Employing fruit juices to hydrolyze edible bird's nest and enhance the antioxidant, anti-tyrosinase, and wound-healing activities of the hydrolysates.

Enzymatic hydrolysis of edible bird's nest (EBN) has attracted great interest in both scientific and commercial fields due to the enhancement of solubility and nutraceutical values. The present study attempted to investigate the hydrolysis of EBN with papaya (Carica papaya L.), pineapple (Ananas comosus (L.) Merr.), and cantaloupe (Cucumis melo L.) juices as well as two commercial enzymes papain and bromelain. Our analysis revealed that EBN hydrolysis with pineapple juice and bromelain produced a degree of hydrolysis (DH) value of approximately 27 % while it was about 25 % for the hydrolysis with cantaloupe juice and 22 % for the hydrolysis with papaya juice and papain after 4 h of treatment. When EBN was digested by fruit juices and enzymes, the protein solubility and free sialic acid content were increased and the highest values were achieved for EBN hydrolysis with pineapple juice and bromelain (estimately 11 mg/mL of soluble protein and 18 g/kg of free sialic acid). The ABTS•+-scavenging, •OH-scavenging, and anti-tyrosinase capacities were higher in the EBN hydrolysates by papaya juice (IC50 of 0.034, 0.108, and 0.419 mg/mL, respectively), pineapple juice (IC50 of 0.025, 0.045, and 0.190 mg/mL, respectively), and cantaloupe juice (IC50 of 0.031 mg/mL, 0.056, and 0.339 mg/mL, respectively) than in the hydrolysates by unhydrolyzed EBN (IC50 of 0.094, 0.366, and 1.611 mg/mL, respectively). An improvement in ABTS•+-scavenging, •OH-scavenging, and anti-tyrosinase abilities was also observed for the hydrolysates by papain (IC50 of 0.041, 0.129, and 0.417 mg/mL, respectively) and bromelain (IC50 of 0.025, 0.069, and 0.336 mg/mL, respectively) but in a lesser extent as compared to the hydrolysates by respective papaya and pineapple juices. Noticeably, the EBN hydrolysates by fruit juices remarkably enhanced the wound closure in human fibroblasts by about 1.4-1.8 times after 24 h of treatment whereas this property was insignificant in the hydrolysates by enzymes. As papaya, pineapple, and cantaloupe juices are easily obtainable and have pleasant flavors, our results provide a possible method to hydrolyze EBN and apply the resultant hydrolysates in functional food products.

Corrigendum to "Polysaccharide and ethanol extracts of Anoectochilus formosanus Hayata: Antioxidant, wound-healing, antibacterial, and cytotoxic activities".

[This corrects the article DOI: 10.1016/j.heliyon.2023.e13559.].

Polysaccharide and ethanol extracts of Anoectochilus formosanus Hayata: Antioxidant, wound-healing, antibacterial, and cytotoxic activities.

Introduction: Polysaccharide and alcohol extracts of Anoectochilus formosanus Hayata have attracted great attention as they exhibit noteworthy properties such as prebiotic and anti-hyperglycemic effects. However, the antioxidant and wound-healing activities of the polysaccharide extract as well as the antibacterial and cytotoxic effects of the ethanol extracts have not been thoroughly uncovered. Therefore, our study investigated these bioactivities of the two extracts prepared from Anoectochilus formosanus to broaden understandings of medical benefits of the plant. Methods: The monosaccharide composition was analyzed by HPAEC-PAD. The antioxidant and wound-healing activities of the polysaccharide extract were evaluated by ABTS and scratch assays, respectively. The broth dilution method was used to determine the antibacterial ability of the ethanol extract. Additionally, the cytotoxic and mechanistic effects of this extract against hepatocellular carcinoma HUH-7 cells was assessed by MTT assay, qRT-PCR and Western blotting methods. Results: The polysaccharide extract possessed an effective free radical scavenging ability in an ABTS assay (IC50 = 44.92 µg/ml). The extract also ameliorated wound recovery in a fibroblast scratch assay. Meanwhile, the ethanol extract was able to inhibit the growth of Staphylococcus aureus (MIC = 2500 µg/ml), Bacillus cereus (MIC = 2500 µg/ml), Escherichia coli (MIC = 2500 µg/ml), and Pseudomonas aeruginosa (MIC = 1250 µg/ml). Additionally, it repressed the viability of HUH-7 cells (IC50 = 53.44 µg/ml), possibly through upregulating the expression of caspase 3 (CASP3), CASP8, and CASP9 at both mRNA and protein levels. Conclusion: The polysaccharide extract of A. formosanus exhibited the antioxidant and wound-healing properties whereas the ethanol extract showed the antibacterial activity and cytotoxicity against HUH-7 cells. These findings specify notable biological effects of the two extracts which could be of potential use in human healthcare.

Specification of leaf dorsiventrality via a prepatterned binary readout of a uniform auxin input.

Developmental boundaries play an important role in coordinating the growth and patterning of lateral organs. In plants, specification of dorsiventrality is critical to leaf morphogenesis. Despite its central importance, the mechanism by which leaf primordia acquire adaxial versus abaxial cell fates to establish dorsiventrality remains a topic of much debate. Here, by combining time-lapse confocal imaging, cell lineage tracing and molecular genetic analyses, we demonstrate that a stable boundary between adaxial and abaxial cell fates is specified several plastochrons before primordium emergence when high auxin levels accumulate on a meristem prepattern formed by the AS2 and KAN1 transcription factors. This occurrence triggers a transient induction of ARF3 and an auxin transcriptional response in AS2-marked progenitors that distinguishes adaxial from abaxial identity. As the primordium emerges, dynamic shifts in auxin distribution and auxin-related gene expression gradually resolve this initial polarity into the stable regulatory network known to maintain adaxial-abaxial polarity within the developing organ. Our data show that spatial information from an AS2-KAN1 meristem prepattern governs the conversion of a uniform auxin input into an ARF-dependent binary auxin response output to specify adaxial-abaxial polarity. Auxin thus serves as a single morphogenic signal that orchestrates distinct, spatially separated responses to coordinate the positioning and emergence of a new organ with its patterning.

Gibberellin Signaling Requires Chromatin Remodeler PICKLE to Promote Vegetative Growth and Phase Transitions.

PICKLE (PKL) is an ATP-dependent chromodomain-helicase-DNA-binding domain (CHD3) chromatin remodeling enzyme in Arabidopsis (Arabidopsis thaliana). Previous studies showed that PKL promotes embryonic-to-vegetative transition by inhibiting expression of seed-specific genes during seed germination. The pkl mutants display a low penetrance of the "pickle root" phenotype, with a thick and green primary root that retains embryonic characteristics. The penetrance of this pickle root phenotype in pkl is dramatically increased in gibberellin (GA)-deficient conditions. At adult stages, the pkl mutants are semidwarfs with delayed flowering time, which resemble reduced GA-signaling mutants. These findings suggest that PKL may play a positive role in regulating GA signaling. A recent biochemical analysis further showed that PKL and GA signaling repressors DELLAs antagonistically regulate hypocotyl cell elongation genes by direct protein-protein interaction. To elucidate further the role of PKL in GA signaling and plant development, we studied the genetic interaction between PKL and DELLAs using the hextuple mutant containing pkl and della pentuple (dP) mutations. Here, we show that PKL is required for most of GA-promoted developmental processes, including vegetative growth such as hypocotyl, leaf, and inflorescence stem elongation, and phase transitions such as juvenile-to-adult leaf and vegetative-to-reproductive phase. The removal of all DELLA functions (in the dP background) cannot rescue these phenotypes in pkl RNA-sequencing analysis using the ga1 (a GA-deficient mutant), pkl, and the ga1 pkl double mutant further shows that expression of 80% of GA-responsive genes in seedlings is PKL dependent, including genes that function in cell elongation, cell division, and phase transitions. These results indicate that the CHD3 chromatin remodeler PKL is required for regulating gene expression during most of GA-regulated developmental processes.

The Arabidopsis RING Domain Protein BOI Inhibits Flowering via CO-dependent and CO-independent Mechanisms.

BOTRYTIS SUSCEPTIBLE1 INTERACTOR (BOI) and its three homologs (BOIs) are RING domain-containing proteins that repress flowering. Here, we investigated how BOIs repress flowering. Genetic analysis of the boiQ quadruple mutant indicates that BOIs repress flowering mainly through FLOWERING LOCUS T (FT). BOIs repress the expression of FT by CONSTANS (CO)-dependent and -independent mechanisms: in the CO-dependent mechanism, BOIs bind to CO, inhibit the targeting of CO to the FT locus, and thus repress the expression of FT; in the CO-independent mechanism, BOIs target the FT locus via a mechanism that requires DELLAs but not CO. This dual repression of FT makes BOIs strong repressors of flowering in both CO-dependent and CO-independent pathways in Arabidopsis thaliana. Our finding that BOIs inhibit CO targeting further suggests that, in addition to modulating CO mRNA expression and CO protein stability, flowering regulation can also modulate the targeting of CO to FT.

DELLA proteins and their interacting RING Finger proteins repress gibberellin responses by binding to the promoters of a subset of gibberellin-responsive genes in Arabidopsis.

DELLA proteins, consisting of GA INSENSITIVE, REPRESSOR OF GA1-3, RGA-LIKE1 (RGL1), RGL2, and RGL3, are central repressors of gibberellin (GA) responses, but their molecular functions are not fully understood. We isolated four DELLA-interacting RING domain proteins, previously designated as BOTRYTIS SUSCEPTIBLE1 INTERACTOR (BOI), BOI-RELATED GENE1 (BRG1), BRG2, and BRG3 (collectively referred to as BOIs). Single mutants of each BOI gene failed to significantly alter GA responses, but the boi quadruple mutant (boiQ) showed a higher seed germination frequency in the presence of paclobutrazol, precocious juvenile-to-adult phase transition, and early flowering, all of which are consistent with enhanced GA signaling. By contrast, BOI overexpression lines displayed phenotypes consistent with reduced GA signaling. Analysis of a gai-1 boiQ pentuple mutant further indicated that the GAI protein requires BOIs to inhibit a subset of GA responses. At the molecular level, BOIs did not significantly alter the stability of a DELLA protein. Instead, BOI and DELLA proteins are targeted to the promoters of a subset of GA-responsive genes and repress their expression. Taken together, our results indicate that the DELLA and BOI proteins inhibit GA responses by interacting with each other, binding to the same promoters of GA-responsive genes, and repressing these genes.