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1.
Cell ; 184(10): 2665-2679.e19, 2021 05 13.
Artigo em Inglês | MEDLINE | ID: mdl-33882274

RESUMO

The bacterial flagellar motor is a supramolecular protein machine that drives rotation of the flagellum for motility, which is essential for bacterial survival in different environments and a key determinant of pathogenicity. The detailed structure of the flagellar motor remains unknown. Here we present an atomic-resolution cryoelectron microscopy (cryo-EM) structure of the bacterial flagellar motor complexed with the hook, consisting of 175 subunits with a molecular mass of approximately 6.3 MDa. The structure reveals that 10 peptides protruding from the MS ring with the FlgB and FliE subunits mediate torque transmission from the MS ring to the rod and overcome the symmetry mismatch between the rotational and helical structures in the motor. The LP ring contacts the distal rod and applies electrostatic forces to support its rotation and torque transmission to the hook. This work provides detailed molecular insights into the structure, assembly, and torque transmission mechanisms of the flagellar motor.


Assuntos
Flagelos/fisiologia , Flagelos/ultraestrutura , Salmonella typhimurium/fisiologia , Microscopia Crioeletrônica , Conformação Proteica , Torque
2.
Cell ; 184(9): 2412-2429.e16, 2021 04 29.
Artigo em Inglês | MEDLINE | ID: mdl-33852913

RESUMO

Cellular versatility depends on accurate trafficking of diverse proteins to their organellar destinations. For the secretory pathway (followed by approximately 30% of all proteins), the physical nature of the vessel conducting the first portage (endoplasmic reticulum [ER] to Golgi apparatus) is unclear. We provide a dynamic 3D view of early secretory compartments in mammalian cells with isotropic resolution and precise protein localization using whole-cell, focused ion beam scanning electron microscopy with cryo-structured illumination microscopy and live-cell synchronized cargo release approaches. Rather than vesicles alone, the ER spawns an elaborate, interwoven tubular network of contiguous lipid bilayers (ER exit site) for protein export. This receptacle is capable of extending microns along microtubules while still connected to the ER by a thin neck. COPII localizes to this neck region and dynamically regulates cargo entry from the ER, while COPI acts more distally, escorting the detached, accelerating tubular entity on its way to joining the Golgi apparatus through microtubule-directed movement.


Assuntos
Vesículas Revestidas pelo Complexo de Proteína do Envoltório/metabolismo , Retículo Endoplasmático/metabolismo , Complexo de Golgi/metabolismo , Microtúbulos/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Transporte Biológico Ativo , Células HeLa , Humanos , Transporte Proteico
3.
Mol Cell ; 83(2): 167-185.e9, 2023 Jan 19.
Artigo em Inglês | MEDLINE | ID: mdl-36577401

RESUMO

The DNA double-strand break repair complex Mre11-Rad50-Nbs1 (MRN) detects and nucleolytically processes DNA ends, activates the ATM kinase, and tethers DNA at break sites. How MRN can act both as nuclease and scaffold protein is not well understood. The cryo-EM structure of MRN from Chaetomium thermophilum reveals a 2:2:1 complex with a single Nbs1 wrapping around the autoinhibited Mre11 nuclease dimer. MRN has two DNA-binding modes, one ATP-dependent mode for loading onto DNA ends and one ATP-independent mode through Mre11's C terminus, suggesting how it may interact with DSBs and intact DNA. MRNs two 60-nm-long coiled-coil domains form a linear rod structure, the apex of which is assembled by the two joined zinc-hook motifs. Apices from two MRN complexes can further dimerize, forming 120-nm spanning MRN-MRN structures. Our results illustrate the architecture of MRN and suggest how it mechanistically integrates catalytic and tethering functions.


Assuntos
Reparo do DNA , DNA , Microscopia Crioeletrônica , DNA/genética , Hidrolases Anidrido Ácido/genética , Quebras de DNA de Cadeia Dupla , Enzimas Reparadoras do DNA/metabolismo , Trifosfato de Adenosina/metabolismo , Proteína Homóloga a MRE11/genética , Proteína Homóloga a MRE11/metabolismo , Proteínas de Ciclo Celular/metabolismo
4.
Mol Cell ; 81(14): 2887-2900.e5, 2021 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-34171298

RESUMO

WhiB7 represents a distinct subclass of transcription factors in the WhiB-Like (Wbl) family, a unique group of iron-sulfur (4Fe-4S] cluster-containing proteins exclusive to the phylum of Actinobacteria. In Mycobacterium tuberculosis (Mtb), WhiB7 interacts with domain 4 of the primary sigma factor (σA4) in the RNA polymerase holoenzyme and activates genes involved in multiple drug resistance and redox homeostasis. Here, we report crystal structures of the WhiB7:σA4 complex alone and bound to its target promoter DNA at 1.55-Å and 2.6-Å resolution, respectively. These structures show how WhiB7 regulates gene expression by interacting with both σA4 and the AT-rich sequence upstream of the -35 promoter DNA via its C-terminal DNA-binding motif, the AT-hook. By combining comparative structural analysis of the two high-resolution σA4-bound Wbl structures with molecular and biochemical approaches, we identify the structural basis of the functional divergence between the two distinct subclasses of Wbl proteins in Mtb.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas Ferro-Enxofre/metabolismo , Mycobacterium tuberculosis/metabolismo , Fatores de Transcrição/metabolismo , Proteínas de Bactérias/genética , Regulação Bacteriana da Expressão Gênica/genética , Proteínas Ferro-Enxofre/genética , Mycobacterium tuberculosis/genética , Regiões Promotoras Genéticas/genética , Fator sigma/genética , Fator sigma/metabolismo , Fatores de Transcrição/genética
5.
J Cell Sci ; 136(20)2023 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-37737012

RESUMO

All endocytosis and exocytosis in the African trypanosome Trypanosoma brucei occurs at a single subdomain of the plasma membrane. This subdomain, the flagellar pocket, is a small vase-shaped invagination containing the root of the single flagellum of the cell. Several cytoskeleton-associated multiprotein complexes are coiled around the neck of the flagellar pocket on its cytoplasmic face. One of these, the hook complex, was proposed to affect macromolecule entry into the flagellar pocket lumen. In previous work, knockdown of T. brucei (Tb)MORN1, a hook complex component, resulted in larger cargo being unable to enter the flagellar pocket. In this study, the hook complex component TbSmee1 was characterised in bloodstream form T. brucei and found to be essential for cell viability. TbSmee1 knockdown resulted in flagellar pocket enlargement and impaired access to the flagellar pocket membrane by surface-bound cargo, similar to depletion of TbMORN1. Unexpectedly, inhibition of endocytosis by knockdown of clathrin phenocopied TbSmee1 knockdown, suggesting that endocytic activity itself is a prerequisite for the entry of surface-bound cargo into the flagellar pocket.


Assuntos
Trypanosoma brucei brucei , Trypanosoma , Trypanosoma/metabolismo , Endocitose/fisiologia , Trypanosoma brucei brucei/metabolismo , Membrana Celular/metabolismo , Cílios/metabolismo , Flagelos/metabolismo , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo
6.
J Biol Chem ; 299(11): 105340, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-37838178

RESUMO

The unicellular protozoan Trypanosoma brucei has a single flagellum that is involved in cell motility, cell morphogenesis, and cell division. Inheritance of the newly assembled flagellum during the cell cycle requires its correct positioning, which depends on the faithful duplication or segregation of multiple flagellum-associated cytoskeletal structures, including the basal body, the flagellum attachment zone, and the hook complex. Along the flagellum attachment zone sites a set of four microtubules termed the microtubule quartet (MtQ), whose molecular function remains enigmatic. We recently reported that the MtQ-localized protein NHL1 interacts with the microtubule-binding protein TbSpef1 and regulates flagellum inheritance by promoting basal body rotation and segregation. Here, we identified a TbSpef1- and NHL1-associated protein named SNAP1, which co-localizes with NHL1 and TbSpef1 at the proximal portion of the MtQ, depends on TbSpef1 for localization and is required for NHL1 localization to the MtQ. Knockdown of SNAP1 impairs the rotation and segregation of the basal body, the elongation of the flagellum attachment zone filament, and the positioning of the newly assembled flagellum, thereby causing mis-placement of the cell division plane, a halt in cleavage furrow ingression, and an inhibition of cytokinesis completion. Together, these findings uncover a coordinating role of SNAP1 with TbSpef1 and NHL1 in facilitating flagellum positioning and cell division plane placement for the completion of cytokinesis.


Assuntos
Flagelos , Microtúbulos , Proteínas de Protozoários , Trypanosoma brucei brucei , Corpos Basais/metabolismo , Divisão Celular , Segregação de Cromossomos , Flagelos/metabolismo , Microtúbulos/metabolismo , Proteínas de Protozoários/metabolismo , Trypanosoma brucei brucei/metabolismo
7.
Plant J ; 114(2): 293-309, 2023 04.
Artigo em Inglês | MEDLINE | ID: mdl-36748183

RESUMO

When covered by a layer of soil, seedling development follows a dark-specific program (skotomorphogenesis). In the dark, seedlings consist of small, non-green cotyledons, a long hypocotyl, and an apical hook to protect meristematic cells. We recently highlighted the role played by mitochondria in the high energy-consuming reprogramming of Arabidopsis skotomorphogenesis. Here, the role played by plastids, another energy-supplying organelle, in skotomorphogenesis is investigated. This study was conducted in dark conditions to exclude light signals so as to better focus on those produced by plastids. It was found that limitation of plastid gene expression (PGE) induced an exaggerated apical hook bending. Inhibition of PGE was obtained at the levels of transcription and translation using the antibiotics rifampicin (RIF) and spectinomycin, respectively, as well as plastid RPOTp RNA polymerase mutants. RIF-treated seedlings also showed expression induction of marker nuclear genes for mitochondrial stress, perturbation of mitochondrial metabolism, increased ROS levels, and an augmented capacity of oxygen consumption by mitochondrial alternative oxidases (AOXs). AOXs act to prevent overreduction of the mitochondrial electron transport chain. Previously, we reported that AOX1A, the main AOX isoform, is a key component in the developmental response to mitochondrial respiration deficiency. In this work, we suggest the involvement of AOX1A in the response to PGE dysfunction and propose the importance of signaling between plastids and mitochondria. Finally, it was found that seedling architecture reprogramming in response to RIF was independent of canonical organelle retrograde pathways and the ethylene signaling pathway.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , Plântula/metabolismo , Hipocótilo , Cloroplastos/metabolismo , Expressão Gênica , Regulação da Expressão Gênica de Plantas , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo
8.
J Cell Sci ; 135(1)2022 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-34897470

RESUMO

In clathrin-independent endocytosis, Hook1, a microtubule- and cargo-tethering protein, participates in sorting of cargo proteins such as CD98 (encoded by SLC3A2) and CD147 (encoded by BSG) into recycling endosomes. However, the molecular mechanism that regulates Hook1-mediated endosomal sorting is not fully understood. In the present study, we found that γ-taxilin is a novel regulator of Hook1-mediated endosomal sorting. γ-Taxilin depletion promoted both CD98-positive tubular formation and CD98 recycling. Conversely, overexpression of γ-taxilin inhibited the CD98-positive tubular formation. Depletion of Hook1, or Rab10 or Rab22a (which are both involved in Hook1-mediated endosomal sorting), attenuated the effect of γ-taxilin depletion on the CD98-positive tubular formation. γ-Taxilin depletion promoted CD147-mediated spreading of HeLa cells, suggesting that γ-taxilin might be a pivotal player in various cellular functions in which Hook1-mediated cargo proteins are involved. γ-Taxilin bound to the C-terminal region of Hook1 and inhibited its interaction with CD98; the latter interaction is necessary for sorting CD98. We suggest that γ-taxilin negatively regulates the sorting of Hook1-mediated cargo proteins into recycling endosomes by interfering with the interactions between Hook1 and the cargo proteins.


Assuntos
Clatrina , Endossomos , Clatrina/metabolismo , Endocitose , Endossomos/metabolismo , Células HeLa , Humanos , Transporte Proteico , Proteínas rab de Ligação ao GTP/genética , Proteínas rab de Ligação ao GTP/metabolismo
9.
Biochem Soc Trans ; 52(2): 603-616, 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38572912

RESUMO

ATP dependent chromatin remodelers have pivotal roles in transcription, DNA replication and repair, and maintaining genome integrity. SWI/SNF remodelers were first discovered in yeast genetic screens for factors involved in mating type switching or for using alternative energy sources therefore termed SWI/SNF complex (short for SWItch/Sucrose NonFermentable). The SWI/SNF complexes utilize energy from ATP hydrolysis to disrupt histone-DNA interactions and shift, eject, or reposition nucleosomes making the underlying DNA more accessible to specific transcription factors and other regulatory proteins. In development, SWI/SNF orchestrates the precise activation and repression of genes at different stages, safe guards the formation of specific cell lineages and tissues. Dysregulation of SWI/SNF have been implicated in diseases such as cancer, where they can drive uncontrolled cell proliferation and tumor metastasis. Additionally, SWI/SNF defects are associated with neurodevelopmental disorders, leading to disruption of neural development and function. This review offers insights into recent developments regarding the roles of the SWI/SNF complex in pluripotency and cell lineage primining and the approaches that have helped delineate its importance. Understanding these molecular mechanisms is crucial for unraveling the intricate processes governing embryonic stem cell biology and developmental transitions and may potentially apply to human diseases linked to mutations in the SWI/SNF complex.


Assuntos
Trifosfato de Adenosina , Linhagem da Célula , Montagem e Desmontagem da Cromatina , Fatores de Transcrição , Humanos , Fatores de Transcrição/metabolismo , Animais , Trifosfato de Adenosina/metabolismo , Proteínas Cromossômicas não Histona/metabolismo
10.
Plant Cell Environ ; 47(4): 1084-1098, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38037476

RESUMO

Beneficial Bacillus subtilis (BS) symbiosis could combat root pathogenesis, but it relies on root-secreted sugars. Understanding the molecular control of sugar flux during colonization would benefit biocontrol applications. The SWEET (Sugar Will Eventually Be Exported Transporter) uniporter regulates microbe-induced sugar secretion from roots; thus, its homologs may modulate sugar distribution upon BS colonization. Quantitative polymerase chain reaction revealed that gene transcripts of SWEET2, but not SWEET16 and 17, were significantly induced in seedling roots after 12 h of BS inoculation. Particularly, SWEET2-ß-glucuronidase fusion proteins accumulated in the apical mature zone where BS abundantly colonized. Yet, enhanced BS colonization in sweet2 mutant roots suggested a specific role for SWEET2 to constrain BS propagation, probably by limiting hexose secretion. By employing yeast one-hybrid screening and ectopic expression in Arabidopsis protoplasts, the transcription factor AHL29 was identified to function as a repressor of SWEET2 expression through the AT-hook motif. Repression occurred despite immunity signals. Additionally, enhanced SWEET2 expression and reduced colonies were specifically detected in roots of BS-colonized ahl29 mutant. Taken together, we propose that BS colonization may activate repression of AHL29 on SWEET2 transcription that would be enhanced by immunity signals, thereby maintaining adequate sugar secretion for a beneficial Bacillus association.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Bacillus subtilis/metabolismo , Raízes de Plantas/metabolismo , Saccharomyces cerevisiae/metabolismo , Açúcares/metabolismo
11.
Pediatr Blood Cancer ; 71(9): e31171, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38961599

RESUMO

The hook effect is a well-described but clinically underappreciated immunoassay interference, where a falsely lowered result is caused by analyte excess. We describe a situation in which ferritin immunoassay results from a 27-year-old female with immune effector cell-associated hemophagocytic lymphohistiocytosis-like syndrome were more than 1000 times lower at a reference laboratory than those determined in-house after dilution. This case underscores the importance for clinical care providers to be aware of the impact of the hook effect on ferritin measurements, and to promptly communicate with the laboratory when there are discrepancies between clinical symptoms and test results.


Assuntos
Ferritinas , Imunoterapia Adotiva , Linfo-Histiocitose Hemofagocítica , Humanos , Feminino , Ferritinas/sangue , Adulto , Linfo-Histiocitose Hemofagocítica/diagnóstico , Linfo-Histiocitose Hemofagocítica/terapia , Linfo-Histiocitose Hemofagocítica/sangue , Imunoterapia Adotiva/efeitos adversos , Imunoterapia Adotiva/métodos , Imunoensaio/métodos , Receptores de Antígenos Quiméricos
12.
Plant Cell Rep ; 43(5): 131, 2024 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-38656568

RESUMO

KEY MESSAGE: The sugar supply in the medium affects the apical hook development of Arabidopsis etiolated seedlings. In addition, we provided the mechanism insights of this process. Dicotyledonous plants form an apical hook structure to shield their young cotyledons from mechanical damage as they emerge from the rough soil. Our findings indicate that sugar molecules, such as sucrose and glucose, are crucial for apical hook development. The presence of sucrose and glucose allows the apical hooks to be maintained for a longer period compared to those grown in sugar-free conditions, and this effect is dose-dependent. Key roles in apical hook development are played by several sugar metabolism pathways, including oxidative phosphorylation and glycolysis. RNA-seq data revealed an up-regulation of genes involved in starch and sucrose metabolism in plants grown in sugar-free conditions, while genes associated with phenylpropanoid metabolism were down-regulated. This study underscores the significant role of sugar metabolism in the apical hook development of etiolated Arabidopsis seedlings.


Assuntos
Arabidopsis , Regulação da Expressão Gênica de Plantas , Plântula , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Arabidopsis/metabolismo , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Plântula/genética , Açúcares/metabolismo , Sacarose/metabolismo , Glucose/metabolismo , Estiolamento , Metabolismo dos Carboidratos , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Cotilédone/metabolismo , Cotilédone/crescimento & desenvolvimento , Cotilédone/genética
13.
J Exp Child Psychol ; 241: 105862, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38320357

RESUMO

Children are strong imitators, which sometimes leads to overimitation of causally unnecessary actions. Here, we tested whether learning from a peer decreases this tendency. First, 65 7- to 10-year-old children performed the Hook task (i.e., retrieve a reward from a jar with tools) with child or adult demonstrators. The overimitation rate was lower after watching a peer versus an adult. Second, we tested whether experiencing peer-to-peer learning versus adult-driven learning (i.e., Montessori or traditional pedagogy) affected overimitation. Here, 66 4- to 18-year-old children and adolescents performed the Hook task with adult demonstrators only. Montessori-schooled children had a lower propensity to overimitate. These findings emphasize the importance of the teaching model across the school years. Whereas peer models favor selective imitation, adult models encourage overimitation.


Assuntos
Comportamento Imitativo , Instituições Acadêmicas , Criança , Adulto , Humanos , Adolescente , Pré-Escolar , Recompensa
14.
Artigo em Inglês | MEDLINE | ID: mdl-39243225

RESUMO

Radial head subluxation, known as pulled elbow or nursemaid's elbow, is a common presentation to the Paediatric Emergency Department (PED). Fifty per cent of children present following an unwitnessed injury or with an atypical history. Furthermore, examination can be difficult in an upset or frightened child meaning the precise location of the injury is not always easily identifiable. Many children receive upper limb x-rays unnecessarily and others undergo attempted reduction for suspected pulled elbow when a fracture is the cause of their pain. OBJECTIVE: To evaluate the ability of point-of-care ultrasound (POCUS) to correctly identify cases of suspected pulled elbow. METHODS: A retrospective review of patients aged 0-5 years presenting to the PED of our large District General Hospital from November 2020 to November 2022. We reviewed the electronic medical records of all patients presenting with a triage category of 'elbow Injury'. We excluded patients presenting with obvious deformity, significant swelling and history of road traffic collision or other major trauma mechanism such as a fall from a height. POCUS was carried out by the authors according to pre-existing clinical protocols. RESULTS: Thirty-seven patients underwent POCUS. Twenty-nine had a typical history and all had a positive POCUS for pulled elbow. Of the remaining eight patients with an atypical history, three were found to have a positive POCUS compared to unaffected elbow. Five patients with an atypical history had negative POCUS findings and were diagnosed as soft tissue injuries after further assessment. All 32 with positive sonographic signs had a final diagnosis of pulled elbow, meaning the specificity was 100% (47.8-100) (P = 0.00003). The sensitivity was 100%. CONCLUSION: Use of POCUS by appropriately trained clinicians, as an extension of a clinical examination, in this retrospective study reliably confirmed pulled elbow. This has the potential to reduce both unnecessary radiation as well as the time in the emergency department for the child and their carers.

15.
World J Surg Oncol ; 22(1): 51, 2024 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-38336734

RESUMO

BACKGROUND: Presurgical computed tomography (CT)-guided localization is frequently employed to reduce the thoracotomy conversion rate, while increasing the rate of successful sublobar resection of ground glass nodules (GGNs) via video-assisted thoracoscopic surgery (VATS). In this study, we compared the clinical efficacies of presurgical CT-guided hook-wire and indocyanine green (IG)-based localization of GGNs. METHODS: Between January 2018 and December 2021, we recruited 86 patients who underwent CT-guided hook-wire or IG-based GGN localization before VATS resection in our hospital, and compared the clinical efficiency and safety of both techniques. RESULTS: A total of 38 patients with 39 GGNs were included in the hook-wire group, whereas 48 patients with 50 GGNs were included in the IG group. There were no significant disparities in the baseline data between the two groups of patients. According to our investigation, the technical success rates of CT-based hook-wire- and IG-based localization procedures were 97.4% and 100%, respectively (P = 1.000). Moreover, the significantly longer localization duration (15.3 ± 6.3 min vs. 11.2 ± 5.3 min, P = 0.002) and higher visual analog scale (4.5 ± 0.6 vs. 3.0 ± 0.5, P = 0.001) were observed in the hook-wire patients, than in the IG patients. Occurrence of pneumothorax was significantly higher in hook-wire patients (27.3% vs. 6.3%, P = 0.048). Lung hemorrhage seemed higher in hook-wire patients (28.9% vs. 12.5%, P = 0.057) but did not reach statistical significance. Lastly, the technical success rates of VATS sublobar resection were 97.4% and 100% in hook-wire and IG patients, respectively (P = 1.000). CONCLUSIONS: Both hook-wire- and IG-based localization methods can effectively identified GGNs before VATS resection. Furthermore, IG-based localization resulted in fewer complications, lower pain scores, and a shorter duration of localization.


Assuntos
Neoplasias Pulmonares , Nódulos Pulmonares Múltiplos , Nódulo Pulmonar Solitário , Humanos , Neoplasias Pulmonares/diagnóstico por imagem , Neoplasias Pulmonares/cirurgia , Verde de Indocianina , Nódulo Pulmonar Solitário/diagnóstico por imagem , Nódulo Pulmonar Solitário/cirurgia , Estudos Retrospectivos , Tomografia Computadorizada por Raios X/métodos , Cirurgia Torácica Vídeoassistida/métodos , Pulmão , Nódulos Pulmonares Múltiplos/diagnóstico por imagem , Nódulos Pulmonares Múltiplos/cirurgia
16.
Proc Natl Acad Sci U S A ; 118(3)2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33419940

RESUMO

In many eukaryotic systems during immune responses, mitogen-activated protein kinases (MAPKs) link cytoplasmic signaling to chromatin events by targeting transcription factors, chromatin remodeling complexes, and the RNA polymerase machinery. So far, knowledge on these events is scarce in plants and no attempts have been made to focus on phosphorylation events of chromatin-associated proteins. Here we carried out chromatin phosphoproteomics upon elicitor-induced activation of Arabidopsis The events in WT were compared with those in mpk3, mpk4, and mpk6 mutant plants to decipher specific MAPK targets. Our study highlights distinct signaling networks involving MPK3, MPK4, and MPK6 in chromatin organization and modification, as well as in RNA transcription and processing. Among the chromatin targets, we characterized the AT-hook motif containing nuclear localized (AHL) DNA-binding protein AHL13 as a substrate of immune MAPKs. AHL13 knockout mutant plants are compromised in pathogen-associated molecular pattern (PAMP)-induced reactive oxygen species production, expression of defense genes, and PAMP-triggered immunity. Transcriptome analysis revealed that AHL13 regulates key factors of jasmonic acid biosynthesis and signaling and affects immunity toward Pseudomonas syringae and Botrytis cinerea pathogens. Mutational analysis of the phosphorylation sites of AHL13 demonstrated that phosphorylation regulates AHL13 protein stability and thereby its immune functions.


Assuntos
Proteínas de Arabidopsis/genética , Cromatina/genética , Fosfoproteínas/genética , Imunidade Vegetal/genética , Motivos AT-Hook/genética , Motivos AT-Hook/imunologia , Arabidopsis/genética , Arabidopsis/imunologia , Regulação da Expressão Gênica de Plantas , Proteínas Quinases Ativadas por Mitógeno/genética , Moléculas com Motivos Associados a Patógenos/imunologia , Moléculas com Motivos Associados a Patógenos/metabolismo , Fosfoproteínas/imunologia , Fosforilação/genética
17.
BMC Musculoskelet Disord ; 25(1): 152, 2024 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-38368342

RESUMO

BACKGROUND: Pedicle screw and lamina hook (PSLH) technique is an effective and popular method for direct pars repair of lumbar spondylolysis. There is a lack of studies to explore factors that may influence the healing of spondylolysis after direct pars repair surgery. The present study aimed to investigate the factors associated with non-fusion after direct pars repair of lumbar spondylolysis with PSLH technique. METHODS: A total of 55 subjects (average age 21.1 ± 6.3 years, a total of 120 pars defects) diagnosed with symptomatic spondylolysis and underwent pars repair surgery with PSLH were followed up and their clinical data were analyzed. Subjects were divided into a non-fusion group and fusion group according to whether the pars defect had bony fusion at last follow-up assessed by CT. Radiographic data, data related to spondylolysis and clinical outcomes were collected and compared between groups. RESULTS: The mean follow-up time of the 55 patients was 24.8 ± 12.0 (12-64) months. Among the 120 pars defects, 101 defects were successfully fused and 19 were not fused according to CT. The fusion rate was 84.2%. Multivariable logistic regression analysis showed the factors correlated with non-fusion after pars repair surgery: whether the spondylolysis segment was associated with spina bifida occulta (SBO) (P = 0.001), stage of the defect (P = 0.047), width of the defect (P = 0.002), and disc degeneration (P = 0.014). CONCLUSION: Direct pars repair by PSHL is a reliable treatment for lumbar spondylolysis with a fusion rate of 84.2%. Association with SBO of the spondylolysis segment, a terminal stage of the defect, a wider defect gap, and grade III disc degeneration may be factors associated with non-fusion after direct pars repair of lumbar spondylolysis with PLSH. Non-fusion patients after pars repair appear to have worse clinical results compared to fusion patients.


Assuntos
Degeneração do Disco Intervertebral , Parafusos Pediculares , Fusão Vertebral , Espondilólise , Humanos , Adolescente , Adulto Jovem , Adulto , Espondilólise/diagnóstico por imagem , Espondilólise/cirurgia , Vértebras Lombares/diagnóstico por imagem , Vértebras Lombares/cirurgia , Fusão Vertebral/efeitos adversos , Fusão Vertebral/métodos , Tomografia Computadorizada por Raios X/métodos , Resultado do Tratamento
18.
BMC Musculoskelet Disord ; 25(1): 174, 2024 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-38409002

RESUMO

PURPOSE: To compare the clinical efficacy of arthroscopic TightRope loop titanium button and clavicular hook plate in the treatment of acromioclavicular joint (ACJ) dislocation of Rockwood III/IV. METHODS: A retrospective analysis of patients with ACJ dislocation in our hospital from January 2018 to December 2020 was conducted. The patients were assigned to be treated with arthroscopic TightRope loop titanium button (TR group) or clavicular hook plate (HP group). The preoperative, intraoperative and postoperative data and imaging findings of the two groups were compared. RESULTS: A total of 58 eligible patients were enrolled in this study. Compared with HP group, TR group had shorter incision length and less blood loss during operation. Postoperative follow-up ranged from 12 to 24 months (mean 15.4 months). At 6 months and 12months postoperatively, compared with HP group, TR group had lower VAS and higher CMS, and the difference was statistically significant. At 12 months postoperatively, compared with HP group, TR group had lower ACJ gap and coracoclavicular joint(CCJ) distance, and the difference was statistically significant.In HP group, there were 3 cases of subacromial impact, 1 case of redislocation, 2 cases of traumatic arthritis and 2 cases of wound infection. There was 1 case of redislocation in TR group. CONCLUSIONS: Compared with clavicular hook plate, arthroscopic TightRope loop titanium button is minimally invasive, safe and effective in the treatment of ACJ dislocation, and has a good trend in clinical application.


Assuntos
Articulação Acromioclavicular , Luxações Articulares , Luxação do Ombro , Humanos , Estudos Retrospectivos , Luxações Articulares/cirurgia , Titânio , Articulação Acromioclavicular/diagnóstico por imagem , Articulação Acromioclavicular/cirurgia , Luxação do Ombro/cirurgia , Placas Ósseas , Resultado do Tratamento
19.
Ren Fail ; 46(1): 2295425, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38178377

RESUMO

AIM: Tripterygium wilfordii Hook F (TwHF) has been shown to substantially reduce proteinuria in patients with diabetic kidney disease (DKD); however, the effect of TwHF on renal outcomes in DKD remains unknown. Accordingly, we aimed to establish the effects of TwHF on renal outcomes in patients with DKD. METHODS: Overall, 124 patients with DKD, induced by type 2 diabetes mellitus, with 24-h proteinuria > 2 g, and an estimated glomerular filtration rate > 30 mL/min/1.73 m2 were retrospectively investigated. The renal outcomes were defined as doubling serum creatinine levels or end-stage kidney disease. Kaplan-Meier curves and Cox regression analyses were performed to analyze prognostic factors for renal outcomes. RESULTS: By the end of the follow-up, renal outcomes were observed in 23 and 11 patients in the non-TwHF and TwHF groups, respectively (p = 0.006). TwHF significantly reduced the risk of renal outcomes (adjusted hazard ratio [HR] 0.271, 95% confidence interval [CI] 0.111-0.660, p = 0.004) in patients with chronic kidney disease (CKD) G3 (adjusted HR 0.274, 95%CI 0.081-0.932, p = 0.039). Based on the Kaplan-Meier analysis, 1- and 3-year proportions of patients without renal outcomes were significantly lower in the non-TwHF group than those in the TwHF group (92.8% vs. 95.5% and 47.2% vs. 76.8%, respectively; p = 0.0018). CONCLUSION: In DKD patients with severe proteinuria, TwHF could prevent DKD progression, especially in patients with CKD G3. A randomized clinical trial is needed to elucidate the benefits of TwHF on renal outcomes in patients with DKD.


Assuntos
Diabetes Mellitus Tipo 2 , Nefropatias Diabéticas , Insuficiência Renal Crônica , Humanos , Nefropatias Diabéticas/complicações , Nefropatias Diabéticas/tratamento farmacológico , Tripterygium , Estudos de Coortes , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/tratamento farmacológico , Estudos Retrospectivos , Proteinúria/tratamento farmacológico , Proteinúria/etiologia
20.
J Shoulder Elbow Surg ; 33(9): 2086-2095, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-38642874

RESUMO

BACKGROUND: Clavicular hook plates are extensively used in the treatment of acromioclavicular dislocation. Subacromial osteolysis is a typical complication following hook plate fixation. We performed a systematic review and meta-analysis to determine the incidence of subacromial osteolysis and analyzed the associated characteristics of subacromial osteolysis to guide surgeons. METHODS: PubMed, EMBASE, and Cochrane Library databases were comprehensively searched for relevant literature. We screened the literature based on the eligibility criteria, extracted relevant data, and assessed the quality of the included studies. Pooled odds ratios or mean differences with 95% confidence intervals (CIs) were calculated by a fixed-effects or random-effects model. Heterogeneity was evaluated by the chi-squared test and I2 statistics. A meta-regression analysis was performed to explore the potential source of heterogeneity. RESULTS: Thirty-two studies met the inclusion criteria. The total pooled incidence of subacromial osteolysis was 29% and the only covariate that could influence the incidence of subacromial osteolysis was the radiological measurement method (P = .017). Patients in the hook plate fixation with coracoclavicular ligament reconstruction group had lower odds of subacromial osteolysis (odds ratio, 2.54, 95% CI 1.54-4.18; P < .001). There were no significant differences in the Constant-Murley scores at the final follow-up between patients with and without subacromial osteolysis (standardized mean difference, -0.17; 95% CI, -0.50 to 0.15; P = .294). CONCLUSIONS: Subacromial osteolysis has a relatively high and variable incidence, and the primary factor influencing the reported incidence is the radiological assessment method. The current analysis suggests coracoclavicular ligament reconstruction as an effective surgical approach for decreasing the incidence of subacromial osteolysis. The presence or absence of subacromial osteolysis did not significantly impact the functional outcomes observed during the final follow-up period.


Assuntos
Articulação Acromioclavicular , Placas Ósseas , Osteólise , Humanos , Articulação Acromioclavicular/cirurgia , Articulação Acromioclavicular/lesões , Osteólise/etiologia , Osteólise/epidemiologia , Placas Ósseas/efeitos adversos , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Luxações Articulares/cirurgia
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