Biological Stability Research Articles

Biological stability and phytotoxicity of sludge generated from extended aeration activated sludge systems

Biological stability and phytotoxicity of sludge generated from extended aeration activated sludge systems

Engineering of Cyclodextrin Glucosyltransferase from Paenibacillus macerans for Improved Regioselectivity and Product Specificity Toward Hydroxyflavone Glycosylation

The regioselective glycosylation and product specificity of hydroxyflavonoid compounds profoundly influences their biological activity and stability, offering significant therapeutic potential. However, most cyclodextrin glucosyltransferases (CGTases) inherently lack regioselectivity and product specificity for flavone glycosylation. Herein, a CGTase from Paenibacillus macerans was engineered for enhanced glycosylation regioselectivity and product specificity by combining molecular docking analysis and saturation mutagenesis strategies. K232L (favoring 4′-and 6-hydroxyflavones) and K232V (favoring 7-hydroxyflavone) were identified with distinct preferences. In addition, H233Y (preferring for 4′-hydroxyflavones), H233T (preferring for 6′-hydroxyflavones), and H233K (preferring for 7′-hydroxyflavones) also demonstrated distinct regioselectivity. These variants further exhibited enhanced hydrolytic activity, enabling the efficient production of short sugar-chain glycosides. Molecular dynamics (MDs) simulations revealed that the variants adopted optimized catalytic conformations with increased loop region flexibility near the binding pocket, enhancing substrate accessibility. These findings underscore the pivotal roles of K232 and H233 in broadening the substrate scope of CGTase and offer valuable guidance for enzyme engineering targeting regioselective glycosylation.

Evolution Driven Microscale Combinatorial Chemistry in Intracellular Mimicking Droplets to Engineer Thermostable RNA for Cellular Imaging.

Fluorescent light-up aptamer/fluorogen pairs are powerful tools for tracking RNA in the cell, however limitations in thermostability and fluorescence intensity exist. Current in vitro selection techniques struggle to mimic complex intracellular environments, limiting in vivo biomolecule functionality. Taking inspiration from microenvironment-dependent RNA folding observed in cells and organelle-mimicking droplets, an efficient system is created that uses microscale heated water droplets to simulate intracellular conditions, effectively replicating the intracellular RNA folding landscape. This system is integrated with microfluidic droplet sorting to evolve RNA aptamers. Through this approach, an RNA aptamer is engineered with improved fluorescence activity by exploring the chemical fitness landscape under biomimetic conditions. The enhanced RNA aptamer named eBroccoli has increased fluorescence intensity and thermal stability, both in vitro and in vivo in bacterial and mammalian cells. In mammalian cell culture conditions, a fluorescence improvement of 3.9-times is observed and biological thermal stability up to 45°C is observed in bacterial systems. eBroccoli enable real-time visualization of nanoscale stress granule formation in mammalian cells during heat shock at 42°C. By introducing the concept of "biomimetic equivalence" based on RNA folding, the platform offers a simple yet effective strategy to mimic intracellular complexity in evolution-based engineering.

Macroporous coating of silver-doped hydroxyapatite/silica nanocomposite on dental implants by EDTA intermediate to improve osteogenesis, antibacterial, and corrosion behavior

Coating a titanium (Ti) implant with hydroxyapatite (HA) increases its bioactivity and biocompatibility. However, implant-related infections and biological corrosion have restricted the success of implant. To address these issues, a modified HA nanocomposite (HA/silica-EDTA-AgNPs nanocomposite) was proposed to take advantage of the sustained release of silver nanoparticles (AgNPs) and silicate ions through the silica-EDTA chelating network. As a result, a uniform layer of nanocomposite, compared to HA as the gold standard, was formed on Ti implants without fracture and with a high level of adhesion, using plasma electrolytic oxidation (PEO). Bioactivity assessment evidenced a shift in the surface phase of the Ti implant to generation of beta-tricalcium phosphate, a more bioresorbable material than HA. Metabolic activity assessments using human dental pulp stem cells revealed that Ti surfaces modified by the new nanocomposite are superior to bare and HA-modified Ti surfaces for cell attachment and proliferationin vitro. In addition, it successfully inhibited bacterial growth and induced osteogenesis on the implant surface. Finally, potentiodynamic polarization behavior of Ti implants before and after coating confirmed that a thick oxide interface layer on the modified Ti surface acts as an electrical barrier and protects the substrate layer from corrosion. Therefore, the HA/silica-EDTA/Ag nanocomposite presented here, compared to HA, can better coat Ti dental implants due to its good biocompatibility and osteoinductive activity, along with improved biological stability.

Effects of Low-Severity Fire on the Composition and Stability of Soil Organic Carbon in Permafrost Peatlands (Northeast China).

Climate change and human activity are increasing the frequency of wildfires in peatlands and threatening permafrost peatland carbon pools. In Northeast China, low-severity prescribed fires are conducted annually on permafrost peatlands to reduce the risk of wildfires. These fires typically do not burn surface peat but lead to the loss of surface vegetation and introduction of pyrogenic carbon. However, the long-term effects of repeated low-severity fires on soil carbon stability in these ecosystems remain unclear. Thus, we conducted low-severity prescribed fire experiments over 3 years in the permafrost peatlands of the Great Khingan Mountains. Our findings showed a gradual decline in the total carbon content, primarily due to the reduction in free particulate organic matter (fPOM). Initially, fPOM was higher in the burned sites but decreased with repeated burning. Chemical analyses revealed a 32% increase in the aromaticity of the fPOM at the burned sites, which diminished the thermal stability of the soil. Furthermore, both prescribed fires and the addition of pyrogenic carbon reduced biological stability while increasing enzyme activity and CO2 production, which was attributed to the introduction of post-fire pyrogenic carbon. These results suggest that low-severity fires compromise the stability of permafrost peatlands, particularly because the pyrogenic carbon input alters the chemical composition of the soil carbon fraction.

Selection of High-Affinity and Selectivity AFB1 Circular Aptamer for Biosensor Application.

Detecting trace amounts of aflatoxin B1 (AFB1), one of the most toxic food contaminants, is crucial for efficiently preventing potential health risks. Circular aptamers are promising candidates for bioanalytical applications due to their enhanced biological and structural stability as well as their compatibility with rolling circle amplification (RCA). Herein, we employed a high-efficiency magnetic chain graphene oxide-based SELEX to generate circular aptamers that bind AFB1 with high affinity and selectivity. Notably, the selected circular aptamer, CAFB1-A-2, exhibited a significant sequence similarity to the classical AFB1 linear aptamer but demonstrated a distinct thermodynamic mechanism for binding. The binding of CAFB1-A-2 was driven by both enthalpy and entropy, whereas the classical linear aptamer exhibited a notable entropy loss that required a greater enthalpy compensation. Utilizing this circular aptamer, we developed a sensitive AFB1 detection system featuring an RCA-assisted allosteric DNAzyme biosensor with a detection limit (LOD) of 265 pM. Furthermore, the constructed aptasensor successfully detected AFB1 in spiked rice and corn, achieving LODs of 11 and 9 μg/kg, respectively, within approximately 4 h. This study provides a sensitive and reliable alternative to the development of an aptasensor for AFB1, holding great potential in the field of food safety detection.

Formulation Factors Affecting the Formation of Visible-Bubbles During the Reconstitution Process of Freeze-Dried Etanercept Formulations: Protein Concentration, Stabilizers, and Surfactants.

Freeze drying is one of the common methods to extend the long-term stability of biologicals. Biological products in solid form have the advantages of convenient transportation and stable long-term storage. However, long reconstitution time and extensive visible bubbles are frequently generated during the reconstitution process for many freeze-dried protein formulations, which can potentially affect the management efficiency of staff, patient compliance, and product quality. The reconstitution time has been extensively studied, but the influence of the formulations on the formation of visible bubbles is often overlooked. This paper investigated the effect of freeze-drying formulation factors (i.e., protein concentrations, surfactant concentrations, and sucrose/mannitol compositions) on product stability and visible bubbles generated during reconstitution of freeze-dried etanercept formulations. The generating and breakup mechanisms of visible bubbles were detected via internal microstructure of cake, surface tension, and viscosity measurement. Under the same protein concentration, the formulation of mannitol mixed with sucrose in a weight ratio of 4:1 produces fewer visible bubbles during the reconstitution process compared to the formulation of sucrose with the same total mass. This has been proven to be due to the large number of smaller radius pores distributed in the pores of the freeze-dried cake of the former, while the average internal structure pores of the latter are much larger than those of the former. As an amorphous stabilizer, sucrose can ensure the long-term stability of protein and greatly reduce the generation and maintenance of foams in the reconstitution process, making it a more robust excipient for freeze-dried protein formulations.

Binding-Site Directed Selection and Large-Scale Click-Synthesis of a Coagulation Factor XIa-Inhibiting Circular DNA Aptamer.

Factor XIa (FXIa) is a plasma protease thatplays a crucial role in the intrinsic pathway of blood coagulation, making it a promising target for antithrombotic therapy.Circular DNA aptamers, with their dramatically enhanced biological and structural stability, hold great potential as new-generation DNA-based anticoagulants. However, the functional selection and large-scale synthesis of them remains a substantial challenge. Here, we explored a selection strategy to enrich circular DNA aptamers that can target the catalytic domain of FXIa and inhibit its function. An asymmetrical dumbbell structured DNA library featuring a hairpin-like primer-binding site was utilized for selection. This approach has resulted in the identification of a circular DNA aptamer, named FICAPT1, which exhibits high binding affinity (Kd= 20 nM), notable stability (half-life of 16 hoursin human plasma) and outstanding anticoagulationactivity (significantly prolonged aPTT), showing great promise as a novel anticoagulant. The hairpin-like constant region further served as a scaffold for the large-scale click-synthesis (~ 2 mg/mL) of the refined circular aptamer. The approach presented in this study enables the efficient generation of functional circular DNA aptamers and their synthesis at a satisfactory scale, making it adaptable for the production of various circular aptamers for therapeutic applications.

Synthesis, Antimalarial Activity and Molecular Dynamics Studies of Pipecolisporin: A Novel Cyclic Hexapeptide with Potent Therapeutic Potential.

Malaria, caused by Plasmodium species and transmitted by Anopheles mosquitoes, continues to pose a significant global health threat. Pipecolisporin, a cyclic hexapeptide isolated from Nigrospora oryzae, has emerged as a promising antimalarial candidate due to its potent biological activity and stability. This study explores the synthesis, antimalarial activity, and computational studies of pipecolisporin, aiming to better understand its therapeutic potential. The peptide was successfully synthesized using Fmoc-based solid-phase peptide synthesis (SPPS) followed by cyclization in solution. The purified compound was characterized using HPLC and mass spectrometry, confirming a molecular ion peak at m/z [M + H]+ 692.4131, which matched the calculated mass. Structural verification through 1H- and 13C-NMR demonstrated strong alignment with the natural product. Pipecolisporin exhibited significant antimalarial activity with an IC50 of 26.0 ± 8.49 nM, highlighting its efficacy. In addition to the experimental synthesis, computational studies were conducted to analyze the interaction of pipecolisporin with key malaria-related enzymes, such as dihydrofolate reductase, plasmepsin V, and lactate dehydrogenase. These combined experimental and computational insights into pipecolisporin emphasize the importance of hydrophobic interactions, particularly in membrane penetration and receptor binding, for its antimalarial efficacy. Pipecolisporin represents a promising lead for future antimalarial drug development, with its efficacy, stability, and binding characteristics laying a solid foundation for ongoing research.

Status analysis of quality control of administered infusion solution with cytotoxic drugs.

The administered infusion solution is a sterile preparation that can be used directly for intravenous infusion in patients by mixing one or more intravenous drugs using aseptic operation technology. The pharmacy intravenous admixture service (PIVAS) center is a professional technical service department in hospitals, where the majority of inpatient-administered infusion solutions are prepared. During the processes of dissolution, dilution, preparation, storage, and use of intravenous drugs, the quality control of the administered infusion solution can be affected by various factors. At present, there are no relevant standards or guidance documents for the quality control of administered infusion solutions. Cytotoxic drugs are still the main treatment option for cancer patients and are mainly prepared in PIVAS centers in most hospitals. In this study, we mainly focused on the quality control of cytotoxic drug-administered infusion solutions and explored associated factors (diluent, container, concentration, temperature, and light), physical stability (visual appearance, pH, osmolality, and particulate matter), chemical stability (content), and biological stability (sterility). Most of the studies reviewed in this paper have insufficient data on the related factors and physicochemical stability of the administered infusion solutions. Research on the sterility of administered infusion solutions is particularly limited, with only one article addressing this aspect. Ensuring the quality of cytotoxic drug-administered infusion solutions is vital for the safe administration of drugs to cancer patients, so it is very important to enhance associated research. This article summarized the relevant literature on the quality control of cytotoxic drug-administered infusion solutions and provided a reference for safer and more efficient use of these drugs in clinical practice.

Synthesis and Evaluation of a Bifunctional Chelator for Thorium-227 Targeted Radiotherapy.

Thorium-227 (227Th) is an α-emitting radionuclide currently under investigation for targeted alpha therapy. Available chelators used for this isotope suffer from challenging multistep syntheses. Here, we present the synthesis and preclinical evaluation of a novel bifunctional chelator, p-SCN-Bn-DOTHOPO, which contains an isothiocyanate group that is suitable for conjugation to biological molecules. This bifunctional chelator was prepared with a 26% overall yield in four steps and conjugated to the human epidermal growth factor receptor 2 targeting antibody, trastuzumab. The resulting immunoconjugate was labeled with [227Th]ThIV (pH 5.5, room temperature, 60 min) with ≥95% radiochemical yield and purity. The conjugate was also labeled with zirconium-89 (89Zr), which can be used for positron emission tomography imaging. The radiometal complexes were subsequently investigated for their biological stability. The results described here provide insight into ligand design strategies and optimization of chelators for the development of the next generation of 89Zr and 227Th radiopharmaceuticals.

An acid polysaccharide from Mentha haplocalyx exerts the antifatigue effect via activating AMPK.

Fatigue is a pathological state that can impair physical and cognitive performance, making the development of effective therapeutic strategies crucial. In this study, an acid polysaccharide (MHa) was isolated from Mentha haplocalyx. Structural analysis showed that MHa (40.7kDa) has a backbone consisting of 4-α-GalAp, 6-α-Galp, and 4,6-α-Galp, with branches at the C6 of 4,6-α-Galp linked to four distinct side chains, including 4-α-Galp, 3,6-β-Manp, t-α-Araf, t-α-Rhap, t-α-Glcp, and t-β-Rhap. MHa possesses a triple-helix conformation with a sheet-like appearance, which may contribute to its biological stability and activity. Functionally, MHa exhibited significant antifatigue effects, with the 400mg/kg dose showing the most potent activity. Compared to the model group, treatment with 400mg/kg of MHa increased the exhaustive swimming time by 1.89-fold in fatigued mice, reduced blood lactate and urea nitrogen levels by 24.21% and 35.57%, respectively, and enhanced liver glycogen, muscle glycogen, and ATP levels by 20.08%, 46.52%, and 50.43%, respectively. MHa improved the activities of Ca2+-Mg2+-ATPase and Na+-K+-ATPase, while also enhancing antioxidant defense. Mechanistically, MHa promotes mitochondrial biogenesis and enhances oxidative defense via activating AMPK. These findings highlight the potential of MHa as a promising candidate for developing antifatigue supplements, offering a novel strategy to mitigate fatigue.

Control mechanism of Escherichia coli invasion by micro-nano bubbles in drinking water distribution system.

Sudden biological contamination in Drinking Water Distribution System (DWDS) significantly threatens the safety of drinking water, with E. coli invasions being particularly hazardous to human health. Traditional disinfection methods (i.e., chlorine, ultraviolet and ozone) provide partial microbial reduction. Micro-nano bubbles (MNBs) offer a promising alternative due to easy preparation, environmental friendliness, and generating ·OH in situ. This study explored the control mechanism of MNBs using different gas sources (i.e., nitrogen, air, oxygen, and ozone) on E. coli invasion in drinking water of the secondary water supply tanks. MNB characteristics, water quality changes, bacterial concentration, and microbial communities were evaluated. Results indicated that E. coli gradually became the dominant bacterium by promoting species interaction and influencing the process of microbial community construction, leading to a 6.25% increase in bacterial counts in water. MNBs generated via a dissolved gas release method exhibited particle sizes ranging from 500 to 800 nm and Zeta of -0.6 to -3.1 mV, and the bubble collapse effect generated a large amount of ·OH (0.11 ∼ 0.40mmol/L), which reduced bacterial abundance by 66.53% and microbial community richness, as revealed by decreases in the Chao (10.53%) and ACE (3.75%) indexes. The oxidative stress induced by ·OH inhibited protein transcription and energy production, which damaged DNA repair mechanisms. Thus, the relative abundance of Gammaproteobacteria, including E. coli as the dominant strain, decreased by 47.6%, leading to a balanced microbial community. Additionally, MNBs showed a complete reduction of bacteria, such as Caldisericia and Fusobacteria, thereby improving the drinking water safety and biological stability. This study highlights the potential of MNBs to address sudden exogenous biological pollution in DWDS, providing critical theoretical support to ensure the safety of drinking water quality.

Real-time monitoring of vancomycin using a split-aptamer surface plasmon resonance biosensor.

Real-time monitoring of therapeutic drugs is crucial for treatment management and pharmacokinetic studies. We present the optimization and affinity tuning of split-aptamer sandwich assay for real-time monitoring of the narrow therapeutic window drug vancomycin, using surface plasmon resonance (SPR). To achieve reversible, label-free sensing of small molecules by SPR, we adapted a vancomycin binding aptamer in a sandwich assay format through the split-aptamer approach. By evaluating multiple split sites within the secondary structure of the original aptamer, we identified position 27 (P27) as optimal for preserving target affinity, ensuring reversibility, and maximizing sensitivity. The assay demonstrated robust performance under physiologically relevant ranges of pH and divalent cations, and the specific ternary complex formation of the aptamer split segments with the analyte was confirmed by circular dichroism spectroscopy. Subsequently, we engineered a series of split-aptamer pairs with increasing complementarity in the stem regions, improving both the affinity and limit of detection up to 10-fold, as compared to the primary P27 pair. The kinetics of the engineered split-aptamer pairs were evaluated, revealing fast association and dissociation rates, confirming improved affinity and detection limits across variants. Most importantly, the reversibility of the assay, essential for real-time monitoring, was maintained in all pairs. Finally, the assay was further validated in complex biological matrices, including the cerebrospinal fluid from dogs and diluted plasma from rats, demonstrating functionality in biological environments and stability exceeding 9 hours. Our study paves the way for applications of split-aptamers in real-time monitoring of small molecules, with potential implications for in vivo therapeutic drug monitoring and pharmacokinetic studies.

Real-life experience with disease-modifying drugs in hereditary transthyretin amyloid polyneuropathy: A clinical and electrophysiological appraisal.

New treatments have dramatically improved the prognosis for Hereditary Transthyretin Amyloid Polyneuropathy (ATTRv-PN). However, there is a lack of routine follow-up studies outside of therapeutic trials. Our aim was to report the long-term clinical and electrophysiological evolution of a cohort of ATTRv-PN patients and to determine which biomarkers are most sensitive to change. We retrospectively collected neuropathy impairment scale (NIS), polyneuropathy disability scale (PND), overall neuropathy limitation scale (ONLS), rash built overall disability scale (RODS), electrodiagnostic data, motor unit number index (MUNIX), troponin and N-terminal pro-brain natriuretic peptide levels. Electrophysiological worsening was defined as a 20% decrease in previous values. Thirty-five patients, with a median age of 58 (interquartile ranges 42-71) years, were followed for a median of 36 (24-48) months. All patients received a transthyretin stabiliser, gene silencer or liver transplant. Overall assessment of the cohort showed clinical, biological and electrophysiological stability. However, on an individual basis, NIS worsened in 45% of patients (14/31), ONLS in 46% (13/28), PND in 28% (9/32) and RODS in 39% (11/28) at the last follow-up. Motor amplitude sum score decreased in 33% (11/33), amplitude recorded on tibialis anterior muscle in 44% (12/27), sensory amplitude sum score in 39% (11/28) and MUNIX sum score in 27% (7/26). Overall effectiveness of ATTRv-PN treatments in routine care is good. However, individual assessments show up to 40% deterioration over time. Electrophysiological measures are valuable monitoring tools but are not more sensitive to change than clinical scores. Results must be confirmed in larger cohorts.

Synthesis of New Polyfluoro Oligonucleotides via Staudinger Reaction.

Nowadays, nucleic acid derivatives capable of modulating gene expression at the RNA level have gained widespread recognition as promising therapeutic agents. A suitable degree of biological stability of oligonucleotide therapeutics is required for in vivo application; this can be most expeditiously achieved by the chemical modification of the internucleotidic phosphate group, which may also affect their cellular uptake, tissue distribution and pharmacokinetics. Our group has previously developed a strategy for the chemical modification of the phosphate group via the Staudinger reaction on a solid phase of the intermediate dinucleoside phosphite triester and a range of, preferably, electron deficient organic azides such as sulfonyl azides during automated solid-phase DNA synthesis according to the conventional β-cyanoethyl phosphoramidite scheme. Polyfluoro compounds are characterized by unique properties that have prompted their extensive application both in industry and in scientific research. We report herein the synthesis and isolation of novel oligodeoxyribonucleotides incorporating internucleotidic perfluoro-1-octanesulfonyl phosphoramidate or 2,2,2-trifluoroethanesulfonyl phosphoramidate groups. In addition, novel oligonucleotide derivatives with fluorinated zwitterionic phosphate mimics were synthesized by a tandem methodology, which involved (a) the introduction of a carboxylic ester group at the internucleotidic position via the Staudinger reaction with methyl 2,2-difluoro-3-azidosulfonylacetate; and (b) treatment with an aliphatic diamine, e.g., 1,1-dimethylethylenediamine or 1,3-diaminopropane. It was further shown that the polyfluoro oligonucleotides obtained were able to form complementary duplexes with either DNA or RNA, which were not significantly differing in stability from the natural counterparts. Long-chain perfluoroalkyl oligonucleotides were taken up into cultured human cells in the absence of a transfection agent. It may be concluded that the polyfluoro oligonucleotides described here can represent a useful platform for designing oligonucleotide therapeutics.

Nanocarrier-Mediated RNA Delivery Platform as a Frontier Strategy for Hepatic Disease Treatment: Challenges and Opportunities.

Hepatic diseases cause serious public health problems worldwide, and there is an urgent need to develop effective therapeutic agents. In recent years, significant progress is made in RNA therapy, and RNA molecules, such as mRNAs, siRNAs, miRNAs, and RNA aptamers, are shown to provide significant advantages in the treatment of hepatic diseases. However, the drawbacks of RNAs, such as their poor biological stability, easy degradation by nucleases in vivo, low bioavailability, and low concentrations in target tissues, significantly limit the clinical application of RNA-based drugs. Therefore, exploring and developing effective nanoscale delivery platforms for RNA therapeutics are of immense value. This review focuses on the different types of hepatic diseases and RNA therapeutics, summarizing various nanoscale delivery platforms and their strengths and weaknesses. Finally, the current status and future prospects of nanoscale delivery systems for RNA therapy are discussed.

An overview of the use of non-titanium MXenes for photothermal therapy and their combinatorial approaches for cancer treatment.

Since the initial publication on the first Ti3C2T x MXene in 2011, there has been a significant increase in the number of reports on applications of MXenes in various domains. MXenes have emerged as highly promising materials for various biomedical applications, including photothermal therapy (PTT), drug delivery, diagnostic imaging, and biosensing, owing to their fascinating conductivity, mechanical strength, biocompatibility and hydrophilicity. Through surface modification, MXenes can mitigate cytotoxicity, enhance biological stability, and improve histocompatibility, thereby enabling their potential use in in vivo biomedical applications. MXenes are also known for their ability to absorb light in the near-infrared (NIR) region and generate heat by localised surface plasmon resonance (LSPR) effects and electron-phonon coupling. Optical excitation laser pulses result in hot photocarrier distribution in MXenes, which quickly transfers surplus energy to the crystal lattice and results in the internal conversion of light into heat with nearly 100% efficiency. The relaxation of hot carrier distribution by electron-phonon interactions leads to the cooling of the lattice by dissipating thermal energy to the surrounding environment. This heating effect of MXenes makes them potential photothermal agents (PTAs), particularly for PTT applications. The adjustable surface of MXenes and their high surface area-to-volume ratios are ideal for the combinatorial approach of PTT along with drug delivery, photodynamic therapy (PDT), bone regeneration and other applications. Since non-Ti MXenes are more biocompatible than Ti MXenes, they are promising candidates for different biomedical applications. This comprehensive review provides a concise overview of the current research patterns, properties, and biomedical applications of non-Ti MXenes, particularly in PTT and its combinatorial approaches.

Deep Learning-Driven Optimization of Antihypertensive Properties from Whey Protein Hydrolysates: A Multienzyme Approach.

This study utilized deep learning to optimize antihypertensive peptides from whey protein hydrolysate. Using the Large Language Models (LLMs), we identified an optimal multienzyme combination (MC5) with an ACE inhibition rate of 89.08% at a concentration of 1 mg/mL, significantly higher than single-enzyme hydrolysis. MC5 (1 mg/mL) exhibited excellent biological stability, with the ACE inhibition decreasing by only 6.87% after simulated digestion. In in vivo experiments, MC5 reduced the systolic and diastolic blood pressure of hypertensive rats to 125.00 and 89.00 mmHg, respectively. MC5 significantly lowered inflammatory markers (TNF-α and IL-6) and increased antioxidant enzyme activity (SOD, GSH-Px, GR, and CAT). Compared to the MC group, the MC5 group showed significantly reduced serum renin and ET-1 levels by 1.25-fold and 1.04-fold, respectively, while serum NO content increased by 3.15-fold. Furthermore, molecular docking revealed four potent peptides (LPEW, LKPTPEGDL, LNYW, and LLL) with high ACE binding affinity. This approach demonstrated the potential of combining computational methods with traditional hydrolysis processes to develop effective dietary interventions for hypertension.

Importance of miRNAs in Cancer Therapy and Therapeutic Delivery Approaches

Uncontrolled cell division and proliferation due to environmental and genetic factors are characteristics of cancer. Despite traditional treatments such as surgery, chemotherapy and radiation therapy, the incidence and mortality rate of cancer increases every year. Therefore, it is necessary to improve new treatment strategies such as microRNA (miRNA). miRNAs are non-coding endogenous RNA molecules with 18-22 nt in length. They regulate important pathways in different cancer types and play active roles in mechanisms such as proliferation, migration, apoptosis and invasion. Due to their functions in the pathogenesis of cancer as oncogenic or tumor suppressors, miRNAs have been currently used as therapeutic agents. However, the limited biological stability, inability to diffuse into the target tissue and their degradability by nucleases of miRNAs are disadvantages of miRNAs. Viral and non-viral miRNA delivery systems provide high transfection to target tissue and avoid nuclease-induced degradation. This study highlights the importance of miRNAs’ regulatory involvement in cancer processes and therapeutic delivery approaches.