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Raspberry ketone is a valuable aromatic compound that occurs only at trace levels in natural sources. In this study, transgenic tomato fruits expressing benzalacetone synthase (BAS) from Rheum palmatum and raspberry ketone/zingerone synthase 1 (RZS1) from Rubus idaeus under the CaMV 35S promoter accumulated raspberry ketone (9.9-19.3 μg g-1 fresh weight) together with minor amounts of rhododenol (2.7-4.4 μg g-1 fresh weight), predominantly as glycosides during fruit ripening. LC-MS/MS analysis using an authentic standard identified raspberry ketone monoglucoside as a major glycosylated form present in transgenic fruits but absent in wild-type controls. Raspberry ketone glycosides were predominantly detected in the fruit peel, with negligible levels in the flesh, seeds, or leaves. Co-expression of the Arabidopsis thaliana R2R3-MYB transcription factor AtMYB12 further enhanced raspberry ketone production, reaching up to 88 μg g-1 fresh weight. These results demonstrate effective metabolic redirection of phenylpropanoid flux in an edible fruit tissue and establish tomato as a viable heterologous host for raspberry ketone production. This metabolic engineering strategy highlights the potential of edible fleshy fruits as sustainable plant-based platforms for the biosynthesis of high-value aromatic phenolics.

Red raspberry (Rubus idaeus L.) cultivation is often constrained by drought stress, which reduces fruit yield and quality. Melatonin (MT) plays an important role in plant stress tolerance. This study examined how exogenous MT improves drought tolerance in red raspberry at the squaring stage. It focused on changes in photosynthetic performance, antioxidant capacity, leaves anatomical structure, gene expression, and hormone homeostasis. Drought stress significantly reduced photosynthetic performance, antioxidant enzyme activities, and hormone homeostasis in red raspberry, and altered leaves anatomical structure. Exogenous MT treatment, particularly at 150 μmol L⁻1, effectively alleviated these drought-induced effects. MT increased net photosynthetic rate and stomatal conductance, reduced oxidative damage, and promoted osmotic accumulation. At the anatomical level, MT-treated leaves showed more compact spongy tissue, increased palisade tissue and leaves thickness under drought stress. Transcriptomic and metabolomic analyses further revealed coordinated molecular changes under MT treatment. These changes mainly involved photosynthesis, carbohydrate and starch metabolism, plant hormone signaling, and amino acid metabolism. Genes related to photosynthetic function (PNSL3 and PNSB4), carbohydrate metabolism (glgc and PYG), antioxidant defense (SOD1 and sodC), and auxin signaling (AUX1 and SAUR) showed marked expression changes under MT treatment. Metabolite profiling also revealed changes in sugars, amino acids, phenolic compounds, and hormone-related compounds. Several transcription factor families, including bHLH, MYB-related, NAC, WRKY, and ERF, showed strong responses to MT. These changes were associated with transcriptional regulation and metabolic adjustment under drought stress. Exogenous MT significantly improved drought tolerance in red raspberry at the squaring stage, with 150 μmol L⁻1 showing the best overall performance. This protective effect was associated with improved photosynthetic performance, enhanced antioxidant capacity, and better maintenance of leaves structure. Integrated transcriptomic and metabolomic analyses further revealed changes in hormone-related and metabolic pathways. These findings indicate that MT enhanced drought tolerance in red raspberry through coordinated physiological, anatomical, and molecular responses.

Metabolism-associated fatty liver disease (MAFLD) has emerged as a severe worldwide public health burden with insufficient available clinical therapeutic strategies, which underscores the urgent demand for safe, natural dietary interventions. Raspberry (Rubus idaeus L.), a typical food-medicine homologous fruit abundant in diverse bioactive components including anthocyanins, flavonoids and polysaccharides, possesses prominent nutritional and medicinal potential. In this study, raspberry aqueous extract (RE) was prepared to comprehensively investigate its ameliorative effects and underlying molecular mechanisms against MAFLD. MAFLD animal model was established in C57BL/6 mice via 12-week high-fat diet (HFD) feeding. From the 9th week, model mice were intragastrically administered with RE at doses of 1 g/kg/d and 2 g/kg/d for continuous intervention. Integrated multi-omics analyses including 16S rRNA microbial sequencing, serum/hepatic biochemical detection, histopathological examination, in vivo microbial colonization assay, and in vitro cellular and metabolomic experiments were performed to systematically clarify the regulatory mechanism. RE treatment markedly improved the core pathological phenotypes of MAFLD mice, and significantly mitigated hepatic steatosis and hepatocellular injury. 16S rRNA sequencing demonstrated that RE remodeled the gut microbial dysbiosis, specifically elevating the abundance of beneficial genus Ileibacterium and suppressing pathogenic microbial taxa. Meanwhile, RE strengthened intestinal mucosal barrier integrity by upregulating tight junction protein expression, and activated hepatic purine metabolic reprogramming to boost the levels of critical metabolites including inosine and ADP. Spearman correlation analysis verified the significantly positive correlation between Ileibacterium abundance and hepatic inosine content, and both factors were closely correlated with the remission of MAFLD pathological indicators. In vivo colonization experiments further validated that Ileibacterium intervention alone remarkably alleviated hepatic lipid deposition and liver damage in MAFLD mice. In vitro strain metabolomics confirmed that Ileibacterium could directly biosynthesize and secrete inosine extracellularly. Furthermore, in vitro AML12 hepatocyte experiments revealed that 100 μM inosine remarkably relieved palmitic acid-induced lipotoxicity via reducing intracellular lipid overload, reactive oxygen species (ROS) accumulation and mitochondrial dysfunction, alongside modulating the expression of lipid metabolism, inflammatory and autophagy-related genes. Collectively, our results elucidate that raspberry aqueous extract alleviates experimental MAFLD through the gut microbiota-purine metabolism-inosine regulatory axis, in which Ileibacterium and inosine act as the core synergistic mediators. This study provides solid preclinical experimental evidence for the development and application of raspberry as a promising functional food for the prevention and nutritional intervention of MAFLD.

Foveolar-type gastric adenomas with a raspberry-like appearance in the stomach have recently been identified. However, their etiology and mechanisms of development and growth remain unknown. We observed a foveolar-type gastric adenoma with a raspberry-like appearance that underwent rapid morphological changes. The first esophagogastroduodenoscopy revealed multiple reddish polyps on the cardia and fundus of the stomach, without Helicobacter pylori infection. The second-largest polyp on the fundus had a red glove appearance and measured 5 mm in diameter. We diagnosed all of them as hyperplastic polyps on the basis of their shape. Given that the patient had received 10 mg of vonoprazan for a long time to treat reflux esophagitis, 20 mg of famotidine was prescribed instead. Eight months later, the largest polyp on the cardia had disappeared.However, a reddish raspberry-like lesion was observed at the site of a previously identified polyp on the fundus and was diagnosed as foveolar-type gastric adenoma by forceps biopsy. The pathological diagnosis was foveolar-type gastric adenoma with high-grade dysplasia as determined by tissue resection with endoscopic mucosal resection (EMR). This is a valuable case report of a lesion evolving from a hyperplastic-like polyp morphology to a raspberry-like lesion within eight months.

Raspberry ketone synthase is the key enzyme for raspberry ketone biosynthesis, yet its activity and thermostability remain suboptimal for industrial use. By combining computational and deep-learning-guided design, we obtained a Rubus idaeus raspberry ketone synthase (RiRZS1) mutant M3 with 12.06-fold higher relative activity and 38.71-fold improved thermostability compared with the wild type. Kinetic analysis showed that M3 exhibited 14.89-fold higher kcat and 5.91-fold higher catalytic efficiency(kcat/Km), indicating markedly enhanced catalytic competence. Structural inspection and molecular dynamics simulations revealed that M3 induced only subtle conformational rearrangements but reduced global flexibility, accounting for its improved thermostability. Meanwhile, tunnel and substrate-pocket reshaping in M3 was associated with increased prereaction-state populations, which may contribute to enhanced catalytic performance. In whole-cell biotransformation, the raspberry ketone titer of M3 was 2.38-fold that of wild-type, and the corresponding space-time yield was 3.57-fold higher. This computational and artificial intelligence-assisted framework enabled a synergistic optimization of RiRZS1 activity and stability, and can serve as a transferable design framework for protein engineering in sustainable bio-manufacturing.

The aim of the study was to modify polylactide with zinc oxide nanoparticles (ZnO), raspberry leaf extract (E), and a combined ZnO/extract system (EZnO) in order to prepare novel packaging materials via a solvent-free method, namely cast extrusion. Physicochemical properties: Morphology (GPC, SEM, FTIR), mechanical (tensile tests, puncture), barrier (WVTR, OTR, UV-Vis) and water contact angle for PLA-based films with two thickness ranges were investigated. Additionally, antimicrobial (antibacterial, antifungal and antiviral) tests were performed. GPC results revealed that the presence of the extract counteracted biopolyester degradation during hot melt processing. The best mechanical properties (TS ca. 50 MPa, EB ca. 18%) were obtained for PLA modified with raspberry leaf extract (PLA/E). EZnO addition led to the highest increase in oxygen (with 25%) and water vapor (up to ca. 28%) barrier properties. The material with EZnO addition was also found to be the only one to demonstrate antibacterial effectiveness, although the activity was insignificant. However, the incorporation of EZnO into the biopolymer matrix enhanced its antiviral properties, resulting in the complete inactivation of Φ6 bacteriophage particles used as a surrogate of SARS-CoV-2 virus.

This study demonstrated the effects of dietary supplementation with 1% raspberry (RO) or 1% strawberry (SO) seed oil from 5 to 12 weeks of age (n = 6/group) on folliculogenesis, hormonal parameters, the ovarian fatty acid profile, and the expression of related genes in juvenile rabbits. After slaughter, ovaries and blood were collected. Ovaries were used for histology, fatty acid profiling, and gene expression analysis, while plasma was used to measure progesterone (P4), testosterone (T), estradiol-17β (E2), follicle-stimulating hormone (FSH), and anti-Müllerian hormone (AMH) concentrations. Both RO and SO reduced the number of primary follicles (p = 0.04), whereas RO increased the number of antral follicles (p = 0.04) compared with the control. In both supplemented groups, FSH (p = 0.04 and p = 0.035) and AMH (p = 0.04) concentrations were higher. RO increased P4 and E2 (p = 0.03 and p = 0.013) concentrations, while SO only increased P4 (p = 0.02) levels. SO altered the ovarian fatty acid profile, increasing selected monounsaturated fatty acids and reducing polyunsaturated fatty acids, likely by increasing the expression of the converting enzyme, stearoyl-CoA desaturase 5 (p = 0.038). Overall, both oils influenced folliculogenesis through hormonal changes, and SO modified ovarian fatty acid composition, which may affect ovarian function in juvenile rabbits.

Foodborne viruses have caused large-scale outbreaks in fresh and frozen berries, such as strawberries, blueberries, and raspberries. Studies have previously evaluated the use of non-thermal technologies (including chemical sanitizers) to inactivate foodborne viruses on the surface of soft berries with varying results. In this study, we investigate a novel model system for utility in characterizing virus attachment and removal, as well as sanitizer inactivation on the surface of berries. A casting method using polydimethylsiloxane (PDMS) was used to create topomimetic artificial surfaces ("replicasts") of strawberries, blueberries, and raspberries to elucidate the impact of berry surface structures (topography) on virus adhesion and inactivation. A human norovirus surrogate, Tulane virus (TV), was inoculated (2 × 105 log10 PFU) on the surface of fresh and replicast berries, and viral recovery levels were compared. Overall, the type of berry impacted viral recovery, with 5.11 log10 PFU recovered from blueberries, 5.04 log10 PFU recovered from strawberries, and 5.66 log10 PFU recovered from raspberries. Surface inoculation of blueberry, strawberry, and raspberry replicasts with TV led to comparable recovery levels of virus as observed in fresh berries, with 4.80, 4.93, and 5.17 log10 PFU recovered, respectively, with a two-step recovery procedure. Sodium hypochlorite (200 ppm) led to less than a one log10 PFU reduction in viral titer in all three types of fresh berries. However, the same sanitizer treatment and two-step recovery procedure led to a 2.45 log10 PFU reduction in blueberry replicasts, 2.24 log10 PFU reduction in strawberry replicasts, and 1.59 log10 PFU reduction in raspberry replicasts. Sodium hypochlorite (50 ppm) and peracetic acid (20 or 80 ppm) had limited efficacy in inactivating viruses inoculated to fresh berry or berry replicast surfaces. However, viral reductions were greater in replicasts compared to fresh berries when observed. These data indicate that while berry replicasts may have utility in modeling surface adhesion of viruses to berries and in viral recovery studies, they are not suitable for studies of viral inactivation by chemical sanitizers.

The tobacco constituent, N'-nitrosonornicotine (NNN) is a potential etiological agent in the development of oral cancer. NNN is metabolically activated to form DNA adducts that release 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB) which was detected at higher levels in the oral cavity of smokers than non-smokers. Black raspberry (BRB) has been proposed as a preventive agent of oral cancer. In a phase 0 clinical trial, 40 active smokers (16 males, 24 females) consumed 5 g of BRB lozenges daily for 8 weeks. Each participant served as his/her own control. Buccal cells and urine were collected at baseline, during BRB administration, and washout period. In all smokers, BRB reduced, but not significantly, the levels of HPB-releasing DNA adducts (Baseline: 0.97 ± 0.51 and End-BRB: 0.61 ± 0.23 HPB/106 Guanine). A sex-specific response to BRB was observed: BRB had no effects in men, but in 16 of 24 women, BRB significantly reduced the levels of HPB. We further tested the hypothesis that detoxification of NNN via N-glucuronidation by BRB may, in part, account for the inhibition of HPB levels. The ratio of NNN-N-glucuronide to free NNN significantly increased (P = 0.028) from 1.29 ± 0.40 (baseline) to 2.77 ± 0.76 (End-BRB) in all 40 women as well as in female responders from 1.24 ± 0.60 (baseline) to 2.87 ± 1.00, but not in non-responders. The metabolic changes of NNN observed in our clinical trial indicate that BRB is a promising candidate for cancer interception and prevention of oral cancer in some, but not in all active smokers.

Accurate time synchronization is essential in distributed electrical signal monitoring, where phase coherence and event correlation depend on precise timing agreement between acquisition nodes. Conventional approaches often rely on a single synchronization source, typically internet-based Network Time Protocol (NTP) or GPS-disciplined clocks, which is impractical in isolated, offline, or cost-sensitive scenarios. This paper introduces an autonomous offline synchronization architecture for multi-node monitoring systems built on Raspberry Pi 5 (RPI5) platforms connected to a private Ethernet network. Instead of depending on one timing method, the system integrates several complementary mechanisms: battery-backed RTC persistence via the J5 interface, deterministic orchestration through systemd services, automated boot time recovery, chrony-managed NTP discipline, and Precision Time Protocol (PTP) hardware timestamping using PTP Hardware Clock (PHC). Synchronization performance is validated through continuous multi-day measurements of long-term stability, inter-node phase coherence, and short-term jitter. Controlled power-loss scenarios are also included to verify recovery behavior. The system maintains sub-microsecond alignment between nodes using only commodity hardware and no external time source. To further confirm inter-node timestamp alignment at the signal level, both hardware-based reference signal injection and software-based synchronized signal emulation are employed, providing ground-truth validation alongside scalable and reproducible evaluation. The results show that low-cost embedded hardware can support reliable, long-duration synchronization in fully offline installations.