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1.
Cell Death Dis ; 13(5): 437, 2022 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-35513389

RESUMO

Aberrantly synchronized neuronal discharges in the brain lead to epilepsy, a devastating neurological disease whose pathogenesis and mechanism are unclear. SAPAP3, a cytoskeletal protein expressed at high levels in the postsynaptic density (PSD) of excitatory synapses, has been well studied in the striatum, but the role of SAPAP3 in epilepsy remains elusive. In this study, we sought to investigate the molecular, cellular, electrophysiological and behavioral consequences of SAPAP3 perturbations in the mouse hippocampus. We identified a significant increase in the SAPAP3 levels in patients with temporal lobe epilepsy (TLE) and in mouse models of epilepsy. In addition, behavioral studies showed that the downregulation of SAPAP3 by shRNA decreased the seizure severity and that the overexpression of SAPAP3 by recombinant SAPAP3 yielded the opposite effect. Moreover, SAPAP3 affected action potentials (APs), miniature excitatory postsynaptic currents (mEPSCs) and N-methyl-D-aspartate receptor (NMDAR)-mediated currents in the CA1 region, which indicated that SAPAP3 plays an important role in excitatory synaptic transmission. Additionally, the levels of the GluN2A protein, which is involved in synaptic function, were perturbed in the hippocampal PSD, and this perturbation was accompanied by ultrastructural morphological changes. These results revealed a previously unknown function of SAPAP3 in epileptogenesis and showed that SAPAP3 may represent a novel target for the treatment of epilepsy.


Assuntos
Epilepsia do Lobo Temporal , Epilepsia , Animais , Epilepsia/metabolismo , Epilepsia do Lobo Temporal/patologia , Hipocampo/metabolismo , Humanos , Camundongos , Proteínas do Tecido Nervoso/genética , Proteínas do Tecido Nervoso/metabolismo , Densidade Pós-Sináptica/metabolismo , Densidade Pós-Sináptica/patologia , Receptores de N-Metil-D-Aspartato/genética , Receptores de N-Metil-D-Aspartato/metabolismo , Convulsões/metabolismo , Sinapses/metabolismo
2.
Cancers (Basel) ; 14(9)2022 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-35565354

RESUMO

In colorectal cancer, somatic mutations have played an important role as prognostic and predictive biomarkers, with some also functioning as therapeutic targets. Another genetic aberration that has shown significance in colorectal cancer is copy number alterations (CNAs). CNAs occur when a change to the DNA structure propagates gain/amplification or loss/deletion in sections of DNA, which can often lead to changes in protein expression. Multiple techniques have been developed to detect CNAs, including comparative genomic hybridization with microarray, low pass whole genome sequencing, and digital droplet PCR. In this review, we summarize key findings in the literature regarding the role of CNAs in the pathogenesis of colorectal cancer, from adenoma to carcinoma to distant metastasis, and discuss the roles of CNAs as prognostic and predictive biomarkers in colorectal cancer.

3.
J Agric Food Chem ; 2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35576531

RESUMO

The present study sought to identify and characterize a novel antimicrobial peptide, named MOp2 from Moringa oleifera seed protein hydrolysates, and elucidate its potential antimicrobial effects on Staphylococcus aureus. MOp2, with the amino acid sequence of His-Val-Leu-Asp-Thr-Pro-Leu-Leu (HVLDTPLL), was characterized as a hydrophobic anionic AMP of the ß-sheet structure. MOp2 exhibited negligible hemolytic activity at 2.0× MIC, suggesting its inhibitory effect on the growth of S. aureus (MIC: 2.204 mM). It maintained more than 90% of antimicrobial activity under 5% salt and about 78% of antimicrobial activity at a high temperature of 115 °C for 30 min. Protease, especially acid protease, reduced its antimicrobial activity to different extents. Moreover, MOp2 caused irreversible membrane damage to S. aureus cells by increasing the membrane permeability, resulting in the release of intracellular nucleotide pools. Additionally, molecular docking revealed that MOp2 could inhibit S. aureus growth by interacting with dihydrofolate reductase and DNA gyrase through hydrogen bonding and hydrophobic interactions. Overall, MOp2 could be a potential novel antimicrobial agent against S. aureus in food processing.

4.
J Colloid Interface Sci ; 622: 443-451, 2022 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-35526408

RESUMO

Engineering of robust nonprecious electrocatalysts toward anodic oxygen evolution reaction (OER) is of great significance for lowering the cost and energy consumption for renewable fuel production. Herein, we report NiFeMoOx nanosheets as high-performance OER electrocatalyst through promoting the thermodynamic-limiting oxidation cycle process in NiFe oxyhydroxide via high-valence Mo doping. The NiFeMoOx nanosheets are prepared by an elaborate in-situ solvothermal etching-depositing process with NiFe alloy framework as substrate and metal precursors. The resultant nanosheets exhibit outstanding alkaline OER activity, requires only 235/282/327 mV overpotentials to achieve current density of 10/100/300 mA cm-2, respectively, with a good long-term stability at 20 mA cm-2 for 72 h. Besides, the Tafel slope low to 28.1 mV dec-1 indicates a favorable OER kinetics. The superior catalytic activity of NiFeMoOx nanosheets should be attributed to the lower oxidation states of Ni and Fe induced by high-valence dopant, leading to easier surface reconstruction at low charge oxidation cycling during OER, thereby effectively reducing the overpotential. The synergy between the electronic effect among multimetallic sites and the unique morphology is expected to inspire the development of robust OER electrocatalyst for industrial application.

5.
Front Plant Sci ; 13: 788828, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35432402

RESUMO

Many factors, including illumination, affect anthocyanin biosynthesis and accumulation in plants. light quality is the key factor affecting the process of photoinduced anthocyanin biosynthesis and accumulation. We observed that the red color of the Upland cotton accession Huiyuan with the R1 mutation turned to normal green color under light-emitting diodes (LEDs), which inspired us to investigate the effect of red and blue lights on the biosynthesis and accumulation of anthocyanins. We found that both red and blue lights elevated accumulation of anthocyanins. Comparative transcriptomic analyses, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and GSEA, revealed that genes differentially expressed under different light conditions were enriched with the pathways of circadian rhythm, phenylpropanoid biosynthesis, anthocyanin biosynthesis, and flavone and flavonol biosynthesis. Not surprisingly, all the major structural genes related to biosynthesis of anthocyanins, including the key regulatory MYB transcription factor (GhPAP1D) and anthocyanin transporter (GhGSTF12), were induced by red or blue light treatment. However, LARs and MATEs related to biosynthesis of proanthocyanidins were more significantly up-regulated by red light radiation than by blue light radiation. Vice versa, the accumulation of anthocyanins under red light was not as high as that under blue light. In addition, we demonstrated a potential role of GhHY5, a key regulator in plant circadian rhythms, in regulation of anthocyanin accumulation, which could be achieved via interaction with GhPAP1D. Together, these results indicate different effect of red and blue lights on biosynthesis and accumulation of anthocyanins and a potential module including GhHY5 and GhPAP1D in regulation of anthocyanin accumulation in cotton. These results also suggest that the substrates responsible the synthesis of anthocyanins under blue light is diverted to biosynthesis of proanthocyanidin under red light.

6.
ACS Appl Mater Interfaces ; 14(16): 18353-18359, 2022 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-35417137

RESUMO

Li-rich layer-structured oxides are considered promising cathode materials for their specific capacities above 250 mAh·g-1. However, the drawbacks such as poor rate performance, fast capacity fading, and the continuous transition metal (TM) migration into the Li layer hinder their commercial applications. To address these issues, surface doping of Ti and Zr was conducted to the Li- and Mn-rich layered oxide (LMR), Li1.2Mn0.54Ni0.13Co0.13O2. The drop of the average discharge potentials of the Ti- and Zr-doped LMR was reduced by 593 and 346 mV in 100 cycles, respectively. With aberration-corrected scanning transmission electron microscopy and electron energy loss spectroscopy, we clarified that Ti4+ and Zr4+ ions are located near the surface of the material, anchor the surface oxygen, and stabilize the LMR structure. The difference in the strengths of the Ti-O and Zr-O bonds and the doping-resultant electronic structures were determined with density functional theory (DFT) calculations and soft X-ray absorption spectroscopy (SXAS), responsible for the electrochemical performance of surface-doped materials. These findings verify our modification strategies to enhance the cycling performances of the promising LMR cathode materials.

7.
Hum Mutat ; 2022 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-35391506

RESUMO

There is growing evidence that synonymous codon variants (SCVs) can cause disease through the disruption of different processes of protein production. The aim of the study is to investigate whether the 14 SCVs reported in the F9 variant database were the pathogenic causes of hemophilia B. The impacts of SCVs on splicing and protein expression were detected using a combination of in silico prediction, in vitro minigene splicing assay and cell expression detection. The splicing transcripts were identified and quantified by co-amplification fluorescent PCR. The mechanism of splicing was verified by a modified pU1snRNA and pU7snRNA approach. Aberrant splicing patterns were found in eight SCVs. Five of the 8 SCVs produced almost all aberrant splicing isoforms, which were expected to truncate protein, three of them presented a partial defect on both splicing and protein secretion, the overall effects were consistent with the residual Factor IX activity of the affected cases. Neither the pre-messenger RNA (mRNA) splicing process nor the protein function was impaired in the rest six SCVs. In conclusion, our study firstly revealed the pathogenic mechanism of the 14 F9 SCVs and highlighted the importance of performing mRNA splicing analysis and protein expression studies of SCVs in inherited disorders.

8.
Sci Total Environ ; 835: 155415, 2022 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-35469876

RESUMO

Trichosporon fermentans can be used to treat refined soybean oil wastewater (RSOW) and produce microbial lipids. Bioflocculation is an effective method to recover Trichosporon fermentans which accumulates intracellular oils from wastewater. During the flocculation, the hydrodynamic distribution and parameters in the reactor are important limiting factors of yeast flocculation performance. In a 0.25 L flocculation device, it was found that the appropriate range of turbulence kinetic energy was within 0.00065-0.00073 m2/s2, the dissipation rate was within 0.119-0.317 m2/s3, and the shear force was less than 0.433 Pa by computational fluid dynamics. In this case, the flocculation rate (Fr) of Trichosporon fermentans could reach more than 90%. The empirical formula associated Fr of Trichosporon fermentans with hydrodynamic parameters was obtained by Matlab, and improved in the enlargement of flocculation device, displaying an error of less than 3.03%. A conical draft tube airlift circulating reactor for flocculation was designed based on the empirical formula, and the Fr reached 91.3%. The study shows that it is feasible to predict Fr of Trichosporon fermentans according to hydrodynamic parameters by numerical simulation, and design the industrial reactor for flocculation harvesting yeasts. It is also helpful for large-scale treatment of RSOW in a safe environment.

10.
Blood ; 2022 Apr 27.
Artigo em Inglês | MEDLINE | ID: mdl-35476848

RESUMO

CD19-directed chimeric antigen receptor (CAR-19)-T cells are groundbreaking immunotherapies approved for use against large B-cell lymphomas. While host inflammatory and tumor microenvironmental markers associate with efficacy and resistance, the tumor-intrinsic alterations underlying these phenomena remain undefined. CD19 mutations associate with resistance but are uncommon, and most patients who relapse retain expression of the wild-type receptor, implicating other genomic mechanisms. We therefore leveraged the comprehensive resolution of whole-genome sequencing to assess 51 tumor samples from 49 CAR-19-treated large B-cell lymphoma patients. We find that pre-treatment presence of complex structural variants, APOBEC mutational signatures, and genomic damage from reactive oxygen species predict CAR-19 resistance. In addition, the recurrent 3p21.31 chromosomal deletion containing the RHOA tumor suppressor was strongly enriched in patients failed by CAR-T therapy. Pretreatment reduced expression or mono-allelic loss of CD19 did not affect responses, suggesting CAR-19 therapy success and resistance are due to multiple mechanisms. Our study shows tumor-intrinsic genomic alterations are key among the complex interplay of factors that underly CAR-19 efficacy and resistance for large B-cell lymphomas.

11.
Mol Ther Nucleic Acids ; 28: 190-201, 2022 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-35402077

RESUMO

Colorectal neoplasia differentially expressed (CRNDE) is an oncogenic long noncoding RNA (lncRNA). Increased CRNDE expression was initially discovered in colorectal cancer and then in a variety of solid tumors and hematological malignancies. CRNDE participates in multiple biological processes, such as cell proliferation, differentiation, migration, and apoptosis. CRNDE has been shown to modulate target gene expression through multiple mechanisms, including transcriptional regulation, post-transcriptional regulation, and competition for microRNA (miRNA) binding. In this review, we summarize the evidence that supports CRNDE in the diagnosis and prognosis predicting of cancers. The functional roles and molecular mechanisms of CRNDE are further described for major types of solid tumors and hematological malignancies. The therapeutic potential of CRNDE as a target for research and development is also discussed.

12.
Anal Biochem ; 647: 114665, 2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35339450

RESUMO

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder. Nevertheless, its accurate mechanisms remain unclear. Metabolomics is a powerful technique to identify small molecules that could be used to discover pathogenesis and therapeutical targets of disease. In the present study, a urinary untargeted metabolomics combined with targeted quantification analysis was performed to uncover metabolic disturbance associated with PCOS. A total of thirty-eight metabolites were obtained between PCOS patients and healthy controls, which were mainly involved in lipids (39.5%), organic acids and derivatives (23.7%), and organic oxygen compounds (18.4%). Based on enrichment analysis, fourteen metabolic pathways were found to be perturbed in PCOS, particularly glycerophospholipid metabolism and tryptophan metabolism. Targeted quantification profiling of tryptophan metabolism demonstrated that seven compounds (tryptophan, kynurenine, kynurenic acid, quinolinic acid, xanthurenic acid, 3-hydroxyanthranilic acid and 3-hydroxykynurenine) were up-regulated in PCOS. And these tryptophan-kynurenine metabolites showed significant correlations with PCOS clinical features, such as positively associated with testosterone, free androgen index, and the ratio of luteinizing hormone to follicle stimulating hormone. Thus, this study disclosed urinary metabolome changes associated with PCOS, and might provide new insights into PCOS pathogenesis elucidation and therapeutical target development.


Assuntos
Síndrome do Ovário Policístico , Feminino , Humanos , Cinurenina/metabolismo , Metaboloma , Metabolômica/métodos , Síndrome do Ovário Policístico/metabolismo , Triptofano/metabolismo
13.
Mikrochim Acta ; 189(4): 145, 2022 03 16.
Artigo em Inglês | MEDLINE | ID: mdl-35296924

RESUMO

A photoelectrochemical (PEC) biosensor based on a multiple signal amplification strategy was established for highly sensitive detection of microRNA (miRNA). TiO2 was prepared on the surface of titanium sheet by laser etching to improve its stability and photoelectrical properties, and CuInS2-sensitized TiO2 was used to form a superior photoelectrical layer, which realized the initial signal amplification. The electron donor dopamine (DA) was modified to H2 as a signal regulator, which effectively increased the photocurrent signal. To further amplify the signal, an enzyme-free hybridization reaction was implemented. When target let-7a and fuel-DNA (F-DNA) were present, the base of H1 specifically recognized let-7a and forced dopamine@AuNPs-H2 away from the electrode surface. Subsequently, the end base of H1 specifically recognized F-DNA, and let-7a was replaced and recycled to participate in the next cycle. Enzyme-free circulation, as a multifunctional amplification method, ensured the recycling of target molecules. This PEC sensor for let-7a detection showed an excellent linear response from 0.5 to 1000 pM with a detection limit of 0.12 pM. The intra-batch RSD was 3.8% and the recovery was 87.74-108.1%. The sensor was further used for clinical biomolecular monitoring of miRNA, showing excellent quantitative detection capability.


Assuntos
Nanopartículas Metálicas , MicroRNAs , Nanocompostos , DNA , Dopamina/química , Técnicas Eletroquímicas/métodos , Ouro/química , Lasers , Limite de Detecção , Nanopartículas Metálicas/química , MicroRNAs/química , Titânio
14.
Theranostics ; 12(5): 2063-2079, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35265199

RESUMO

Background: Dietary tyrosine regulating melanoma progression has been well-recognized. However, whether tyrosine-based melanin anabolism contributes to pulmonary and cerebral organotropic colonization of melanoma remains elusive. Furthermore, approaches based on targeting tyrosinase activity to inhibiting multi-organ metastasis of melanoma cells need to be designed and validated. Methods: Patients derived melanoma cells and mouse B16 melanoma cells with different pigmentation were employed in this investigation. Tyrosine content dynamics in tumors and multiple organs during the melanoma progression was monitored, and tyrosine-based melanin synthesis of melanoma cells derived from multi-organ was determined. Additionally, we also adopted RNA-seq, flow cytometry, real-time PCR and composite metastasis mouse model to analyze organotropic colonization and to validate designed therapeutic strategies. Results: B16 melanoma cells with high activity of tyrosinase and sensitivity of tyrosine utilization for melanin synthesis (Tyr-H cells) easily colonized in the lung, while B16 melanoma cells lacking above characteristics (Tyr-L cells) exhibited potent proliferation in the brain. Mechanistically, Tyr-H cells recruited and trained neutrophils and macrophages to establish pulmonary metastatic niche dependent on highly secreted CXCL1 and CXCL2 and an excessive melanosome accumulation-induced cell death. Tyr-L cells enhanced PD-L1 expression in tumor-infiltrated macrophages when they are progressing in the brain. Accordingly, intervention of tyrosinase activity (2-Ethoxybenzamide or hydroquinone) in combination with inhibitors of phagocytosis (GSK343) or chemotaxis (SB225002) suppressed organotropic colonization and significantly improved the survival of melanoma- bearing mice treated with immune checkpoint blockade (PD1 antibody). Conclusions: The heterogeneity of melanoma cells in utilization of tyrosine is associated with organotropic colonization, providing the basis for developing new strategies to combat melanoma.


Assuntos
Melaninas , Melanoma Experimental , Animais , Linhagem Celular Tumoral , Humanos , Pulmão/patologia , Camundongos , Monofenol Mono-Oxigenase/metabolismo , Microambiente Tumoral , Tirosina
15.
Res Pract Thromb Haemost ; 6(2): e12670, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35284778

RESUMO

Background: Emicizumab is a subcutaneously administered humanized, bispecific, monoclonal antibody approved for prophylaxis in people with hemophilia A. Methods: HAVEN 5 (NCT03315455) is a randomized, open-label, phase 3 study of individuals aged ≥12 years with severe hemophilia A without factor VIII (FVIII) inhibitors, or hemophilia A of any severity with FVIII inhibitors, across the Asia-Pacific region. Participants were randomly assigned (2:2:1) to receive emicizumab 1.5 mg/kg once weekly (arm A), emicizumab 6 mg/kg every 4 weeks (arm B), or no prophylaxis (arm C). The primary end point was annualized bleeding rate (ABR) for treated bleeds; ABRs were compared between people receiving emicizumab prophylaxis versus those with no prophylaxis. Secondary end points included ABR for treated target joint bleeds. Safety was also evaluated. Results: From April 26, 2018, to January 4, 2019, 70 of 76 screened participants were enrolled and randomized (arm A, n = 29; arm B, n = 27; arm C, n = 14). ABRs (95% confidence interval) for treated bleeds and treated target joint bleeds, respectively, were: arm A, 1.0 (0.53-1.85) and 0.4 (0.18-1.09); arm B, 1.0 (0.50-1.84) and 0.3 (0.12-0.85); arm C, 27.0 (13.29-54.91) and 8.6 (3.15-23.42). The most common adverse event, upper respiratory tract infection, was reported for 14 of 56 (25.0%; emicizumab) and 2 of 14 (14.3%; no prophylaxis) participants. No thrombotic events, thrombotic microangiopathies, or deaths were reported. Conclusion: Emicizumab 1.5 mg/kg once weekly and 6 mg/kg every 4 weeks demonstrated bleed control in this study population, was well tolerated, and could improve use of prophylaxis in people with hemophilia A.

16.
Mol Plant Pathol ; 2022 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-35279937

RESUMO

Huanglongbing (HLB), associated with "Candidatus Liberibacter asiaticus" (CLas), is a globally devastating plant disease. The highly reduced genome of CLas encodes a number of secretory proteins. The conserved prophage-encoded protein AGH17470 is herein identified as a nonclassical secretory protein. We confirmed that the N-terminal and C-terminal sequences jointly determine the secretion of AGH17470. The transient expression of AGH17470 protein in Nicotiana benthamiana caused hypersensitive response (HR) cell death in infiltrated leaves and systemically infected leaves as well as the dwarfing of the entire plant, suggesting that AGH17470 is involved in the plant immune response, growth, and development. Overexpression of AGH17470 in N. benthamiana and citrus plants up-regulated the transcription of pathogenesis-related and salicylic acid (SA)-signalling pathway genes and promoted SA accumulation. Furthermore, transient expression of AGH17470 enhanced the resistance of sweet orange to Xanthomonas citri subsp. citri. To our knowledge, AGH17470 is the first prophage-encoded secretory protein demonstrated to elicit an HR and induce a strong plant immune response. The findings have increased our understanding of prophage-encoded secretory protein genes, and the results provide clues as to the plant defence response against CLas.

17.
Org Lett ; 24(10): 2059-2063, 2022 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-35262358

RESUMO

A photocatalyzed transformation from sulfinic acids to sulfoxides under visible-light irradiation in the presence of N-heterocyclic carbene is established. Various alkyl groups from four-substituted Hantzsch esters or Meyer nitriles are smoothly converted to the corresponding sulfoxides through a radical coupling pathway in the presence of 1,1-carbonyldiimidazole. This method allows sulfoxide synthesis to refrain from relying on the oxidation of sulfides and provides an alternative route for the preparation of sulfoxides.

18.
J Phys Chem Lett ; 13(13): 2976-2985, 2022 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-35343699

RESUMO

Two-dimensional electron gas (2DEG) formed at the heterointerface between two oxide insulators hosts plenty of emergent phenomena and provides new opportunities for electronics and photoelectronics. However, despite being long sought after, on-demand properties controlled through a fully optical illumination remain far from being explored. Herein, a giant tunability of the 2DEG at the interface of γ-Al2O3/SrTiO3 through a fully optical gating is discovered. Specifically, photon-generated carriers lead to a delicate tunability of the carrier density and the underlying electronic structure, which is accompanied by the remarkable Lifshitz transition. Moreover, the 2DEG can be optically tuned to possess a maximum Rashba spin-orbit coupling, particularly at the crossing region of the sub-bands with different symmetries. First-principles calculations essentially well explain the optical modulation of γ-Al2O3/SrTiO3. Our fully optical gating opens a new pathway for manipulating emergent properties of the 2DEGs and is promising for on-demand photoelectric devices.

19.
Front Microbiol ; 13: 799819, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35308338

RESUMO

Citrus Huanglongbing (HLB) is the most devastating disease of citrus caused by the Gram-negative phloem-limited bacterium "Candidatus Liberibacter asiaticus" (CLas). It can be transmitted by the Asian citrus psyllid "Diaphorina citri," by grafting, and by the holoparasitic dodder. In this study, the non-natural host periwinkle (Catharanthus roseus) was infected via dodder (Cuscuta campestris) from CLas-infected citrus plants, and the asymptomatic leaves (AS) were subjected to transcriptomic and small-RNA profiling. The results were analyzed together with a transcriptome dataset from the NCBI repository that included leaves for which symptoms had just occurred (S) and yellowing leaves (Y). There were 3,675 differentially expressed genes (DEGs) identified in AS, and 6,390 more DEGs in S and further 2109 DEGs in Y. These DEGs were commonly enriched in photosystem, chloroplast, membrane, oxidation-reduction process, metal/zinc ion binding on GO. A total of 14,974 DEGs and 336 DE miRNAs (30 conserved and 301 novel) were identified. Through weighted gene co-expression network and nested network analyses, two critical nested miRNA-mRNA regulatory networks were identified with four conserved miRNAs. The primary miR164-NAC1 network is potentially involved in plant defense responses against CLas from the early infection stage to symptom development. The secondary network revealed the regulation of secondary metabolism and nutrient homeostasis through miR828-MYB94/miR1134-HSF4 and miR827-ATG8 regulatory networks, respectively. The findings discovered new potential mechanisms in periwinkle-CLas interactions, and its confirmation can be done in citrus-CLas system later on. The advantages of periwinkle plants in facilitating the quick establishment and greater multiplication of CLas, and shortening latency for disease symptom development make it a great surrogate for further studies, which could expedite our understanding of CLas pathogenesis.

20.
J Dairy Sci ; 105(5): 3770-3781, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35181141

RESUMO

A cysteine peptidase was previously identified from germinated Moringa oleifera seeds, but its milk-clotting properties on bovine caseins was still unclear. In this study, this novel cysteine peptidase (MoCP) showed preferential activity on κ-casein (κ-CN), with greater hydrolytic activity compared with calf rennet, whereas weak hydrolysis of α-casein and ß-casein made MoCP suitable for application in cheesemaking and may yield various functional peptides. All 3 evaluated caseins were hydrolyzed to form relatively stable peptide bands within 3 h of proteolysis with MoCP. Cleavage sites were determined by gel electrophoresis, liquid chromatography mass spectrometry/mass spectrometry, and peptide sequencing, which revealed that cleavage of κ-CN by MoCP occurred at residue Ile129-Pro130 and generated a 14,895.37-Da peptide. The flocculation reaction between MoCP and κ-CN determined by 3-dimensional microscopy with super-depth of field revealed that the initial 30 min of reaction were key for milk coagulation, which may affect curd yield. Overall, the findings presented herein suggest that the cysteine peptidase from germinated M. oleifera seeds can be considered a promising plant-derived rennet alternative for use in cheese manufacture.


Assuntos
Queijo , Cisteína Proteases , Moringa oleifera , Animais , Caseínas/química , Bovinos , Queijo/análise , Cisteína/análise , Leite/química , Moringa oleifera/química , Peptídeos/análise , Sementes/química
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