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45 pages, 6630 KB  
Article
Metabolomic–Metabolite Profiling: Progressive Insight and Biochemical Pathway in Crude Oil Waste Sludge Co-Composting Bioremediation
by Onyedikachi Ubani and Veronica M. Ngole-Jeme
Metabolites 2026, 16(9), 605; https://doi.org/10.3390/metabo16090605 - 25 Aug 2026
Viewed by 161
Abstract
Background: Crude oil refinery waste sludge (COWS) ranks among the most compositionally complex and ecotoxicologically hazardous industrial residues. Although bulk total petroleum hydrocarbon (TPH) and summed polycyclic aromatic hydrocarbon (PAH) removal are routinely reported, the metabolite-level biochemical fate of individual petrogenic compounds, spanning [...] Read more.
Background: Crude oil refinery waste sludge (COWS) ranks among the most compositionally complex and ecotoxicologically hazardous industrial residues. Although bulk total petroleum hydrocarbon (TPH) and summed polycyclic aromatic hydrocarbon (PAH) removal are routinely reported, the metabolite-level biochemical fate of individual petrogenic compounds, spanning ring dihydroxylation, catechol cleavage, and entry into central carbon metabolism, remains largely unmapped under co-composting with diverse animal manures. Objectives: This study aimed to construct a metabolite-resolved, microbially anchored biochemical fate map of crude oil sludge during co-composting. Methods: Aerobic microcosms combining crude oil sludge, garden soil, and a wood-chip bulking agent were amended separately with poultry, horse, cow, or swine/pig manure alongside an unamended control and then incubated at 22 °C for 300 days. Analyses integrated untargeted gas chromatography-mass spectrometry (GC-MS) metabolomics, targeted PAH quantification (EPA Methods 3541/8270), 16S rRNA gene amplicon sequencing (Illumina MiSeq, V1–V3, paired-end 300 bp), physicochemical monitoring, and culture-dependent isolation, with National Institute of Standards and Technology (NIST) Mass Spectral library annotation. Results: GC-MS resolved 1169 metabolite features across 17 samples, comprising 538 annotated compounds within 11 chemical classes and 631 unknowns, of which 151 recurred in at least 10 samples. Petrogenic markers (n-alkanes C14–C36, hopanoids, steranes, and alkylated dibenzothiophenes) and ring-cleavage intermediates (2-hydroxyfluorene, 1,4-naphthoquinone, phenanthrene-methanol, benzenediols, butanedioic acid, fatty alcohols C16–C20) elucidated a four-stage degradation cascade consistent with Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways map01220 and map00624. PAH mean-removal ranked swine/pig (88.0%) > horse (87.0%) > poultry (80.5%) > cow (79.1%) > control (68.2%). Sequencing recovered 2969 operational taxonomic units (OTUs) enriched in Pseudomonas, Achromobacter, Stutzerimonas, Dietzia, Gordonia, and Mycobacterium, with Pseudomonas dominating high-removal systems; respiration peaked at 18.7 mg CO2-C g−1 in poultry treatments. Conclusions: This work establishes a metabolite-resolved map linking hydrocarbonoclastic taxa to separate degradation steps. The co-occurrence of oxygenated PAH intermediates with decreasing parent PAH concentrations serves as an indicator of transformation processes and may assist in identifying potential residual-risk signals, thereby supporting remediation evaluation and process optimization. Full article
(This article belongs to the Section Advances in Metabolomics)
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31 pages, 96845 KB  
Article
Characterization of the New Pentafluorosulfanyl-Substituted Chalcone 246TMP-3SF5 as a Potential New Treatment Option Against Hepatocellular Carcinoma
by Alessandra Viperino, Linda Hammerich, Bernhard Biersack, Supriya Pradhan, Ion Andronache, Isabel Groth, Nicole Edel, Michael Hoepfner and Bianca Nitzsche
Cancers 2026, 18(16), 2640; https://doi.org/10.3390/cancers18162640 - 16 Aug 2026
Viewed by 399
Abstract
Background/Objectives: Advanced-stage hepatocellular carcinoma is characterized by a very poor prognosis; thus, highly effective medication is still needed. Often overexpressed heat shock protein 90 is a promising target due to its pivotal role in carcinogenesis. Methods: Antiproliferative effects of 246TMP-3SF5 on [...] Read more.
Background/Objectives: Advanced-stage hepatocellular carcinoma is characterized by a very poor prognosis; thus, highly effective medication is still needed. Often overexpressed heat shock protein 90 is a promising target due to its pivotal role in carcinogenesis. Methods: Antiproliferative effects of 246TMP-3SF5 on HepG2 and HuH-7 cells were assessed by crystal violet staining. Apoptosis was evaluated via subG1 peak, caspase-3 activity and PARP cleavage, and ferroptosis via ROS, glutathione and malondialdehyde levels. Migration was assessed by scratch assay, and in ovo models were used to study angiogenesis and drug effects on microtumors. CAM vascular networks were quantified by semi-automatic segmentation, morphometric and box-counting fractal analysis. Molecular docking and molecular dynamics simulation of heat shock protein 90 were carried out using Autodock Vina and Gromacs respectively. Results: Profound dose- and time-dependent antiproliferative effects of 246TMP-3SF5 against HCC cell lines were observed, revealing low micromolecular IC50 values and selectivity for carcinoma cells with selectivity indices > 1. A significant increase in the sub-G1 peak, key effector caspase-3 activity, as well as cleavage of PARP strongly suggested apoptosis playing a crucial role in the antiproliferative effects. Additionally, HuH-7 cells revealed an elevation of reactive oxygen species and both cell lines showed significant glutathione depletion concomitant with an increase in malondialdehyde concentration upon treatment. The observed effect could be partially reversed by applying ferrostatin-1, suggesting ferroptosis as an additional relevant mode of action. Changes in the cell cycle as well as impaired tumor cell migration were observed. Upon treatment, angiogenesis was impaired and mass of microtumors was significantly reduced. Quantitative CAM analysis showed that vascularized area fraction increased in controls but decreased under both 17-AAG and 246TMP-3SF5. Likewise global mean vessel width decreased relative to controls, while box-counting dimension was reduced under 246TMP-3SF5. Molecular docking and molecular dynamics simulation analysis predicts 246TMP-3SF5 to be binding in catalytic site of heat shock protein 90. Conclusions: 246TMP-3SF5 is a promising novel inhibitor meriting further research as a potential treatment against hepatocellular carcinoma. Full article
(This article belongs to the Special Issue Updates on Anti-Cancer Drug Research)
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14 pages, 3150 KB  
Article
Exploring Co-Occurring Clinical and Treatment Characteristics Associated with In Vitro Fertilization Failure in Polycystic Ovary Syndrome: An Association Rule Mining Approach
by Xuehong Zhu, Lina Ge, Guanghui Dong, Yanlin Tang, Zhong Lin and Feng Han
Healthcare 2026, 14(16), 2508; https://doi.org/10.3390/healthcare14162508 - 12 Aug 2026
Viewed by 159
Abstract
Background: Polycystic ovary syndrome (PCOS) is a leading cause of anovulatory infertility and a frequent indication for in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). Multivariable regression estimates adjusted associations, whereas association rule mining (ARM) can describe frequently co-occurring attributes; however, ARM outputs are unadjusted [...] Read more.
Background: Polycystic ovary syndrome (PCOS) is a leading cause of anovulatory infertility and a frequent indication for in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). Multivariable regression estimates adjusted associations, whereas association rule mining (ARM) can describe frequently co-occurring attributes; however, ARM outputs are unadjusted and may be sensitive to analytic specification. Objectives: We used ARM alongside multivariable logistic regression and internal robustness analyses to describe combinations of clinical and treatment-related variables that co-occurred with clinical pregnancy failure among women with PCOS undergoing IVF/ICSI. Methods: We retrospectively analyzed 733 deidentified patients with PCOS from the Reproductive Hospital of Guangxi. The original one-hot-encoded analytic file was linked to a supplemental clinical record file containing continuous age, BMI, infertility duration, anti-Müllerian hormone, total gonadotropin dose, fertilization method, embryo-transfer day, number of embryos transferred, and embryo score. Embryo quality was derived from D3 cleavage-stage and blastocyst morphology records. Multivariable logistic regression was used to estimate adjusted associations. ARM results were assessed using Fisher’s exact test, Benjamini–Hochberg false discovery rate (FDR) correction, 1000 bootstrap resamples, fivefold cross-validation, and threshold/cutoff sensitivity analyses. Results: In the multivariable model (n = 730), age remained associated with clinical pregnancy failure (adjusted odds ratio [aOR] = 1.04 per year, 95% CI 1.00–1.08, p = 0.031), whereas BMI was not independently associated with failure (aOR = 1.00 per kg/m2, 95% CI 0.96–1.05, p = 0.860). Transfer of two embryos (aOR = 0.55, 95% CI 0.35–0.85, p = 0.008), D5 blastocyst transfer (aOR = 0.49, 95% CI 0.32–0.75, p = 0.001), and transfer of at least one high-quality embryo (aOR = 0.63, 95% CI 0.43–0.93, p = 0.020) were associated with lower odds of failure. Under the primary host-factor specification, age > 35 years + BMI ≥ 24 kg/m2 + pelvic adhesion defined 48 patients, 30 of whom had clinical pregnancy failure (support = 0.07; confidence = 0.63; lift = 1.38; FDR q = 0.098). In the secondary treatment-inclusive analysis, infertility duration > 5 years + no high-quality embryo transferred defined 43 patients with 30 failures (support = 0.06; confidence = 0.70; lift = 1.54; FDR q = 0.026). Lift represents the subgroup failure proportion divided by the overall failure proportion and is not an adjusted risk ratio. The rule composition and the number of FDR-supported rules changed under alternative cutoffs and confidence thresholds. Conclusions: ARM described transparent but specification-sensitive co-occurrence profiles that complement, rather than compete with, regression. Treatment-inclusive profiles combine baseline and downstream cycle characteristics and are therefore especially exploratory. These findings require prospective multicenter validation before any clinical decision-support use. Full article
(This article belongs to the Section Women’s and Children’s Health)
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35 pages, 4205 KB  
Review
Advancements in CRISPR/Cas Technologies for Sensitive Cancer Detection: Mechanisms, Platforms, and Clinical Translation Roadmap
by Rokeya Akter, Sook Won Ryu and Jong-Han Lee
Diagnostics 2026, 16(16), 2531; https://doi.org/10.3390/diagnostics16162531 - 11 Aug 2026
Viewed by 320
Abstract
Early cancer detection is critical for improving patient outcomes, yet current diagnostic approaches often fail to identify malignancies when tumor-specific biomarkers are relatively scarce. Emerging CRISPR/Cas-based technologies have revolutionized the ultra-sensitive detection of cancer biomarkers in liquid biopsies, overcoming the inherent limitations of [...] Read more.
Early cancer detection is critical for improving patient outcomes, yet current diagnostic approaches often fail to identify malignancies when tumor-specific biomarkers are relatively scarce. Emerging CRISPR/Cas-based technologies have revolutionized the ultra-sensitive detection of cancer biomarkers in liquid biopsies, overcoming the inherent limitations of traditional diagnostic approaches such as tissue biopsies and imaging, which frequently fail to detect early-stage malignancies with sufficient sensitivity. This review explores recent advances in CRISPR/Cas diagnostics (CRISPR/Cas-Dx) that employ programmable CRISPR effectors, including Cas9, Cas12, Cas13, and Cas14. In particular, the collateral (trans-) cleavage activities of Cas12, Cas13, and Cas14 enable signal amplification for highly sensitive detection of circulating tumor DNA, microRNAs, exosomes, and other multi-omics biomarkers, often without the need for extensive nucleic acid amplification. Representative CRISPR/Cas-Dx platforms include amplification-coupled assays, amplification-free frameworks, biosensing and multiplexing capabilities, and new digital or droplet-based configurations that include artificial intelligence to improve analytical precision. These technologies demonstrate single-molecule resolution and adaptability for point-of-care testing in malignancies such as non-small cell lung, colorectal, and breast carcinoma. Finally, we outline a clinical translation roadmap encompassing manufacturability, regulatory and standardization requirements, and real-world implementation challenges. This perspective offers a blueprint for CRISPR-powered, ultra-sensitive liquid biopsy diagnostics that can enable population-scale early cancer screening and truly preventive oncology by bridging molecular insights, engineering innovation, and clinical imperatives. Full article
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21 pages, 5167 KB  
Article
Temporal Multi-Omics Reveals Microbial and Metabolic Succession Following Astilbin Treatment in a Human Colonic Model
by Tingwei Wang, Chang Liu, Jian Ji, Shuang Zhang, Shengfang Wu, Bangen Xia, Ruowei Xia, Nian Qu, Maiqiu Wang, Lei Zhang and Yongli Ye
Nutrients 2026, 18(16), 2616; https://doi.org/10.3390/nu18162616 - 10 Aug 2026
Viewed by 242
Abstract
Background: Astilbin is a bioactive flavonoid with documented anti-inflammatory properties; however, its sustained interactions with the gut microbiota remain poorly understood. Fecal samples from three healthy donors were pooled and fermented with astilbin in an in vitro human colonic model over 7 days. [...] Read more.
Background: Astilbin is a bioactive flavonoid with documented anti-inflammatory properties; however, its sustained interactions with the gut microbiota remain poorly understood. Fecal samples from three healthy donors were pooled and fermented with astilbin in an in vitro human colonic model over 7 days. This exploratory study aimed to characterize the temporal ecological shifts associated with prolonged astilbin exposure. Methods: Time-resolved 16S rRNA gene sequencing, PICRUSt2 functional prediction, BugBase phenotypic inference, and pseudo-targeted metabolomics were integrated to track microbial-metabolic dynamics. Results: Astilbin exposure was associated with a highly coordinated, three-stage microbial succession. Day 3 (D3) emerged as a putative inflection point, where the enrichment of pioneer degraders (Flavonifractor, Bacteroides) was temporally correlated with the appearance of polyphenol cleavage intermediates. This transition featured an early decrease in markers of proteolytic fermentation alongside a transient in vitro lipid-stress response. By D7, the community shifted toward a stable configuration enriched in butyrogenic taxa (Roseburia, Subdoligranulum, Megamonas), with progressive depletion of potentially opportunistic pathogens (Escherichia-Shigella). Multi-omics integration suggested that these structural successions were strongly associated with marked metabolic shifts. Inflammatory lipid markers (e.g., leukotriene B4) showed a characteristic “D3-burst/D7-clearance” pattern, whereas potentially barrier-protective metabolites, particularly 3-indolepropionic acid (3-IPA, log2FC = 2.00) and urolithin B (log2FC = 1.18), accumulated substantially. Conclusions: This exploratory study provides valuable high-resolution insights into astilbin’s potential as a dynamic ecological modulator. It outlines a temporal framework illustrating how the gut microbiota may shift from proteolytic fermentation toward 3-IPA-associated homeostasis. Although limited by a pooled fecal model and the absence of a vehicle control, these hypothesis-generating findings offer a solid foundation for future in vivo studies and mechanistic validations across diverse human cohorts. Full article
(This article belongs to the Section Proteins and Amino Acids)
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16 pages, 2909 KB  
Article
Altered Embryonic SIRT6 and SIRT7 Expression Is Associated with Impaired Developmental Competence and Oocyte-Dependent Embryo Quality in Embryos Derived from Subfertile Males
by Alicja Kowalczyk, Mercedes Camiña García, José Ángel Hernández Malagón, Jose Pedro Araujo, Ivona Žura Žaja, Marko Pećin, Dražen Đuričić, Elżbieta Gałęska, Zbigniew Dobrzański and Marko Samardžija
Int. J. Mol. Sci. 2026, 27(16), 7115; https://doi.org/10.3390/ijms27167115 - 8 Aug 2026
Viewed by 260
Abstract
Paternal contributions to preimplantation embryo development are increasingly recognized as an important component of reproductive success, but the associations between male reproductive phenotype, early embryonic competence, and embryo molecular characteristics remain insufficiently defined. Using bovine in vitro embryo production as a translational large-mammalian [...] Read more.
Paternal contributions to preimplantation embryo development are increasingly recognized as an important component of reproductive success, but the associations between male reproductive phenotype, early embryonic competence, and embryo molecular characteristics remain insufficiently defined. Using bovine in vitro embryo production as a translational large-mammalian model, this study evaluated whether paternal reproductive phenotype and oocyte origin were associated with embryo developmental performance and embryonic sirtuin expression. Oocytes obtained from mature cows or heifers were fertilized with semen from bulls differing in reproductive status, including bulls characterized by libido disturbances and irregular semen collection. Fertilization, cleavage, blastocyst formation, and hatching were assessed, while embryonic SIRT2, SIRT3, SIRT6, and SIRT7 expression was quantified by immunofluorescence. Early developmental endpoints were relatively preserved, whereas differences were more evident at later stages of preimplantation development. Cow-derived oocytes were associated with higher hatching rates than heifer-derived oocytes. Embryos generated using semen from bulls with an impaired reproductive phenotype showed lower developmental competence and lower SIRT6 and SIRT7 expression. SIRT6 expression showed the strongest positive correlation with hatching, and positive correlations were also observed among the analyzed sirtuins. These findings indicate that paternal reproductive phenotype and oocyte origin are associated with bovine embryo developmental competence and embryonic sirtuin profiles. Embryonic SIRT6 and SIRT7 expression may represent candidate molecular indicators of embryo competence; however, causal relationships cannot be inferred from the present study design. Full article
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22 pages, 12664 KB  
Article
Chemical Composition, Microstructure, and Mechanical Properties of Steel Rails Produced by China Hanyang Iron Works
by Tengshi Liu, Gangsheng Xie, Han Yi, Di Zhang, Zhouyan Cai, Yulin Xia and Han Dong
Metals 2026, 16(8), 862; https://doi.org/10.3390/met16080862 - 5 Aug 2026
Viewed by 342
Abstract
The history of steel rail manufacturing at the Hanyang Iron Works in China is briefly summarized first in this paper. The chemical composition, microstructure, inclusions, and mechanical properties of the manufactured rails were then systematically analyzed. The progress of manufacturing process evolution and [...] Read more.
The history of steel rail manufacturing at the Hanyang Iron Works in China is briefly summarized first in this paper. The chemical composition, microstructure, inclusions, and mechanical properties of the manufactured rails were then systematically analyzed. The progress of manufacturing process evolution and the enhancement of product quality are elucidated through this study. The chemical composition of steel rails produced by Hanyang Iron Works in its initial stage (before 1904) was characterized by low carbon content (0.13–0.22 wt.%) and high phosphorus levels (P ≥ 0.15 wt.%). Inclusions were primarily identified as sulfides (MnS) and composite inclusions of sulfides and silicates (MnS·SiO2). The microstructure consisted of a large amount of ferrite and pearlite. Following the technical transformation from 1905 to 1908, dephosphorization was achieved and the composition control of the steel rails was optimized. The carbon content of the rails was increased to above 0.48 wt.%, while the phosphorus content was significantly reduced (P ≤ 0.10 wt.%). The inclusions were identified as sulfides (MnS) and composite inclusions consisting of sulfides and aluminum oxides (MnS·Al2O3). The microstructure was transformed into a combination of a small amount of proeutectoid network ferrite and pearlite. The mechanical performance of the steel rails was substantially improved via the implementation of technological upgrades at the Hanyang Iron Works. A tensile strength of 800 MPa grade was achieved in some rails, which constitutes a 200 MPa increment over the strength of rails from the early production period. A transition in the fracture morphology of tensile specimens was observed, shifting from large and shallow dimples with a small amount of cleavage fracture to small, shallow dimples combined with predominant cleavage fracture. Full article
(This article belongs to the Topic Advances in Manufacturing and Mechanics of Materials)
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14 pages, 1077 KB  
Article
Outcomes of Single Blastocyst Transfer Versus Double Cleavage-Stage Embryo Transfer in Young Women with Recurrent Implantation Failure: A Secondary Analysis of a Multicenter Randomized Trial
by Yuan Wang, Changyang Xu, Yaqiong He, Jiaan Huang, Yun Sun, Yao Lu and Zhe Wei
J. Clin. Med. 2026, 15(14), 5544; https://doi.org/10.3390/jcm15145544 - 15 Jul 2026
Viewed by 396
Abstract
Objectives: Although blastocyst transfer may improve embryo selection and early pregnancy outcomes, its impact on live birth in women with recurrent implantation failure (RIF) remains uncertain. This study compared reproductive and perinatal outcomes between single blastocyst transfer (SBT) and double cleavage-stage embryo transfer [...] Read more.
Objectives: Although blastocyst transfer may improve embryo selection and early pregnancy outcomes, its impact on live birth in women with recurrent implantation failure (RIF) remains uncertain. This study compared reproductive and perinatal outcomes between single blastocyst transfer (SBT) and double cleavage-stage embryo transfer (DET) among women younger than 38 years with RIF. Methods: Secondary observational analysis of a multicenter, randomized, double-blind, placebo-controlled clinical trial conducted across eight academic fertility centers in China. In the original trial, patients with RIF under 38 years of age were enrolled and randomized to receive either prednisone or placebo. For the present analysis, a total of 582 patients were categorized according to the embryo transfer strategy. The SBT-versus-DET comparison was not randomized, and the two strategies differed in both embryo developmental stage and number of embryos transferred. Results: No statistically significant difference in live birth rate was observed between the SBT and DET groups (41.8% vs. 38.4%; p = 0.471), despite a higher biochemical pregnancy rate in the SBT group (59.9% vs. 45.7%, p = 0.003). Pregnancy loss was more frequent in the SBT group than in the DET group (30.2% vs. 15.9%; p = 0.018), particularly biochemical pregnancy loss (15.5% vs. 5.8%; p = 0.036), whereas twin live birth was less frequent (0.7% vs. 9.9%; p < 0.001). After adjustment, no significant difference in live birth rate was observed between DET and SBT (adjusted OR 1.04, 95% CI 0.68–1.57). In addition, Day 5 SBT showed a higher live birth rate than Day 6 SBT, whereas pregnancy loss rates were similar between Day 5 and Day 6 SBT cycles. Other exploratory sensitivity and stratified analyses showed broadly consistent findings. Conclusions: Although SBT is generally preferred in clinical practice, our study found no statistically significant difference in live birth rates between SBT and DET among women younger than 38 years with RIF. Given that pregnancy loss was observed more frequently in the SBT group, whereas twin live births were less frequent, our findings support individualized counseling rather than uniform use of either transfer strategy. However, given the non-randomized nature of this comparison, these findings should be interpreted with caution. Full article
(This article belongs to the Section Reproductive Medicine & Andrology)
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17 pages, 2420 KB  
Article
Research on Optimization of Horizontal Well Fracturing Parameters Based on Numerical Simulation
by Yutong Fu, Congyu Zhong, Jingjing Liu, Ruosi Zhao, Jishi Geng, Jinting Xiong, Chao Yang and Luyi Wang
Energies 2026, 19(14), 3309; https://doi.org/10.3390/en19143309 - 14 Jul 2026
Viewed by 196
Abstract
The deep coalbed methane in the SSL block is characterized by strong in situ stresses, elevated thermal conditions, and over-pressured reservoirs. These conditions lead to poor adaptability of vertical-well fracturing technology for CBM extraction. Due to the high investment cost of staged fracturing [...] Read more.
The deep coalbed methane in the SSL block is characterized by strong in situ stresses, elevated thermal conditions, and over-pressured reservoirs. These conditions lead to poor adaptability of vertical-well fracturing technology for CBM extraction. Due to the high investment cost of staged fracturing of horizontal wells, it is necessary to optimize the well type and fracturing process parameters before the experiment, in order to find the optimal horizontal section length and fracturing process parameters. On the basis of in-depth analysis of the characteristics of deep coalbed methane reservoirs, numerical simulation technology was used to carry out optimization research on segmented fracturing horizontal well technology from the aspects of horizontal section length, number of fracturing fractures, fracturing conductivity, fracturing length, and fracturing pattern. Research has shown that segmented fracturing of horizontal wells can communicate the coal seam cleavage fracture system, with a large drainage area and good pressure reduction and gas production effects. The optimal wellbore structure parameters for segmented fracturing of horizontal wells are a horizontal section length of 1000–1200 m, divided into 4–6 sections for fracturing, with a spacing of 120–150 m and uniform distribution of fractures. This well type has been applied in adjacent blocks and has shown good development benefits. Full article
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17 pages, 454 KB  
Article
Embryo-Derived Sample Categories Show Differing Assay Evaluability and Apparent MSY-Classification Patterns in a Low-Input Bovine Embryo PCR Workflow
by Lilla Sándorová, Katalin Nagy, Szilárd Bodó, Ákos Bodnár, István Egerszegi, Dániel Fodor, Ferenc Pajor and Viktor Stéger
Ruminants 2026, 6(3), 55; https://doi.org/10.3390/ruminants6030055 - 8 Jul 2026
Viewed by 461
Abstract
Reliable preimplantation embryo sexing is relevant in cattle breeding; however, PCR-based Y-marker detection may be sensitive to limited DNA input and sample-type-specific properties of embryo-derived material. In this observational study, we examined assay-output patterns in a low-input multiplex PCR workflow targeting a male-specific [...] Read more.
Reliable preimplantation embryo sexing is relevant in cattle breeding; however, PCR-based Y-marker detection may be sensitive to limited DNA input and sample-type-specific properties of embryo-derived material. In this observational study, we examined assay-output patterns in a low-input multiplex PCR workflow targeting a male-specific Y-chromosome marker (MSY) together with an autosomal internal control targeting cyclin-dependent kinase 1 (CDK1). A total of 141 bovine embryo-derived samples were analyzed across three sample categories: whole embryos, cleavage-stage blastomere biopsies, and blastocyst-stage trophectoderm biopsies. Assay evaluability differed across sample categories, being highest in blastocyst-stage biopsy samples (90.6%, 29/32), intermediate in whole embryos (82.9%, 58/70), and lowest in blastomere-biopsy samples (64.1%, 25/39). Among technically evaluable CDK1-positive samples, apparent MSY-positive classification also differed across categories, with rates of 55.2% (32/58) in whole embryos, 52.0% (13/25) in blastomere-biopsy samples, and 6.9% (2/29) in blastocyst-stage biopsy samples. In a same-lysate repeat-amplification subset, overall evaluability concordance was 92.5% and overall apparent MSY-based classification concordance was 88.2%, although subgroup sizes were limited. Because no independent reference method for true embryo sex was available and sample category was structurally confounded with developmental stage, semen type, sire, and partly PCR-date structure, the findings should be interpreted as assay-output observations rather than measures of biological sex ratio or diagnostic sexing accuracy. Overall, embryo-derived sample categories showed differing assay evaluability and apparent MSY-classification patterns within this low-input PCR setting, supporting the need for sample-type-specific validation when low-cell embryo-derived samples are used. Full article
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11 pages, 282 KB  
Article
Embryological and Clinical Outcomes of Oocytes Retrieved from the Pouch of Douglas During Transvaginal Oocyte Pick-Up
by Selçuk Yetkinel, Gülşen Doğan Durdağ, Didem Alkaş Yağınç, Pınar Çağlar Aytaç and Erhan Şimşek
J. Clin. Med. 2026, 15(13), 5129; https://doi.org/10.3390/jcm15135129 - 1 Jul 2026
Viewed by 295
Abstract
Background/Objectives: Oocytes retrieved from the pouch of Douglas during transvaginal oocyte pick-up (OPU) have historically been described mainly in isolated case reports and small series. Their developmental competence and clinical relevance remain incompletely characterized. This study aimed to evaluate the embryological and [...] Read more.
Background/Objectives: Oocytes retrieved from the pouch of Douglas during transvaginal oocyte pick-up (OPU) have historically been described mainly in isolated case reports and small series. Their developmental competence and clinical relevance remain incompletely characterized. This study aimed to evaluate the embryological and clinical outcomes of oocytes recovered from pouch of Douglas fluid and to compare developmental outcomes between ovarian-derived and pouch of Douglas-derived oocytes. Methods: This retrospective observational study was conducted at a tertiary referral university hospital IVF center. Clinical and embryological data collected between February 2023 and June 2025 were retrospectively analyzed. A total of 2423 OPU cycles were screened, and 49 cycles with ≥2 cm of free fluid in the pouch of Douglas before OPU met the inclusion criteria. Following completion of ovarian follicular aspiration, free peritoneal fluid from the pouch of Douglas was aspirated separately and assessed for the presence of oocytes. Cycles were categorized according to whether oocytes were identified in the pouch of Douglas. Baseline cycle characteristics, ovarian response parameters, embryological outcomes, and clinical outcomes were evaluated. Developmental outcomes of ovarian-derived and pouch of Douglas-derived oocytes were compared within the same cycles. Results: Oocytes were recovered from pouch of Douglas fluid in 30 of 49 cycles (61.2%). Baseline characteristics, trigger-day hormonal parameters, follicle numbers, and ovarian embryological outcomes were similar between cycles with and without pouch of Douglas oocytes. A total of 44 oocytes were retrieved from pouch of Douglas fluid, of which 36 (81.8%) were metaphase II (MII) oocytes. Twenty-six oocytes demonstrated normal fertilization (2PN), with subsequent cleavage-stage and blastocyst-stage embryo development observed. No significant differences were observed between ovarian-derived and pouch of Douglas-derived oocytes in fertilization or embryo development rates. The proportion of MII oocytes was significantly higher among pouch of Douglas-derived oocytes (81.8% vs. 66.2%, p = 0.021). Five embryo transfers involving embryos derived from pouch of Douglas oocytes resulted in clinical pregnancy and live birth, including three transfers exclusively involving pouch of Douglas-derived embryos. No procedure-related complications were observed. Conclusions: Oocytes recovered from pouch of Douglas fluid demonstrated fertilization capacity, embryo developmental potential, and the ability to contribute to clinical pregnancy and live birth. These findings suggest that free peritoneal fluid identified before OPU may contain developmentally competent oocytes that would otherwise remain unrecovered. Given the retrospective design, limited number of embryo transfers, and uncertainty regarding the precise origin of these oocytes, the findings should be considered exploratory and require confirmation in larger prospective studies. Full article
(This article belongs to the Section Reproductive Medicine & Andrology)
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17 pages, 5051 KB  
Article
Multi-Field Coupled Cyclic Degradation Mechanisms of Alumina Ceramic Fiber Ropes
by Hongkai Guo, Lei Shang, Hanlei Zhai, Chunlin Wang, Zhihong Han, Jiajin Xu, Jiahui Zhou, Zhiqiang Luan, Xing Peng and Wenbo Han
Nanomaterials 2026, 16(13), 812; https://doi.org/10.3390/nano16130812 - 30 Jun 2026
Viewed by 481
Abstract
Continuous alumina (Al2O3) fibers are critical reinforcement materials for ceramic matrix composites (CMCs) utilized in extreme high-temperature environments. While their baseline thermal and mechanical properties are well-documented, their long-term service reliability in complex, multi-field environments—specifically coupled thermal, hygral, and [...] Read more.
Continuous alumina (Al2O3) fibers are critical reinforcement materials for ceramic matrix composites (CMCs) utilized in extreme high-temperature environments. While their baseline thermal and mechanical properties are well-documented, their long-term service reliability in complex, multi-field environments—specifically coupled thermal, hygral, and atmospheric conditions—remains insufficiently quantified. This study systematically investigates the degradation mechanisms of alumina ceramic fiber ropes subjected to simulated engine exhaust atmospheres and cyclic rain exposure. By integrating macroscopic tensile testing with rigorous multi-scale microstructural characterizations (SEM, XRD, TGA, and advanced surface chemical state analyses via EDS and XPS), a comprehensive degradation model is proposed. Our findings reveal a pronounced two-stage mechanical degradation behavior: an initial catastrophic strength collapse followed by a stabilization phase. We elucidate that the initial embrittlement is governed not merely by thermal damage, but fundamentally by the hydrothermal volatilization and depletion of the surface amorphous SiO2 binder, which annihilates the inter-fiber cooperative load-sharing capability. Concurrently, quantitative XPS and XRD analyses strongly suggest that the internal amorphous grain-boundary films undergo rapid structural rearrangement and crystallization, effectively homogenizing the microstructure and shifting the fracture mechanics from energy-dissipative crack deflection to unhindered brittle cleavage. After the preferential depletion of the amorphous silicate phase, the exposed α-Al2O3 core dictates a stabilized mechanical response. This research provides critical theoretical frameworks and experimental evidence for the life-cycle assessment and microstructural optimization of advanced oxide ceramic fibers in next-generation aerospace applications. Full article
(This article belongs to the Special Issue Advanced Carbon/Ceramic Nanocomposites: Microstructure and Properties)
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32 pages, 9441 KB  
Article
Architecture-Dependent Thermal Decomposition of RAFT-Modified Polypropylene Glycol Maleate-Acrylic Acid Copolymers: Results of TG–MS and Kinetic Analysis
by Akmaral Zh. Sarsenbekova, Almagul S. Makhmutova, Meruyert S. Zhunissova, Nazigul S. Remetova, Meruyert B. Issabayeva, Gulnissa K. Kurmantayeva, Mussa E. Zholdasbayev and Bibigul B. Ashirbekova
Polymers 2026, 18(13), 1599; https://doi.org/10.3390/polym18131599 - 26 Jun 2026
Cited by 1 | Viewed by 552
Abstract
The effect of reversible addition–fragmentation chain transfer (RAFT) polymerization on the structure, morphology, and thermal degradation behavior of polypropylene glycol maleate–acrylic acid copolymers (p-PGM:AA) was investigated using 2-cyano-2-propyl dodecyl trithiocarbonate (CPDT) as the RAFT agent. Copolymers synthesized at different CPDT concentrations were characterized [...] Read more.
The effect of reversible addition–fragmentation chain transfer (RAFT) polymerization on the structure, morphology, and thermal degradation behavior of polypropylene glycol maleate–acrylic acid copolymers (p-PGM:AA) was investigated using 2-cyano-2-propyl dodecyl trithiocarbonate (CPDT) as the RAFT agent. Copolymers synthesized at different CPDT concentrations were characterized by 1H/13C NMR spectroscopy, gel permeation chromatography (GPC), transmission electron microscopy (TEM), thermogravimetric analysis coupled with mass spectrometry (TG–MS), isoconversional kinetic methods, and density functional theory (DFT) calculations. 1H NMR spectroscopy revealed a progressive decrease in the relative intensity of vinyl proton signals with increasing CPDT concentration, indicating enhanced conversion of unsaturated fragments during copolymerization. Alkaline hydrolysis followed by 1H NMR and GPC analysis of the degradation products confirmed cleavage of polyester segments and yielded low-molecular-weight fragments with Mn = 1370 g mol−1 and narrow dispersity (Đ = 1.035), providing additional information on the architecture of the vinyl-polymerized segments. Increasing CPDT concentration resulted in lower molecular weights and narrower molecular weight distributions of the soluble copolymer fractions. TEM analysis demonstrated broader domain size distributions and increased morphological heterogeneity in RAFT-modified samples, accompanied by an increase in swelling degree. Thermogravimetric analysis showed that RAFT-modified systems undergo multi-stage thermal degradation with the appearance of an additional low-temperature stage associated with thermolabile fragments. TG–MS revealed earlier evolution of CO2 and oxygen-containing species and changes in the distribution of volatile products. DFT calculations indicated a decrease in the HOMO–LUMO energy gap and suggested the participation of RAFT-derived fragments in the energetic characteristics of decarboxylation processes. Isoconversional and nonlinear kinetic analyses demonstrated increased kinetic heterogeneity for branched copolymer s synthesized at elevated CPDT concentrations, whereas cross-linked systems exhibited more uniform degradation behavior. The combined experimental and theoretical results demonstrate that RAFT polymerization provides an effective route for tuning the macromolecular architecture, morphology, and thermal degradation pathways of p-PGM:AA copolymers. Full article
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22 pages, 32128 KB  
Article
Atomistic Mechanisms of Silicone Rubber Degradation Under Coupled Temperature–Humidity–Electric Field Conditions
by Yiheng Zhou, Zhijun An, Yixin He, Cong Qian, Qiuhua Zhou, Wentian Zeng, Xinhan Qiao and Wenyu Ye
Polymers 2026, 18(12), 1530; https://doi.org/10.3390/polym18121530 - 19 Jun 2026
Viewed by 631
Abstract
Silicone rubber is an important external insulating material for composite bushings, composite insulators, and other power equipment. During long-term service, it is inevitably exposed to coupled environmental and electrical stresses, such as elevated temperature, moisture ingress, strong electric fields, and partial discharge, which [...] Read more.
Silicone rubber is an important external insulating material for composite bushings, composite insulators, and other power equipment. During long-term service, it is inevitably exposed to coupled environmental and electrical stresses, such as elevated temperature, moisture ingress, strong electric fields, and partial discharge, which may lead to hydrophobicity loss, surface chalking, crack propagation, and particle shedding. To reveal the microscopic degradation mechanism of silicone rubber under complex operating conditions, a molecular model of methyl vinyl silicone rubber was constructed using Materials Studio. A stable silicone rubber molecular structure was obtained through crosslinking, geometry optimization, and ensemble relaxation. Subsequently, a reactive molecular dynamics simulation system under coupled temperature–humidity–electric field conditions was established using LAMMPS and the ReaxFF reactive force field. Different temperature gradients, electric field intensities, and aging–recovery stages were designed to investigate the degradation behavior of silicone rubber. The evolution of the maximum carbon content, maximum silicon content, carbon-containing decomposition products, and typical small-molecule products, including H2, H2O, CH4, C2H2, C2H4, and C2H6, was statistically analyzed. In addition, atomic trajectory tracking was performed to clarify the processes of methyl group detachment, Si-O bond cleavage, water molecule participation, and molecular chain reconstruction. The results show that high temperature mainly promotes methyl group detachment from side chains and fracture of the siloxane main chain, while a strong electric field accelerates the decomposition process and induces the transformation of long siloxane chains into shorter chains. Water molecules can react with broken siloxane chains to form hydroxyl-containing structures, making the structural degradation partially irreversible. The degradation process of silicone rubber under coupled temperature–humidity–electric field stress can be summarized as side-chain detachment, main-chain scission, water-assisted reactions, free-radical recombination, and local molecular aggregation. This study provides a molecular-level theoretical basis for aging mechanism analysis, condition assessment, and lifetime prediction of composite external insulating materials. Full article
(This article belongs to the Section Polymer Analysis and Characterization)
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24 pages, 841 KB  
Article
Spatial and Molecular Parameters of Pre-OPU Ovarian Follicles and Their Association with Embryo Developmental Competence in Assisted Reproductive Technology
by Patrycja Strączyńska, Aleksandra Pytel, Emilia Morawiec, Zenon Czuba and Anna Bednarska-Czerwińska
Int. J. Mol. Sci. 2026, 27(12), 5280; https://doi.org/10.3390/ijms27125280 - 10 Jun 2026
Viewed by 451
Abstract
Advanced maternal age is a significant social and clinical issue associated with the natural decline in a woman’s ovarian reserve. This prospective, single-center study included women with primary infertility who presented to the Gyncentrum Clinic in Katowice and analyzed 77 ovarian follicles. The [...] Read more.
Advanced maternal age is a significant social and clinical issue associated with the natural decline in a woman’s ovarian reserve. This prospective, single-center study included women with primary infertility who presented to the Gyncentrum Clinic in Katowice and analyzed 77 ovarian follicles. The study group consisted of patients of advanced reproductive age with diminished ovarian reserve, who underwent hormonal stimulation in preparation for oocyte retrieval. Each metaphase II (MII) oocyte was fertilized in vitro and cultured individually in a time-lapse incubator. Follicular fluid obtained during oocyte retrieval was collected separately from each follicle and used for non-invasive biochemical analysis of prognostic factors using a Multiplex assay. The concentrations of interleukin 10 (IL-10), granulocyte colony-stimulating factor (G-CSF), granulocyte–macrophage colony-stimulating factor (GM-CSF), and C-type natriuretic peptide (CNP) were evaluated. The analysis showed that lower concentrations of GM-CSF and CNP were associated with an increased probability of oocyte fertilization, whereas higher levels of IL-10 and G-CSF had the greatest impact on blastocyst formation. This model was supported by continuous embryo monitoring. An eightfold increase in the likelihood of blastocyst formation was observed when early embryo cleavage occurred between 25 and 27 h after insemination. Furthermore, prolonged duration of the first cytokinesis reduced the probability of blastocyst development, while an extended cell cycle at the two-blastomere stage significantly affected further embryo development. These findings may support non-invasive embryo selection strategies. Full article
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