Carob (Ceratonia siliqua) seeds are rich in diverse bioactive compounds, including galactomannan, β-sitosterol, unsaturated fatty acids and proteins with bioactive peptides in their sequence. This study delineates the compositional characterization of six carob seed flour types derived from different production processes, providing valuable insights for designing tailored nutraceutical products based on desired bioactive compound profiles. Our analysis indicated that a higher purity of galactomannan resulted in a greater mannose/galactose ratio, which increased the linearity of the galactomannan polymer and could enhance interchain interaction, thereby increasing aggregation capacity. A higher viscosity could potentially increase the capacity of galactomannan to create satiety and lower cholesterol levels. Among the different tested flours, those whose main compound was the endosperm were optimal for containing high galactomannan content, whereas those derived from the germ were ideal for having high concentrations of fatty acids (i.e., oleic and linoleic acids) and β-sitosterol. The presence of these lipids in carob flours could offer cardiovascular and metabolic health benefits, contributing synergistically. Additionally, flours that contain the germ have beneficial peptides included in proteins like glycinin and conglutin with potential anticholesterolemic and antidiabetic properties. This work provides different methods for obtaining carob flours rich in bioactive compounds, offering the nutraceutical industry a framework to select the best option for industrial-scale production.

Proteolytic products are extensively used in the nutraceutical sector to improve protein digestion and muscle quality in target populations (e.g., athletes or elderly). These products are processed using techniques that often lead to low purity but competitive pricing. Despite their widespread use and well-established production methods, the industry lacks standardized analytical methods for assessing these products and detecting potential fraud. This study proposes a comprehensive and harmonized pipeline for their analysis, which includes quantifying total soluble protein and proteolytic activity, as well as the determination of product stability and protein profile using SDS-PAGE and proteomic techniques. Despite the fact that protease extracts from pineapple had the highest protein content, most of the bromelain remained inactive, unlike in kiwi and papaya. SDS-PAGE revealed partial protein degradation of pineapple extracts, whereas kiwi extracts reflected a lower purification level but a higher protein integrity. The application of proteomic approaches strengthened the identification and origin tracing of the proteases. This study contributes to the development of a robust framework for analyzing proteolytic extracts, spanning from soluble protein quantification to protein profiling and activity determination. It may also ensure reliable supplier selection, high-quality manufacturing practices, and the implementation of optimal storage and formulation strategies in the nutraceutical industry.

Background/Objectives: Sexual dysfunction is a prevalent and multifactorial condition affecting a large proportion of the global population, with limited therapeutic options beyond pharmacological approaches primarily targeting erectile dysfunction. This has increased interest in botanical supplements for sexual health, although mechanistic evidence and clear links between phytochemical composition and biological activity remain scarce. The present study provides an integrative evaluation of a commercial Turnera diffusa extract (Liboost^®) formulated to support sexual health by combining detailed phytochemical characterization with targeted in vitro mechanistic assays. Methods: The extract was characterized by HPLC-DAD-HRMS, enabling the identification and semi-quantification of its major constituents. A total of 49 compounds were detected, predominantly flavonoids, including luteolin- and apigenin-derived glycosides, flavonols, methoxyflavones, flavanones, and coumaroyl derivatives, with a total quantified flavonoid content of 15.9 mg·g⁻¹. Biological activity was evaluated in human cell models without cytotoxic effects at the tested concentrations. Results: Liboost^® significantly reduced PDE5 expression, inhibited aromatase activity, and moderately increased nitric oxide production. These complementary effects suggest a multi-target modulation of pathways involved in sexual function, integrating vascular, endocrine, and nitrergic mechanisms. Conclusions: Although limited to in vitro models, the findings provide mechanistic support for the biological activity of T. diffusa extracts and highlight the importance of linking phytochemical composition with functional evidence when evaluating botanical supplements.

Protein utilization efficiency is a key determinant of metabolic health, body composition, and muscle function, particularly under high-protein dietary conditions and when using plant-based protein sources with lower digestibility. However, the extent to which enhancing early gastric proteolysis translates into improved whole-body protein utilization remains unclear. The present in vivo study investigated whether supplementation with an actinidin-enriched kiwifruit concentrate (Kwd+^®) modulates protein utilization efficiency and physiological outcomes under normoproteic and hyperproteic dietary conditions using casein or pea protein as dietary sources. Rats were fed normoproteic casein (NP), high-protein casein (CHP), or high-protein pea (PHP) diets with or without Kwd+^® supplementation for 8 weeks. Gastric protein hydrolysis patterns were evaluated by SDS–PAGE, while metabolic outcomes were assessed through body composition, plasma amino acid profiling, skeletal muscle morphology, and contractile function. Kwd+^® supplementation enhanced gastric proteolysis in a protein source-dependent manner. Despite the absence of significant changes in circulating amino acid concentrations after multiple comparison correction, Kwd+^® improved markers of protein utilization efficiency depending on dietary protein source. In animals fed a high-protein pea diet, supplementation significantly reduced fat mass relative to accumulated protein intake, indicating improved nutrient partitioning. In contrast, in animals fed a high-protein casein diet, Kwd+^® increased muscle mass relative to protein intake, suggesting enhanced anabolic efficiency. Under normoproteic conditions, Kwd+^® supplementation was associated with an increased muscle fiber cross-sectional area and improved fatigue resistance without alterations in fiber type composition or contractile protein abundance. These findings demonstrate that modulation of early gastric proteolysis through actinidin produces protein source-dependent effects on protein utilization efficiency, nutrient partitioning, and muscle function. This work highlights a novel nutritional strategy to improve metabolic outcomes and muscle performance, particularly in the context of high-protein and plant-based diets.

Protein utilization efficiency is a key determinant of metabolic health, body composition, and muscle function, particularly under high-protein dietary conditions and when using plant-based protein sources with lower digestibility. However, the extent to which en-hancing early gastric proteolysis translates into improved whole-body protein utilization remains unclear. The present in vivo study investigated whether supplementation with an actinidin-enriched kiwifruit concentrate (Kwd+®) modulates protein utilization efficiency and physiological outcomes under normoproteic and hyperproteic dietary conditions us-ing casein or pea protein as dietary sources. Rats were fed normoproteic casein (NP), high-protein casein (CHP), or high-protein pea (PHP) diets with or without Kwd+® sup-plementation for 8 weeks. Gastric protein hydrolysis patterns were evaluated by SDS–PAGE, while metabolic outcomes were assessed through body composition, plasma amino acid profiling, skeletal muscle morphology, and contractile function. Kwd+® supplemen-tation enhanced gastric proteolysis in a protein source-dependent manner. Despite the absence of significant changes in circulating amino acid concentrations after multiple comparison correction, Kwd+® improved markers of protein utilization efficiency de-pending on dietary protein source. In animals fed a high-protein pea diet, supplementa-tion significantly reduced fat mass relative to accumulated protein intake, indicating im-proved nutrient partitioning. In contrast, in animals fed a high-protein casein diet, Kwd+® increased muscle mass relative to protein intake, suggesting enhanced anabolic efficiency. Under normoproteic conditions, Kwd+® supplementation was associated with increased muscle fiber cross-sectional area and improved fatigue resistance without alterations in fiber type composition or contractile protein abundance. These findings demonstrate that modulation of early gastric proteolysis through actinidin produces protein source-dependent effects on protein utilization efficiency, nutrient partitioning, and mus-cle function. This work highlights a novel nutritional strategy to improve metabolic out-comes and muscle performance, particularly in the context of high-protein and plant-based diets.