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2.
Adv Sci (Weinh) ; : e2402732, 2024 Jun 23.
Artículo en Inglés | MEDLINE | ID: mdl-38923364

RESUMEN

The development of in situ techniques to quantitatively characterize the heterogeneous reactions is essential for understanding physicochemical processes in aqueous phase. In this work, a new approach coupling in situ UV-vis spectroscopy with a two-step algorithm strategy is developed to quantitatively monitor heterogeneous reactions in a compact closed-loop incorporation. The algorithm involves the inverse adding-doubling method for light scattering correction and the multivariate curve resolution-alternating least squares (MCR-ALS) method for spectral deconvolution. Innovatively, theoretical spectral simulations are employed to connect MCR-ALS solutions with chemical molecular structural evolution without prior information for reference spectra. As a model case study, the aqueous adsorption kinetics of bisphenol A onto polyamide microparticles are successfully quantified in a one-step UV-vis spectroscopic measurement. The practical applicability of this approach is confirmed by rapidly screening a superior adsorbent from commercial materials for antibiotic wastewater adsorption treatment. The demonstrated capabilities are expected to extend beyond monitoring adsorption systems to other heterogeneous reactions, significantly advancing UV-vis spectroscopic techniques toward practical integration into automated experimental platforms for probing aqueous chemical processes and beyond.

3.
Plants (Basel) ; 13(11)2024 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-38891297

RESUMEN

Salt stress is one of the major adverse factors affecting plant growth and crop production. Rapeseed is an important oil crop, providing high-quality edible oil for human consumption. This experiment was conducted to investigate the effects of salt stress on the phenotypic traits and physiological processes of rapeseed. The soil salinity was manipulated by setting three different levels: 0 g NaCl kg-1 soil (referred to as S0), 1.5 g NaCl kg-1 soil (referred to as S1), and 3.0 g NaCl kg-1 soil (referred to as S2). In general, the results indicated that the plant height, leaf area, and root neck diameter decreased with an increase in soil salinity. In addition, the biomass of various organs at all growth stages decreased as soil salinity increased from S0 to S2. The increasing soil salinity improved the distribution of biomass in the root and leaf at the seedling and flowering stages, indicating that rapeseed plants subjected to salt stress during the vegetative stage are capable of adapting their growth pattern to sustain their capacity for nutrient and water uptake, as well as leaf photosynthesis. However, as the soil salinity increased, there was a decrease in the distribution of biomass in the pod and seed at the maturity stage, while an increase was observed in the root and stem, suggesting that salt stress inhibited carbohydrate transport into reproductive organs. Moreover, the C and N accumulation at the flowering and maturity stages exhibited a reduction in direct correlation with the increase in soil salinity. High soil salinity resulted in a reduction in the C/N, indicating that salt stress exerted a greater adverse effect on C assimilation compared to N assimilation, leading to an increase in seed protein content and a decrease in oil content. Furthermore, as soil salinity increased from S0 to S2, the activity of superoxide dismutase (SOD) and catalase (CAT) and the content of soluble protein and sugar increased by 58.39%, 33.38%, 15.57%, and 13.88% at the seedling stage, and 38.69%, 22.85%, 12.04%, and 8.26% at the flowering stage, respectively. In summary, this study revealed that salt stress inhibited C and N assimilation, leading to a suppressed phenotype and biomass accumulation. The imbalanced C and N assimilation under salt stress contributed to the alterations in the seed oil and protein content. Rapeseed had a certain degree of salt tolerance by improving antioxidants and osmolytes.

4.
Nature ; 630(8017): 736-743, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38839956

RESUMEN

Phagocytosis is the process by which myeloid phagocytes bind to and internalize potentially dangerous microorganisms1. During phagocytosis, innate immune receptors and associated signalling proteins are localized to the maturing phagosome compartment, forming an immune information processing hub brimming with microorganism-sensing features2-8. Here we developed proximity labelling of phagosomal contents (PhagoPL) to identify proteins localizing to phagosomes containing model yeast and bacteria. By comparing the protein composition of phagosomes containing evolutionarily and biochemically distinct microorganisms, we unexpectedly identified programmed death-ligand 1 (PD-L1) as a protein that specifically enriches in phagosomes containing yeast. We found that PD-L1 directly binds to yeast upon processing in phagosomes. By surface display library screening, we identified the ribosomal protein Rpl20b as a fungal protein ligand for PD-L1. Using an auxin-inducible depletion system, we found that detection of Rpl20b by macrophages cross-regulates production of distinct cytokines including interleukin-10 (IL-10) induced by the activation of other innate immune receptors. Thus, this study establishes PhagoPL as a useful approach to quantifying the collection of proteins enriched in phagosomes during host-microorganism interactions, exemplified by identifying PD-L1 as a receptor that binds to fungi.


Asunto(s)
Antígeno B7-H1 , Macrófagos , Fagosomas , Proteínas Ribosómicas , Fagosomas/metabolismo , Antígeno B7-H1/metabolismo , Animales , Ratones , Macrófagos/metabolismo , Macrófagos/inmunología , Macrófagos/microbiología , Proteínas Ribosómicas/metabolismo , Unión Proteica , Interleucina-10/metabolismo , Fagocitosis , Saccharomyces cerevisiae/metabolismo , Proteínas Fúngicas/metabolismo , Ligandos , Humanos , Femenino , Inmunidad Innata
5.
Liver Int ; 2024 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-38847599

RESUMEN

BACKGROUND AND AIMS: Metabolic dysfunction-associated steatotic liver disease (MASLD) represents the foremost cause of chronic liver disease, yet its underlying mechanisms remain elusive. Our group previously discovered a novel long non-coding RNA (lncRNA) in rats, termed lncHC and its human counterpart, LNCHC. This study aimed to explore the role of LNCHC in the progression of MASLD. METHODS: RNA-binding proteins bound to LNCHC were searched by mass spectrometry. The target genes of LNCHC and Y-Box binding protein 1 (YBX1) were identified by RNA-seq. MASLD animal models were utilised to examine the roles of LNCHC, YBX1 and patatin-like phospholipase domain containing 3 (PNPLA3) in MASLD progression. RESULTS: Here, we identified LNCHC as a native restrainer during MASLD development. Notably, LNCHC directly binds YBX1 and prevents protein ubiquitination. Up-regulation of YBX1 then stabilises PNPLA3 mRNA to alleviate lipid accumulation in hepatocytes. Furthermore, both cell and animal studies demonstrate that LNCHC, YBX1 and PNPLA3 function to improve hepatocyte lipid accumulation and exacerbate metabolic dysfunction-associated steatohepatitis development. CONCLUSIONS: In summary, our findings unveil a novel LNCHC functionality in regulating YBX1 and PNPLA3 mRNA stability during MASLD development, providing new avenues in MASLD treatment.

6.
Nucleic Acids Res ; 2024 Jun 27.
Artículo en Inglés | MEDLINE | ID: mdl-38932701

RESUMEN

Androgen receptor- (AR-) indifference is a mechanism of resistance to hormonal therapy in prostate cancer (PC). Here we demonstrate that ONECUT2 (OC2) activates resistance through multiple drivers associated with adenocarcinoma, stem-like and neuroendocrine (NE) variants. Direct OC2 gene targets include the glucocorticoid receptor (GR; NR3C1) and the NE splicing factor SRRM4, which are key drivers of lineage plasticity. Thus, OC2, despite its previously described NEPC driver function, can indirectly activate a portion of the AR cistrome through epigenetic activation of GR. Mechanisms by which OC2 regulates gene expression include promoter binding, enhancement of genome-wide chromatin accessibility, and super-enhancer reprogramming. Pharmacologic inhibition of OC2 suppresses lineage plasticity reprogramming induced by the AR signaling inhibitor enzalutamide. These results demonstrate that OC2 activation promotes a range of drug resistance mechanisms associated with treatment-emergent lineage variation in PC and support enhanced efforts to therapeutically target OC2 as a means of suppressing treatment-resistant disease.

7.
Mol Biol Evol ; 2024 Jun 06.
Artículo en Inglés | MEDLINE | ID: mdl-38842255

RESUMEN

The origins and extreme morphological evolution of the modern dog breeds are poorly studied because the founder populations are extinct. Here, we analyse eight 100-200 years old dog fur samples obtained from traditional North Swedish clothing, to explore the origin and artificial selection of the modern Nordic Lapphund and Elkhound dog breeds. Population genomic analysis confirmed the Lapphund and Elkhound breeds to originate from the local dog population, and showed a distinct decrease in genetic diversity in agreement with intense breeding. We identified eleven genes under positive selection during the breed development. In particular, the MSRB3 gene, associated with breed-related ear morphology, was selected in all Lapphund and Elkhound breeds, and functional assays showed that a SNP mutation in the 3'UTR region suppresses its expression through miRNA regulation. Our findings demonstrate analysis of near-modern dog artifacts as an effective tool for interpreting the origin and artificial selection of the modern dog breeds.

8.
Nano Lett ; 24(20): 6124-6130, 2024 May 22.
Artículo en Inglés | MEDLINE | ID: mdl-38717388

RESUMEN

The identification of nanoparticles within heterogeneous mixtures poses significant challenges due to the similarity in physical properties among different nanomaterials. Here, we present electrochemically assisted high-resolution plasmonic scattering interferometric microscopy (HR-PSIM). This technique allows for the high-throughput identification of nanoparticles by accurately measuring the refractive index of individual nanoparticles without interference from background signals. Through elimination of parabolic scattering interference and employing electrochemical modulation, HR-PSIM demonstrates high spatial resolution and stability against background noise, enabling the differentiation of nanoparticles with closely matched refractive indices, such as Au and Ag nanoparticles. The efficacy of this method is demonstrated through its application in real-time, label-free imaging of nanoparticle electrochemical activity, providing a platform for the precise and high-throughput characterization of nanomaterials. The robustness of our approach against electrochemical interference and its high spatial resolution mark a significant advancement in the field of nanomaterial analysis, promising wide-ranging applications in nanoparticle research and beyond.

9.
J Adv Res ; 2024 May 04.
Artículo en Inglés | MEDLINE | ID: mdl-38710468

RESUMEN

BACKGROUND: Arachidonic acid (AA), one of the most ubiquitous polyunsaturated fatty acids (PUFAs), provides fluidity to mammalian cell membranes. It is derived from linoleic acid (LA) and can be transformed into various bioactive metabolites, including prostaglandins (PGs), thromboxanes (TXs), lipoxins (LXs), hydroxy-eicosatetraenoic acids (HETEs), leukotrienes (LTs), and epoxyeicosatrienoic acids (EETs), by different pathways. All these processes are involved in AA metabolism. Currently, in the context of an increasingly visible aging world population, several scholars have revealed the essential role of AA metabolism in osteoporosis, chronic obstructive pulmonary disease, and many other aging diseases. AIM OF REVIEW: Although there are some reviews describing the role of AA in some specific diseases, there seems to be no or little information on the role of AA metabolism in aging tissues or organs. This review scrutinizes and highlights the role of AA metabolism in aging and provides a new idea for strategies for treating aging-related diseases. KEY SCIENTIFIC CONCEPTS OF REVIEW: As a member of lipid metabolism, AA metabolism regulates the important lipids that interfere with the aging in several ways. We present a comprehensivereviewofthe role ofAA metabolism in aging, with the aim of relieving the extreme suffering of families and the heavy economic burden on society caused by age-related diseases. We also collected and summarized data on anti-aging therapies associated with AA metabolism, with the expectation of identifying a novel and efficient way to protect against aging.

11.
AORN J ; 119(6): e1-e9, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38804729

RESUMEN

Minimally invasive surgery can involve the use of robotics to improve patient outcomes. Some robotic systems require special instruments with a designated number of uses. In China, during the reprocessing of the robotic instruments, health care personnel determined that the existing tracking processes were inadequate. They conducted a quality improvement project with the goal of establishing a barcode-based standardized process for tracking robotic instruments. They implemented technology that generated a unique identifier each time a robotic instrument was reprocessed after use. Nurses scanned the identifier when surgeons used the instrument. The findings included the increased accuracy of use documentation and decreases in untraceable sterilization and use records, charging concerns, and average daily and monthly inventory times. An increase in adverse event reports associated with robotic instruments also was noted. The use of barcode technology for robotic instrument tracking continues at the facility and may be expanded for additional specialty instruments.


Asunto(s)
Procesamiento Automatizado de Datos , Procesamiento Automatizado de Datos/métodos , Humanos , Robótica/instrumentación , Robótica/normas , Mejoramiento de la Calidad , Procedimientos Quirúrgicos Robotizados/métodos , Procedimientos Quirúrgicos Robotizados/instrumentación , China
12.
Plants (Basel) ; 13(10)2024 May 14.
Artículo en Inglés | MEDLINE | ID: mdl-38794423

RESUMEN

The degree of forage lignification is a key factor affecting its digestibility by ruminants such as cattle and sheep. Sudan grass (Sorghum sudanense S.) is a high-quality sorghum forage, and its lignocellulose is mostly stored in the secondary cell wall. However, the secondary cell wall synthesis mechanism of Sudan grass has not yet been studied in depth. To further study the secondary cell wall synthesis mechanism of Sudan grass using established transcriptome data, this study found that CcNAC6, a homologous gene of Arabidopsis AtSND2, is related to the secondary cell wall synthesis of Sudan grass. Accordingly, we constructed a CcNAC6-overexpressing line of Arabidopsis to investigate the function of the CcNAC6 gene in secondary cell wall synthesis. The results showed that the overexpression of the CcNAC6 gene could significantly increase the lignin content of Arabidopsis. Based on subcellular localization analysis, CcNAC6 is found in the nucleus. In addition, yeast two-hybridization screening showed that CcCP1, associated with secondary cell wall synthesis, can interact with CcNAC6. Therefore, the above results indicate that CcNAC6 has a positive regulatory effect on the secondary cell wall synthesis of Sudan grass, and it is speculated that CcNAC6 may be the main regulator of the secondary cell wall synthesis of Sudan grass through its interaction with another regulatory protein, CcCP1. This study provides a theoretical basis and new genetic resources for the creation of new Sudan grass germplasm with a low lignin content.

13.
Artículo en Inglés | MEDLINE | ID: mdl-38809743

RESUMEN

Self-supervised Learning (SSL) including the mainstream contrastive learning has achieved great success in learning visual representations without data annotations. However, most methods mainly focus on the instance level information (ie, the different augmented images of the same instance should have the same feature or cluster into the same class), but there is a lack of attention on the relationships between different instances. In this paper, we introduce a novel SSL paradigm, which we term as relational self-supervised learning (ReSSL) framework that learns representations by modeling the relationship between different instances. Specifically, our proposed method employs sharpened distribution of pairwise similarities among different instances as relation metric, which is thus utilized to match the feature embeddings of different augmentations. To boost the performance, we argue that weak augmentations matter to represent a more reliable relation, and leverage momentum strategy for practical efficiency. The designed asymmetric predictor head and an InfoNCE warm-up strategy enhance the robustness to hyper-parameters and benefit the resulting performance. Experimental results show that our proposed ReSSL substantially outperforms the state-of-the-art methods across different network architectures, including various lightweight networks (eg, EfficientNet and MobileNet).

14.
mBio ; : e0013324, 2024 May 30.
Artículo en Inglés | MEDLINE | ID: mdl-38814088

RESUMEN

Botrytis cinerea is a typical necrotrophic plant pathogenic fungus which can deliberately acidify host tissues and trigger oxidative bursts therein to facilitate its virulence. The white collar complex (WCC), consisting of BcWCL1 and BcWCL2, is recognized as the primary light receptor in B. cinerea. Nevertheless, the specific mechanisms through which the WCC components, particularly BcWCL2 as a GATA transcription factor, control virulence are not yet fully understood. This study demonstrates that deletion of BcWCL2 results in the loss of light-sensitive phenotypic characteristics. Additionally, the Δbcwcl2 strain exhibits reduced secretion of citrate, delayed infection cushion development, weaker hyphal penetration, and decreased virulence. The application of exogenous citric acid was found to restore infection cushion formation, hyphal penetration, and virulence of the Δbcwcl2 strain. Transcriptome analysis at 48 h post-inoculation revealed that two citrate synthases, putative citrate transporters, hydrolytic enzymes, and reactive oxygen species scavenging-related genes were down-regulated in Δbcwcl2, whereas exogenous citric acid application restored the expression of the above genes involved in the early infection process of Δbcwcl2. Moreover, the expression of Bcvel1, a known regulator of citrate secretion, tissue acidification, and secondary metabolism, was down-regulated in Δbcwcl2 but not in Δbcwcl1. ChIP-qPCR and electrophoretic mobility shift assays revealed that BcWCL2 can bind to the promoter sequences of Bcvel1. Overexpressing Bcvel1 in Δbcwcl2 was found to rescue the mutant defects. Collectively, our findings indicate that BcWCL2 regulates the expression of the global regulator Bcvel1 to influence citrate secretion, tissue acidification, redox homeostasis, and virulence of B. cinerea.IMPORTANCEThis study illustrated the significance of the fungal blue light receptor component BcWCL2 protein in regulating citrate secretion in Botrytis cinerea. Unlike BcWCL1, BcWCL2 may contribute to redox homeostasis maintenance during infection cushion formation, ultimately proving to be essential for full virulence. It is also demonstrated that BcWCL2 can regulate the expression of Bcvel1 to influence host tissue acidification, citrate secretion, infection cushion development, and virulence. While the role of organic acids secreted by plant pathogenic fungi in fungus-host interactions has been recognized, this paper revealed the importance, regulatory mechanisms, and key transcription factors that control organic acid secretion. These understanding of the pathogenetic mechanism of plant pathogens can provide valuable insights for developing effective prevention and treatment strategies against fungal diseases.

15.
Influenza Other Respir Viruses ; 18(4): e13291, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38653953

RESUMEN

BACKGROUND: Nonpharmaceutical interventions (NPIs) targeted at SARS-CoV-2 have remarkably affected the circulation of other respiratory pathogens, including respiratory syncytial virus (RSV). This study aimed to assess the changes in epidemiological and clinical characteristics of RSV infections in hospitalized children before and during the pandemic in Suzhou, China. METHODS: We prospectively enrolled children aged < 18 years who were hospitalized in Soochow University Affiliated Children's Hospital with acute lower respiratory infection (ALRIs) from January 2018 to July 2022. Changes in epidemiological and clinical characteristics of RSV infections were analyzed. RESULTS: Compared with the same period in 2018-2019, the difference in the overall positive rate of RSV was not statistically significant in 2020, while it increased significantly in 2021 (11.8% [662/5621] vs. 20.8% [356/1711], p < 0.001) and 2022 (9.0% [308/3406] vs. 18.9% [129/684], p < 0.001). Specifically, the positive rates declined considerably from October to December 2020 but sharply increased during the summer of 2021. Compared to prepandemic period, RSV infections were more frequently observed in older children during the pandemic. RSV-positive children exhibited milder clinical characteristics during the COVID-19 pandemic, including decreased proportion of patients with hospital stay ≥ 11 days (10.3% vs. 6.7%, p < 0.05), less requirement for oxygen therapy (13.7% vs. 6.9%, p < 0.001), and fewer cases of polypnea (12.2% vs. 9.7%, p < 0.05) and wheeze (50.1% vs. 42.9%, p < 0.001). CONCLUSIONS: The implementation of multilayered NPIs targeted at COVID-19 has affected the activity of RSV. Ongoing monitoring of RSV is warranted as the changing RSV epidemiology can provide valuable insights for future healthcare system planning.


Asunto(s)
COVID-19 , Hospitalización , Infecciones por Virus Sincitial Respiratorio , SARS-CoV-2 , Humanos , Infecciones por Virus Sincitial Respiratorio/epidemiología , Infecciones por Virus Sincitial Respiratorio/prevención & control , COVID-19/epidemiología , COVID-19/prevención & control , Preescolar , Masculino , Femenino , Lactante , Niño , China/epidemiología , Estudios Prospectivos , Hospitalización/estadística & datos numéricos , Adolescente , Virus Sincitial Respiratorio Humano , Niño Hospitalizado/estadística & datos numéricos , Recién Nacido
16.
Phys Rev Lett ; 132(11): 116701, 2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-38563939

RESUMEN

Cavity magnonics is an emerging research area focusing on the coupling between magnons and photons. Despite its great potential for coherent information processing, it has been long restricted by the narrow interaction bandwidth. In this Letter, we theoretically propose and experimentally demonstrate a novel approach to achieve broadband photon-magnon coupling by adopting slow waves on engineered microwave waveguides. To the best of our knowledge, this is the first time that slow wave is combined with hybrid magnonics. Its unique properties promise great potentials for both fundamental research and practical applications, for instance, by deepening our understanding of the light-matter interaction in the slow wave regime and providing high-efficiency spin wave transducers. The device concept can be extended to other systems such as optomagnonics and magnomechanics, opening up new directions for hybrid magnonics.

17.
Behav Brain Res ; 466: 114990, 2024 May 28.
Artículo en Inglés | MEDLINE | ID: mdl-38582411

RESUMEN

Previous studies have shown that alertness is closely related to executive control function, but its impact on components of post-error adjustment is unknown. This study applied the Attentional Networks Test and the Four-choice Flanker task with three response stimulus intervals (RSIs) to explore the correlation between alertness and post-error adjustment. The linear mixed-effects model of alertness and RSI on the post-error processing indicators showed a significant negative correlation between the alertness and post-error slowing (PES) under 200 ms RSI , as well as between alertness and post-error improvement in accuracy (PIA) under both 700 ms RSI and 1200 ms RSI. Participants with lower alertness showed larger post-error slowing in the early stages, while those with higher alertness had smaller PIA in later stages. This study revealed the effects of alertness on different processing components of post-error adjustment. The control strategies utilized by individuals with high and low levels of alertness differed in preparation for performance monitoring. Alertness improved post-error response speed in a task-unspecific manner, but not post-error adaptation.


Asunto(s)
Atención , Función Ejecutiva , Desempeño Psicomotor , Tiempo de Reacción , Humanos , Masculino , Femenino , Atención/fisiología , Adulto Joven , Tiempo de Reacción/fisiología , Función Ejecutiva/fisiología , Adulto , Desempeño Psicomotor/fisiología , Adaptación Psicológica/fisiología
19.
ACS Nano ; 18(13): 9704-9712, 2024 Apr 02.
Artículo en Inglés | MEDLINE | ID: mdl-38512797

RESUMEN

Label-free probing of the material composition of (bio)nano-objects directly in solution at the single-particle level is crucial in various fields, including colloid analysis and medical diagnostics. However, it remains challenging to decipher the constituents of heterogeneous mixtures of nano-objects with high sensitivity and resolution. Here, we present deep-learning plasmonic scattering interferometric microscopy, which is capable of identifying the composition of nanoparticles automatically with high throughput at the single-particle level. By employing deep learning to decode the quantitative relationship between the interferometric scattering patterns of nanoparticles and their intrinsic material properties, this technique is capable of high-throughput, label-free identification of diverse nanoparticle types. We demonstrate its versatility in analyzing dynamic surface chemical reactions on single nanoparticles, revealing its potential as a universal platform for nanoparticle imaging and reaction analysis. This technique not only streamlines the process of nanoparticle characterization, but also proposes a methodology for a deeper understanding of nanoscale dynamics, holding great potential for addressing extensive fundamental questions in nanoscience and nanotechnology.

20.
J Cell Biol ; 223(5)2024 May 06.
Artículo en Inglés | MEDLINE | ID: mdl-38448164

RESUMEN

The microtubule motor dynein plays a key role in cellular organization. However, little is known about how dynein's biosynthesis, assembly, and functional diversity are orchestrated. To address this issue, we have conducted an arrayed CRISPR loss-of-function screen in human cells using the distribution of dynein-tethered peroxisomes and early endosomes as readouts. From a genome-wide gRNA library, 195 validated hits were recovered and parsed into those impacting multiple dynein cargoes and those whose effects are restricted to a subset of cargoes. Clustering of high-dimensional phenotypic fingerprints revealed co-functional proteins involved in many cellular processes, including several candidate novel regulators of core dynein functions. Further analysis of one of these factors, the RNA-binding protein SUGP1, indicates that it promotes cargo trafficking by sustaining functional expression of the dynein activator LIS1. Our data represent a rich source of new hypotheses for investigating microtubule-based transport, as well as several other aspects of cellular organization captured by our high-content imaging.


Asunto(s)
Dineínas , Microtúbulos , Humanos , Dineínas/genética , Microtúbulos/genética , Peroxisomas/genética , Sistemas CRISPR-Cas , Técnicas Genéticas
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