Medical biotechnology serves as a foundational platform for identifying disease markers and developing specific diagnostic assays. Complementing this, biosensors and medical devices translate these biomarkers into quantifiable signals enabling real-time health assessment. Artificial intelligence (AI) augments this framework by processing vast datasets to recognize patterns and generate predictive insights thereby aiding clinicians in decision-making and streamlining personalized disease management. The integration of molecular-based companion diagnostics, biosensor technology, and medical AI forms the cornerstone of contemporary, data-driven precision medicine. This review explores the synergistic convergence of these fields specifically examining how AI-based approaches enhance molecular diagnostics and biosensor devices to optimize clinical outcomes. This interdisciplinary collaboration facilitates advancing precision medicine. Consequently, these discovery technologies profoundly impact disease prevention, early diagnosis, and targeted therapeutic strategies. Despite challenges such as data privacy and technology accessibility, this cross-disciplinary approach holds tremendous potential for transforming the future of precision healthcare.
Letter recognition is assumed to involve several levels of analysis, including coarse tuning for category and novelty and more fine tuning for specific features, related to letter orientation. We employed an oddball fast periodic visual stimulation (FPVS) paradigm with magnetoencephalography (Elekta VectorView, 306 sensors) to study neural discrimination responses in the source space. Using contrasts between native and foreign letters, digits, or inverted native letters, we aimed to isolate the neural responses to visual novelty, category, and orientation during character analysis. The study was conducted with a cohort of 25 adults. The response topography demonstrated bilateral organization, including language-related brain regions such as the ventral occipitotemporal cortex, inferior parietal cortex, and middle temporal areas. Comparing conditions, we revealed right lateralized parietal clusters, associated with novelty tuning, and left lateralized occipitotemporal clusters exhibiting higher activity for letters among digits discrimination, supporting the role of this area in letter processing. No distinct spatial patterns specific to orientation tuning were observed in comparison to novelty and category tuning. We propose that expertise-dependent orientation-specific tuning mechanisms may operate within an embedded neural framework that spatially overlaps with coarse tuning systems, but are characterized by specific spatiotemporal patterns.
Glycoprotein hormones, including follicle-stimulating hormone (FSH), luteinizing hormone (LH), and thyroid-stimulating hormone (TSH), play essential roles in the reproductive endocrinology of teleosts. However, research on these hormones in non-model species such as the Japanese eel (Anguilla japonica) has been constrained by the lack of specific immunological detection tools. Recombinant A. japonica FSH, LH, and TSH were expressed using a Bombyx mori-based silkworm-baculovirus system. Polyclonal antibodies were then generated using two distinct immunogen strategies: (i) chimeric constructs fusing β-subunits with a murine α-subunit, and (ii) synthetic peptides representing unique, non-conserved β-subunit epitopes. Both approaches induced antibody responses, but only peptide-based immunogens yielded polyclonal antisera with high specificity and sensitivity. These antibodies distinguished homologous glycoprotein hormones without cross-reactivity and retained target selectivity in mixtures simulating physiological serum conditions. Peptide-derived antibodies provide a robust and precise tool for the immunological discrimination of FSH, LH, and TSH in A. japonica. This strategy establishes a scalable framework for species-specific hormone assay development and offers translational potential for endocrine monitoring, reproductive management, and diagnostic applications in aquaculture biotechnology.
Bacteria exhibit extraordinary evolutionary and ecological diversity. They range from dominant, well-characterized phyla to rare lineages that are known only through environmental sequencing. This chapter reviews four key bacterial phyla, including Pseudomonadota, Bacillota, Actinomycetota, and Bacteroidota. These phyla are widely distributed, metabolically versatile, and play a central role in ecosystem functioning and human health. We discuss unique phyla within the PVC superphylum (Planctomycetota, Verrucomicrobiota, Chlamydiota) for their unusual cell biology, compartmentalization, and host associations. We also highlight hyperthermophilic phyla, such as Thermotogota, Aquificota, and Thermodesulfobacteriota, that thrive in geothermal ecosystems and drive sulfur and carbon cycling. We consider less-cultivated lineages, including Deinococcota, Acidobacteriota, Nitrospirota, Fusobacteriota, Fibrobacterota, Synergistota, Deferribacterota, and Chrysiogenota, in terms of their ecological niches, metabolic specializations, and roles in biogeochemical cycles, symbiosis, and disease. Collectively, these examples demonstrate the remarkable metabolic flexibility and ecological impact of bacteria, ranging from host-associated commensals and pathogens to free-living autotrophs in extreme environments. Despite advances in genomics and cultivation-independent methods, vast portions of bacterial diversity remain uncultured and poorly understood. Continued exploration of both dominant phyla and rare lineages promises to refine bacterial taxonomy, expand our understanding of microbial evolution, and reveal novel metabolic pathways with implications for ecology, medicine, and biotechnology.
We investigated the effectiveness of hydrated deep eutectic solvents (DESs) to achieve tunable seaweed cellulose microstructures from Ulva lactuca. Cellulose was extracted via a sequential chemical protocol, then treated with 30% DESs (choline chloride (ChCl) or betaine as hydrogen bond acceptors and urea, citric acid, or oxalic acid as donors) combined with mechanical shearing. While most DESs combinations yielded spherical seaweed cellulose microparticles (dry diameter of 605-777 nm), the ChCl:urea formulation successfully produced seaweed cellulose microfibers (SCMFs). These SCMFs exhibited superior quality (dry diameter of 372 nm), and water dispersibility with a hydrodynamic diameter of 134 nm and a polydispersity index of 0.23. Crucially, the DES composition dictated cellulose structure: ChCl:urea-treated SCMF remained predominantly amorphous, whereas other DESs treatments increased microparticles crystallinity. Furthermore, ChCl:oxalic acid introduced carboxyl functional groups. Selecting appropriate hydrated DESs thus offers a sustainable biotechnology tool to tune cellulose morphology, crystallinity, and surface chemistry.
This study evaluated real-world treatment patterns and 1-year outcomes in patients with neovascular age-related macular degeneration (nAMD) initiating faricimab. FARETINA-AMD was a retrospective study using data from the IRIS® Registry for patients diagnosed with nAMD initiating faricimab from February 2022 to March 2023. Included in the study were 2,025 treatment-naive patients (2,184 eyes) and 22,253 patients (26,851 eyes) previously treated with anti-vascular endothelial growth factor (anti-VEGF) therapy. Visual acuity improved by 2.0 ± 15.0 (mean ± SD) letters in treatment-naive eyes (P < .001) and was maintained in previously treated eyes at injection 7. Central subfield thickness (CST) improved by -53.1 ± 64.8 μm in treatment-naive and -28.5 ± 79.7 μm in previously treated eyes (both P < .0001); 78.4% and 66.8%, respectively, had achieved/maintained CST ≤ 280 μm at injection 7. Dosing frequency was reduced in the second 6 months (mean 2.4-3.2 injections) versus the first 6 months (4.0-4.2) of treatment. Outcomes among patients with nAMD receiving faricimab over 1-year follow-up support the real-world effectiveness and extended durability of treatment.
Bacillus subtilis is a key industrial microorganism widely used to produce enzymes, vitamins, antibiotics, and recombinant proteins. Its non-pathogenic nature, efficient protein secretion systems, and genetic tractability make it an ideal candidate for industrial biotechnology. Genome-scale metabolic modeling has emerged as a key systems biology approach to understand, explore and manipulate the metabolism of B. subtilis for improved strain performance. To date, the modeling efforts have resulted in fourteen published genome-scale metabolic reconstructions for B. subtilis; however, substantial opportunities for improvement remain, especially in integrating regulatory mechanisms for refining model accuracy. Furthermore, there is a growing trend toward modelling the cell as a whole system. This article is the first comprehensive and up-to-date review that highlights the genome sequencing, annotation efforts and the current state of genome-scale metabolic modeling for B. subtilis, emphasizing future directions.
Xylanases have attracted considerable attention due to their strong potential for industrial use. In this study, a xylanase-producing strain isolated from soil was identified as Trichoderma semiorbis Tsejk8, and the conditions for xylanase production were optimized. Additionally, two xylanase-related genes were cloned, and their functions were analyzed. The optimal conditions for xylanase production included maltose as the carbon source, peptone as the nitrogen source, an optimal pH of 6.0, and an incubation time of 120 h, yielding an enzyme activity of 40.7 U/mL. Following the purification of xylanase via ammonium sulfate precipitation and ion exchange chromatography, four distinct protein bands were observed. Mass spectrometry analysis of these bands identified 14 associated proteins. Bioinformatics analysis revealed that two of these proteins belong to GH3 (Glycoside Hydrolase family 3) β-xylosidase. In summary, the newly isolated strain Tsejk8 exhibits xylanase activity, offering an effective and eco-friendly means of converting biomass into raw materials for industrial applications.
Laccase is an environmentally friendly biocatalyst widely used in wastewater treatment, the food industry, and biosensors. However, free laccase is susceptible to environmental factors such as pH and temperature. Immobilizing it on nanomaterials can significantly mitigate these issues. This approach also enhances the reusability of laccase, demonstrating broad application prospects. Four different morphologies of MnO2 were compared, with δ-MnO2 (sheet) demonstrating the most effective immobilization effect as a carrier. Under conditions of pH=5, 30 °C, and laccase concentration of 1 mg/L, the reaction achieves optimal immobilization after 4 h of adsorption. Characterization of δ-MnO2 and immobilized laccase was performed using Fourier transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM). The enzymatic properties of immobilized laccase were also investigated. The results indicate that the optimal temperature for immobilized laccase is 75 °C, with an optimal pH of 3. Compared to free laccase, stability has been significantly enhanced. Furthermore, even after 30 days of storage at -4 °C the relative enzyme activity of the immobilized laccase remained as high as 74.51%. The kinetic constants demonstrate that immobilized laccase not only enhances the maximum reaction rate but also significantly improves substrate affinity. Compared to free laccase, the relative enzyme activity of MnO2-immobilized laccase (MnO2@Lac) is significantly enhanced. This may be attributed to the synergistic effect between MnO2 nanoparticles and laccase during substrate conversion. This study creates favorable conditions for the further application of immobilized laccase in the treatment of organic pollutants.
Immobilization of lipase enzyme is a promising approach towards cost-effective production of biodiesel to protect the enzyme from denaturation during the transesterification process. In the present study, hydrogen titanate nanotubes (HTNTs), synthesized by simple hydrothermal method, was used to immobilize lipase from Aspergillus niger via physical adsorption with the following ratios of HTNTs and lipase; 1 T: 1 L, 1 T: 0.75 L, 1 T: 0.5 L, and 1 T: 0.25 L. All prepared samples were characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM). The catalytic activities of free and immobilized lipases were evaluated for biodiesel production using sunflower oil, methanol to oil molar ratio of 4: 1 at 40 °C for 90 min. The biodiesel yields (0% water content, 0.877 g/cm3 density, and 0.041 mg acid value) were 79.2 ± 0.01, 82.3 ± 0.9, 79.1 ± 1.2, 81.97 ± 1.4, 78 ± 0.1 percent for 1 T: 1 L, 1 T: 0.75 L, 1 T: 0.5 L, 1 T: 0.25 L, and free lipase (1 L), respectively. Compared with the control (1 L), immobilizing lipase using (HTNTs) enabled a 50- 75% reduction in lipase required quantity while maintaining or even increasing biodiesel production levels. The work establishes a promising method for lipase immobilization in biodiesel production that can be evaluated for large-scale application in further studies.
The aldehyde dehydrogenase (ALDH) superfamily plays a critical role in acetaldehyde detoxification. The mitochondrial ALDH2 isoenzyme serves as the core enzyme in alcohol metabolism because of its high affinity for acetaldehyde. Loss-of-function mutation (such as ALDH2*2) of the gene is highly prevalent in East Asian populations. These mutations cause acetaldehyde accumulation and significantly increase the risk of alcohol-related diseases. Therefore, it is necessary to develop efficient recombinant ALDH2 supplementation therapies. However, its heterologous expression faces three major bottlenecks: (1) catalytic turnover leading to NAD⁺ depletion; (2) difficulty in functional tetramer assembly; and (3) cytotoxicity of the substrate acetaldehyde, which inhibits cell growth. The review covers rational host selection and multi-level synergistic optimization of transcription, translation, folding, and metabolism. Examples include promoter optimization, codon optimization, terminator optimization, and chaperone co-expression. Ultimately, we explore essential stabilization formulations and innovative delivery strategies, such as nano-encapsulation and engineered probiotics, needed to realize the therapeutic potential of ALDH2. The goal is to promote the industrial production and clinical translation of recombinant ALDH2.
Hemp (Cannabis sativa L.) is one of the oldest cultivated plants in human history. It is a valuable plant that has economic value for all industrial branches. The hemp plant has fibers of the highest quality. Accordingly, research aiming to enhance the fiber yield of hemp has significant agronomic and industrial relevance. To this end, three soilless media treatments were applied to the modified Hoagland solution. The effects of N: P:K/255:45:267 (A1), A1 + 10 mg/L silicon (A2), and A1 + 1 mg/L indole butyric acid (A3) on plant growth and fiber yield were investigated in Narlı hemp (Cannabis sativa L.), a registered local variety (Türkiye), in deep water culture and soil media (A4). The analysis determined that the soilless medium was more productive than the soil medium in terms of root fresh-dry weight, stem fresh-dry weight, stem length, stem thickness, leaf length and width, relative water content (RWC), photosynthetic pigment content, fiber content, and total cellulose content. According to these results, the lowest values were obtained in the soilless medium in terms of malondialdehyde (MDA) and leaf secondary metabolite (cannabidiol: CBD, tetrahydrocannabinol: THC, and cannabinol: CBN) contents, which indicate stress in the plant. CBD was measured %0.0213, %0.0134, %0.0395, %0.0326, THC was measured %0.002, %0.0017, %0.0032, %0.0033, CBN was measured %0.0003, %0.0002, %0.0002, %0.0009 (A1, A2, A3, A4 respectively). Considering all the parameters studied, the hemp plant grown in the deep water culture medium containing Si yielded the best results among the treatments. The fiber content was 29.12% in A2, 24.02% in A1, 19.99% in A4, and 13.74% in A3. The cellulose content was determined to range from 61% to 82% on average in the treatments tested. The highest total cellulose content of 82% was obtained from treatment A2. The current study revealed that silicon (Si) treatment significantly increased fiber and cellulose contents. Incorporating such treatments into production models is regarded as a cost-effective and efficient strategy.
Phospholipase C enzymes (plcA, plcB, plcC) represent critical virulence determination in mycobacterium tuberculosis pathogenesis. These enzymes play pivotal roles in disrupting phagosomal maturation, inducing macrophage necrosis, and facilitating immune evasion mechanisms. Protein sequences of phospholipase C gene plcA, plcB,plcC were collected and screened against human to exclude homologous matches and minimize cross-reactivity. Linear B-cell epitopes and T cell epitopes were identified and evaluated for the ability to produce strong antigenicity, solubility, toxicity and allergenicity. Suitable segments were linked using EAAK for adjuvant fusion, GPGPG between T-cell epitopes, and AAY between cytotoxin T-cell epitopes, with the L7/L12 ribosomal proteins at N-terminus as immunostimulatory adjuvant, The full vaccine structures were modelled in 3-D and improved for accuracy, and interaction of tuberculosis related proteins were analysed. Immunological potent epitopes for all 3 phospholipase C enzymes with favourable physiochemical properties and structural stability were identified. Immune simulation predicted effective stimulation of both humoral and cell-mediated responses. Molecular docking revealed promising interactions with key targets, characterized by favourable binding energies and stable complex formation dominated by hydrogen bonds and electrostatic interactions, suggesting potential functional efficacy of the designed vaccine. The study presents a strong potential of rationally designed multi-epitope vaccine candidate targeting phospholipase C virulence factor of M. tuberculosis. The computational workflow established a rigorous selection process for immunologically relevant epitopes assembled into chimeric construct with predicted vaccine potential. Further experimental validation through invitro antigenicity assay and in-vivo immunization studies is needed to assess the translational potential of this computationally designed vaccine.
Adeno-associated virus (AAV), a 4.7 kb single-stranded DNA virus, is widely used as a gene therapy vector, but its limited packaging capacity poses challenges for delivering large genes, always resulting in truncation during packaging. Quantifying truncation is difficult because both strands of the plasmid can be packaged from the 3' end. In this article, we aim to first produce single-polarity AAV and then explore its truncation pattern. To address this, we modified one of the ITRs and created an oversized self-complementary AAV, which functions as a single-polarity vector to some extent. Using this modified backbone, we generated a series of oversized single-polarity AAV (spAAV) vectors of varying lengths and sequences, analyzing DNA truncation patterns via quantitative PCR (qPCR). The results show that as the distance from the 3'-ITR increased, less DNA was detected. At 3000 bp from the 3'-ITR, 70% of the genomic DNA remained; this dropped to 50% at 4000 bp, 20% at 4500 bp, and almost none beyond 5000 bp. Additionally, reporter gene expression significantly decreased when the expression cassette extended to 4.5 kb compared to 2.7 kb under identical in vitro conditions. Our results show that DNA will be truncated far earlier before 4.5 kb during the packaging of very large genomes. This study provides important insights into the truncation patterns of AAV genomes, which is crucial for optimizing AAV vector design in gene therapy.
Starch's versatility inspires biomaterials for biomedical uses, customized through modifications, blending, or substituents. Diligent efforts have been dedicated to the development of starch-based biomaterials, leveraging the material's inherent biocompatibility and biodegradability, while aligning with environmentally sustainable considerations. While promising, most studies lack in vivo data and scalability assessments. In many cases, the reported advances are restricted to in vitro evaluations with limited information on long-term performance, clinical translation, and large-scale manufacturing feasibility in both economic and operational terms. This review furnishes an up-to-date synthesis of information available in the literature concerning recent breakthroughs in utilizing starch as a biomaterial, primarily focusing on advancements in areas such as wound dressings, drug delivery systems, the creation of scaffolds for regenerative medicine, and applications in tissue engineering. Advances have been made, with biomaterials presenting adequate biodegradability rates, active functions, good biocompatibility, and mechanical properties. However, it is noted that most research has not yet reached in vivo evaluations and lacks notions of large-scale production, in both economic and operational terms.
Insect resistance is a predominant trait in the globally approved genetically modified (GM) crops. The cry1Ab transgene is featured in over 25% of globally approved insect resistant GM events including both single as well as stacked trait events. GM maize notably contributes for 87% among the approved GM events with cry1Ab gene. GM detection of cry1Ab gene in maize seeds and food products is important for authentication purpose in the countries where GM maize events with cry1Ab gene such as Bt11, Bt176, MON810 are approved, and for surveillance of unauthorized occurrence of such events in the countries where these are not approved. Cost-efficient and rapid GM detection methods are essential for decentralized monitoring for unauthorized GM events across agricultural fields, border inspections, and food supply chains, and for regulatory compliance enabling quick decision-making. A rapid GM detection method employing loop-mediated isothermal amplification (LAMP) was developed for visual analysis of cry1Ab gene in GM maize. The developed assay showed acceptable specificity, which could reliably detect as low as 0.005% of cry1Ab within 60 min. Practical applicability of this assay was also verified for GM detection in maize containing products. The developed method offers a convenient, rapid and cost-effective tool for checking the cry1Ab gene either for regulatory purpose in the countries where the GM crops with cry1Ab gene are restricted or for confirmatory purpose for the samples with cry1Ab gene whenever required.
Great mass of tea tree seeds (TTS) are naturally rotted away as agricultural waste because there is no suitable technology used for extracting oil from TTS. The fermentation method is a new process and that can simultaneously recover oil, starch and saponins from TTS. In this study, a key bacterial strain named JJZ21 was isolated from fermented TTS milk (ground TTS and water) by the serial dilution method and identified as Lactiplantibacillus plantarum strain through molecular analyses. The production of cellulase, pectinase, amylase and protease of JJZ21 were visualized on agar plates containing specific enzyme substrates. Compared with the control, addition treatment with JJZ21 dramatically improved the activities of cellulase, pectinase, amylase and protease, reduced content of soluble sugar, protein and dry matter and pH in the TTS milk, shortened the fermentation time and increased the yield of TTS oil. Meanwhile, the addition of JJZ21 had no significant effect on quality of TTS oil. However, the yield of TTS oil decreased with excessive fermentation. Response surface methodology was used to evaluate the optimum conditions for fermentation process to obtain the maximum oil yield.
The growing demand for biopharmaceuticals has increased the need for efficient and cost-effective recombinant protein production platforms. Transcriptional gene silencing after random genomic integration of expression vectors remains a major limitation in mammalian expression systems and often reduces protein yield. Incorporation of ubiquitous chromatin opening elements (UCOEs) into expression cassettes has emerged as a promising strategy to mitigate position effects and enhance transgene expression. This study examined the effect of a UCOE-containing expression system on erythropoietin (EPO) production in Chinese hamster ovary (CHO) DG44 cells. A codon-optimized EPO expression cassette was introduced into CHO DG44 cells using either a conventional pOptiVEC vector and a UCOE-containing vector following random genomic integration. Recombinant EPO expression was assessed at the transcriptional and protein levels using reverse transcription quantitative polymerase chain reaction (RT-qPCR), western blotting, and enzyme-linked immunosorbent assay (ELISA). The UCOE-containing cell pool exhibited a significant enhancement in recombinant EPO expression, with approximately a 3.8-fold increase in mRNA levels and a sevenfold increase in secreted protein levels compared to the non-UCOE control cell pool. These results show that incorporating a UCOE reduces position-dependent gene silencing and increases recombinant mRNA and protein expression in CHO DG44 cell pools. This strategy supports improved efficiency during the early stages of cell line development for recombinant protein production.
The increasing incidence of breast cancer is leading researchers to investigate new treatment approaches. Targeted therapy approaches are particularly attractive because they minimize the detrimental effects of therapeutic agents on healthy tissues and cells by focusing on tumor sites. This study focuses on synthesizing mPEG-modified triblock copolymers as carrier materials for drug delivery applications, enabling the efficient encapsulation of DOX, and evaluating the cytotoxic effects of the resulting nanocarriers on breast cancer cell lines. In this study, mPEG-poly(butylene adipate)-mPEG and mPEG-poly(ethylene adipate)-mPEG triblock copolymers were synthesized by a step-growth polycondensation polymerization method. Firstly, poly(butylene adipate) (pBAd) and poly(ethylene adipate) (pEAd) were synthesized to form the body of the triblock copolymer, and their chemical structures were characterized using Fourier transform infrared (FT-IR) and 1H NMR spectroscopy. The end-group analysis method was applied to determine the average molecular weights of the pBAd and pEAd polymers before their modification with mPEG-500. The nanocarriers produced by the double emulsion method were analyzed using the dynamic light scattering (DLS) method, while encapsulation efficiency and the DOX release profile were measured using a spectrofluorometer. The antiproliferative effects and cellular uptake capacities of the resulting nanocarriers were subsequently examined in MCF-7 and MDA-MB-231 cells. The cytotoxicity of DBANP and DEANP nanocarriers was lower than that of free DOX, demonstrating that encapsulation reduces drug-associated toxicity and may enhance safety. These findings suggest that the nanocarrier systems developed in this study show strong potential as promising candidates for breast cancer therapy.
Achieving maternal health care coverage by 2030 requires strengthening the midwifery workforce. To ensure accessible, culturally appropriate, and high-quality care, midwives must receive education aligned with global standards. This study aimed to evaluate the effect of a 20 hours Simulation-Based Mastery Learning (SBML) combined with Deliberate Practice (DP) and the Objective Structured Clinical Examination (OSCE) on midwifery students' knowledge, skills, and self-confidence in managing the third stage of labor. Sixty midwifery students were randomly allocated to experimental (n = 30) and control (n = 30) groups. Following the intervention, all students met or surpassed the minimum passing standard (MPS), achieving at least 90% in skills (Z=-4.791; p<.001; r~0.87), 80% in knowledge (Z=-4.814; p<.001, r~0.87 ), and an increased score in self-confidence (Z=-4.792; p<.001; r~0.87 ). These improvements were maintained after the clinical experience. The performance of students trained through the traditional approach was significantly lower than that of those trained with SBML, and none of them achieved the MPS. This study highlights the need to structure nursing and midwifery training programs according to the Theory-SBML-Clinical placement model to strengthen students' competencies. La réalisation de la couverture universelle des soins de santé maternelle d’ici 2030 passe par un renforcement substantiel de la profession de sage-femme. Pour assurer des soins accessibles, culturellement adaptés et de qualité optimale, il est essentiel que les sages-femmes bénéficient d’une formation conforme aux normes internationales. Cette étude avait pour objectif d’évaluer l’impact de 20 heures d’apprentissage par simulation fondé sur la pédagogie de la maîtrise, la pratique délibérée, et l’Examen Clinique Objectif Structuré (ECOS) sur les connaissances, les compétences et la confiance en soi des étudiantes sages-femmes dans la prise en charge de la troisième phase de l’accouchement. Un total de 60 étudiantes sages-femmes ont été réparties de façon aléatoire en un groupe expérimental (n = 30) et un groupe témoin (n = 30). À la suite de l’intervention, toutes les étudiantes ont atteint ou dépassé la norme minimale de réussite (MPS), obtenant au moins 90 % en compétences (Z=-4.791; p<.001 ; r~0.87), 80% en connaissances (Z=-4.814; p<.001 ; r~0.87), ainsi qu’une amélioration de leur score de confiance en soi (Z=-4.792; p<.001 ; r~0.87). Ces progrès ont été conservés après l’expérience clinique. Les performances des étudiantes formées selon l’approche traditionnelle étaient significativement inférieures, et aucune d’entre elles n’a atteint le Seuil Minimal de Réussite (SMR). L’apprentissage par maîtrise, la pratique délibérée et l’ECOS apparaissent comme des éléments essentiels au développement d’un programme de formation par simulation de haute qualité, favorisant un transfert efficace des acquis vers la pratique clinique.