Search Results (212)

Carbohydrate-active enzymes (CAZymes) encoded by the human gut microbiome are central mediators of dietary glycan metabolism and host–microbe biochemical homeostasis. Among these, β-glucuronidases represent functionally pivotal hydrolases implicated in metabolism, intestinal physiology, and therapeutic modulation. The present study performs an integrative in silico structural and functional interrogation of β-glucuronidases derived from Acidobacterium capsulatum (3VNY), Bacteroides ovatus (6D8K), and Faecalibacterium prausnitzii (6ED2). An integrated computational framework encompassing physicochemical parameters profiling, hierarchical structural prediction, tertiary-structure validation, salt-bridge energetics, functional domain and motif annotation, protein–protein interaction reconstruction, ligand-binding thermodynamics via molecular docking, and residue-resolved non-covalent interaction network mapping using the Protein Contacts Atlas (PCA) was employed. Physicochemical analyses indicated that all enzymes are thermostable, intracellular, and hydrophilic, while secondary-structure organization revealed a functional balance between helix-mediated rigidity and coil-driven flexibility. Structural validation metrics identified 6ED2 as the most conformationally stable architecture, whereas 6D8K displayed enhanced functional complexity, including enriched motif composition, membrane-associated features, and superior ligand-binding affinity. Docking simulations highlighted castanospermine and calcium saccharate as the most favorable interacting ligands across enzyme variants. Importantly, PCA-based interaction analysis revealed distinct ligand-centered atomic contact networks, with immediate contact counts of 57 (3VNY), 32 (6D8K), and 41 (6ED2), providing residue-level insight into stabilization mechanisms and interaction topology beyond conventional docking metrics. Collectively, these findings establish a multidimensional computational framework linking structural stability, functional diversification, ligand recognition, and atomic interaction networks in gut microbial β-glucuronidases, thereby supporting future biochemical validation, microbiome-targeted therapeutics, and biotechnological or cosmeceutical applications. Full article

Marine sponges represent a rich source of lectins with diverse biological activities and biotechnological potential. In this study, we report the purification and comprehensive biochemical and structural characterization of a lectin (CspL) from the marine sponge Cinachyrella sp. and evaluate its effects on bacterial agglutination, planktonic bacterial growth, and biofilm formation. CspL was isolated using classical chromatographic approaches and identified as a galectin-like protein based on sequence similarity, conserved carbohydrate-recognition motifs, and a predominance of β-sheet structures revealed by circular dichroism. Oligomeric analysis indicated a homotetrameric organization, consistent with the quaternary structure described for other sponge proto-type galectins. Carbohydrate-binding assays demonstrated that CspL preferentially recognizes galactoside-containing motifs, showing strong inhibition by mucin-type glycoproteins, while displaying lower affinity toward more complex glycan structures. This binding profile suggests a preference for accessible carbohydrate epitopes, likely associated with its canonical galectin architecture. Regarding antibacterial activity, CspL also exhibited selective, carbohydrate-dependent bacterial agglutination, particularly against Staphylococcus aureus strains. In addition, CspL exhibited antibiofilm activity against S. aureus and Escherichia coli, significantly reducing biofilm biomass and viable cell counts. Additionally, the lectin modulated antibiotic activity, showing synergistic effects with tetracycline and strain-dependent interactions with oxacillin. Together, these findings highlight CspL as a structurally conserved yet functionally relevant member of sponge galectins and reinforce the role of structural diversity in shaping glycan recognition and antimicrobial activity in marine lectins. Full article

Background/Objectives: Automated insulin delivery (AID) systems have improved the management of type 1 diabetes (T1D), but exercise and nutrition remain challenging because they rapidly alter glucose flux, substrate oxidation, hepatic glucose output, insulin requirements, and fuel availability. This narrative review aimed to synthesize current evidence on the interaction between AID systems, physical activity, and nutritional strategies from a metabolism-oriented perspective. Methods: A narrative bibliographic approach was used to integrate evidence from clinical trials, observational studies, technical studies, consensus statements, and reviews involving people with T1D across different life stages, including pediatric, adolescent, adult, and pregnancy-related contexts, when available. The review focused on AID systems, exercise physiology, nutritional strategies, meal announcement, bolus adjustment, dual-hormone systems, metabolic biomarkers, and emerging metabolomic approaches. Results: AID systems generally improve time in range and reduce hypoglycemia across several user groups, although most exercise- and nutrition-specific evidence comes from adult and pediatric/adolescent cohorts rather than pregnancy-specific exercise studies. Exercise-related glucose responses remain highly dependent on user input, exercise modality, insulin on board, meal timing, and metabolic state. Planned exercise announcement, prandial bolus reduction before postprandial activity, and individualized carbohydrate intake remain key strategies. Biomarkers such as lactate, ketone bodies, non-esterified fatty acids, and counter-regulatory hormones may help explain interindividual variability and support future personalization. Conclusions: Nutrition and exercise management in AID users should be interpreted as a dynamic metabolic interface among exogenous insulin, endogenous counter-regulation, substrate availability, and algorithmic control. Emerging approaches, including activity sensors, adaptive algorithms, dual-hormone systems, digital twins, and metabolomics-informed personalization, may improve safety and reduce user burden, but several remain exploratory and require further validation in diverse free-living conditions. Full article

Background: Cutaneotrichosporon jirovecii is an under-characterized basidiomycetous yeast within the family Trichosporonaceae. Its taxonomic placement, ecological distribution, and functional potential remain incompletely understood because genome-scale resources for C. jirovecii and closely related lineages are limited. Methods: We characterized strain H0426_7, a C. jirovecii-related yeast recovered from a human fecal sample, using ITS-based type-strain comparison, ITS phylogenetic analysis, whole-genome sequencing, average nucleotide identity analysis, read-level assessment of public C. jirovecii-labeled datasets, and comparative functional annotation. Antifungal susceptibility was assessed using the Sensititre YeastOne plate. Results: The ITS sequence of H0426_7 closely matched type-strain material of C. jirovecii, including CBS 6864 and its equivalent deposits. The ITS-based tree placed H0426_7 adjacent to CBS 6864 with bootstrap support of 87%. The final draft genome comprised 38.66 Mb in 1974 contigs, with a GC content of 63.76% and BUSCO completeness of 80.0%. ANI analysis showed that H0426_7 was genomically distinct from the recognized Cutaneotrichosporon species included in the ANI analysis but highly similar to two unclassified feces-derived strains, P10-008 and PK4640, with ANI values exceeding 98.8%. Two public datasets labeled as C. jirovecii showed anomalously low ANI values with H0426_7; read-level taxonomic profiling indicated low target-fungal read proportions, suggesting that these datasets are unsuitable as definitive genome-level references. CAZyme annotation identified 285 family assignments in H0426_7, representing 278 non-redundant predicted proteins, including relatively high GH5 and GH31 counts, suggesting candidate carbohydrate-utilization features shared with the H0426_7/P10-008/PK4640 lineage. Conclusions: H0426_7 is best described as a C. jirovecii-related Cutaneotrichosporon isolate pending availability of a high-quality genome assembly from the C. jirovecii type strain. This study expands genome-scale resources for underrepresented basidiomycetous yeasts and provides a comparative framework for future taxonomic, ecological, and functional studies of feces-associated Cutaneotrichosporon lineages. Full article

Colombian palm wine is a traditional fermented beverage produced from the sap of Attalea butyracea, whose microbiota and biochemical features remain poorly characterized. A comprehensive analytical framework was applied to palm wine samples from three Andean producers. This included the determination of proximate composition, total phenolic content, and antioxidant activity, alongside a dual microbiological approach: traditional plate counting and high-throughput 16S rRNA/ITS metabarcoding. PICRUSt2 was employed to predict KEGG-based metabolic pathways to elucidate the microbial functional potential. The wines exhibited a low pH (3.35–3.65), a variable ethanol content (1.62–8.40 g/L), high residual sugars, moderate microbial loads, and limited antioxidant activity (as measured using the ABTS and DPPH assays). Analysis using high-throughput sequencing revealed high bacterial community diversity, dominated by Liquorilactobacillus nagelii, Limosilactobacillus fermentum, Limosilactobacillus panis, Lacticaseibacillus casei, and Zymomonas mobilis alongside the yeast Saccharomyces cerevisiae. Functional profiling revealed a significant enrichment in metabolic pathways related to carbohydrates, amino acids, and cofactors/vitamins, as well as xenobiotic biodegradation and metabolism. These findings provide the first integrated microbiological and physicochemical characterization of Colombian palm wine and highlight its biotechnological potential. Full article

Background: Gallbladder carcinoma (GBC) represents one of the most aggressive malignancies of the hepatobiliary system, evolving along a continuum from chronic inflammation to preneoplastic lesions and invasive cancer. This progression is frequently associated with gallstones and chronic cholecystitis and shares common pathogenic mechanisms with systemic inflammatory and metabolic disorders. Despite its relatively low incidence, GBC is characterized by poor prognosis, largely due to late-stage diagnosis and limited understanding of its molecular underpinnings. Methods: We conducted an observational study including 60 adult patients with radiologically suspected gallbladder cancer (GBC). Patients with disseminated disease, ongoing oncologic treatment, or synchronous malignancies were excluded. Fasting venous blood samples were collected to evaluate tumor markers and biochemical parameters, including carcinoembryonic antigen (CEA) and carbohydrate antigen CA 19-9. Surgical specimens were analyzed histopathologically and staged according to the European Society for Medical Oncology TNM classification system. Statistical analysis was performed using SPSS software (version 26.0), with appropriate parametric or non-parametric tests applied based on data distribution, and a p-value < 0.05 considered statistically significant. Results: Based on histological findings, patients were stratified into benign gallbladder disease (GBD) and GBC groups. CA 19-9 demonstrated higher mean serum levels with lower variability compared to CEA, suggesting superior sensitivity and diagnostic stability for gallbladder adenocarcinoma. In contrast, CEA levels exhibited greater fluctuation, limiting its reliability as a standalone biomarker. Importantly, the combined use of CA 19-9 and CEA improved diagnostic accuracy, supporting a multimarker approach for better clinical stratification. Our findings highlight the diagnostic value of CA 19-9 as a robust biomarker in GBC and support the integration of combined biomarker panels. Beyond tumor markers, the study identified a strong interplay between systemic inflammation and metabolic comorbidities, with obesity and hypertension significantly associated with chronic gallbladder pathology, and diabetes mellitus contributing to increased risk of acute inflammatory episodes. Elevated inflammatory markers, leukocytosis, and cholestatic enzyme alterations further supported the presence of a systemic inflammatory milieu. Multivariate analysis revealed that C-reactive protein (CRP), as a marker of systemic inflammation, was significantly influenced by a combination of clinical and biochemical variables, including age, hemoglobin, hypertension, amylase, CA 19-9, and CEA, explaining over 50% of its variability and up to 85% in advanced fibrotic changes. Additionally, platelet counts were significantly reduced in adenocarcinoma and correlated specifically with CA 19-9 levels, suggesting a potential link between tumor burden, inflammation, and platelet dynamics. Conclusions: Therefore, the observed associations between chronic inflammation, metabolic dysregulation, and tumor marker expression suggest a potential link between gallbladder carcinogenesis and systemic cardiometabolic pathways, opening new perspectives for early detection and targeted therapeutic strategies. Full article

Background: Effective glycemic control in diabetes management relies heavily on dietary carbohydrate knowledge. This study aimed to assess carbohydrate knowledge in individuals with type 1 diabetes (T1D) and insulin-treated type 2 diabetes (itT2D) using the GluciQuizz tool. Methods: A total of 465 persons (96 with T1D, 153 with itT2D; 89 and 127 matched controls without diabetes, respectively) from the French NutriNet-Santé prospective cohort were included. Participants completed the GluciQuizz questionnaire, which evaluates carbohydrate knowledge across five domains: carbohydrate food recognition; carbohydrate food content; nutrition label reading; glycemic targets and hypoglycemia prevention and treatment; and carbohydrate content of meals. Results: The mean age ± standard deviation of participants with diabetes was 65.8 ± 11.2 years, 44.2% male, with a diabetes duration of 23.3 ± 12.9 years. T1D participants scored significantly higher on the GluciQuizz compared to those with itT2D (23.9 ± 5.0 vs. 17.5 ± 5.6, p < 0.001). In secondary analysis, T1D participants showed superior knowledge to their matched controls without diabetes, whereas itT2D participants showed similar knowledge to their matched controls without diabetes. Conclusions: T1D participants demonstrated the best carbohydrate knowledge compared to those with itT2D. Targeted educational interventions in itT2D populations may improve dietary management and clinical outcomes. Full article

Background: Optimized nitrogen (N) application and planting density can enhance sweet potato yield. However, the agronomic mechanisms underlying their effects on photosynthetic efficiency and carbohydrate metabolism in sweet potato remain unclear. Methods: To address this, a two-year field experiment was conducted using a split-plot design with two varieties (YS-25 and GX-14), three N levels (60, 90, and 120 kg/ha; designated N60, N90, and N120, respectively), and three planting densities (D1–D3: 50,000, 62,500, and 83,333 plants/ha). Each treatment was replicated three times. Results: The results showed that the N60D2 treatment (60 kg/ha N; 62,500 plants/ha) optimized canopy light distribution by significantly increasing IPAR, light transmission rate, and extinction coefficient (K). This treatment enhanced individual plant photosynthetic capacity (higher photosynthetic rate: Pn, Ci, Gs, and Tr) and light energy use efficiency (Fv/Fm, Y(II), ETR, and qP), and promoted carbohydrate metabolism (sucrose, starch, fructose, and glucose) by increasing enzyme activities (Rubisco, SuSy, SPS, NI, SSS, and AGPase) in functional leaves and roots. These effects improved source–sink coordination, ultimately increasing storage root yield by 63.27–95.47% compared with the control plants (N120D1). Correlation analysis revealed that single-plant root weight and medium-sized root count were important yield determinants for both varieties. Conclusions: These results indicate that reducing nitrogen fertilizer combined with dense planting shapes a reasonable canopy structure for light distribution at the population level and optimizes light and carbon use efficiency at the individual plant level, thereby improving storage root yield and commercial characteristics of sweet potato. Full article

Consumers believe that expired products are unsafe, and, in most cases, misinterpreting the information on food labels often leads to large amounts of food waste. Yogurt is among the most widely eaten dairy products that can still be consumed after its expiration date, even though most consumers throw it away the very day it expires. The aim of this study was to determine whether commercial yogurts currently available on the market remain safe for consumption after their expiration date, with a view to reducing the amount of food waste generated in households. Therefore, the quality, stability, and edible safety of 10 commercial yogurts (two plain with 2% and 4% fat and the others with fruit, such as apricots, strawberries, bananas, blueberries, berries and strawberries, blackberries and raspberries, and cherries) stored at 4 °C before and at the expiration date were investigated. Physicochemical, textural, microbiological, and sensory analyses were performed to evaluate changes in functionality, safety, and acceptability of these yogurts. The results showed that, prior to their expiration date, certain yogurt samples (with apricots, strawberries, and blueberries, as well as plain yogurt with 4% fat) tested positive for total coliform bacteria, with values ranging from 20 to 50 CFU/g, suggesting substandard hygiene practices and insufficient sanitary conditions during and following the production process. No Escherichia coli, Listeria, Salmonella, Enterobacter spp., or Enterococcus spp. were detected in any of the yogurt samples that were within their expiration date. Blueberry, berry, and strawberry yogurts change their physical and chemical properties less than other types of yogurts analyzed after expiration. Yogurts containing berries and strawberries, blackberries, and raspberries remain safe at the expiration date, as they do not show the presence of harmful microorganisms such as coliform bacteria, Escherichia coli, Enterobacter spp., Enterococcus spp., Listeria, or Salmonella. Yogurt with berries and strawberries appears to be the most suitable from a microbiological point of view at expiration, as it has a low total mesophilic bacteria count and lactic acid bacteria exceeding 1 × 10⁶ CFU/g. At the time of expiration, this fruit yogurt type (with berries and strawberries) had a total solids content of 21.29%, 5.22% protein, 2.11% fat, 13.19% carbohydrates, 4.07 pH, 26.79% syneresis, 73.21% water retention capacity, 64.78% total phenolic content, and 10.55% DPPH (inhibition percentage). Nevertheless, at the time of expiration, from a sensory perspective (only appearance and consistency, odor, and color, without taste), the yogurt samples that were most appreciated contained blackberries and raspberries. The obtained results indicate that only certain types of fruit yogurts stored unopened at 4 °C may remain safe and edible after the expiration date, but further studies are needed to help the dairy industry and policymakers promote the reduction in food waste in households. Full article

Background: Pancreatic ductal adenocarcinoma (PDAC) has one of the poorest prognoses among malignant tumors, mainly due to the difficulty of early diagnosis. Therefore, it is crucial to identify reliable blood markers for a highly sensitive diagnostic system. We previously developed a highly sensitive extracellular vesicle (EV)-counting system, which can quantify the absolute number of specific EVs in serum. In this study, a multiplex assay using lectins that recognize specific glycans on EVs in the serum of PDAC patients was performed to select the optimal lectin combination. Methods: The glycan alteration signature of serum EVs from patients with PDAC was analyzed using a lectin-based multiplex assay combined with the EV-counting system. The optimal lectin combination that recognizes PDAC-specific changes was selected using machine learning analyses (support vector machine) for high diagnostic performance across independent patient cohorts. Results: An optimal lectin combination, Jacalin and Agaricus bisporus agglutinin (ABA), for PDAC detection was identified using machine learning analysis. This lectin-based system, reflecting changes in Jacalin/ABA binding, showed significantly higher diagnostic performance (area under the curve [AUC] = 0.890 and 0.971) than that of the conventional diagnostic marker carbohydrate antigen 19-9 (CA19.9; AUC = 0.752). Notably, the system achieved an AUC of 0.870 in patients with the stage I disease. Conclusions: These findings highlight the potential of a serum EV-based diagnostic system leveraging Jacalin and ABA glycan recognition for the early detection of PDAC. Full article

Background/Objectives: Effective glycemic management from the time of diagnosis is essential in the care of children and adolescents with type 1 diabetes (T1D), as early glycemic patterns can influence long-term health outcomes. Methods: This exploratory study evaluated a one-month interactive, group- and play-based education program designed to enhance food and carbohydrate counting skills among families of children and adolescents with newly diagnosed (ND) T1D (<1 year since diagnosis) or suboptimal glycemic control (SGC) (hemoglobin A1c (HbA1c) > 7.5% (58 mmol/mol)). The intervention included hands-on learning activities in food and carbohydrate counting, and peer interaction to support development of diabetes self-management skills. Data were collected at baseline, post-intervention, and at six-months follow-up through medical records, glucose sensor data, and a questionnaire assessing diabetes self-management skills, dietary practices, and carbohydrate counting. Results: Between September 2022 and April 2024, 55 children and adolescents were enrolled in the ND group and 22 in the SGC group. Post-intervention, carbohydrate counting skills improved, particularly in the ND group. Participants reported greater confidence and independence in carbohydrate counting and insulin dosing, with parents noting sustained benefits at six-months follow-up. No significant changes were observed in glycemic control, including time-in-range and postprandial glucose profiles. Conclusions: In this exploratory study, early interactive and play-based group education was associated with improvements in carbohydrate counting skills and self-care confidence in children and adolescents with newly diagnosed T1D. These improvements were not accompanied by changes in glycemic outcomes. The findings occurred during a complex and transitional phase following diagnosis. Further research is needed to examine sustainability and long-term clinical impact. Full article

This study evaluated whether body size could serve as a quality and traceability marker for cage-reared butter catfish (Ompok bimaculatus) from the Pak Phanang Basin, Nakhon Si Thammarat, Thailand, in support of Geographical Indication (GI) certification. Fish were classified into three commercial size grades—small (12–15 fish/kg), medium (6–10 fish/kg), and large (3–5 fish/kg)—corresponding to fish harvested after 6, 8, and 12 months of rearing, respectively, with mean body weight and total length of 75 ± 7 g and 19.8 ± 1.1 cm (small), 120 ± 9 g and 25.8 ± 2.1 cm (medium), and 260 ± 10 g and 32.2 ± 2.8 cm (large). Dorsal muscle samples were comparatively analyzed to assess size-related differences in physicochemical properties, nutritional composition, and microbiological quality. Proximate analysis showed that moisture, lipid, ash, and total energy contents increased significantly with fish size (p < 0.05), whereas protein and carbohydrate contents did not differ significantly among size groups (p > 0.05). Small fish exhibited slightly higher muscle pH (7.02 ± 0.18) than medium and large fish (6.65 ± 0.11 to 6.66 ± 0.25) (p < 0.05). Flesh color was characterized by a pale whitish to slightly yellow appearance with high lightness (L*; p < 0.05), while redness (a*) and yellowness (b*) did not differ significantly among size groups (p > 0.05). Mineral composition varied with size, with large fish containing significantly higher levels of calcium and magnesium (p < 0.05). Amino acid profiling identified 17 amino acids, including eight essential amino acids (EAA) and nine non-essential amino acids (NEAA), with EAA being more abundant in large fish—particularly leucine and lysine—indicating improved protein quality, while NEAA were dominated by glutamic and aspartic acids across all size groups. Fatty acid analysis revealed higher proportions of polyunsaturated fatty acids (PUFA) and greater unsaturated fatty acid/saturated fatty acid (UFA/SFA) ratios (1.7–1.8) in medium and large fish. Microbiological assessment showed a decrease in total viable counts with increasing fish size, while Escherichia coli, Staphylococcus aureus, and Salmonella spp. were not detected in any size group, as these microorganisms are commonly used as key hygiene and food-safety indicators in fish products (E. coli for fecal/handling hygiene, S. aureus for human-handling contamination, and Salmonella spp. as a major foodborne pathogen). Overall, body size was associated with consistent variations in physicochemical characteristics, nutritional composition, and microbial quality of Pak Phanang Basin butter catfish. These findings provide baseline compositional and safety markers that can support product specification development and GI documentation. Full article

In the Trentino Alto Adige region, disposal of grape stalks (GS) represents a major cost for wineries, despite their content of phenolic and tannin-rich compounds with potential functional value in ruminant nutrition. This study evaluated whether dietary GS supplementation could influence milk microbiota and cheese quality, supporting local circular-economy strategies. GS from three red cultivars (L-GS, CS-GS, M-GS) were dried, milled, and assessed for safety; their average total polyphenol content was approximately 15 g/kg DM. 3 Holstein cows underwent a 7-week trial consisting of alternating control (CTRL) and GS-supplemented periods (2% DM). Weekly milk samples (n = 21) and corresponding mini-cheeses (n = 21) were analyzed. GS supplementation did not affect milk coliforms (2.3–2.5 log CFU/mL), while total mesophilic counts were significantly lower in M-GS (2.8 ± 0.46 log CFU/mL) than in CTRL (4.5 ± 0.71; p < 0.05). Acinetobacter dominated the milk microbiota but decreased from 34.0% in CTRL to 18.0% in L-GS. Cheese total polyphenols were highest in CS-GS (224 ± 34 mg/kg). ¹H-NMR and VOCs profiling indicated cultivar-dependent shifts in carbohydrate-related metabolites and short-chain fatty acids. Overall, GS supplementation subtly modulated cheese biochemical and aromatic traits without impairing technological performance, supporting the valorization of winery by-products within integrated dairy-viticulture systems. Full article

Although plant-based convenience foods have gained significant market share, many are high in fat, salt, and sugar while low in nutrients. The current study aimed to develop a vegan oyster mushroom soup powder enriched with moringa, mung bean, and pumpkin. These ingredients were chosen for their high nutritional value and availability. Four soup formulas, each containing varying amounts of moringa (0%, 1%, 2%, and 3%), were prepared, and a sensory evaluation, proximate analysis, and total aerobic plate count were carried out. The 1% moringa formulation showed the highest consumer acceptance. In this formula, moisture, ash, protein, fat, fiber, carbohydrate, and energy content were reported as 13.6%, 7.6%, 16.3%, 2.2%, 9.8%, 50.5%, and 287 kcal/100 g, respectively. The novel powdered soup product had higher amounts of phenolic compounds, total antioxidants, and iron compared to local, commercially available equivalents. Total aerobic plate counts remained below 10⁵ CFU/g; a common acceptability limit for dried soups, throughout the 4-month storage study under ambient conditions. Overall, the developed soup powder demonstrated superior nutritional quality and could support consumers in meeting their daily nutrient requirements. With further refinement, particularly by optimizing the drying process to better retain heat-sensitive nutrients, this product shows potential as an affordable and nutritious option to address inadequate protein intake and iron deficiency in Sri Lanka. Full article

This study investigates a mechanically aerated, yeast-free bread technology incorporating apple-derived plant ingredients (juice, purée, and powder) in response to the growing demand for clean-label bakery products. The global bakery sector represents one of the largest food markets worldwide, with the baking yeast segment alone accounting for several billion USD annually, while interest in yeast-free and yeastless-dough products continues to expand. To address technological limitations associated with yeast exclusion, dough aeration was achieved using a two-stage whipping protocol (1000 rpm for 4 min, followed by 500 rpm for 1 min and stabilization at 500 rpm for 1 min under 4.0 ± 0.1 MPa gauge pressure), forming a stable protein–carbohydrate foam system. Rheological evaluation using Mixolab 2 showed that formulations containing 3–5% apple purée exhibited the most favorable dough development characteristics, with stability increasing from 3.30 ± 0.15 min in the control to 8.90 ± 0.20 min. Texture profiling using a CT-2 analyzer equipped with a cylindrical probe (50% compression, 60 mm/min, slices 25 mm thick, n = 5) revealed a significant reduction in crumb firmness, from 3.01 ± 0.15 N in the control to 2.12 ± 0.10 N in the purée- and powder-enriched samples (p < 0.05). Nutritional assessment indicated improvements in vitamin C content (up to 2.23 mg/100 g) and protein quality: the amino acid score, calculated according to FAO/WHO reference patterns on a mg/g-protein basis, increased from 76.5 ± 1.8% to 89.2 ± 2.3%. Microbiological analysis showed reduced total aerobic mesophilic counts after 72 h of storage—4.7 × 10³ CFU/g in the control versus 1.8–3.4 × 10³ CFU/g in apple-enriched breads. Overall, the results demonstrate that mechanical aeration combined with apple-derived ingredients enhances the structural, nutritional, and microbiological quality of yeast-free bread, offering a promising clean-label approach for functional bakery products. Full article