Search Results (228)

Background/Objectives: Baobab (Adansonia digitate L.) is an underutilized species and edible parts (fruits, leaves and seeds) contribute to food security and human health in tropical areas. Although the fruits have attracted greater research interest and have recently been approved for consumption in EU countries, the leaves are traditionally consumed but they have yet to be studied from an interventional perspective. The aim of this study was to propose a protocol for a dietary intervention using baobab leaves (BLs) to achieve the recommended reference values for proteins and minerals (K, Ca, Mg, Na, Fe, Mn) for different target groups of the Sudanese population. Methods: Dry matter, crude fat, protein and ash content, mineral content (Na, Mg, K, Ca, Fe, Mn), total phenolic, and flavonoid compounds were determined in BLs from six different areas. To assess the health and nutrition status in Sudan, time-series data (2013–2023) from the DataBank Health Nutrition and Population Statistics database were used. The reference values for nutrients recommended by the European Food Safety Authority were used to estimate the amount of baobab leaf intake (BLI, g/day). Results: For each nutrient, the study area with the lowest amount of BLs to be consumed is recommended. Leaves from the area of El Gari (BN3) 18.312 g/day and 30.712 g/day are recommended for K and Ca, which are particularly beneficial for children aged 1–3 years and lactating women. Leaves from Kor Tagat (KR1) are suitable for sodium intake, requiring approximately 13–23 g/day across all age groups. Leaves from Kazgil (KR2) (46–81 g/day), (35–66 g/day), (0.48–0.68 g/day), and (4–6 g/day) are optimal for fulfilling the daily requirements of magnesium, iron, manganese, and protein in this order. Conclusions: The systematic inclusion of BLs in the diet can positively support the nutritional status of various demographics. Moreover, the findings of this study demonstrated the foundation for public health and nutritional policy-makers on how they will tackle malnutrition and food insecurity worldwide by incorporating naturally available diets and nutritious alternatives. Recommendation: Further research should focus on assessing the nutritional composition factors that could affect the absorption of nutrients such as phytates and oxalates and investigating the in vitro bioavailability of the elements. Full article

Background: Preclinical cell tracking is enhanced with a multimodal imaging approach. Bioluminescence imaging (BLI) is a highly sensitive optical modality that relies on engineering cells to constitutively express a luciferase gene. Magnetic particle imaging (MPI) is a newer imaging modality that directly detects superparamagnetic iron oxide (SPIO) particles used to label cells. Here, we compare BLI and MPI for imaging cells in vitro and in vivo. Methods: Mouse 4T1 breast carcinoma cells were transduced to express firefly luciferase, labeled with SPIO (ProMag), and imaged as cell samples after subcutaneous injection into mice. Results: For cell samples, the BLI and MPI signals were strongly correlated with cell number. Both modalities presented limitations for imaging cells in vivo. For BLI, weak signal penetration, signal attenuation, and scattering prevented the detection of cells for mice with hair and for cells far from the tissue surface. For MPI, background signals obscured the detection of low cell numbers due to the limited dynamic range, and cell numbers could not be accurately quantified from in vivo images. Conclusions: It is important to understand the shortcomings of these imaging modalities to develop strategies to improve cellular detection sensitivity. Full article

Turbomachinery shock wave patterns occur as a natural result of operating at off-design points and are accountable for some of the loss in performance. In some cases, shock wave–boundary layer (SW-BLIs) interactions may even lead to map restrictions. The current paper refers to experimental findings on a transonic linear cascade specifically designed to mitigate shock waves using porous walls on the blades. Schlieren visualization reveals two phenomena: Firstly, the shock waves were dissipated in all bladed passages, as predicted by the CFD studies. Secondly, a lower-pressure wave pattern was observed upstream of the blades. It is this phenomenon that the paper reports and attempts to describe. Attempts to replicate this pattern using Reynolds-averaged Navier–Stokes (RANS) calculations indicate that the numerical method may be too dissipative to accurately capture it. The experimental campaign demonstrated a 4% increase in flow rate, accompanied by minimal variations in pressure and temperature, highlighting the potential of this approach for enhancing turbomachinery performance. Full article

The introduction of antibiotics in the beginning of the 20th century was one of the most important scientific breakthroughs in history. However, in recent decades, the growing threat of antimicrobial resistance (AMR) has shown the limitations of the current research and development programs for new antimicrobial drugs. In the last decade, 20 antibiotics, 7 β-lactam/β-lactamase inhibitor (BL/BLI) combinations and 4 non-traditional antibacterial drugs have been launched worldwide. Methods: This study aimed to assess the time to patient access for new antibacterial drugs in countries in the European Union and the European Economic Area (EU/EEA). Time differences in marketing authorization from the U.S. Food and Drug Agency (FDA) and the European Medicines Agency (EMA) were also described, as well as the availability of each drug in the countries in the EU/EEA according to the national competent authorities. Results: Substantial differences between countries were observed, with no or only one new drug available in some countries. Conclusions: Improving pricing and reimbursement timelines and fostering collaboration between national health authorities and market authorization holders can enhance timely and equitable patient access to new antibacterial treatments in Europe. Equitable and sustainable access to antibacterial drugs is a cornerstone in the battle against AMR. Full article

This study investigates the efficacy of Streptococcus salivarius K12 in preventing upper respiratory tract infections (URTIs) in healthy adults. URTIs are a common issue, particularly in physically active individuals, leading to significant disruptions in daily life. Probiotics, such as S. salivarius K12, have emerged as a potential preventive strategy for these infections. This research was conducted as a randomized, double-blind, placebo-controlled trial involving 112 participants aged between 19 and 25. Participants were randomly divided into two groups: one group received a daily dose of S. salivarius K12, marketed as Bactoblis^®, while the other received a placebo. The trial lasted for four months, during which adherence to the treatment protocol was closely monitored. The primary goal was to measure the incidence of URTIs using the Jackson Scale and the Wisconsin Upper Respiratory Symptom Survey (WURSS-11). The results indicated that higher adherence to the S. salivarius K12 treatment was associated with an increased number of days without URTI symptoms. Although the overall severity of symptoms did not differ significantly between the treatment and control groups, those with high adherence to S. salivarius K12 (greater than 90%) reported more days free from illness. In conclusion, S. salivarius K12 demonstrated potential as a preventive measure against URTIs, especially in individuals who adhered strictly to the treatment regimen. However, further research involving larger populations and longer follow-up periods is needed to fully confirm these findings and better understand the role of S. salivarius K12 in preventing respiratory infections. Full article

Proteins are the foundation of life, and designing functional proteins remains a key challenge in biotechnology. Before the development of AlphaFold2, the focus of design was primarily on structure-centric approaches such as using the well-known open-source software Rosetta3. Following the development of AlphaFold2, deep-learning techniques for protein design gained prominence. This study proposes a new method to generate functional proteins using the diffusion model and ESM2 protein language model. Diffusion models, which are widely used in image and natural language generation, are used here for protein design, facilitating the controlled generation of new sequences. The ESM2 model, trained on the basis of large-scale protein sequence data, provides a deep understanding of the context of the sequence, thus improving the model’s ability to generate biologically relevant proteins. In this study, we used the Protein A-like peptide as a model study object, combined the diffusion model and the ESM2 model to generate new peptide sequences from minimal input data, and verified their biological activities through experiments such as the BLI affinity test. In conclusion, we developed a new method for protein design that provides a novel strategy to meet the challenges of generic protein generation. Full article

Charcot-Marie-Tooth disease type 2N (CMT2N) is an inherited nerve disorder caused by mutations in the alanyl-tRNA synthetase (AlaRS) gene, resulting in muscle weakness and sensory issues. Currently, there is no cure for CMT2N. Here, we found that all five AlaRS mutations in the aminoacylation domain can interact with neuropilin-1 (Nrp1), which is consistent with our previous findings. Interestingly, three of these mutations did not affect alanine activation activity. We then performed a high-throughput screen of 2000 small molecules targeting the prevalent R329H mutant. Using thermal stability assays (TSA), biolayer interferometry (BLI), ATP consumption, and proteolysis assays, we identified Tanshinone I as a compound that binds to and modifies the conformation of the R329H mutant and other CMT-related AlaRS mutants interacting with Nrp1. Molecular docking and dynamic simulation studies further clarified Tanshinone I’s binding mode, indicating its potential against various AlaRS mutants. Furthermore, co-immunoprecipitation (Co-IP) and pull-down assays showed that Tanshinone I significantly reduces the binding of AlaRS mutants to Nrp1. Collectively, these findings suggest that Tanshinone I, by altering the conformation of mutant proteins, disrupts the pathological interaction between AlaRS CMT mutants and Nrp1, potentially restoring normal Nrp1 function. This makes Tanshinone I a promising therapeutic candidate for CMT2N. Full article

The Sam (Sterile alpha motif) domain of the lipid phosphatase Ship2 (Ship2-Sam) is engaged by the Sam domain of the receptor tyrosine kinase EphA2 (EphA2-Sam) and, this interaction is principally linked to procancer effects. Peptides able to hinder the formation of the EphA2-Sam/Ship2-Sam complex could possess therapeutic potential. Herein, by employing the FoldX software suite, we set up an in silico approach to improve the peptide targeting of the so-called Mid Loop interface of Ship2-Sam, representing the EphA2-Sam binding site. Starting from a formerly identified peptide antagonist of the EphA2-Sam/Ship2-Sam association, first, the most stabilizing mutations that could be inserted in each peptide position were predicted. Then, they were combined, producing a list of potentially enhanced Ship2-Sam ligands. A few of the in silico generated peptides were experimentally evaluated. Interaction assays with Ship2-Sam were performed using NMR and BLI (BioLayer Interferometry). In vitro assays were conducted as well to check for cytotoxic effects against both cancerous and healthy cells, and also to assess the capacity to regulate EphA2 degradation. This study undoubtedly enlarges our knowledge on how to properly target EphA2-Sam/Ship2-Sam associations with peptide-based tools and provides a promising strategy that can be used to target any protein–protein interaction. Full article

Luciferase (luc) bioluminescence (BL) is the most used light-emitting protein that has been engineered to be expressed in multiple cancer cell lines, allowing for the detection of tumor nodules in vivo as it can penetrate most tissues. The goal of this study was to develop an oncolytic adenovirus (OAd)-resistant human triple-negative breast cancer (TNBC) that could express luciferase. Thus, when combining an OAd with chemotherapies or targeted therapies, we would be able to monitor the ability of these compounds to enhance OAd antitumor efficacy using BL in real time. The TNBC cell line HCC1937 was stably transfected with the plasmid pGL4.50[luc2/CMV/Hygro] (HCC1937/luc2). Once established, HCC1937/luc2 was orthotopically implanted in the 4th mammary gland fat pad of NSG (non-obese diabetic severe combined immunodeficiency disease gamma) female mice. Bioluminescence imaging (BLI) revealed that the HCC1937/luc2 cell line developed orthotopic breast tumor and lung metastasis over time. However, the integration of luc plasmid modified the HCC1937 phenotype, making HCC1937/luc2 more sensitive to OAdmCherry compared to the parental cell line and blunting the interferon (IFN) antiviral response. Testing two additional luc cell lines revealed that this was not a universal response; however, proper controls would need to be evaluated, as the integration of luciferase could affect the cells’ response to different treatments. Full article

Brevetoxins (PbTxs) are very potent marine neurotoxins that can cause an illness clinically described as neurologic shellfish poisoning (NSP). These toxins are cyclic polyether in chemistry and have increased their geographical distribution in the past 2 decades. However, the ethical problems as well as technical difficulties associated with currently employed analysis methods for marine toxins have spurred the quest for suitable alternatives to be applied in a regulatory monitoring regime. In this work, we reported the first instance of concurrent aptamer selection of Brevetoxin-1 (PbTx-1) and Brevetoxin-2 (PbTx-2) and constructed a biolayer interferometry (BLI) biosensor utilizing PbTx-1 aptamer as a specific recognition element. Through an in vitro selection process, we have, for the first time, successfully selected DNA aptamers with high affinity and specificity to PbTx-1 and PbTx-2 from a vast pool of random sequences. Among the selected aptamers, aptamer A5 exhibited the strongest binding affinity to PbTx-1, with an equilibrium dissociation constant (K_D) of 2.56 μM. Subsequently, we optimized aptamer A5 by truncation to obtain the core sequence (A5-S3). Further refinement was achieved through mutations based on the predictions of a QGRS mapper, resulting in aptamer A5-S3G, which showed a significant increase in the K_D value by approximately 100-fold. Utilizing aptamer A5-S3G, we fabricated a label-free, real-time optical BLI aptasensor for the detection of PbTx-1. This aptasensor displayed a broad detection range from 100 nM to 4000 nM PbTx-1, with a linear range between 100 nM and 2000 nM, and a limit of detection (LOD) as low as 4.5 nM. Importantly, the aptasensor showed no cross-reactivity to PbTx-2 or other marine toxins, indicating a high level of specificity for PbTx-1. Moreover, the aptasensor exhibited excellent reproducibility and stability when applied for the detection of PbTx-1 in spiked shellfish samples. We strongly believe that this innovative aptasensor offers a promising alternative to traditional immunological methods for the specific and reliable detection of PbTx-1. Full article

The imperative for energy conservation and environmental protection has led to the development of innovative aircraft designs. This study explored a novel thrust control configuration for blended-wing-body (BWB) aircraft with distributed electric boundary-layer ingestion (BLI) propulsors, addressing the issues of sagging and altitude loss during landing. The research focused on a small-scale BWB demonstrator equipped with six BLI fans, each with a 90 mm diameter. Various thrust control configurations were evaluated to achieve significant thrust reduction while maintaining lift, including dual-layer sleeve, separate flap-type, single-stage linkage flap-type, and dual-stage linkage flap-type configurations. The separate flap-type configuration was tested through ground experiments. Control experiments were conducted under three different experimental conditions as follows: deflection of the upper cascades only, deflection of the lower cascades only, and symmetrical deflection of both cascades. For each condition, the deflection angles tested were 0°, 10°, 20°, 30°, 40°, 50°, and 60°. The thrust reductions observed for these three conditions were 0%, 37.5%, and 27.5% of the maximum thrust, respectively, without additional changes in the pitch moment. A combined thrust adjustment method maintaining a zero pitch moment demonstrated a linear thrust reduction to 20% of its initial value. The experiment concluded that the novel thrust control configuration effectively adjusted thrust without altering the BLI fans’ rotation speed, solving the coupled lift–thrust problem and enhancing BWB landing stability. Full article

Background: In vitro-transcribed (IVT) mRNA has been established as a promising platform for therapeutics and vaccine development. Double-stranded RNA (dsRNA) is a major impurity of IVT mRNA and can trigger unfavored immune responses, potentially causing adverse events in patients. Existing dsRNA detection and quantitation methods, such as gel electrophoresis, ELISA, or homogeneous time-resolved fluorescence (HTRF), have low sensitivity or are time-consuming. A recently published lateral flow immunoassay (LFSA) was shown to be fast, but it lacks the sensitivity for dsRNA with uridine modifications. Methods: In this study, we provided a possible explanation for the reduced sensitivity of existing quantitation methods for dsRNA with modified uridines by characterizing the binding affinities of commonly used anti-dsRNA antibodies. Then, a rapid and sensitive biolayer interferometry (BLI) dsRNA detection assay utilizing Flock House Virus (FHV) B2 protein was developed to overcome the challenges in dsRNA detection and the reduced sensitivity. Results: This assay allows the detection of dsRNA with different uridine modifications (ψ, m1ψ, 5 moU) with similar sensitivity as dsRNA without modification. Furthermore, we demonstrated this method can be used to quantify both short and long dsRNA, as well as hairpin-structured dsRNA, providing a more comprehensive detection for dsRNA impurities. Moreover, we applied this assay to monitor dsRNA removal through a purification process. Conclusions: Taken together, this BLI method could enable real-time monitoring of impurities in IVT mRNA production to prevent immunogenicity stemming from dsRNA. Full article

Synbiotics are mixtures of prebiotics and probiotics that enhance the activity of probiotic bacteria when co-administered to provide greater benefits to the host. Traditionally, the synbiotics that have been discovered enhance gut probiotic strains and are nutritionally complex molecules that survive digestive breakdown until they reach the later stages of the intestinal tract. Here, we screened and identified sugars or sugar substitutes as synbiotics for the oral probiotic strains Streptococcus salivarius BLIS K12 and BLIS M18. Using a modified deferred antagonism assay, we found that 0.5% (w/v) galactose and 2.5% (w/v) raffinose were the best candidates for use as synbiotics with BLIS K12 and M18, as they trigger enhanced antimicrobial activity against a range of bacteria representing species from the mouth, gut, and skin. Using reverse transcriptase quantitative PCR, we found that this enhanced antimicrobial activity was caused by the upregulation of the lantibiotic genes salA, salB, and sal9 in either K12 or M18. This led to the conclusion that either 2.5% (w/v) raffinose or 0.5% (w/v) galactose, respectively, are suitable synbiotics for use in conjunction with BLIS K12 and M18 to enhance probiotic performance. Full article

This study evaluates the utilization of in vitro embryo production (IVEP) technology for the conservation and breeding of the Auliekol cattle breed, a primary beef breed in Kazakhstan facing population decline due to the cessation of breeding programs and the incursion of transboundary diseases. We assessed the effect of consecutive ovum pick-up (OPU) procedures on oocyte yield and embryo production in Auliekol and Aberdeen Angus cows. A total of 2232 and 3659 oocytes were aspirated from Auliekol and Aberdeen Angus donors, respectively, with significantly higher yields and embryo production observed in Aberdeen Angus cows. The application of a meiotic block using Butyrolactone I (BLI) and subsequent in vitro fertilization (IVF) protocols was employed, with embryo development monitored up to the morula/blastocyst stage. Results indicated that Auliekol cows exhibited lower oocyte recovery, cleavage, and blastocyst rates compared to Aberdeen Angus cows, likely due to genetic characteristics. Despite the challenges, IVEP presents a valuable tool for the preservation and future propagation of the Auliekol breed, highlighting the need for further research to enhance reproductive outcomes and conservation strategies. Full article

Tailings ponds are recognized as significant sources of potential man-made debris flow and major environmental disasters. Recent frequent tailings dam failures and growing trends in fine tailings outputs underscore the critical need for innovative monitoring and safety management techniques. Here, we propose an approach that integrates UAV photogrammetry with convolutional neural networks (CNNs) to extract beach line indicators (BLIs) and conduct enhanced dam safety evaluations. The significance of real 3D geometry construction in numerical analysis is investigated. The results demonstrate that the optimized You Only Look At CoefficienTs (YOLACT) model outperforms in recognizing the beach boundary line, achieving a mean Intersection over Union (mIoU) of 72.63% and a mean Pixel Accuracy (mPA) of 76.2%. This approach shows promise for future integration with autonomously charging UAVs, enabling comprehensive coverage and automated monitoring of BLIs. Additionally, the anti-slide and seepage stability evaluations are impacted by the geometry shape and water condition configuration. The proposed approach provides more conservative seepage calculations, suggesting that simplified 2D modeling may underestimate tailings dam stability, potentially affecting dam designs and regulatory decisions. Multiple numerical methods are suggested for cross-validation. This approach is crucial for balancing safety regulations with economic feasibility, helping to prevent excessive and unsustainable burdens on enterprises and advancing towards the goal of zero harm to people and the environment in tailings management. Full article