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15 pages, 2277 KB  
Article
A Comparison of Flow Cytometry-based versus ImmunoSpot- or Supernatant-based Detection of SARS-CoV-2 Spike-specific Memory B Cells in Peripheral Blood
by Georgia Stylianou, Sharon Cookson, Justin T. Nassif, Greg A. Kirchenbaum, Paul V. Lehmann and Stephen M. Todryk
Vaccines 2026, 14(1), 20; https://doi.org/10.3390/vaccines14010020 - 24 Dec 2025
Abstract
Background: Memory B cells (Bmem) facilitate the generation of renewed and rapid antigen-specific antibody responses long after the initial antigen exposure, at a time when circulating serum antibodies may have declined. As the generation and/or recruitment of Bmem is at [...] Read more.
Background: Memory B cells (Bmem) facilitate the generation of renewed and rapid antigen-specific antibody responses long after the initial antigen exposure, at a time when circulating serum antibodies may have declined. As the generation and/or recruitment of Bmem is at the core of most vaccination strategies, the assessment of antigen-specific Bmem is highly informative for forecasting and profiling the elicited B cell immune response. Methods: The two prevalent techniques used to detect antigen-specific Bmem cells at single-cell resolution are probe-based flow cytometry and B cell ImmunoSpot, while the measurement of B cell-derived antibodies in culture supernatants of stimulated B cells offers a semi-quantitative alternative. To the best of our knowledge, a direct side-by-side comparison of these assay systems has not yet been reported using the same starting PBMC material in a blinded fashion to test all three assays simultaneously. Results: These three assay systems were run in parallel to detect SARS-CoV-2 Wuhan-1 strain Spike-specific IgG+ Bmem in peripheral blood mononuclear cell (PBMC) samples obtained from well-defined cohorts comprising pre-COVID-19 era “naïve” individuals (negative controls), individuals shortly after recovery from a PCR-verified SARS-CoV-2 infection (positive controls), and a cohort of donor PBMCs isolated in 2024 (the experimental group). Each assay was able to discern Spike-exposed individuals from naïve , with ImmunoSpot suggesting superior sensitivity and specificity. ImmunoSpot and flow cytometry results were closely correlated. Conclusions: The study demonstrates that all three assays are suited for the detection of specific Bmem in antigen-primed individuals when such Bmem occur in the mid- to high-frequency range, and that they broadly concur. Strengths and weaknesses of the three test systems are discussed. Full article
(This article belongs to the Special Issue Immune Monitoring in 2026)
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19 pages, 6012 KB  
Article
Donor- and Acceptor-Side Protection Against Photosystem I Photoinhibition in Arabidopsis thaliana
by Marina Kozuleva
Int. J. Mol. Sci. 2026, 27(1), 9; https://doi.org/10.3390/ijms27010009 - 19 Dec 2025
Viewed by 166
Abstract
Photosystem I (PSI) photoinhibition (PI(I)) is gaining traction as a potentially more significant threat to plant performance than photoinhibition of photosystem II (PSII). The increased focus is facilitated by the implementation of specific protocols that induce PI(I), such as artificial fluctuating light (FL) [...] Read more.
Photosystem I (PSI) photoinhibition (PI(I)) is gaining traction as a potentially more significant threat to plant performance than photoinhibition of photosystem II (PSII). The increased focus is facilitated by the implementation of specific protocols that induce PI(I), such as artificial fluctuating light (FL) and repetitive short saturating pulses (rSPs). rSPs were long considered a specific sub-case of FL. However, recent evidence suggests that PI(I) proceeds via at least two distinct, treatment-dependent mechanisms, leading to damage at the donor or acceptor side of PSI. This discovery suggests that rSPs and FL represent distinct photoinhibitory stresses and that different mechanisms protect PSI against FL and rSPs. This study comparatively analyzed the effects of FL and rSPs on PSI activity in Arabidopsis thaliana wild-type plants and a selection of mutants (pgr5, pgrl1, stn7, tap38/pph1, and pgr1), previously noted or hypothesized to have altered PI(I) sensitivity relative to the wild type. The results of this work, particularly the contrasting sensitivity of tap38/pph1 compared to the wild type under FL and rSP conditions, strongly suggest that pulsed illumination and fluctuating light are distinct photoinhibitory treatments, and different mechanisms protect PSI against them. Full article
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12 pages, 497 KB  
Review
Temporary Portocaval Shunts During Liver Transplantation: A Narrative Review of Technical Solutions and Post-Transplant Outcomes
by Elisa Schirra, Alberto Mauro, Giuseppe Bianco, Marco Maria Pascale, Francesco Frongillo, Erida Nure, Alfonso Wolfango Avolio, Salvatore Agnes and Gabriele Spoletini
J. Clin. Med. 2025, 14(24), 8723; https://doi.org/10.3390/jcm14248723 - 9 Dec 2025
Viewed by 210
Abstract
Background: Liver transplantation (LT) continues to evolve with techniques aimed at minimizing perioperative complications associated with caval and portal vein clamping. Caval-sparing approaches, such as the piggyback technique, preserve hemodynamic stability; however, portal clamping remains necessary and may trigger postreperfusion syndrome, endotoxemia, [...] Read more.
Background: Liver transplantation (LT) continues to evolve with techniques aimed at minimizing perioperative complications associated with caval and portal vein clamping. Caval-sparing approaches, such as the piggyback technique, preserve hemodynamic stability; however, portal clamping remains necessary and may trigger postreperfusion syndrome, endotoxemia, and hepatic microcirculatory disturbances. Temporary portocaval shunts (PCSs) have been developed to maintain portal flow during LT, mitigating these adverse effects and allowing for hemodynamic stability and a reduced intraoperative bleeding. Portocaval Shunts: Various PCS techniques—including end-to-side, right-branch, portosaphenous, mesenterico-saphenous, iliac-venous conduit interposition, portoumbilical, and Rex-saphenous shunts—allow an individualized approach based on patient anatomy and surgical complexity. Review of Evidence: Available evidence demonstrates that PCS improves intraoperative hemodynamic stability, reduces blood transfusion requirements, and preserves renal function, particularly in patients with high portal flow or severe portal hypertension. PCS may also shorten warm ischemia time, facilitate hepatectomy, and enhance outcomes in extended criteria donor grafts or marginal organs. Meta-analyses and randomized studies support its role in reducing intraoperative blood loss, improving early graft function, and accelerating postoperative recovery. However, the effect of PCS on long-term survival and major postoperative morbidity remains variable, likely due to heterogeneity in patient populations, donor types, and perioperative management. Conclusions: Overall, PCS represents a safe and feasible adjunct in LT, offering significant hemodynamic and technical advantages. Its use should be individualized based on patient risk factors, intraoperative hemodynamics, and anticipated intraoperative challenges. PCS provides a practical strategy to preserve portal flow, minimizing intraoperative complications and facilitating the hepatectomy. However, the decision to create a PCS during LT still depends on the surgeon’s preference. Postoperative outcomes and impact on long-term survival require further investigation. Full article
(This article belongs to the Special Issue Liver Transplantation: Current Hurdles and Future Perspectives)
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15 pages, 2096 KB  
Article
Reducing Storage-Related Bias in Reproductive Vitamin D Research: Towards a Stable and Reliable Biomarker of Ovarian Vitamin D Status
by Evelin E. Lara-Molina, Jason M. Franasiak, Almudena Devesa-Peiro, Marina López-Nogueroles, Alberto Vázquez, David Amorós, Agustín Ballesteros, Antonio Pellicer, Patricia Sebastian-Leon and Patricia Diaz-Gimeno
Nutrients 2025, 17(23), 3783; https://doi.org/10.3390/nu17233783 - 2 Dec 2025
Viewed by 326
Abstract
Background/Objectives: One of the main reasons for discrepancies in the role of vitamin D in ART could be the measurement of the conventional biomarker 25(OH)D3. It is known that this value is affected by multiple factors, such as tissue origin, assay [...] Read more.
Background/Objectives: One of the main reasons for discrepancies in the role of vitamin D in ART could be the measurement of the conventional biomarker 25(OH)D3. It is known that this value is affected by multiple factors, such as tissue origin, assay variability, classification criteria, and potential storage-related degradation. In this study, we investigate 24,25(OH)2D3 as a new biomarker to improve vitamin D assessment in women’s reproductive health, particularly regarding oocyte development. Methods: A prospective cohort study including 35 oocyte donors undergoing controlled ovarian stimulation, who were recruited between October and November 2017, was conducted. Vitamin D metabolites were measured at the baseline and after seven months of storage at −80 °C. Paired serum and pooled follicular fluid (FF) samples were collected at oocyte retrieval. 25(OH)D3 and 24,25(OH)2D3 were quantified by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). Statistical analyses included paired tests (serum vs. FF; baseline vs. stored) and Pearson’s correlations (two-sided α = 0.05). Results: At the baseline, the mean serum 25(OH)D3 concentration was 91.56 ± 39.01 nmol/L and the mean FF concentration was 58.13 ± 19.55 nmol/L (p < 0.0001). Serum 24,25(OH)2D3 averaged 15.62 ± 10.99 nmol/L, compared with 11.26 ± 6.09 nmol/L in FF (p = 0.004). In both fluids, 25(OH)D3 and 24,25(OH)2D3 were strongly correlated (serum R2 = 0.92; FF R2 = 0.91). Across fluids, the serum–FF correlation was stronger for 24,25(OH)2D3 (R2 = 0.77, p <0.0001) than for 25(OH)D3 (R2 = 0.69, p < 0.0001). After seven months of storage, 25(OH)D3 concentrations decreased significantly (serum −32%; FF −38%; both p < 0.0001), whereas 24,25(OH)2D3 levels remained stable (serum p = 0.24; FF p = 0.36). Conclusions: Serum 24,25(OH)2D3 is a more reliable and minimally invasive biomarker for assessing ovarian vitamin D status than the current gold standard, 25(OH)D3. Incorporating this metabolite into research studies and storage quality control may improve the reliability of retrospective analyses based on cryopreserved material, contributing to a better understanding of the role of vitamin D in human reproduction. Full article
(This article belongs to the Special Issue Assessment of Vitamin D Status and Intake in Human Health)
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11 pages, 340 KB  
Article
EZ Lyn: A Confirmed Period-Bouncer Cataclysmic Variable Below the Period Minimum
by Nadezhda L. Vaidman, Almansur T. Agishev, Serik A. Khokhlov and Aldiyar T. Agishev
Galaxies 2025, 13(6), 121; https://doi.org/10.3390/galaxies13060121 - 30 Oct 2025
Viewed by 718
Abstract
We model the short-period cataclysmic variable EZ Lyn with MESA binary evolution and infer its present-day parameters through a staged statistical search. First, we compute a coarse grid of tracks in (M1,0,P0) at fixed [...] Read more.
We model the short-period cataclysmic variable EZ Lyn with MESA binary evolution and infer its present-day parameters through a staged statistical search. First, we compute a coarse grid of tracks in (M1,0,P0) at fixed M2,0 and rank snapshots by a profile likelihood. We then resample the neighbourhood of the minimum to build a refined Δχ2 surface. Finally, we sample this surface with an affine-invariant MCMC to obtain posteriors, using a likelihood that treats the one-sided constraint on the donor temperature and the ambiguity of component roles in the binary output. The best-fit snapshot reproduces the observables and identifies EZ Lyn as a period bouncer with a substellar donor. We infer MWD=0.850±0.019M, M2=0.0483±0.0137M, RWD=0.0092±0.0001R, R2=0.099±0.005R, TWD=11,500±20K, and T2=1600±50K. The instantaneous mass-transfer rate at the best-fit snapshot is M˙=3.66×1011Myr1, consistent with the secular range implied by the white-dwarf temperature. Independent checks from the Roche mean-density relation, surface gravities, and the semi-empirical donor sequence support the solution. In population context, EZ Lyn lies in the period-minimum spike and on the low-mass tail of the donor mass–period plane. The classification is robust to modest displacements along the shallow Δχ2 valley. We release inlists, tracks, and analysis scripts for reproducibility. Full article
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26 pages, 3173 KB  
Article
Enhancement of Photosynthetic Efficiency and Antioxidant Response in Wheat Under Drought Stress by Quercetin–Copper Complex
by Marta Jańczak-Pieniążek, Dagmara Migut, Tomasz Piechowiak and Maciej Balawejder
Int. J. Mol. Sci. 2025, 26(21), 10365; https://doi.org/10.3390/ijms262110365 - 24 Oct 2025
Viewed by 483
Abstract
One way to counteract the effects of environmental stresses, including drought, is to use products with growth-promoting properties for plants. Such agents include quercetin, which is known for its antioxidant and photosynthesis-enhancing properties. In the conducted experiment, the influence of the quercetin–copper complex [...] Read more.
One way to counteract the effects of environmental stresses, including drought, is to use products with growth-promoting properties for plants. Such agents include quercetin, which is known for its antioxidant and photosynthesis-enhancing properties. In the conducted experiment, the influence of the quercetin–copper complex (Q-Cu (II)) treatment, characterized by strong high solubility in water and strong antioxidant properties, was investigated. The pot experiment demonstrated the effect of spraying with Q-Cu (II) solutions (0.01, 0.05 and 0.1%) on wheat plants growing under drought stress conditions. Two treatments of Q-Cu (II) solutions were applied, and chlorophyll content and chlorophyll fluorescence (the maximum quantum yield of photosystem II (PSII) photochemistry (Fv/Fm), the efficiency of the water-splitting complex on the donor side of PSII (Fv/Fo), and the photosynthetic efficiency index (PI)), as well as gas exchange (photosynthetic network intensity (PN), transpiration rate (E), stomatal conductance (gs) and intercellular CO2 concentration (Ci)), were measured 1 and 7 days after each treatment. In addition, antioxidant enzyme activity (catalase (CAT), peroxidase (SOD) and guaiacol peroxidase (GPOX)) and reactive oxygen species (ROS) levels were determined. Drought stress caused a decrease in chlorophyll content, and values of parameters Fv/Fm, Fv/Fo, PI and PN, E, gs, Ci, as well as an increase in ROS levels and antioxidant enzyme activity. Exogenous Q-Cu (II) improved photosynthetic indices and modulated redox status in a dose-dependent manner: 0.01–0.05% reduced ROS, whereas 0.1% increased ROS while concomitantly enhancing antioxidant enzyme activities and photosynthetic performance, consistent with ROS-mediated priming. The conducted research indicates the possibility of using Q-Cu (II) as a product to enhance the efficiency of the photosynthetic process under drought stress. Full article
(This article belongs to the Special Issue Molecular Mechanisms of Plant Abiotic Stress Tolerance: 2nd Edition)
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31 pages, 2915 KB  
Review
A Review of the Progress in the Microbial Biosynthesis of Prenylated Aromatic Compounds
by Min Tang, Wanze Zhang, Yanjie Tian, Jianjun Qiao, Xiaobing Li, Weiguo Li and Qinggele Caiyin
Molecules 2025, 30(19), 3931; https://doi.org/10.3390/molecules30193931 - 30 Sep 2025
Viewed by 1625
Abstract
Prenylated aromatic compounds (PACs) are widely distributed in nature and have important applications in medicine, cosmetics, and food due to their antioxidant, anticancer, and anti-inflammatory activities as well as role in the prevention of neurological diseases. Traditional methods of PAC production such as [...] Read more.
Prenylated aromatic compounds (PACs) are widely distributed in nature and have important applications in medicine, cosmetics, and food due to their antioxidant, anticancer, and anti-inflammatory activities as well as role in the prevention of neurological diseases. Traditional methods of PAC production such as plant extraction and chemical synthesis remain constrained by the low content of these compounds in plants and the complexity of the chemical processes. PACs are synthesized from aromatic compound receptors and prenyl side chain donors, which are in turn synthesized via the shikimate pathway and 2-C-methyl-D-erythritol-4-phosphate/mevalonic acid pathways, respectively. Increasing exploration and research on prenyltransferases (PTs), the key enzymes involved in PAC biosynthesis, have facilitated the emergence of microbial synthesis of PACs as a promising alternative to industrial production. The microbial biosynthesis of PACs is summarized herein, mainly from the perspective of screening and modification of the key enzymes PTs, selection of suitable host systems, and engineering the modification of microbial cell factories to enhance the yields of PACs. The future prospects and challenges of PAC biosynthesis are also discussed. Full article
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13 pages, 784 KB  
Article
Real-Life Experience with Cytomegalovirus Hyperimmune Globulin in a Lung Transplant Unit: Long-Term Outcomes
by Raquel Sanabrias Fernández de Sevilla, Sarela García-Masedo Fernández, Rosalía Laporta Hernández, Myriam Aguilar Pérez, Christian García Fadul, María Teresa Lázaro Carrasco de la Fuente, Enrique Rodríguez Rubio, Amelia Sánchez Guerrero, Carlos Almonacid Sánchez and María Piedad Ussetti Gil
Therapeutics 2025, 2(4), 17; https://doi.org/10.3390/therapeutics2040017 - 30 Sep 2025
Viewed by 783
Abstract
Background/Objectives: Cytomegalovirus (CMV) infection is a frequent complication after lung transplantation, especially in high-risk donor-positive/recipient-negative (D+/R−) patients. CMV-specific hyperimmunoglobulin (CMV-HIG), administered either with antivirals or as monotherapy, may be beneficial for preventing or treating CMV infection in selected clinical scenarios. This study [...] Read more.
Background/Objectives: Cytomegalovirus (CMV) infection is a frequent complication after lung transplantation, especially in high-risk donor-positive/recipient-negative (D+/R−) patients. CMV-specific hyperimmunoglobulin (CMV-HIG), administered either with antivirals or as monotherapy, may be beneficial for preventing or treating CMV infection in selected clinical scenarios. This study evaluated CMV-HIG indications and their impact on clinical outcomes in our lung transplant unit. Methods: We retrospectively analyzed adult lung transplant recipients (2010–2023) who received ≥2 doses of CMV-HIG for universal prophylaxis, monotherapy prophylaxis, preemptive therapy, or treatment of invasive disease. Results: CMV-HIG was administered to 204 out of 336 recipients (61%). CMV-HIG was well tolerated, with no treatment-related adverse events. Indications were preemptive therapy (63%), universal prophylaxis (24%), monotherapy prophylaxis (7%), and treatment of invasive disease (6%). CMV-HIG was well tolerated, with no treatment-related adverse events. No patients developed invasive disease during combination prophylaxis or preemptive treatment. The combination treatment of patients with invasive disease was also effective, and no cases of VGC resistance were detected. CMV-HIG monoprophylaxis has allowed us to delay or prevent viral replication in recipients who developed VGC side effects. Rates of acute rejection, Chronic Lung Allograft Dysfunction (CLAD), and overall survival were similar across CMV risk groups. Conclusions: Our results showed that the combined use of CMV-HIG and antiviral agents is effective in preventing CMV infection and disease in high-risk lung transplant recipients. This combination is also useful in treating invasive disease and preventing VGC resistance. Additionally, CMV-HIG monoprohylaxis can delay or prevent viral replication in recipients experiencing VGC-related side effects. These findings support the use of CMV-HIG in selected clinical settings, although prospective studies are needed to define its potential benefits within the current therapeutic armamentarium. Full article
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10 pages, 1190 KB  
Technical Note
Efficacy of a Modified Superficial Temporal Artery–Middle Cerebral Artery Bypass Using Superficial Temporal Artery Side-Branch Donors in Adult Moyamoya Disease: A Technical Note
by Shintaro Arai, Tatsuya Sugiyama, Tohru Mizutani, Kenji Sumi, Masaki Matsumoto, Kouzou Murakami, Ryo Irie and Yoichi Morofuji
J. Clin. Med. 2025, 14(19), 6904; https://doi.org/10.3390/jcm14196904 - 29 Sep 2025
Viewed by 985
Abstract
Background: Adult moyamoya disease (MMD) is a progressive steno-occlusive cerebrovascular disorder for which surgical revascularization is the primary treatment. The standard direct superficial temporal artery–middle cerebral artery (STA-MCA) bypass uses the frontal and/or parietal branch of the STA as the donor. However, in [...] Read more.
Background: Adult moyamoya disease (MMD) is a progressive steno-occlusive cerebrovascular disorder for which surgical revascularization is the primary treatment. The standard direct superficial temporal artery–middle cerebral artery (STA-MCA) bypass uses the frontal and/or parietal branch of the STA as the donor. However, in some patients, conventional STA-MCA bypass may be suboptimal because of a large mismatch in caliber between the STA branch and the recipient artery, increasing the risk of cerebral hyperperfusion. This study aimed to investigate the impact of a modified STA-MCA bypass on MMD treatment. Methods: We retrospectively reviewed adult cases of MMD at our institution (2012–2025) for patients who underwent modified direct STA-MCA bypass using a small side branch of the STA as the donor artery. Surgical techniques and clinical outcomes of these cases were analyzed descriptively. Results: Five cases (five hemispheres in four patients) underwent side-branch STA-MCA bypass. All procedures were completed successfully, with 100% graft patency confirmed by intraoperative indocyanine green angiography, and a mild increase in cerebral blood flow confirmed by postoperative single-photon emission computed tomography. No patients developed postoperative cerebral hyperperfusion syndrome or wound healing complications. Clinically, all patients experienced a stable or improved neurological status, with no reported new ischemic or hemorrhagic events during follow-up. Conclusions: In this small feasibility series, the side-branch STA–MCA bypass was technically feasible and safe, with no cerebral hyperperfusion syndrome observed. Any risk-mitigating effect on hyperperfusion remains theoretical and requires confirmation in comparative studies. Full article
(This article belongs to the Section Clinical Neurology)
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24 pages, 3497 KB  
Article
Substituent Effect in Histamine and Its Impact on Interactions with the G Protein-Coupled Human Receptor H1 Modelled by Quantum-Chemical Methods
by Anna Jezuita, Małgorzata Makowska-Janusik, Krzysztof Ejsmont and Wojciech Marczak
Molecules 2025, 30(18), 3736; https://doi.org/10.3390/molecules30183736 - 15 Sep 2025
Viewed by 864
Abstract
Neutral and protonated histamine tautomers, mono-substituted with twelve functional groups, were studied theoretically as isolated molecules and complexes with the H1 receptor. Geometry and energy of tautomers were optimized using the DFT method with the B3LYP functional and the aug-cc-pVTZ basis set. [...] Read more.
Neutral and protonated histamine tautomers, mono-substituted with twelve functional groups, were studied theoretically as isolated molecules and complexes with the H1 receptor. Geometry and energy of tautomers were optimized using the DFT method with the B3LYP functional and the aug-cc-pVTZ basis set. The approach was based on the charge of the substituent active region (cSAR) parameters and the Harmonic Oscillator Model of Aromaticity (HOMA) indices. The cSAR parameters characterized the electron density better than the conventional Hammett’s constants σ. In general, the cSAR parameters correlate with other characteristics of the charge distribution, particularly those for substituents at the carbon atom in the ring adjacent to the side chain. Substituents at this atom affected the aromaticity less strongly than those located between two nitrogen atoms, which confirmed recent reports. Our results suggest that the 3H tautomer isomerizes into the 1H one after binding to the H1 receptor. Moreover, the electron structure of the molecule hydrogen-bonded to the receptor may significantly depend on the electron donor-acceptor properties of the substituent. The strong electron-accepting substituents, e.g., NO2, favor the imidazole configuration of the ring in the bonded molecule, while the strong electron-donating ones, e.g., NH2, promote the imidazolium one. Full article
(This article belongs to the Section Computational and Theoretical Chemistry)
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12 pages, 4469 KB  
Article
Clinical and Radiological Outcomes of Revision Anterior Cruciate Ligament Reconstruction Using a Quadriceps Tendon Autograft with a Bone Block: A Single-Center Case Series
by Dhong Won Lee, Sung Gyu Moon, Ji Hee Kang, Seung Ik Cho and Woo Jong Kim
Medicina 2025, 61(9), 1634; https://doi.org/10.3390/medicina61091634 - 10 Sep 2025
Viewed by 1251
Abstract
Background and Objectives: Revision anterior cruciate ligament reconstruction (ACLR) is demanding and yields inferior outcomes compared with primary procedures. The quadriceps tendon (QT) autograft with bone block has biomechanical and biological advantages though clinical evidence in revision remains limited. This study evaluated the [...] Read more.
Background and Objectives: Revision anterior cruciate ligament reconstruction (ACLR) is demanding and yields inferior outcomes compared with primary procedures. The quadriceps tendon (QT) autograft with bone block has biomechanical and biological advantages though clinical evidence in revision remains limited. This study evaluated the clinical and radiological outcomes of revision ACLR using bone-block QT autograft in young, active patients. Materials and Methods: A case series with a level of evidence of 4. Thirty-four patients (28 men, 6 women; mean age, 27.2 ± 5.8 years) who underwent revision ACLR with a bone-block QT autograft between 2021 and 2023 were retrospectively reviewed. The mean follow-up was 37.4 ± 3.2 months. Clinical assessments included the Lysholm, International Knee Documentation Committee (IKDC) subjective, and Tegner activity scores, along with isokinetic strength testing. Objective stability was evaluated using pivot shift grading and Telos stress radiography. Radiological analyses included 3D computed tomography for tunnel positioning and magnetic resonance imaging for tunnel widening. Perioperative and postoperative complications were recorded. Results: All clinical outcomes improved significantly from baseline to 2-year follow-up: Lysholm (62.7 ± 9.6 to 87.1 ± 10.3), IKDC (59.0 ± 10.8 to 79.5 ± 11.1), and Tegner (3.5 ± 1.2 to 5.6 ± 1.3; all p < 0.001). However, the Tegner score remained lower than the pre-injury level (6.1 ± 1.4; p = 0.035). At the final follow-up, 91.2% of the patients had returned to sports, with 59% resuming sports at their pre-injury level or higher. Side-to-side anterior laxity decreased from 8.5 ± 1.7 mm to 1.4 ± 1.1 mm on Telos stress radiography (p < 0.001). Preoperatively, 82% of patients demonstrated high grade pivot shift (≥grade 2), which improved to 91% graded as negative or grade 1 at final follow-up (p < 0.001). Isokinetic evaluation showed improvements in quadriceps (28.7% ± 12.5% to 12.4% ± 8.1%) and hamstring (18.3% ± 9.7% to 8.9% ± 6.5%) deficit (both p < 0.001). MRI demonstrated minimal tunnel widening (tibia, +1.3 ± 0.9 mm, p = 0.012; femur, +0.3 ± 0.6 mm, p = 0.148). Three complications (8.8%) were observed: one cyclops lesion, one transient extension deficit, and one graft rupture. No patellar fractures, septic arthritis, or revision procedures occurred during the follow-up period. Conclusions: Bone-block QT autografts provide a reliable option for revision ACLR, yielding functional improvement, restored stability, and minimal donor-site morbidity, with low complications. These findings support their consideration as the preferred graft choice for young active patients needing revision reconstruction. Full article
(This article belongs to the Special Issue Anterior Cruciate Ligament (ACL) Injury)
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12 pages, 212 KB  
Article
Management of the Venous Anastomoses of a Tertiary Referral Centre in Reconstructive Microvascular Surgery Using Fasciocutaneous Free Flaps in the Head and Neck
by Nocini Riccardo, Muneretto Carlotta, Lobbia Guido, Zatta Esmeralda, Athena Eliana Arsie, Molteni Gabriele, Arietti Valerio and Barbera Giorgio
J. Clin. Med. 2025, 14(17), 6171; https://doi.org/10.3390/jcm14176171 - 1 Sep 2025
Cited by 1 | Viewed by 803
Abstract
Objectives: The application of fasciocutaneous free flaps for reconstruction of head and neck defects following oncological surgery has increased since the 1970s, coinciding with developments in microvascular techniques. Although reported success rates are between 90% and 99%, flap failure continues to occur, [...] Read more.
Objectives: The application of fasciocutaneous free flaps for reconstruction of head and neck defects following oncological surgery has increased since the 1970s, coinciding with developments in microvascular techniques. Although reported success rates are between 90% and 99%, flap failure continues to occur, most frequently due to venous congestion. This study examines the rates of re-exploration and flap failure according to the number of venous anastomoses in patients receiving free flap reconstruction after head and neck cancer surgery. Materials and Methods: This retrospective analysis included 163 patients who underwent head and neck reconstruction with free flaps (radial forearm free flap [RFFF] and anterolateral thigh flap [ALTF]) at the University Hospital of Verona between January 2019 and June 2024. Variables examined comprised the type of flap performed, donor and recipient vessels utilized, and number of venous anastomoses, as well as the type (end-to-end [ETE] versus end-to-side [ETS]) and site (internal jugular vein versus external jugular vein) of venous anastomosis. Results: The overall success rate was 93.3%, with no significant difference between single and dual venous anastomosis groups. Prompt re-exploration upon detecting signs of flap failure is critical, as approximately one-third of such failures may be prevented through timely intervention. Conclusions: Single venous end-to-end anastomosis utilizing the internal jugular vein system is typically effective. Further research is warranted to clarify the indications for dual anastomosis involving the external jugular vein system. Full article
(This article belongs to the Special Issue Innovations in Head and Neck Surgery)
14 pages, 1757 KB  
Article
Photosynthetic Electron Transport in Winter Wheat: Responses to Low-Temperature and Weak-Light Condition
by Cheng Yang, Minghan Liu, Simeng Du, Deqi Zhang, Xiangdong Li, Liting Wu, Yanhua Shi, Baoting Fang, Ge Yan and Fang Wei
Cells 2025, 14(16), 1275; https://doi.org/10.3390/cells14161275 - 18 Aug 2025
Viewed by 960
Abstract
Spring low temperatures are a serious natural threat to wheat production in the Huang-Huai wheat region, and they are often accompanied by weak light environments during the day. To elucidate the response patterns and adaptation mechanisms of winter wheat leaves to low-temperature and [...] Read more.
Spring low temperatures are a serious natural threat to wheat production in the Huang-Huai wheat region, and they are often accompanied by weak light environments during the day. To elucidate the response patterns and adaptation mechanisms of winter wheat leaves to low-temperature and weak-light environments, we simultaneously measured prompt chlorophyll a fluorescence, delayed chlorophyll a fluorescence, and modulated 820 nm light reflection; moreover, we analyzed the effects of low temperature and weak light treatment for different duration (2 h and 4 h) on the donor-side activity of photosystem II (PSII), the degree of PSII unit dissociation, the efficiency of light energy absorption and capture by PSII, electron transfer to QA and PSI terminal, PSI activity and cyclic electron transport activity in isolated wheat leaves under controlled conditions. The results, which were corroborated using the three methods, revealed that in low-temperature and weak-light environments, the degree of PSII unit dissociation, and the efficiency of light energy absorption, capture, and electron transfer to QA decreased, while the donor-side activity remained unaffected. In contrast, the efficiency of electron transfer to the PSI terminal and the overall performance of photosynthetic electron transport increased. Comprehensive analysis suggests that the increase in the electron receptor pool at the PSI terminal under low-temperature stress is a crucial factor contributing to the enhanced electron transfer efficiency to the PSI terminal and the improved overall performance of the photosynthetic electron transport chain, which is also a crucial factor in the high cold tolerance of winter wheat. Full article
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17 pages, 7479 KB  
Article
Development and Validation of a Custom-Built System for Real-Time Monitoring of In Vitro Rumen Gas Fermentation
by Zhen-Shu Liu, Bo-Yuan Chen, Jacky Peng-Wen Chan and Po-Wen Chen
Animals 2025, 15(15), 2308; https://doi.org/10.3390/ani15152308 - 6 Aug 2025
Viewed by 728
Abstract
While the Ankom RF system facilitates efficient high-throughput in vitro fermentation studies, its high cost and limited flexibility constrain its broader applicability. To address these limitations, we developed and validated a low-cost, modular gas monitoring system (FerME), assembled from commercially available components. To [...] Read more.
While the Ankom RF system facilitates efficient high-throughput in vitro fermentation studies, its high cost and limited flexibility constrain its broader applicability. To address these limitations, we developed and validated a low-cost, modular gas monitoring system (FerME), assembled from commercially available components. To evaluate its performance and reproducibility relative to the Ankom RF system (Ankom Technology, Macedon, NY, USA), in vitro rumen fermentation experiments were conducted under strictly controlled and identical conditions. Whole rumen contents were collected approximately 2 h post-feeding from individual mid- or late-lactation dairy cows and immediately transported to the laboratory. Each fermenter received 50 mL of processed rumen fluid, 100 mL of anaerobically prepared artificial saliva buffer, and 1.2 g of the donor cow’s diet. Bottles were sealed with the respective system’s pressure sensors, flushed with CO2, and incubated in a 50 L water bath maintained at 39 °C. FerME (New Taipei City, Taiwan) and Ankom RF fermenters were placed side-by-side to ensure uniform thermal conditions. To assess the effect of filter bag use, an additional trial employed Ankom F57 filter bags (Ankom Technology, Macedon, NY, USA; 25 μm pore size). Trial 1 revealed no significant differences in cumulative gas production, volatile fatty acids (VFAs), NH3-N, or pH between systems (p > 0.05). However, the use of filter bags reduced gas output and increased propionate concentrations (p < 0.05). Trial 2, which employed filter bags in both systems, confirmed comparable results, with the FerME system demonstrating improved precision (CV: 4.8% vs. 13.2%). Gas composition (CH4 + CO2: 76–82%) and fermentation parameters remained consistent across systems (p > 0.05). Importantly, with 12 pressure sensors, the total cost of FerME was about half that of the Ankom RF system. Collectively, these findings demonstrate that FerME is a reliable, low-cost alternative for real-time rumen fermentation monitoring and could be suitable for studies in animal nutrition, methane mitigation, and related applications. Full article
(This article belongs to the Section Animal System and Management)
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15 pages, 2095 KB  
Article
T-Lymphocyte Phenotypic and Mitochondrial Parameters as Markers of Incomplete Immune Restoration in People Living with HIV+ on Long-Term cART
by Damian Vangelov, Radoslava Emilova, Yana Todorova, Nina Yancheva, Reneta Dimitrova, Lyubomira Grigorova, Ivailo Alexiev and Maria Nikolova
Biomedicines 2025, 13(8), 1839; https://doi.org/10.3390/biomedicines13081839 - 28 Jul 2025
Cited by 1 | Viewed by 1023
Abstract
Background/Objectives: Restored CD4 absolute counts (CD4AC) and CD4/CD8 ratio in the setting of continuous antiretroviral treatment (ART) do not exclude a low-level immune activation associated with HIV reservoirs, microbial translocation, or the side effects of ART itself, which accelerates the aging of [...] Read more.
Background/Objectives: Restored CD4 absolute counts (CD4AC) and CD4/CD8 ratio in the setting of continuous antiretroviral treatment (ART) do not exclude a low-level immune activation associated with HIV reservoirs, microbial translocation, or the side effects of ART itself, which accelerates the aging of people living with HIV (PLHIV). To delineate biomarkers of incomplete immune restoration in PLHIV on successful ART, we evaluated T-lymphocyte mitochondrial parameters in relation to phenotypic markers of immune exhaustion and senescence. Methods: PLHIV with sustained viral suppression, CD4AC > 500 and CD4/CD8 ratio >0.9 on ART (n = 39) were compared to age-matched ART-naïve donors (n = 27) and HIV(–) healthy controls (HC, n = 35). CD4 and CD8 differentiation and effector subsets (CCR7/CD45RA and CD27/CD28), activation, exhaustion, and senescence markers (CD38, CD39 Treg, CD57, TIGIT, and PD-1) were determined by flow cytometry. Mitochondrial mass (MM) and membrane potential (MMP) of CD8 and CD4 T cells were evaluated with MitoTracker Green and Red flow cytometry dyes. Results: ART+PLHIV differed from HC by increased CD4 TEMRA (5.3 (2.1–8.8) vs. 3.2 (1.6–4.4), p < 0.05), persistent TIGIT+CD57–CD27+CD28– CD8+ subset (53.9 (45.5–68.9) vs. 40.1 (26.7–58.5), p < 0.05), and expanding preapoptotic TIGIT–CD57+CD8+ effectors (9.2 (4.3–21.8) vs. 3.0 (1.5–7.3), p < 0.01) in correlation with increased CD8+ MMP (2527 (1675–4080) vs.1477 (1280–1691), p < 0.01). These aberrations were independent of age, time to ART, or ART duration, and were combined with increasing CD4 T cell MMP and MM. Conclusions: In spite of recovered CD4AC and CD4/CD8 ratio, the increased CD8+ MMP, combined with elevated markers of exhaustion and senescence in ART+PLHIV, signals a malfunction of the CD8 effector pool that may compromise viral reservoir latency. Full article
(This article belongs to the Special Issue Emerging Insights into HIV)
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