía Liquida/métodos , Glucósidos , Monoterpenos , Ratas , Ratas Sprague-Dawley , Espectrometría de Masas en Tándem/métodos , Distribución Tisular
10.
Artículo en Chino | WPRIM (Pacífico Occidental) | ID: wpr-928026

RESUMEN

This study aims to establish a rapid and sensitive UPLC-MS/MS method for simultaneously determining the content of strychnine and paeoniflorin in plasma and brain tissue of rats, and compare the pharmacokinetic behavior and brain tissue distribution of paeoniflorin combined with normal and toxic doses of strychnine in rats after percutaneous administration. Compared with those in the toxic-dose strychnine group, the AUC_(0-t), AUC_(0-∞), and C_(max) of strychnine decreased by 51.51%, 45.68%, and 46.03%, respectively(P<0.01), and the corresponding values of paeoniflorin increased by 91.41%, 102.31%, and 169.32%, respectively(P<0.01), in the compatibility group. Compared with the normal-dose strychnine group, the compatibility group showed insignificantly decreased C_(max), AUC_(0-t), and AUC_(0-∞) of strychnine, increased C_(max) and T_(max) of paeoniflorin(P<0.01), 66.88% increase in AUC_(0-t), and 70.55% increase in AUC_(0-∞) of paeoniflorin. In addition, the brain tissue concentration of strychnine decreased and that of paeoniflorin increased after compatibility. The combination of paeoniflorin with normal dose and toxic dose of strychnine can inhibit the percutaneous absorption of strychnine, and greatly promote the percutaneous penetration of paeoniflorin, whereas the interaction mechanism remains to be explored. The UPLC-MS/MS method established in this study is easy to operate and has good precision. It is suitable for in vivo study of pharmacokinetic behavior and brain tissue distribution of paeoniflorin and strychnine after percutaneous administration in rats, which provides reference for the safe and rational clinical use of strychnine and the combined use of drugs, and lays a solid foundation for the development of external preparations containing Strychni Semen.


Asunto(s)
Animales , Ratas , Administración Cutánea , Encéfalo , Hidrocarburos Aromáticos con Puentes/farmacología , Cromatografía Liquida/métodos , Glucósidos , Monoterpenos , Ratas Sprague-Dawley , Estricnina , Espectrometría de Masas en Tándem/métodos , Distribución Tisular
11.
BMC Musculoskelet Disord ; 22(1): 743, 2021 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-34461864

RESUMEN

BACKGROUND: To determine an optimized treatment protocol during the COVID-19 epidemic for patients with closed fracture and delayed surgery. METHODS: The epidemic data of three hospitals, randomly selected from different administrative regions of Wuhan, were analyzed retrospectively from 23 January to 31 March 2020. Changes in the number of confirmed cases per day (cumulative and new) of each region were tracked as a reflection of changing epidemic risk levels. The risk level map was drawn. The epidemic status, treatment protocols, and treatment efficiencies for patients with closed fracture in the three hospitals were compared. RESULTS: Overall, 138 patients with closed fracture were admitted. Each hospital had established its own protocol, according to the initial perceived risk. Based on the risk level map, over the study period, the risk levels of the three regions changed independently and were not in sync. All patients recovered and were timely discharged. No staff member was detected with COVID-19. CONCLUSIONS: The COVID-19 risk level of each area is dynamic. To optimize medical resources, avoid cross-infection, and improve efficiency, changes in epidemic risk should be monitored. For patients with closed fracture, treatment protocols should be adjusted according to changes in epidemic risk.


Asunto(s)
COVID-19 , Fracturas Cerradas , China , Protocolos Clínicos , Hospitales , Humanos , Estudios Retrospectivos , SARS-CoV-2
12.
Sci Adv ; 7(27)2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-34215584

RESUMEN

We report a role for the mitochondrial single-stranded DNA binding protein (mtSSB) in regulating mitochondrial DNA (mtDNA) replication initiation in mammalian mitochondria. Transcription from the light-strand promoter (LSP) is required both for gene expression and for generating the RNA primers needed for initiation of mtDNA synthesis. In the absence of mtSSB, transcription from LSP is strongly up-regulated, but no replication primers are formed. Using deep sequencing in a mouse knockout model and biochemical reconstitution experiments with pure proteins, we find that mtSSB is necessary to restrict transcription initiation to optimize RNA primer formation at both origins of mtDNA replication. Last, we show that human pathological versions of mtSSB causing severe mitochondrial disease cannot efficiently support primer formation and initiation of mtDNA replication.


Asunto(s)
Replicación del ADN , Proteínas de Unión al ADN , Animales , ADN Mitocondrial/genética , ADN Mitocondrial/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Células HeLa , Humanos , Mamíferos/genética , Ratones , Mitocondrias/genética , Mitocondrias/metabolismo
13.
Nat Commun ; 12(1): 3588, 2021 Jun 11.
Artículo en Inglés | MEDLINE | ID: mdl-34117250

RESUMEN

Catastrophic accidents caused by fatigue failures often occur in engineering structures. Thus, a fundamental understanding of cyclic-deformation and fatigue-failure mechanisms is critical for the development of fatigue-resistant structural materials. Here we report a high-entropy alloy with enhanced fatigue life by ductile-transformable multicomponent B2 precipitates. Its cyclic-deformation mechanisms are revealed by real-time in-situ neutron diffraction, transmission-electron microscopy, crystal-plasticity modeling, and Monte-Carlo simulations. Multiple cyclic-deformation mechanisms, including dislocation slips, precipitation strengthening, deformation twinning, and reversible martensitic phase transformation, are observed in the studied high-entropy alloy. Its improved fatigue performance at low strain amplitudes, i.e., the high fatigue-crack-initiation resistance, is attributed to the high elasticity, plastic deformability, and martensitic transformation of the B2-strengthening phase. This study shows that fatigue-resistant alloys can be developed by incorporating strengthening ductile-transformable multicomponent intermetallic phases.

14.
PLoS Genet ; 16(12): e1009242, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-33315859

RESUMEN

Deletions and duplications in mitochondrial DNA (mtDNA) cause mitochondrial disease and accumulate in conditions such as cancer and age-related disorders, but validated high-throughput methodology that can readily detect and discriminate between these two types of events is lacking. Here we establish a computational method, MitoSAlt, for accurate identification, quantification and visualization of mtDNA deletions and duplications from genomic sequencing data. Our method was tested on simulated sequencing reads and human patient samples with single deletions and duplications to verify its accuracy. Application to mouse models of mtDNA maintenance disease demonstrated the ability to detect deletions and duplications even at low levels of heteroplasmy.


Asunto(s)
ADN Mitocondrial/genética , Eliminación de Gen , Duplicación de Gen , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , Análisis de Secuencia de ADN/métodos , Animales , ADN Mitocondrial/química , Secuenciación de Nucleótidos de Alto Rendimiento/normas , Ratones , Reproducibilidad de los Resultados , Análisis de Secuencia de ADN/normas
15.
Cogn Neurodyn ; 14(5): 643-656, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-33014178

RESUMEN

Facial attractiveness is an important research direction of genetic psychology and cognitive psychology, and its results are significant for the study of face evolution and human evolution. However, previous studies have not put forward a comprehensive evaluation system of facial attractiveness. Traditionally, the establishment of facial attractiveness evaluation system was based on facial geometric features, without facial skin features. In this paper, combined with big data analysis, evaluation of face in real society and literature research, we found that skin also have a significant impact on facial attractiveness, because skin could reflect age, wrinkles and healthful qualities, thus affected the human perception of facial attractiveness. Therefore, we propose a comprehensive and novel facial attractiveness evaluation system based on face shape structural features, facial structure features and skin texture feature. In order to apply face shape structural features to the evaluation of facial attractiveness, the classification of face shape is the first step. Face image dataset is divided according to face shape, and then facial structure features and skin texture features that represent facial attractiveness are extracted and fused. The machine learning algorithm with the best prediction performance is selected in the face shape structural subsets to predict facial attractiveness. Experimental results show that the facial attractiveness evaluation performance can be improved by the method based on classification of face shape and multi-features fusion, the facial attractiveness scores obtained by the proposed system correlates better with human ratings. Our evaluation system can help people project their cognition of facial attractiveness into artificial agents they interact with.

16.
Biosci Biotechnol Biochem ; 84(6): 1153-1159, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32019421

RESUMEN

Peptidic inhibitors of proteases are attracting increasing interest not only as drug candidates but also for studying the function and regulation mechanisms of these enzymes. Previously, we screened out a cyclic peptide inhibitor of human uPA [Formula: see text] and found that Ala substitution of P2 residue turns upain-1 to a substrate. To further investigate the effect of P2 residue on the peptide behavior transformation, we constructed upain-1-W3F, which has Phe replacement in the P2 position. We determined KD and Ki of upain-1-W3F and found that upain-1-W3F might still exist as an inhibitor. Furthermore, the high-resolution crystal structure of upain-1-W3F·uPA reveals that upain-1-W3F indeed stays as an intact inhibitor bind to uPA. We thus propose that the P2 residue plays a nonnegligible role in the conversion of upain-1 to a substrate. These results also proposed a strategy to optimize the pharmacological properties of peptide-based drug candidates by hydrophobicity and steric hindrance.Abbreviations : uPA: urokinase-type plasminogen activator; SPD: serine protease domain; S1 pocket: specific substrate-binding pocket.


Asunto(s)
Péptidos Cíclicos/química , Saccharomycetales/genética , Saccharomycetales/metabolismo , Inhibidores de Serina Proteinasa/química , Activador de Plasminógeno de Tipo Uroquinasa/química , Dominio Catalítico , Cristalografía por Rayos X , Humanos , Enlace de Hidrógeno , Hidrólisis , Unión Proteica , Estructura Secundaria de Proteína
17.
Res Pract Thromb Haemost ; 3(3): 412-419, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31294329

RESUMEN

BACKGROUND: Blood coagulation factor VIIa (FVIIa) plays its critical physiological role in the initiation of hemostasis. Even so, recombinant FVIIa is successfully used as a bypassing agent for factor VIII or IX in the treatment of bleeds in patients with severe hemophilia with inhibitors. To investigate the utility of more potent FVIIa variants with enhanced intrinsic activity, molecules such as V21D/E154V/M156Q-FVIIa (FVIIaDVQ) were designed. METHODS: Surface plasmon resonance was used to characterize the binding of mAb4F5 to FVIIaDVQ and related variants. X-ray crystallography was used to determine the structure of the Fab fragment of mAb4F5 (Fab4F5). Molecular docking and small angle X-ray scattering led to a model of FVIIaDVQ:Fab4F5 complex. RESULTS: The binding experiments, functional effects on FVIIaDVQ and structure of mAb4F5 (originally intended for quantification of FVIIaDVQ in samples containing FVII(a)) pinpointed the epitope (crucial role for residue Asp21) and shed light on the role of the N-terminus of the protease domain in FVIIa allostery. The potential antigen-combining sites are composed of 1 hydrophobic and 1 negatively charged pocket formed by 6 complementarity-determining region (CDR) loops. Structural analysis of Fab4F5 shows that the epitope interacts with the periphery of the hydrophobic pocket and provides insights into the molecular basis of mAb4F5 recognition and tight binding of FVIIaDVQ. CONCLUSION: The binary complex explains and supports the selectivity and functional consequences of Fab4F5 association with FVIIaDVQ and illustrates the potentially unique antigenicity of this FVIIa variant. This will be useful in the design of less immunogenic variants.

18.
Front Microbiol ; 10: 1232, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31244791

RESUMEN

Intramembrane proteases hydrolyze peptide bonds within the cell membrane as the decision-making step of various signaling pathways. Sporulation factor IV B protease (SpoIVB) and C-terminal processing proteases B (CtpB) play central roles in cellular differentiation via regulated intramembrane proteolysis (RIP) process which activates pro-σK processing at the σK checkpoint during spore formation. SpoIVB joins CtpB in belonging to the widespread family of PDZ-proteases, but much remains unclear about the molecular mechanisms and structure of SpoIVB. In this study, we expressed inactive SpoIVB (SpoIVBS378A) fused with maltose binding protein (MBP)-tag and obtained the solution structure of SpoIVBS378A from its small angle X-ray scattering (SAXS) data. The fusion protein is more soluble, stable, and yields higher expression compared to SpoIVB without the tag. MBP-tag not only facilitates modeling of the structure in the SAXS envelope but also evaluates reliability of the model. The solution structure of SpoIVBS378A fits closely with the experimental scattering data (χ2= 1.76). Comparing the conformations of PDZ-proteases indicates that SpoIVB adopts a PDZ-protease pattern similar to the high temperature requirement A proteases (HtrAs) rather than CtpB. We not only propose that SpoIVB uses a more direct and simple way to cleave the substrates than that of CtpB, but also that they work together as signal amplifiers to activate downstream proteins in the RIP pathway.

19.
J Avian Med Surg ; 33(1): 29-37, 2019 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-31124609

RESUMEN

To compare the bending strength of a locking plate (LP), nonlocking plate (NLP), and an external skeletal fixator intramedullary pin (ESF-IM) tie-in fixation applied by a dorsal approach in an avian humerus fracture model, 5 left humeri obtained from pigeon (Columba livia) cadavers were randomly assigned to each repair technique (n = 15). The ESF-IM group was repaired with a 0.062-inch intramedullary pin tied-in with two 0.035-inch positive profile transfixation pins using acrylic filled plastic tubing. The LP group was repaired with a dorsally applied titanium 1.6-mm screw 7-hole locking plate (1 bicortical and 2 monocortical screws in each segment). The NLP group was repaired with a dorsally applied 6-hole stainless steel 1.5-mm dynamic compression plate (all bicortical screws). All constructs were applied before complete ostectomy to allow perfect reconstruction. Constructs were cyclically tested nondestructively for 1000 cycles in four-point bending before being tested to failure. Outcome measures included stiffness, strength, and strain energy. All specimens cycled without failure. The ESF-IM specimens were significantly stiffer and stronger than the plated repair groups. Plated constructs had significantly higher strain energies than ESF-IM. LP and NLP were of equal stiffness, strength, and strain energies. This study demonstrated that bending biomechanical properties of the ESF-IM configuration were superior to those of the dorsal plate fixation. Exact properties of fixation required to facilitate avian fracture healing are largely unknown. Further study, including assessments of optimal plate position and configuration, and torsional and in vivo studies in avian species are warranted.


Asunto(s)
Placas Óseas/veterinaria , Columbidae/lesiones , Columbidae/cirugía , Fijación de Fractura/veterinaria , Fracturas Óseas/veterinaria , Húmero/lesiones , Análisis de Varianza , Animales , Animales Salvajes , Fenómenos Biomecánicos , Clavos Ortopédicos/veterinaria , Placas Óseas/clasificación , Placas Óseas/normas , Cadáver , Fijación de Fractura/instrumentación , Fijación de Fractura/métodos , Curación de Fractura , Fracturas Óseas/cirugía , Húmero/cirugía , Distribución Aleatoria
20.
Med Sci Monit ; 25: 1679-1693, 2019 Mar 05.
Artículo en Inglés | MEDLINE | ID: mdl-30833538

RESUMEN

BACKGROUND This study aimed to identify significantly altered circRNAs/lncRNAs/miRNAs/mRNAs pathways in preeclampsia (PE), investigate their target relationships, and determine their biological functions. MATERIAL AND METHODS Base on RNA-seq technique and the GEO database, expression profiles of circRNAs/lncRNAs/miRNAs/mRNAs related to PE were obtained. Differentially expressed RNAs were determined using the Limma package in R. Gene set enrichment analysis (GSEA) was performed using GSEA software (v. 3.0) and illustrated by ClusterProfiler and ggplot2 package in R. DAVID database (v. 6.8) was implemented to analyze functional categories and the association between genes and the corresponding Gene Ontology (GO) classification. The R visualization package GOPlot was used to get a better visualization of the relationships between genes and the selected functional categories. CeRNA networks which visualized the correlations between circRNA/lncRNA-miRNA-mRNA were constructed using Cytoscape software (v. 3.6.0). Targetscan and miRanda database were used to predict target relationships between circRNA/lncRNA-miRNA-mRNA. QRT-PCR and luciferase reporter assay were used to verify the expression and target relationship of has_circ_0088196/LINC01492/miR-100-5p/LIF (leukemia inhibitory factor). RESULTS The jak-stat signaling pathway was activated and miR-100-5p was downregulated in PE compared with normal tissues both in collected placental tissue samples and GEO database. Upregulated LIF, LINC01492, and hsa_circ_0088196 were negatively correlated with miR-100-5p expression and had a targeted relationship with miR-100-5p. CONCLUSIONS miR-100-5p may suppress PE development, while LIF, LINC01492, and hsa_circ_0088196 may promote it though inhibiting miR-100-5p. The jak-stat signaling pathway was activated and involved in PE progression.


Asunto(s)
Biología Computacional/métodos , Preeclampsia/genética , ARN/genética , China , Femenino , Ontología de Genes , Humanos , MicroARNs/análisis , MicroARNs/genética , Placenta , Placentación/genética , Embarazo , ARN/análisis , ARN Circular , ARN Largo no Codificante/análisis , ARN Largo no Codificante/genética , ARN Mensajero/análisis , ARN Mensajero/genética
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