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The trihelix transcription factor (GT) gene family members play vital roles in plant growth and development, responses to abiotic or biotic stress, and fruit ripening. However, its role in banana fruit ripening remains unclear. Here, 59 MaGT gene members were identified in banana and clustered into five subfamilies, namely GT1, GT2, GTγ, SIP1, and SH4. This classification is completely supported by their gene structures and conserved motif analysis. Transcriptome data analysis indicated that MaGT14, MaGT21, and MaGT27 demonstrated significant differential expression during fruit ripening. Quantitative real-time PCR analysis revealed that these three genes were highly induced by ethylene treatment, responded to cold and heat stress, and had a high expression abundance in ripe fruit. Subcellular localization demonstrated that MaGT21 and MaGT27 functioned as nuclear proteins, while MaGT14 functioned as a nuclear and cell membrane protein. Further investigation indicated MaGT21 could positively stimulate the transcription of the key ethylene biosynthesis gene MaACO1 by directly targeting the GT motif in the promoter. MaGT21 transient overexpression in banana fruit upregulated MaACO1 and accelerated fruit ripening. Our findings provide comprehensive and valuable information for further functional studies of MaGT genes in banana, help to understand the roles of MaGTs during banana fruit ripening.

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OBJECTIVE: Balloon catheters have shown promising results in the induction of labor (IOL); however, evidence regarding the optimal time of balloon catheter placement is still lacking. Thus, this study aims to evaluate the efficacy and safety of six-hour placement compared to 12 hours. DATA SOURCE: We conducted a comprehensive search through a search strategy across "Embase, SCOPUS, PubMed (via MEDLINE), and Cochrane Central Register of Controlled Trials (CENTRAL)" from inception until April 20, 2024. STUDY ELIGIBILITY CRITERIA: We included the randomized controlled trials (RCTs) evaluating the efficacy and safety of six-hour balloon catheter placement compared with 12 hours for cervical ripening in IOL. Covidence was used to screen eligible articles. METHODS: All relevant outcome data were synthesized using risk ratios (RR) or mean differences (MD) with 95% confidence intervals (CIs) in meta-analysis models within RevMan 5.4. PROSPERO ID: CRD42024540935 RESULTS: We included five RCTs with 960 women undergoing the IOL with balloon catheter placement. Six-hour placement was associated with significant lower insertion to overall delivery interval (MD: -4.25, 95% CI [-5.34, -3.15], P< 0.00001) and insertion to vaginal delivery interval (MD: -4.65, 95% CI [-6.08, -3.23], P< 0.00001) without significant difference in BISHOP score change (MD: -0.02, 95% CI [-0.23, 0.20], P= 0.88), catheter removal to delivery interval (MD: 0.72, 95% CI [-0.39, 1.83], P= 0.20) and total duration of oxytocin infusion (MD: -0.36, 95% CI [-0.85, 0.14], P= 0.16) compared to twelve hours. Also, significantly lower overall cesarean delivery (CD) rate (RR: 0.81, 95% CI [0.68, 0.96], P= 0.01) and CD due to malpresentation (RR: 0.39, 95% CI [0.16, 0.93], P= 0.03) were observed with six-hour placement. CONCLUSION: A planned six-hour balloon catheter placement reduced insertion to delivery intervals and CD rate and has equal efficacy in BISHOP score change and catheter removal to delivery interval compared to twelve hours.

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Long non-coding RNAs (lncRNAs) play crucial roles in various biological processes in plants. However, the functional mechanism of lncRNAs in fruit ripening, particularly the transition from unripe to ripe stages, remains elusive. One such lncRNA1840, reported by our group, was found to have important role in tomato fruit ripening. In the present study, we gain insight into its functional role in fruit ripening. CRISPR-Cas9 mediated lncRNA1840 mutants caused the delayed tomato fruit ripening. Notably, loss function of lncRNA1840 did not directly impact ethylene signaling but rather delay ethylene synthesis. Transcriptomic analysis revealed differences in the expression of ripening related genes in lncRNA1840 mutants, suggesting that it is involved in gene regulation of fruit ripening. We used Chromatin Isolation by RNA Purification (ChIRP)-Seq to identify lncRNA1840 binding sites on chromatin. ChIRP-seq suggested that lncRNA1840 had occupancy on 40 genes, but none of them is differentially expressed genes in transcriptomic analysis, which indicated lncRNA1840 might indirectly modulate the gene expression. ChIRP-mass spectrometry analysis identified potential protein interactors of lncRNA1840, Pre-mRNA processing splicing factor 8, highlighting its involvement in post-transcriptional regulatory pathways. In summary, lncRNA1840 is key player in tomato plant growth and fruit ripening, with multifaceted roles in gene expression and regulatory networks.

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Background: Previous studies that suggest a shorter time from cervical ripening balloon placement to delivery with shorter total balloon placement time have excluded patients with prior cesarean deliveries. Objective: To evaluate, in patients with a prior history of cesarean delivery undergoing cervical ripening with a double-balloon catheter, whether planned removal of device after 6 vs 12 hours would result in shorter time to vaginal delivery. Study Design: A before-and-after study was performed after a practice change occurred November 2020, shortening the planned time of double-balloon catheter placement for cervical ripening from 12 to 6 hours. Data were collected via retrospective electronic chart review. Primary outcome was time from balloon placement to vaginal delivery. Secondary outcomes included rates of cesarean delivery, maternal intraamniotic infection, and uterine rupture. Kaplan-Meier curves compared median times to delivery between the groups. A Cox proportional-hazards model was used to adjust for time of balloon placement, number of previous vaginal deliveries, and co-medications used. Results: From November 2018 to November 2022, 189 analyzable patients with a prior history of cesarean delivery received a double-balloon catheter for cervical ripening during their trial of labor. Patients were separated into pre- and postpolicy change groups (n=91 and 98, respectively). The median time to vaginal delivery for the pregroup was 28 hours (95% CI: 26, 35) and 25 hours (95% CI: 23, 29) for those in the postgroup (P value .052). After adjusting for dilation at time of balloon placement, number of previous vaginal deliveries, and co-medication, the estimated hazard ratio for successful vaginal delivery postpolicy change was 1.89 (95% CI: 1.27, 2.81). There were no differences in rates of secondary outcomes. Conclusion: In patients with prior cesarean delivery undergoing mechanical cervical ripening with a double-balloon catheter, planned removal at 6 hours compared to 12 hours may result in higher chances of successful vaginal delivery and possibly a shorter time to delivery, without increasing rates of cesarean delivery and intraamniotic infection.

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The aroma is critical in the reproductive biology of truffles and in their commercial quality. However, previous research has almost exclusively focused on characterizing ripe ascocarps. We characterized the volatilome of the highly-prized black truffle (Tuber melanosporum) ascocarps from July, in an early development stage, to March, in the late harvesting season, and investigated the relationships among aroma, ascocarp growth and morphogenetic development. The aroma profile was analyzed using a head space gas chromatography technique coupled with mass spectrometer. Seventy-one volatile compounds were identified and three development stages were clearly distinguished according to the volatile profile. In unripe ascocarps of July-September, the profile was dominated by methanethiol (19 %), 4-penten-2-ol (11 %) and acetone (11 %), the monthly mean weight of ascocarps ranged 2-20 g, and morphogenetic stages 4-6a were prevalent. In unripe ascocarps of October-December, the most abundant volatiles were 4-penten-2-ol (21 %), methanethiol (20 %) and ethanol (13 %), the monthly mean ascocarp weight ranged 28-43 g, and morphogenetic stages 6a, 6b-c were prevalent. In ripe ascocarps (December-March), the most abundant volatiles were 4-penten-2-ol (17 %), dimethyl sulfide (16 %) and ethanol (10 %), ascocarp weight did not increase significantly, and 6b-c was practically the sole morphogenetic stage. Thirty volatiles were associated to one of these three development stages. Amongst those with higher occurrence, 4-penten-2-ol, dimethyl sulfide, ethyl acetate, 2-pentanol and 2-butanone were associated to ripe truffles, whereas methanethiol, isobutyl isobutyrate, butanedione and 3-methylanisole were associated to unripe truffles.

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Rapeseed (Brassica napus L.) is the second largest globally cultivated oil crop, but the effects of post-harvested ripening on rapeseed quality is unclear and unpredictable. This study reveals the relationship between post-harvest ripening periods (PHR) and physicochemical quality of different rapeseed cultivars using comprehensive physicochemical indicators analysis. The results indicate that PHR led to a gradual decrease in chlorophyll, carotenoid and moisture content but continually increased oil and total phenol content (TPC). Besides, 295 lipid molecules from 13 lipid subclasses were identified, revealing that the relative content of triacylglycerol (TG) was progressively increased while diacylglycerol (DG) demonstrated a consistent decline throughout the PHR. Correlation analysis, hierarchical cluster analysis (HCA) and principal component analysis (PCA) were employed to construct and verify the comprehensive quality evaluation model for rapeseeds in PHR. This paper develops a comprehensive quality evaluation model for post-harvest ripening rapeseeds and advances the development of agricultural products.

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Utilizing different chymosin and pepsin ratios in cheesemaking may represent a potential strategy to shape the sensory profile of hard cheeses. This study investigated the impact of rennet with varying chymosin and pepsin ratios on the chemical profile and sensory attributes of Grana Padano PDO cheese at different ripening times (10 to 20 months). The research involved the analysis of hard cheese manufactured with distinct calf chymosin percentages (99 %, 95 %, and 83 %), exploiting sensory analyses and untargeted metabolomics to identify marker compounds correlating with specific sensory traits. The results demonstrated that varying the rennet composition significantly affected sensory profile; in particular, the rennet made by 83 % chymosin and 17 % pepsin generated a more complex sensory profile starting from 12 months. AMOPLS and ASCA analysis on untargeted metabolomics signatures revealed that ripening time was the only significant factor when compared with rennet type and the interaction ripening x rennet. Finally, at more advanced ripening times, 3-methylbutanoic acid and homoethone were significantly up-accumulated in cheese samples manufactured with higher pepsin percentages, likely explaining sensory outcomes. This study provides valuable insights into using rennet to tailor the sensory qualities of hard cheeses, underscoring the importance of enzyme selection in cheese manufacturing to drive innovation in the dairy industry.

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Abscisic acid (ABA) and gibberellic acid (GA3) are regulators of fruit color and sugar levels, and the application of these hormones is a common practice in commercial vineyards dedicated to the production of table grapes. However, the effects of exogenous ABA and GA3 on wine cultivars remain unclear. We investigated the impact of ABA and GA3 application on Malbec grapevine berries across three developmental stages. We found similar patterns of berry total anthocyanin accumulation induced by both treatments, closely associated with berry H2O2 levels. Quantitative proteomics from berry skins revealed that ABA and GA3 positively modulated antioxidant defense proteins, mitigating H2O2. Consequently, proteins involved in phenylpropanoid biosynthesis were downregulated, leading to decreased anthocyanin content at the almost ripe stage, particularly petunidin-3-G and peonidin-3-G. Additionally, we noted increased levels of the non-anthocyanins E-viniferin and quercetin in the treated berries, which may enhance H2O2 scavenging at the almost ripe stage. Using a linear mixed-effects model, we found statistical significance for fixed effects including the berry H2O2 and sugar contents, demonstrating their roles in anthocyanin accumulation. In conclusion, our findings suggest a common molecular mechanism by which ABA and GA3 influence berry H2O2 content, ultimately impacting anthocyanin dynamics during ripening.

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The garden strawberry (Fragaria x ananassa Duch.) is cultivated and consumed worldwide because of the pleasant flavor and health-promoting phytochemicals of its false fruits. Monocrop cultivars produce fully ripe strawberries in about one month post-anthesis throughout the spring, while everbearing cultivars undergo additional strawberry production in autumn. In this work, we evaluated the impact of different season-dependent environmental conditions on the ripening program of an everbearing field-gown strawberry variety from autumn 2015 to spring 2016. We combined ad hoc sampling and environmental data collection with LC-MS-based untargeted metabolomics to dissect the effects of cumulative temperature and solar irradiation on fruit quality parameters and secondary metabolism during ripening. Different dynamics in specific sub-groups of metabolites were observed in strawberries experiencing distinct amounts of cumulative temperature and solar irradiation during spring and autumn. The integration of statistical analyses on collected data revealed that solar irradiation mainly affected fruit fresh weight and organic acid levels, whereas temperature had a more selective effect on the accumulation of specific flavonols, anthocyanins, and soluble sugar. These findings are of suitable interest to design further approaches for the study of the complex interactions among environmental conditions and ripening in strawberries grown in a real-world scenario.

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Collapsing and degradation of active materials caused by the electrode/electrolyte interface instability in aqueous batteries are one of the main obstacles that mitigate the capacity. Herein by reversing the notorious side reactions include the loss and dissolution of electrode materials: as we applied Ostwald ripening (OR) in the electrochemical cycling of a copper hexacyanoferrate electrode in a hydronium-ion batteries, the dissolved Cu and Fe ions undergo a crystallization process that creates a stable interface layer of cross-linked cubes on the electrode surface. The layer exposed the low-index crystal planes (100) and (110) through OR-induced electrode particle growth, supplemented by vacancy-ordered (100) superlattices that facilitated ion migration. Our design stabilized the electrode-electrolyte interface considerably, achieving a cycle life of one million cycles with capacity retention of 91.6%, and a capacity retention of 91.7% after 3000 cycles for a full battery.

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Transcription factors (TFs) are crucial for regulating fruit ripening in tomato (Solanum lycopersicum). The GRAS (GAI, RGA, and SCR) TFs are involved in various physiological processes, but their role in fruit ripening has seldom been reported. We have previously identified a gene encoding GRAS protein named SlFSR (Fruit Shelf-life Regulator), which is implicated in fruit ripening by regulating cell wall metabolism; however, the underlying mechanism remains unclear. Here, we demonstrate that SlFSR proteins are localized to the nucleus, where they could bind to specific DNA sequences. SlFSR acts downstream of the master ripening regulator RIN and could collaborate with RIN to control the ripening process by regulating expression of ethylene biosynthesis genes. In SlFSR-CR (CRISPR/Cas9) mutants, the initiation of fruit ripening was not affected but the reduced ethylene production and a delayed coloring process occurred. RNA-sequencing (RNA-seq) and promoter analysis reveal that SlFSR directly binds to the promoters of two key ethylene biosynthesis genes (SlACO1 and SlACO3) and activates their expression. However, SlFSR-CR fruits displayed a significant down-regulation of key rate-limiting genes (SlDXS1 and SlGGPPS2) in the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, which may account for the impaired lycopene synthesis. Altogether, we propose that SlFSR positively regulates ethylene biosynthesis and lycopene accumulation, providing valuable insights into the molecular mechanisms underlying fruit ripening.

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The transition to sustainable materials and eco-efficient processes in commercial electronics is a driving force in developing green electronics. Iron-catalyzed laser-induced graphitization (IC-LIG) has been demonstrated as a promising approach for rendering biomaterials electrically conductive. To optimize the IC-LIG process and fully exploit its potential for future green electronics, it is crucial to gain deeper insights into its catalyzation mechanism and structural evolution. However, this is challenging due to the rapid nature of the laser-induced graphitization process. Therefore, multiscale preparation techniques, including ultramicrotomy of the cross-sectional transition zone from precursor to fully graphitized IC-LIG electrode, are employed to virtually freeze the IC-LIG process in time. Complementary characterization is performed to generate a 3D model that integrates nanoscale findings within a mesoscopic framework. This enabled tracing the growth and migration behavior of catalytic iron nanoparticles and their role during the catalytic laser-graphitization process. A three-layered arrangement of the IC-LIG electrode is identified including a highly graphitized top layer with an interplanar spacing of 0.343 nm. The middle layer contained γ-iron nanoparticles encapsulated in graphitic shells. A comparison with catalyst-free laser graphitization approaches highlights the unique opportunities that IC-LIG offers and discuss potential applications in energy storage devices, catalysts, sensors, and beyond.

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The study focuses on the comprehensive analysis of glutamyl dipeptides in cheese, particularly their formation during the cheese ripening process and the influence of various factors, such as origin, the use of various mold cultures, and cheese types. For the first time, all three subgroups of glutamyl dipeptides, namely α-Glu-X, X-Glu, and γ-Glu-X, are covered in a comprehensive analytical LC-MS/MS method offering robust quantitation of all 56 glutamyl dipeptides. The workflow includes a simplified extraction protocol and an optimized separation of the analytes on the stationary phase. Validation experiments demonstrate the method's reliability, including repeatability, detection limits, and recovery. The comprehensive analysis of all glutamyl dipeptides in 122 cheese samples with ripening times between 2 weeks and 15 years shows a strong increase in all peptide classes with prolonged ripening and particularly in the presence of mold.

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Fleshy fruit metabolism is intricately influenced by environmental changes, yet the hormonal regulations underlying these responses remain poorly elucidated. ABA and ethylene, pivotal in stress responses across plant vegetative tissues, play crucial roles in triggering fleshy fruit ripening. Their actions are intricately governed by complex mechanisms, influencing key aspects such as nutraceutical compound accumulation, sugar content, and softening parameters. Both hormones are essential orchestrators of significant alterations in fruit development in response to stressors like drought, salt, and temperature fluctuations. These alterations encompass colour development, sugar accumulation, injury mitigation, and changes in cell-wall degradation and ripening progression. This review provides a comprehensive overview of recent research progress on the roles of ABA and ethylene in responding to drought, salt, and temperature stress, as well as the molecular mechanisms controlling ripening in environmental cues. Additionally, we propose further studies aimed at genetic manipulation of ABA and ethylene signalling, offering potential strategies to enhance fleshy fruit resilience in the face of future climate change scenarios.

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Background: The increased demand for induction of labour (IOL) at 39 weeks' gestation in normal-risk nulliparous patients creates significant logistical challenges for busy maternity units. A potential innovation is commencing induction by means of outpatient cervical ripening, using either a vaginal prostaglandin preparation (Propess) or an osmotic cervical dilator (Dilapan-S). Methods: A Phase III, open label, single centre non-inferiority trial (EudraCT number 2019-004697-25) randomised healthy nulliparous women who chose elective IOL at 39 weeks to one of three methods of initial cervical ripening, specifically 12 h of Dilapan-S(D12), 24 h of Dilapan-S(D24), or 24 h of Propess(P24) between November 2020 and July 2023. After initial administration of the IOL agent in the hospital, participants returned home for 12 or 24 h, before readmission to complete delivery. The primary outcome was vaginal delivery achieved at any time, and this was compared in a non-inferiority analysis of Dilapan-S compared to Propess, within a 10% non-inferiority margin. Secondary outcomes included pairwise comparisons for each induction agent, and a range of logistical factors, such as time to delivery, the need for an additional cervical ripening agent, and length of hospital stay. Findings: Of the 327 women randomised at 38 weeks, 271 (83%) completed the induction intervention. The D24 and P24 groups showed similarly high rates of vaginal delivery, 75% and 76% respectively. D12 had a lower vaginal delivery rate of 64% and consequently the overall comparison of Dilapan-S to Propess did not demonstrate non-inferiority (difference = -6%, 95% CI = -17%, 5%) because the lower 95% CI exceeded the -10% threshold of non-inferiority. The majority of participants across all groups were delivered by any means within 72 h of starting the induction process, inclusive of time spent at home (89% of the D24 group, 98% of the D12 group, 95% of the P24 group). There were no differences in rates of adverse events between groups. Interpretation: There were similarly high vaginal delivery rates for D24 and P24, with at least 75% of patients successfully delivering vaginally following outpatient cervical ripening, with no significant adverse maternal or neonatal outcomes. Funding: The Rotunda Foundation, Medicem Technology s.r.o.

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The objective of this study was to examine the impact of stage of lactation (early, mid and late) and proportion of pasture in the cows diet (high: GRS, medium: PMR and no: TMR) on the composition and quality of Cheddar cheese. Triplicate trials were carried out in each stage of lactation, and milk protein and fat contents were standardized for Cheddar cheese manufacture at pilot scale. As cheese milks were standardized for milk fat and protein contents, gross composition did not differ as a result of diet. Fatty acid profiles of GRS cheese were significantly different from TMR, while PMR profiles were less distinct and more similar to both GRS and TMR profiles, as illustrated by partial least squares discriminatory analysis. Fatty acids including CLA C18:2 cis-9, trans-11, C22:1 n-9 and C18:3 n-3 were most influential in this separation of profiles. Fatty acid profiling revealed that GRS derived cheese contained higher proportions of nutrients considered beneficial for human health including higher proportions of unsaturated fatty acids and omega-3 fatty acids. A biomarker model utilizing the proportions of 5 fatty acids was constructed and was effective at distinguishing between cheese of GRS, TMR and PMR feeding systems. Proportions of ρ-κ-casein, αs2-casein and αs1-casein in cheese also differed between diets while proportions of ρ-κ-casein, αs1-casein and ß-casein were lowest in late lactation cheese. The impact of diet was less influential compared with that of stage of lactation on the ripening characteristics of cheese. An index of primary proteolysis was highest in late lactation cheese. The peptides derived from the proteolysis of κ-casein and ß-casein and levels of secondary proteolysis, in particular, the proportions of 12 free amino acids were most influenced by stage of lactation. Overall this study demonstrated the effects of increasing pasture allowance and stage of lactation on the nutritional quality and ripening properties of Cheddar cheese.

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HYPOTHESIS: Polyglycerol esters of fatty acids are generated via the esterification of a polydisperse mixture of polyglycerol with naturally derived fatty acids. The polymerization process of polyglycerol results in the production of various oligomers, ranging from di-, tri-, and higher-order forms, which contribute to the complexity of final products. The combination of complementary experimental techniques and adequate theoretical interpretations can reveal the wide variety of their physicochemical properties. EXPERIMENTS: The colloid and interface properties of polyglyceryl mono-laurate, mono-stearate, mono-oleate, and a mixture of mono-caprylate and mono-caprate esters solutions were characterized by measurements of the electrolytic conductivity, static and dynamic surface tension, aggregate and micelle sizes and distributions, thin liquid film stability and stratification, and solubility in aqueous and in oil phases. The formation, stability, and bubble size distribution of foams generated from polyglycerol esters aqueous solutions were systematically investigated. FINDINGS: The low concentrations of double-tail molecules and fatty acids in polyglycerol esters affect considerably their micellar, aggregation, and vesicle formations in aqueous solutions. The theoretical data interpretation of polyglycerol esters isotherms and thin liquid films data provide information on the adsorption energies, excluded areas per molecule, interaction parameters of molecules at interfaces, surface electrostatic potential, and the size of micelles. Polyglyceryl mono-oleate exhibits spontaneous emulsification properties. Short chain length polyglycerol esters have excellent foaming ability but relatively low foam stability. The optimal weight fractions of the short-chain polyglyceryl esters and polyglyceryl mono-stearate mixtures with respect to good foaminess and foam stability upon Ostwald ripening are obtained. The reported physicochemical characterization of the water-soluble polyglycerol esters could be of interest to increase the range of their applicability in practice.

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Lactobacillus delbrueckii ssp. bulgaricus M58 (M58) and Streptococcus thermophilus S10 (S10) are 2 dairy starter strains known for their favorable fermentation characteristics. Therefore, this research aimed to study the effects of 1-d low-temperature ripening on the physicochemical properties and metabolomics of fermented milk. Initially, the performance of single (M58 or S10) and dual (M58+S10) strain fermentation was assessed, revealing that the M58+S10 combination resulted in a shortened fermentation time, a stable gel structure, and desirable viscosity, suggesting positive strain interactions. Subsequently, non-targeted metabolomics analyses using LC-MS and GC-MS were performed to comparatively analyze M58+S10 fermented milk samples collected at the end of fermentation and after 1-d low-temperature ripening. The results showed a significant increase in almost all small peptides and dodecanedioic acid in the samples after one day of ripening, while there was a substantial decrease in indole and amino acid metabolites. Moreover, notable increases were observed in high-quality flavor compounds, such as geraniol, delta-nonalactone, 1-hexanol,2-ethyl-, methyl jasmonate, and undecanal. This study provides valuable insights into the fermentation characteristics of the dual bacterial starter consisting of M58 and S10 strains and highlights the specific contribution of the low-temperature ripening step to the overall quality of fermented milk.

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The classification of carambola, also known as starfruit, according to quality parameters is usually conducted by trained human evaluators through visual inspections. This is a costly and subjective method that can generate high variability in results. As an alternative, computer vision systems (CVS) combined with deep learning (DCVS) techniques have been introduced in the industry as a powerful and an innovative tool for the rapid and non-invasive classification of fruits. However, validating the learning capability and trustworthiness of a DL model, aka black box, to obtain insights can be challenging. To reduce this gap, we propose an integrated eXplainable Artificial Intelligence (XAI) method for the classification of carambolas at different maturity stages. We compared two Residual Neural Networks (ResNet) and Visual Transformers (ViT) to identify the image regions that are enhanced by a Random Forest (RF) model, with the aim of providing more detailed information at the feature level for classifying the maturity stage. Changes in fruit colour and physicochemical data throughout the maturity stages were analysed, and the influence of these parameters on the maturity stages was evaluated using the Gradient-weighted Class Activation Mapping (Grad-CAM), the Attention Maps using RF importance. The proposed approach provides a visualization and description of the most important regions that led to the model decision, in wide visualization follows the models an importance features from RF. Our approach has promising potential for standardized and rapid carambolas classification, achieving 91 % accuracy with ResNet and 95 % with ViT, with potential application for other fruits.

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Flesh firmness is a critical breeding trait that determines consumer selection, shelf life, and transportation. The genetic basis controlling firmness in apple (Malus×domestica Borkh.) remains to be fully elucidated. We aimed to decipher genetic variance for firmness at harvest and develop potential molecular markers for marker-assisted breeding. Maturity firmness for 439 F1 hybrids from a cross of 'Cripps Pink' and 'Fuji' was determined in 2016 and 2017. The phenotype segregated extensively, with a Gaussian distribution. In a combined bulked segregant analysis (BSA) and RNA-sequencing analysis, eighty-four differentially expressed genes were screened from the 10 QTL regions. Interestingly, next-generation re-sequencing analysis revealed a Harbinger-like transposon element insertion upstream of the candidate gene PECTATE LYASE5 (MdPL5); the genotype was associated with flesh firmness at harvest. The presence of this transposon repressed MdPL5 expression and was closely linked to the extra-hard phenotype. MdPL5 was demonstrated to promote softening in apples and tomatoes. Subsequently, using the MdPL5 promoter as bait, MdNAC1-L was identified as a transcription activator that positively regulates ripening and softening in the developing fruit. We also demonstrated that MdNAC1-L could induce the up-regulation of MdPL5, MdPG1, and the ethylene-related genes MdACS1 and MdACO1. Our findings provide insight into TE-related genetic variation and the PL-mediated regulatory network for the firmness of apple fruit.