Search Results (1,412)

Bartonella henselae is a flea-borne zoonotic bacterium for which domestic cats constitute the principal reservoir. However, contemporary molecular epidemiological data from Portugal remain scarce. This retrospective laboratory study analysed EDTA-stabilised blood samples from apparently healthy cats submitted for routine screening by 74 veterinary centres across mainland Portugal and autonomous regions over an 11-year period (2015–2025). DNA extracts were tested using a species-specific TaqMan qPCR assay for B. henselae with an internal extraction control, and a subset of samples was subsequently confirmed by nested PCR followed by Sanger sequencing (ribC). Among 270 cats, 47 tested positive, yielding a qPCR prevalence of 17.4% (95% confidence interval [CI] 13.1–22.5). Submissions were predominantly from Northern Portugal, and infection status was not statistically associated with the Nomenclature of Territorial Units for Statistics (NUTS) level 2 region (p = 0.478). Infection was more frequent in younger cats (median age 2 years, interquartile range [IQR] 1–5; p = 0.037), while sex (p = 0.103) and breed (p = 0.730) were not significantly associated with infection status. These findings support endemic circulation of B. henselae in Portuguese cats at levels comparable to other temperate European regions. The detection of subclinical infection in apparently healthy cats is relevant to transfusion medicine and supports the inclusion of B. henselae qPCR screening in donor selection protocols. Full article

Age-related osteoporosis is driven in part by senescence-associated rewiring of bone marrow mesenchymal stem cells (MSCs) from osteogenic toward adipogenic fates. Accumulating evidence indicates that epigenetic drift and reduced autophagy are not isolated lesions but are mechanistically coupled through a bidirectional DNA methylation and autophagy axis. Here, we summarize how promoter hypermethylation of genes involved in autophagy and osteogenesis suppresses autophagic flux and osteoblast lineage transcriptional programs. Conversely, autophagy insufficiency reshapes the methylome by limiting methyl donor availability, most notably S-adenosylmethionine (SAM), and by reducing the turnover of key epigenetic regulators, including DNA methyltransferases (DNMTs), ten-eleven translocation (TET) dioxygenases, and histone deacetylases (HDACs). This self-reinforcing circuitry exacerbates mitochondrial dysfunction, oxidative stress, and inflammation driven by the senescence-associated secretory phenotype (SASP), thereby stabilizing adipogenic bias and progressively impairing marrow niche homeostasis and bone remodeling. We further discuss therapeutic strategies to restore balance within this axis, including selective modulation of epigenetic enzymes; activation of AMP-activated protein kinase (AMPK) and mechanistic target of rapamycin (mTOR) signaling with downstream engagement of Unc-51-like autophagy-activating kinase 1 (ULK1) and transcription factor EB (TFEB); targeting sirtuin pathways; mitochondria- and autophagy-supportive natural compounds; and bone-targeted delivery approaches or rational combination regimens. Full article

Background/Objectives: Donor kidney function measured by glomerular filtration rate (GFR) is widely used as a selection criterion in kidney transplantation (KT). This study addresses the knowledge gap regarding the relationship between donor GFR at organ procurement and graft function in deceased donor KT. Methods: We retrospectively analyzed 918 deceased donor KTs and compared donor GFRs at procurement and recipient GFRs after KT at hospital discharge and in the one-year follow-up. The Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) formula was used to estimate and compare GFRs. Donor baseline GRF was defined as the last available estimated GRF prior to organ procurement. The Kaplan–Meier analysis was used to estimate recipient and graft survival. Results: The median donor GFR was 92.8 mL/min/1.73 m², while the median recipient GFR at hospital discharge was 37.5 mL/min/1.73 m² (−60% to donor baseline, p < 0.001), increasing to 51.4 mL/min/1.73 m² (+37%, p < 0.001) at one-year follow-up. One-year graft and patient survival rates were 95.3% and 98.1%, respectively. Except for grafts from donors with a GFR < 15 mL/min/1.73 m² due to acute renal failure that resulted in a significantly higher delayed graft function (DGF) rate and inferior graft survival (71.4%), no correlation was observed between baseline GFRs and DGF occurrence nor graft survival. Conclusions: Excellent results can be achieved in KT with subnormal donor GFR. The decision to refuse a kidney offer for KT should not solely be based on donor GFR. Kidneys from donors with very low GFR (<15 mL/min/1.73 m²) may be transplanted, but our observation is based on a very small sample (n = 7) and should therefore be interpreted with caution, particularly given the associated higher risk of DGF and lower graft survival. Full article

Adipose tissue plays a crucial role in breast cancer (BC) progression by actively modulating the tumor microenvironment. We investigated how tumor proximity modifies adipose tissue by analyzing selected adipose-related and prognosis-associated markers in explants from BC patients and healthy donors. Explants were categorized by proximity to the tumor as adjacent (less than 2 cm), distant (over 2 cm), alongside normal explants (controls). FABP4 and vimentin expression was increased in proximity to the tumor, while caveolin-1, CD44, MMP9, and adiponectin showed minimal or no changes. Conditioned media (CM) from adjacent and normal explants were then assessed for their effects on tumorigenic traits in hormone-receptor-positive breast cancer (HR+ BC) and triple-negative breast cancer (TNBC) cell lines. Adjacent-CM enhanced migration, induced cytoskeletal remodeling, reduced adhesion, and promoted an elongated, motile phenotype in T47D cells. Poor-prognosis markers (caveolin-1, vimentin, CD44) were upregulated in at least one HR+ BC model, whereas Nanog and KLF4 showed modest variation. In TNBC cells, both normal- and adjacent-CM partially shifted MDA-MB-231 morphology toward a more epithelial-like state, decreasing caveolin-1 levels, while adjacent-CM increased MMP9 expression. Overall, these results reveal that adipose tissue-derived soluble factors exert significant and subtype-dependent effects on BC tumorigenicity. Full article

Oxidative stress (OS) is a major cause of defective sperm function. During laboratory handling, gametes are exposed to OS, potentially mitigated by in vitro antioxidant supplementation. This study evaluates the protective role of caffeic acid (CAF) on basal human semen and under induced OS. First, six semen samples from normozoospermic donors were incubated with CAF concentrations ranging from 50 to 500 µM at 37 °C for 2 h. Sperm motility and DNA integrity (acridine orange) were evaluated. Then, ten semen samples were divided into four aliquots and incubated, respectively, with CAF at 100 µM, H₂O₂ at 2 mM, or H₂O₂ at 2 mM + CAF at 100 µM, or untreated. Motility, DNA integrity, acrosome status (Pisum sativum agglutinin), OS quantified by F₂-isoprostanes (ELISA), and expression of Nrf2, Keap1, and HO-1 (qRT-PCR) were assessed. CAF at 100 µM improved progressive motility without damaging DNA and was selected for subsequent experiments. CAF showed protective effects on sperm damage induced by H₂O₂ treatment, restoring motility, DNA integrity, and acrosome status and reducing F₂-isoprostane levels. Nrf2 and HO-1 expression were upregulated by CAF, downregulated by H₂O₂, and restored by the co-treatment. CAF supplementation may protect human spermatozoa during in vitro handling by reducing OS, improving several sperm parameters, with a possible mechanism of action involving the Nrf2 pathway. Full article

Background/Objectives: Antimicrobial resistance (AMR) surveillance is a cornerstone of national AMR strategies but requires sustained, cross-sectoral financing. While the need for such financing is well recognized, its quantification remains scarce in low- and middle-income countries. This study aimed to estimate the full costs of AMR surveillance across the human health, animal health, and food sectors (2021–2030) in selected facilities in Nepal and generate evidence to inform sustainable financing. Methods: A bottom-up micro-costing approach was used to analyze data from five sites. Costs were adjusted for inflation using projected gross domestic product deflators, and probabilistic sensitivity analyses were conducted to assess uncertainty in laboratory sample volumes under four scenarios. Results: The total cost of AMR surveillance in Nepal was $6.7 million: $3.4 million for human health (50.3% out of the aggregated costs), $2.7 million for animal health (39.8%), and $0.7 million for the food sector (9.9%). Laboratories accounted for >90% of total costs, with consumables and personnel as the main cost drivers. Average cost per sample was $150 (animal), $64 (food), and $6 (human). Conclusions: This study offers the first robust, multi-sectoral 10-year cost estimates of AMR surveillance in Nepal. The findings highlight that sustaining AMR surveillance requires predictable domestic financing, particularly to cover recurrent laboratory operations as donor support declines. These results provide cost evidence to support future budgeting and policy planning toward sustainable, nationally financed AMR surveillance in Nepal. Full article

Background: Takotsubo cardiomyopathy (TTC) has been historically considered a contraindication for heart donation due to its transient left ventricular dysfunction. However, emerging evidence supports that hearts from donors with fully recovered Takotsubo Cardiomyopathy can be safely transplanted. Methods: This case series describes seven heart transplantations performed between January 2022 and September 2025 using donors with previously diagnosed Takotsubo cardiomyopathy. Donor characteristics, intraoperative data, echocardiography data and postoperative outcomes were analyzed. Results: The mean donor age was 33.5 years (range 18–58), with a male-to-female ratio of 6:1. All donors exhibited echocardiographic evidence of Takotsubo Cardiomyopathy at the time of brain death, with full or partial recovery before procurement. Coronary angiography excluded obstructive coronary disease. Echocardiographic follow-up demonstrated the mean LVEF increased to 52 ± 6%, reaching 58 ± 4% at 12 months, global longitudinal strain (GLS) improved progressively (from −14.2 ± 2.8% to −18.5 ± 1.9%), confirming normalization of myocardial deformation and the right ventricular function, assessed by TAPSE, rose from 15 ± 3 mm at discharge to 20 ± 2 mm at 12 months. All patients transplanted with donors who had Takotsubo cardiomyopathy are alive at the 12-month follow-up. Conclusions: Hearts from donors with resolved Takotsubo Cardiomyopathy can be safely used for transplantation without compromising early- or mid-term outcomes. Expanding donor eligibility criteria to include selected TTC donors may contribute to mitigating organ shortages in advanced heart failure patients. Full article

Acetyl phosphate (AcP) is a microbial metabolite acting as a link between cell metabolism and signaling, providing the survival of bacteria in the host. AcP was also identified as an intermediate of pyruvate oxidation in mammalian mitochondria and was found in the human blood in some severe pathologies. The possible contribution of circulating AcP to the maintenance of the physiological or pathological states of the body has not been studied. Since AcP can function as a donor of phosphate groups, we have examined in vitro the influence of AcP on calcium signaling in mitochondria and cells by measuring the membrane potential and the calcium retention capacity of mitochondria by selective electrodes and by assaying the cell calcium signaling by Fura-2AM fluorescent radiometry. AcP was shown to induce a concentration-dependent increase in the mitochondrial resistance to calcium ion loading both in the control and in the presence of ADP. This effect was especially pronounced when mitochondria were incubated in a phosphate-free medium; under these conditions, AcP strongly raised the membrane potential and increased the rate of calcium uptake and the calcium retention capacity several times. Moreover, AcP induced similar changes in human cells when calcium signaling was activated by ATP, to a greater extent in neuroblastoma cells than in astrocytes. In the presence of AcP, a tendency for an increase in the amplitude and a decrease in the continuance of the ATP-induced calcium response was observed. These changes are probably associated with the activation of calcium buffering by mitochondria due to the delivery of phosphate during the hydrolysis of AcP. The results show that AcP is involved in the regulation of the Ca²⁺ balance in cells by activating the accumulation of calcium ions by mitochondria, especially under phosphate deficiency. A shift in calcium signaling mediated by AcP supplementation may be caused by hyperphosphatemia, which is now considered as one of basic contributors to cellular dysfunction and progression of various diseases, including sepsis. Full article

Dientamoeba fragilis is a protozoan of the human digestive tract, yet its transmission and pathogenic role remain poorly understood. This study aimed to evaluate its impact on the efficacy and safety of fecal microbiota transplantation (FMT) in treating recurrent Clostridioides difficile infection (rCDI). This longitudinal cohort study analyzed stool samples from FMT donors and recipients pre-treatment and at 2 and 8 weeks post-FMT. All samples were retrospectively tested using real-time PCR. Shotgun metagenomics was also performed on selected donor–recipient pairs to explore transmission. CDI cure rates, gastrointestinal adverse events (AEs), and serious adverse events (SAEs) were assessed prospectively. A total of 53 FMT were analyzed (179 samples), with 23 (43%) derived from D. fragilis-positive donor stool (4 of 10 donors, 40%). Four of 52 recipients (18.2%), initially negative and who received treatment from positive donors, tested positive post-FMT. Shotgun metagenomics could not definitely confirm transmission due to the lack of a good reference genome. No significant differences in efficacy, AE, or SAE were observed between FMT from D. fragilis-positive versus -negative donors, even in immunocompromised patients. No SAEs were attributed to FMT. D. fragilis may be transmitted via FMT without evidence of short-term clinical impact. Consequently, RT-PCR detection should be interpreted cautiously in the context of donor exclusion decisions. Full article

Apple breeding programs focus on enhancing yield, quality, and disease resistance, with a strong emphasis on evaluating phenological traits like flowering time and pomological traits such as fruit size and flavour, which are crucial for commercial success and consumer preference. Twenty-four families were obtained by crossing six apple varieties selected as pollen receptors and four apple genotypes resistant to scab selected as pollen donors. Data related to bud burst date, flowering date, harvest date, lengths of the periods between bud burst and flowering and from flowering to harvest (developmental period), fruit equatorial and polar diameter, fruit polar/diameter ratio, soluble solid content (SSC) and flesh firmness were analysed as a genetic partial diallel design. The study’s ANOVA on 24 fruit families across two years revealed significant genotype–environment interactions affecting flowering date, harvest date, and developmental periods, with some variables like fruit weight and soluble solids showing consistent variation. During each year, temperature influenced phenological phases, with earlier budbreak and flowering in warmer, less variable conditions in 2019. Analysis of genetic effects indicated high heritability for phenological traits and moderate heritability for fruit morphology and quality, with parental genetic contributions varying over years. Principal component and Procrustes analyses identified key variable groupings and parent profiles, highlighting genotypes such as ‘Granny Smith’, ‘McIntosh’, and ‘HM100’ with consistent additive effects, and certain families with notable heterotic performance. Overall, genetic and environmental interactions significantly shape phenological and fruit quality traits, guiding breeding strategies. Full article

Background/Objectives: Individual responses to CFTR modulators vary widely among people with cystic fibrosis (pwCF), underscoring the need for functional approaches that provide biological context alongside genotype-based therapy selection. Nasal epithelial cultures provide an individual-specific model for theratyping, but most studies rely on freshly isolated cells, restricting repeated testing and long-term sample use. In this study, we tested whether CFTR modulator responses measured in biobanked nasal cells were associated with real-world clinical outcomes. Methods: Cryopreserved nasal epithelial cells from 23 pwCF were differentiated at the air–liquid interface and assessed for CFTR modulator-responsive ion transport using Ussing chambers. In vitro responses were correlated with 6-month changes in sweat chloride concentration (SCC), FEV₁, and BMI. Results: Cryopreserved cultures retained donor-specific CFTR modulator responsiveness. Modulator-induced forskolin/IBMX-stimulated currents correlated with changes in SCC (R = −0.512). CFTR inhibitor-sensitive currents correlated with FEV₁ (R = 0.564). Associations between forskolin/IBMX-stimulated currents and FEV₁ were positive but did not reach statistical significance using two-tailed analysis. BMI changes showed no significant association. Conclusions: Biobanked nasal epithelial cultures preserve clinically relevant CFTR modulator responses at the cohort level, supporting their use as functional assays for population-level assessment in cystic fibrosis. This cryopreservation-based strategy enables repeated testing and may expand access to theratyping beyond freshly obtained samples. Full article

Per- and polyfluoroalkyl substances (PFASs), as a class of “permanent chemicals” with high environmental persistence and bioaccumulation, have attracted much attention. In this study, we focused on the molecular mechanism of the interaction between perfluoroalkyl acids (PFAAs) and peroxisome proliferator-activated receptor δ (PPARδ). Using molecular docking, binding free energy calculation, and structural analysis, we systematically investigated the binding modes, key amino acid residues, and binding energies of 20 structurally diverse PFAAs with PPARδ. The results showed that the binding energies of PFAAs with PPARδ were significantly affected by the molecular weight, the number of hydrogen bond donors, and the melting point of PFAAs. PFAAs with smaller molecular weights and fewer hydrogen bond donors showed stronger binding affinity. The binding sites were concentrated in high-frequency amino acid residues such as TRP-256, ASN-269, and GLY-270, and the interaction forces were dominated by hydrogen and halogen bonds. PFAAs with branched structure of larger molecular weight (e.g., 3m-PFOA, binding energy of −2.92 kcal·mol⁻¹; 3,3m₂-PFOA, binding energy of −2.45 kcal·mol⁻¹) had weaker binding energies than their straight-chain counterparts due to spatial site-blocking effect. In addition, validation group experiments further confirmed the regulation law of binding strength by physicochemical properties. In order to verify the binding stability of the key complexes predicted by molecular docking, and to investigate the dynamic behavior under the conditions of solvation and protein flexibility, molecular dynamics simulations were conducted on PFBA, PFOA, 3,3m₂-PFOA, and PFHxA. The results confirmed the dynamic stability of the binding of the high-affinity ligands selected through docking to PPARδ. Moreover, the influence of molecular weight and branched structure on the binding strength was quantitatively verified from the perspectives of energy and RMSD trajectories. The present study revealed the molecular mechanism of PFAAs interfering with metabolic homeostasis through the PPARδ pathway, providing a theoretical basis for assessing its ecological and health risks. Full article

Background: Hepatocellular carcinoma (HCC) remains a major cause of cancer-related mortality. Transarterial radioembolization (TARE) has emerged as a locoregional therapy to downstage tumors and expand surgical eligibility. Methods: This retrospective study included patients with HCC who underwent TARE as a bridging treatment. The primary outcomes assessed were the efficacy of TARE in facilitating curative surgery and long-term oncological outcomes, specifically overall survival (OS) and disease-free survival (DFS). Results: This study included 25 patients. 17 patients subsequently underwent surgical resection and eight underwent living-donor liver transplantation (LDLT). At a median follow-up of 33.4 months, the median disease-free survival (DFS) was 11.2 months. Patients with recurrence had a median DFS of 3.65 months, and those without recurrence had a median DFS of 27.1 months. The median overall survival (OS) for the cohort was 33.4 months. At the last follow-up, 76% of patients were alive and disease-free. Kaplan–Meier analysis demonstrated sustained OS in the LDLT group, while resection patients gradually declined within the first two years. Conclusions: TARE is an effective bridging strategy that enables curative-intent surgery in selected patients with HCC and supports favorable long-term oncological outcomes. Careful patient selection and multidisciplinary management remain essential to optimize survival benefits. Full article

Background/Objectives: Glucose Transporter 1 Deficiency Syndrome (GLUT1-DS) is a neurodevelopmental disorder caused by mutations in the gene encoding glucose transporter 1 (GLUT1), which leads to impaired glucose transport into the brain and is characterized by drug-resistant epilepsy. Limited glucose supply disrupts neuronal and astrocytic energy homeostasis, but how hypometabolism translates into network hyperexcitability remains poorly understood. Here, we used induced pluripotent stem cells (iPSCs)-derived brain organoids to examine how reduced metabolic substrate availability shapes epileptiform dynamics in human neuronal circuits from GLUT1-DS. Methods: Brain organoids were generated from a healthy donor or a GLUT1-DS patient and interfaced with multielectrode arrays (MEA) for recording of neuronal activity. A unified Python (v3.10)-based analytical pipeline was developed to quantify spikes, bursts, and power spectral density (PSD) across frequency bands of neuronal activity. Organoids were challenged with reduced glucose, pentylenetetrazol (PTZ), potassium chloride (KCl), and tetrodotoxin (TTX) to assess metabolic and pharmacological modulation of excitability. Results: GLUT1-DS organoids exhibited elevated baseline hyperexcitability compared to healthy control, characterized by increased spike rates, prolonged bursts, increased spikes per burst, and elevated PSD. Reduced glucose availability further amplified these features selectively in GLUT1-DS. Conclusions: Human brain organoids reproduce the pathological coupling between hypometabolism and hyperexcitability in GLUT1-DS. Our platform provides a mechanistic model and quantification tool for evaluating metabolic and anti-epileptic therapeutic strategies. Full article

Liver graft shortage remains a major limiting factor in contemporary liver transplantation, particularly in the setting of increasing waiting list pressure and constrained donor availability. While the biological quality of donor organs cannot be modified surgically, several operative strategies have been developed to optimize liver utilization and compensate for insufficient graft volume. These include split liver transplantation (SLT), dual-graft living donor liver transplantation (DGLT), auxiliary procedures, and selected multi-graft or hybrid configurations. This review provides an updated and structured overview of surgical concepts aimed at maximizing effective liver mass for transplantation. We discuss indications, technical considerations, and reported outcomes of split, dual, and combined graft approaches, with particular emphasis on graft-to-recipient weight ratio (GRWR), portal inflow modulation, and prevention of small-for-size syndrome. The role of machine perfusion technologies—including normothermic and hypothermic approaches—as enabling tools for graft assessment and safer utilization of partial grafts is also examined. Finally, we address ethical and logistical challenges associated with complex graft strategies and outline future directions in which advances in perfusion, graft assessment, and staged transplantation concepts may further refine patient selection and procedural safety. Collectively, these strategies represent complementary solutions for extending liver transplantation beyond conventional single-graft paradigms in highly selected settings. Full article