Search Results (52)

Replacing partial chemical fertilizers with organic fertilizer can increase organic carbon input, change soil nutrient stoichiometry and microbial metabolism, and then affect soil organic carbon (SOC) storage. A 6-year field experiment was used to explore the mechanism of SOC storage under different ratios of manure substitution in northeast China, with treatments including chemical fertilizer application alone (nitrogen, phosphorus, and potassium, NPK) and replacing 1/4 (1/4M), 2/4 (2/4M), 3/4 (3/4M), and 4/4 (4/4M) of chemical fertilizer N with manure N. Soil nutrients, enzymatic activity, and SOC fractions were analyzed to evaluate the effect of different manure substitution ratios on SOC storage. A high ratio of manure substitution (>1/4) significantly increased soil total N, total P, total K, and available nutrients (NO₃⁻-N, available P, and available K), and the 4/4M greatly decreased the C/N ratio compared to the NPK. Manure incorporation increased microbial biomass carbon (MBC) by 18.3–53.0%. Treatments with 50%, 75%, and 100% manure substitution (2/4M, 3/4M, and 4/4M) enhanced bacterial necromass carbon (BNC), fungal necromass carbon (FNC), and total microbial necromass carbon (MNC) by 31.9–63.5%, 25.5–107.1%, and 27.4–94.2%, respectively, compared to the NPK treatment. Notably, the increase in FNC was greater than that of BNC as the manure substitution ratio increased. The increasing manure substitution significantly enhanced particulate organic C (POC) and total SOC but did not affect mineral-associated organic C (MAOC). High soil N and P supplies decreased leucine aminopeptidases (LAPs) and alkaline phosphatase activities but increased the activity ratio of β-glucosidase (BG)/(N-acetyl-glucosaminidase (NAG) + LAP). Treatments with 25% manure substitution (1/4M) maintained maize and soybean yield, but with increasing manure rate, the maize yield decreased gradually. Overall, the high ratio of manure substitution enhanced SOC storage via increasing POC and MNC, and decreasing the decomposition potential of manure C and soil C resulting from low N- and P-requiring enzyme activities under high nutrient supplies. This study provides empirical evidence that the rational substitution of chemical fertilizers with manure is an effective measure to improve the availability of nutrients, and its effect on increasing crop yields still needs to be continuously observed, which is still a beneficial choice for enhancing black soil fertility. Full article

Background/Objectives: Diabetic kidney disease (DKD) is a major complication of type 2 diabetes mellitus (T2DM), and recent research highlights that amino acid composition—rather than total protein intake alone—may influence DKD risk. This study aimed to evaluate the associations between dietary protein intake, specific amino acid profiles, and the risk of DKD among adults with T2DM. Methods: A total of 378 T2DM patients were enrolled in this cross-sectional study. Dietary intake was assessed via a 24 h recall and a validated semi-quantitative food frequency questionnaire. Nutrient analysis was based on the Taiwanese Food Composition Database. Participants were categorized into three protein intake groups: Group 1 (≤0.8 g/kg), Group 2 (0.9–1.2 g/kg), and Group 3 (≥1.3 g/kg). Cox proportional hazards models were used to evaluate the associations of crude protein, branched-chain amino acids to aromatic amino acids (BCAA/AAA) ratio, and ketogenic amino acid intake with DKD risk. Adjustments were made for age, sex, diabetes duration, and blood pressure. Results: While crude protein intake showed no significant association with DKD risk, higher intake of ketogenic amino acids (e.g., leucine and lysine) was consistently and significantly associated with reduced DKD risk (adjusted HR range = 0.698–0.716, p < 0.01). Our findings highlight the protective potential of ketogenic amino acids such as leucine and lysine, which were significantly associated with lower DKD risk. The BCAA/AAA ratio also showed a downward trend in DKD risk, though not statistically significant. Kaplan–Meier analysis revealed that moderate protein intake (0.9–1.2 g/kg) corresponded to the most favorable DKD-free survival. Conclusions: Our findings suggest that, beyond total protein quantity, the intake of ketogenic amino acids may play a protective role in DKD prevention. Moderate protein consumption combined with higher leucine and lysine intake appears beneficial. These results support incorporating amino acid profiling in dietary strategies for DKD risk reduction. Further longitudinal and interventional studies are recommended to validate these associations. Full article

Background/Objectives: This study aimed to develop a dry powder inhalation (DPI) formulation of rivaroxaban (RVX) using a combination of bead milling (BM) and jet milling (JM) to enhance lung-targeted delivery for the effective treatment of pulmonary embolism while minimizing systemic exposure. Methods: A carrier-free DPI formulation of RVX was developed using sequential BM and JM, with L-leucine incorporated at various concentrations (1%, 5%, and 10%) as a force control agent. The formulations were characterized for particle morphology, size distribution, crystallinity, and thermal properties. The in-vitro aerodynamic performance was evaluated using a next-generation impactor, while ex-vivo studies assessed anticoagulant activity. Pharmacokinetic and tissue distribution studies were carried out in Sprague Dawley rats following intratracheal administration, and the effects of inhaled RVX were compared with those of oral administration. Results: The optimized BM-JM-5L formulation achieved a Dv50 of 2.58 ± 0.01 µm and a fine particle fraction of 72.10 ± 2.46%, indicating suitability for pulmonary delivery. The two-step milling effectively reduced particle size and enhanced dispersibility without altering RVX’s physicochemical properties. Ex-vivo anticoagulation tests confirmed maintained or improved activity. In-vivo studies showed that pulmonary administration (5 mg/kg) led to a 493-fold increase in lung drug concentration and 2.56-fold higher relative bioavailability vs. oral dosing, with minimal heart tissue accumulation, confirming targeted lung delivery. Conclusions: The two-step milled RVX DPI formulations, particularly BM-JM-5L with 5% leucine, demonstrated significant potential for pulmonary administration by achieving high local drug concentrations, rapid onset, and improved bioavailability at lower doses. These findings highlight the feasibility of RVX as a DPI formulation for pulmonary delivery in treating pulmonary embolism. Full article

Background: Nanocrystals, a carrier-free nanotechnology, offer significant advantages for pulmonary drug delivery by enhancing the dissolution and solubility of poorly soluble drugs while maintaining favorable biological properties and low toxicity. This study aims to investigate the aerodynamic performance and stability of nanocrystal-based dry powders (NC-DPs). Methods: Nanocrystalline suspensions were produced via wet media milling and subjected to stability studies before undergoing nano spray drying. A factorial design was employed to optimize the process parameters. The influence of mannitol and leucine, individually and in combination, was evaluated in terms of aerodynamic properties (Aerodynamic Particle Sizer (APS), in silico modeling) and the physicochemical stability at room temperature (in a desiccator) and accelerated conditions (40 ± 2 °C, 75 ± 5% relative humidity). Results: APS analysis revealed that leucine-containing powders (K-NC-Ls) exhibited the smallest median (1.357 µm) and geometric mean (1.335 µm) particle sizes, enhancing dispersibility. However, in silico results indicated the highest exhaled fraction for K-NC-L, highlighting the need for optimized excipient selection. Although mannitol showed the lowest exhaled fraction, it was mainly deposited in the extra-thoracic region in silico. The mannitol/leucine combination (K-NC-ML) revealed a low exhaled fraction and high lung deposition in silico. Also, K-NC-ML demonstrated superior stability, with a 6% reduction in D[0.5] and a 5% decrease in span overtime. Furthermore, no significant changes in crystallinity, thermal behavior, drug release, or mass median aerodynamic diameter were observed under stress conditions. Conclusions: These findings confirm that combined incorporation of mannitol and leucine in NC-DP formulations enhances stability and aerodynamic performance, making it a promising approach for pulmonary drug delivery. Full article

The family Potamanthidae is widely distributed across the Holarctic and Oriental regions, with nymphs often utilized as bioindicators for water pollution and human-induced environmental disturbances. However, limited mitochondrial genomes (mitogenomes) have been reported for this family. This study presents the first complete mitochondrial genome sequences of two Potamanthidae species, Potamanthus longitibius and Rhoenanthus youi. The mitogenome sizes were 15,430 bp and 15,232 bp, with A + T contents of 68.79% and 66.75%, respectively. The most abundant amino acids were phenylalanine (Phe), isoleucine (Ile), glycine (Gly), and alanine (Ala), with leucine (Leu2) showing the highest relative synonymous codon usage (RSCU) value. The analysis of tRNA secondary structures revealed high conservation among the tRNAs encoded on the H-strand. Phylogenetic reconstruction, incorporating 40 species from 13 families of Ephemeroptera, confirmed the monophyly of all the families and supported a sister group relationship between Potamanthidae and Ephemeridae. Additionally, the sequence previously identified as Rhoenanthus sp. JZ-2021 was reassigned to the genus Potamanthus. This study provides valuable insights into the evolutionary relationships within Potamanthidae and lays a foundation for future phylogenetic and taxonomic research. Full article

This study explores the production and valorization of hemp seed cake protein concentrate (HPC) as a functional ingredient to enhance the nutritional quality and sensory attributes of choux pastry products, specifically éclairs. By integrating varied concentrations of HPC (0%, 1%, 5%, 10%, 15%, and 20%) into traditional formulations, the physicochemical properties, proximate composition, amino acid profile, and sensory characteristics of the resulting pastries were assessed. Sensory attributes were assessed using the check-all-that-apply (CATA) method, where a trained panel selected applicable descriptors from a predefined list. Results indicated that the incorporation of HPC significantly increased protein content from 8.23% in the control sample (HPC0%) to 11.32% in the HPC20% formulation and improved moisture retention, leading to greater exterior and interior éclairs volume, increasing from 42.15 cm³ to 51.5 cm³ and from 18.34 cm³ to 38.47 cm³, respectively. Furthermore, sensory evaluation revealed pronounced differences in attributes such as flavor, appearance, and mouthfeel, with optimal sensory profiles noted at 10% HPC inclusion. The amino acid analysis demonstrated a balanced composition, particularly of essential amino acids, emphasizing HPC’s potential as a valuable protein source, with significant contributions from leucine (8.17 g/100 g protein), isoleucine (5.56 g/100 g protein), and phenylalanine (6.31 g/100 g protein), as well as notable levels of immunoactive amino acids such as arginine (10.92 g/100 g protein) and glutamic acid (20.16 g/100 g protein). These findings highlight the significant nutritional benefits of HPC enrichment, supporting the development of healthier bakery products and contributing to sustainable food practices within the industry. Full article

Royal jelly and medical grade honey are traditionally used in treating wounds and infections, although their effectiveness is often variable and insufficient. To overcome their limitations, we created novel amphiphiles by modifying the main reparative and antimicrobial components, queen bee acid (hda) and 10-hydroxyl-decanoic acid (hdaa), through peptide bonding with specific tripeptides. Our molecular design incorporated amphiphile targets as being biocompatible in wound healing, biodegradable, non-toxic, hydrogelable, prohealing, and antimicrobial. The amphiphilic molecules were designed in a hda(hdaa)-aa1-aa2-aa3 structural model with rational selection criteria for each moiety, prepared via Rink/Fmoc-tBu-based solid-phase peptide synthesis, and structurally verified by NMR and LC–MS/MS. We tested several amphiphiles among those containing moieties of hda or hdaa and isoleucine–leucine–aspartate (ILD-amidated) or IL-lysine (ILK-NH₂). These tests were conducted to evaluate their prohealing and antimicrobial hydrogel properties. Our observation of their hydrogelation and hydrogel-rheology showed that they can form hydrogels with stable elastic moduli and injectable shear-thinning properties, which are suitable for cell and tissue repair and regeneration. Our disc-diffusion assay demonstrated that hdaa-ILK-NH₂ markedly inhibited Staphylococcus aureus. Future research is needed to comprehensively evaluate the prohealing and antimicrobial properties of these novel molecules modified from hda and hdaa with tripeptides. Full article

Coffee pulp, a by-product of wet coffee processing, shows significant potential in the food and health domains, but its real applications remain underexplored. This work investigated the chemical composition and bioactive properties of coffee pulp from São Miguel Island (Azores, Portugal). The studied coffee pulp exhibited high fiber content (52% dw), mostly insoluble; notable mineral levels (10.6%), mainly K, Ca, and Mg; and 6% dw of total amino acids, with hydroxyproline, aspartic acid, glutamic acid, and leucine in higher amounts. Despite containing low fat (1.6% dw), mainly saturated, it also showed considerable amounts of polyunsaturated fatty acids with a favorable n6/n3 ratio (1.40) and vitamin E (α-, β-, and γ-tocopherols). Its antioxidant capacity can be partially explained by the chlorogenic acid content (9.2 mg/g dw), and caffeine (0.98%) was present in similar amounts to those observed in some arabica coffee beans. A decrease in glucose uptake in Caco-2 cells was found, but not in fructose, suggesting selective inhibition of SGLT1 and potential antidiabetic effects. These results show that Azorean coffee pulp has potential as a sustainable and bioactive ingredient for incorporation into functional foods or dietary supplements. Full article

The genetic basis controlling the content of two essential amino acids (isoleucine and leucine) in rapeseed meal was investigated through a replicated trial of the two BC₁F₁ populations from a two-way backcross between 202 TN DH population strains and their parents (‘Tapidor’ and ‘Ningyou7′). Given the impact of rapeseed embryos and maternal plants on seed qualities, a multi-genetic-system QTL mapping method was employed, incorporating both genetic main effects and environmental interaction effects. The results demonstrated the presence of nine QTLs associated with isoleucine and leucine content in the A1, A4, A5, A7, A9, and C2 linkage groups. These included six QTLs controlling isoleucine content and three QTLs controlling leucine content, which collectively explained 55.49% and 56.06% of the phenotypic variation, respectively. Of these, four QTLs were identified as the main QTL, which collectively explained over 10% of the phenotypic variation. All of the identified QTLs exhibited a highly significant additive and dominant effects on seed embryos. Additionally, one of the QTLs demonstrated had a particularly significant additive effect derived from the maternal genome. QTLs controlling isoleucine and leucine were identified in the A1, A4, and C2 linkage groups. Moreover, two QTL clusters influencing these essential amino acid contents were identified in the A4 and C2 linkage groups, situated between molecular markers HS-K02-2 and HBR094 and between EM18ME6-220 and NA12C03, respectively. Full article

Unconventional protein sources are currently extensively studied as food ingredients. This study aimed to evaluate the effect of 1.5% and 3% field cricket powder (GB) and 2–8% of its mixture (1:1) with spirulina powder (S) on the nutritional value, physicochemical properties, and sensory characteristics of corn extrudates. Additionally, 2% baking powder (BP) was added to assess its impact on the properties of the enriched extrudates. The results showed that both GB and GB + S improved nutritional value, with protein content increasing by up to 46% and higher levels of essential amino acids, particularly leucine and valine. However, these ingredients decreased the expansion ratio (by up to 15%), colour lightness (by up to 30%), and yellowness (by up to 47%) and increased the hardness (by up to 25%) of the corn extrudates. The S addition positively influenced product storage stability but decreased its sensory acceptance, especially aroma and taste. The BP addition mitigated the negative effects of higher GB and GB + S concentrations, particularly on sensory characteristics. In conclusion, incorporating up to 6% of the GB + S mixture provides a higher protein content with only minor changes to the product’s characteristics compared to GB. Ratings exceeding 4.2 points indicate the good acceptability of these snacks. Full article

The vertical stratification of oxygen concentration in deep reservoirs impacts nutrient cycling and ecosystem biodiversity. The Serra da Mesa reservoir, the largest in Brazil, was studied to evaluate the structure and production of the prokaryote community at five depths. Using ³H leucine incorporation and fluorescent in situ hybridization (FISH), the study focused on different depths near the dam, particularly within the euphotic zone. The water column was characterized into oxic, transitional, and hypoxic layers based on dissolved oxygen concentration. The highest densities and biomasses of prokaryotes were found at the euphotic zone’s depth limit, where bacterial production was low, suggesting inactive or slow-growing bacteria. Cell size differences and filamentous bacteria presence near the surface were observed, likely due to varying predation pressures. Prokaryote community composition differed across depths. At the subsurface level, with high dissolved organic carbon, alphaproteobacteria, betaproteobacteria, and Cytophaga–Flavobacter had similar densities, but the lowest bacterial biomass was recorded. The highest dissolved oxygen concentration depth had the lowest bacterial density, dominated by alphaproteobacteria and gammaproteobacteria. The study revealed that prokaryotic community structure and production vary with depth, indicating that microbial participation in layer dynamics is differentiated, with variations in abundance and distribution linked to oxygen concentrations. Full article

Microalgae are exceptional organisms from a nutritional perspective, boasting an array of bioactive compounds that have long justified their incorporation into human diets. In this study, we explored the potential of five microalgae species: Nannochloropsis sp., Tetraselmis chuii, Chaetoceros muelleri, Thalassiosira weissflogii, and Tisochrysis lutea. We conducted comprehensive analyses of their nutritional profiles, encompassing protein content, individual amino acid composition, mineral and trace element levels, fatty acid profiles (including saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs), and polyunsaturated fatty acids (PUFAs)), polyphenol compositions, and vitamin B content. The antioxidant activity of the ethanolic extracts was evaluated using two methods: ABTS and DPPH radical scavenging assay. The total protein content of the microalgae ranged from 34.09 ± 0.39% to 42.45 ± 0.18%, with the highest concentration observed in T. weissflogii. Essential amino acids such as histidine, threonine, lysine, valine, isoleucine, leucine, phenylalanine, and methionine were present in concentrations ranging from 0.53 ± 0.02 to 12.55 ± 2.21 g/16 g N. Glutamic acid emerged as the most abundant amino acid, with concentrations ranging from 6.73 ± 0.82 to 12.55 ± 2.21 g/16 g N. Among the microalgae species, T. chuii exhibited the highest concentrations of calcium (Ca) and manganese (Mn), while C. muelleri showed prominence in magnesium (Mg), sodium (Na), and iron (Fe). T. weissflogii stood out for its potassium (K) content, and T. lutea contained notable amounts of copper (Cu), zinc (Zn), and lead (Pb). Regarding fatty acid profiles, Nannochloropsis sp. and T. chuii were predominantly composed of SFA, while C. muelleri and T. weissflogii were rich in MUFA. PUFAs dominated the fatty acid profile of T. lutea, which also exhibited the most diverse range of polyphenolic substances. We also analyzed the B vitamin content, with T. lutea displaying the highest concentrations of niacin (B₃) and riboflavin (B₂). Antioxidant activity was confirmed for all microalgae tested using DPPH and ABTS radical IC₅₀ (mg/mL) converted to Trolox equivalent (TEAC). These findings underscore the substantial potential of the examined microalgae species as sources of biologically valuable substances characterized by rapid growth and relatively undemanding cultivation conditions. Full article

Dry-powder inhalers (DPIs) are valued for their stability but formulating them is challenging due to powder aggregation and limited flowability, which affects drug delivery and uniformity. In this study, the incorporation of L-leucine (LEU) into hot-melt extrusion (HME) was proposed to enhance dispersibility while simultaneously maintaining the high aerodynamic performance of inhalable microparticles. This study explored using LEU in HME to improve dispersibility and maintain the high aerodynamic performance of inhalable microparticles. Formulations with crystalline itraconazole (ITZ) and LEU were made via co-jet milling and HME followed by jet milling. The LEU ratio varied, comparing solubility, homogenization, and aerodynamic performance enhancements. In HME, ITZ solubility increased, and crystallinity decreased. Higher LEU ratios in HME formulations reduced the contact angle, enhancing mass median aerodynamic diameter (MMAD) size and aerodynamic performance synergistically. Achieving a maximum extra fine particle fraction of 33.68 ± 1.31% enabled stable deep lung delivery. This study shows that HME combined with LEU effectively produces inhalable particles, which is promising for improved drug dispersion and delivery. Full article

Alternanthera philoxeroides and Hypophthalmichthys molitrix offer significant nutritional benefits. This study evaluates the proximate composition, amino acid profile, GC-MS analysis, FT-IR spectroscopy, SEM and EDX, and color values of edible paper sheets (EPSs) derived from Alternanthera philoxeroides incorporating different levels of Hypophthalmichthys molitrix flesh. The protein content in the EPSs varied based on fish flesh incorporation, peaking at 52.66% in Ap100/Hm300 (Non-boil). Protein and carbohydrate contents showed an inverse correlation across EPSs, with the highest carbohydrate content of 60.89% in sample Ap400/Hm0 (Boil). Lipid content was also found to correlate with H. molitrix flesh content in EPSs, ranging from 1.59% to 18.41%. Amino acid analysis identified 11 types, with methionine as the most prevalent, followed by leucine, phenylalanine, and lysine. GC-MS analysis revealed 51 bioactive compounds, including carbonic acid, hentriacontane, and various fatty acids. FT-IR analysis showed characteristic bonds, while color analysis displayed L* values ranging from 24.37 to 30.97. SEM analyses depicted the microstructure, surface view, and elemental composition of the EPSs, and EDX showed an abundance of Ca, N, K, O, C, Mg, Na, P, Cl, Mn, and Fe. Therefore, EPSs prepared from A. philoxeroides and H. molitrix could offer a promising approach for effectively utilizing aquatic biomass and providing both plant and animal nutrients to consumers. Full article

Ralstonia eutropha strain H16 is a chemoautotrophic bacterium that oxidizes hydrogen and accumulates poly[(R)-3-hydroxybutyrate] [P(3HB)], a prominent polyhydroxyalkanoate (PHA), within its cell. R. eutropha utilizes fructose or CO₂ as its sole carbon source for this process. A PHA-negative mutant of strain H16, known as R. eutropha strain PHB⁻4, cannot produce PHA. Strain 1F2, derived from strain PHB⁻4, is a leucine analog-resistant mutant. Remarkably, the recombinant 1F2 strain exhibits the capacity to synthesize 3HB-based PHA copolymers containing 3-hydroxyvalerate (3HV) and 3-hydroxy-4-methyvalerate (3H4MV) comonomer units from fructose or CO_2. This ability is conferred by the expression of a broad substrate-specific PHA synthase and tolerance to feedback inhibition of branched amino acids. However, the total amount of comonomer units incorporated into PHA was up to around 5 mol%. In this study, strain 1F2 underwent genetic engineering to augment the comonomer supply incorporated into PHA. This enhancement involved several modifications, including the additional expression of the broad substrate-specific 3-ketothiolase gene (bktB), the heterologous expression of the 2-ketoacid decarboxylase gene (kivd), and the phenylacetaldehyde dehydrogenase gene (padA). Furthermore, the genome of strain 1F2 was altered through the deletion of the 3-hydroxyacyl-CoA dehydrogenase gene (hbdH). The introduction of bktB-kivd-padA resulted in increased 3HV incorporation, reaching 13.9 mol% from fructose and 6.4 mol% from CO₂. Additionally, the hbdH deletion resulted in the production of PHA copolymers containing (S)-3-hydroxy-2-methylpropionate (3H2MP). Interestingly, hbdH deletion increased the weight-average molecular weight of the PHA to over 3.0 × 10⁶ on fructose. Thus, it demonstrates the positive effects of hbdH deletion on the copolymer composition and molecular weight of PHA. Full article