Search Results (499)

Systemic lupus erythematosus (SLE) is a severe autoimmune disease characterized by autoantibody production and multi-organ involvement. Anifrolumab, a monoclonal antibody targeting the type I interferon (IFN) receptor, has been approved for the treatment of SLE. Our aim was to investigate the long-term effects of inhibited type I IFN signaling on circulating follicular helper T subsets (T_FH), follicular regulatory T cells (T_FR), and B lymphocyte subpopulations, reflecting the ongoing germinal center reactions in SLE patients. Peripheral blood samples were obtained from ten SLE patients before the initiation of anifrolumab treatment, and at months 6 and 12 of the intervention period. Flow cytometry analysis was performed to assess the frequencies of circulating T_FH cell subsets, T_FR cells, and certain B cell subpopulations. Serological parameters, including autoantibody levels and complement components, were determined as part of the routine diagnostic evaluation. We observed a significant and sustained reduction in the percentage of activated circulating T_FH cells. Notably, the frequency of CXCR3⁻CCR6⁺ T_FH17 cells decreased, whereas the proportion of CXCR3⁺CCR6⁻ T_FH1 cells increased significantly. Furthermore, the proportion of the IgD⁻CD27⁻ double-negative B lymphocytes was also significantly reduced. These findings suggest that anifrolumab therapy attenuates T_FH cell activation, which may contribute to its clinical efficacy by modulating germinal center responses in SLE. Full article

The role of on-site wastewater treatment (OSWT) is increasingly important for water reuse and local sustainability, but treatment efficiency is highly dependent on hydraulic behavior and mixing. This study used validated CFD simulations and tracer experiments to analyze flow patterns and mixing performance in a six-zone OSWT unit under different operational scenarios, including inflow, aeration, recirculation, combined mechanisms, and closed-loop operation without inflow. The results show that influent flow is essential for maintaining convective transport and system-wide momentum, while aeration and recirculation enhance local mixing, but cannot fully overcome geometric dead zones. The combined use of inflow, aeration, and recirculation achieved the highest mixing efficiency and minimized the dead volume, whereas scenarios lacking inflow exhibited severe stagnation and expanded dead zones. These findings highlight the need to integrate hydraulic interventions with thoughtful reactor design to ensure effective and resilient small-scale wastewater treatment systems. Full article

Reproductive efficiency in pigs is regulated by hormonal pathways that control follicular development at Day 15 of the estrous cycle, during corpus luteum regression. Miniature pigs are extensively employed as human-relevant models in biomedical research, yet their reproductive characteristics during mid-luteal regression remain inadequately characterized, limiting assessments of their translational reliability. Differences in follicular morphology, hormonal signaling, and vascular development may underlie their lower fertility compared to conventional pigs. In this study, follicular development after corpus luteum formation was compared between conventional pigs and minipigs using histological staining, immunofluorescence, hormonal assays, and transcriptomic profiling. The expression of VEGF, mTOR, LH, FSH, PAPP-A, and apoptosis markers was evaluated across the granulosa and thecal regions. Differential gene expression was analyzed using microarray data followed by GO categorization. Minipigs exhibited smaller follicles, reduced vascularization, and lower VEGF and MMP activity compared to conventional pigs. Expression of LH and PAPP-A was higher in conventional pigs, while minipigs showed relatively elevated E2 and FSH levels. Transcriptomic data revealed greater upregulation of cell-survival- and angiogenesis-related genes in conventional pigs, including genes involved in IGF pathways. Apoptosis and poor extracellular matrix remodeling were more pronounced in minipigs. Minipigs demonstrated impaired follicular remodeling and weaker hormonal signaling after corpus luteum formation, which likely contributed to their reduced reproductive efficiency. Understanding these species differences can guide breeding strategies and fertility management in biomedical and agricultural settings. Full article

Background/Objectives: The oral cavity represents a convenient route of administration for drugs that exhibit significant hepatic first-pass extraction. In this study, the mucosal permeation properties of selected active pharmaceutical ingredients (APIs) incorporated into oral cavity drug products that are approved by the U.S. Food and Drug Administration were quantified using the human-derived sublingual HO-1-u-1 and buccal EpiOral™ in vitro tissue models. Methods: Epithelial barrier properties were monitored using propranolol and Lucifer Yellow as prototypic transcellular and paracellular markers. APIs were dissolved in artificial saliva, pH 6.7, and transepithelial flux from the apical to the basolateral compartment was quantified using HPLC. Results: Apparent permeability coefficients (Papp) calculated for these APIs in the sublingual HO-1-u-1 tissue model varied from Papp = 2.72 ± 0.06 × 10⁻⁵ cm/s for asenapine to Papp = 6.21 ± 2.60 × 10⁻⁵ cm/s for naloxone. In contrast, the buccal EpiOral™ tissue model demonstrated greater discrimination power in terms of permeation properties for the same APIs, with values ranging from Papp = 3.31 ± 0.83 × 10⁻⁷ cm/s for acyclovir to Papp = 2.56 ± 0.68 × 10⁻⁵ cm/s for sufentanil. The tissue-associated dose fraction recovered at the end of the transport experiment was significantly increased in the buccal EpiOral™ tissue model, reaching up to 8.5% for sufentanil. Conclusions: Experimental permeation data collected for selected APIs in FDA-approved oral cavity products will serve as a training set to aid the development of predictive computational models for improving algorithms that describe drug absorption from the oral cavity. Following a robust in vitro–in vivo correlation analysis, it is expected that such innovative in silico modeling strategies will the accelerate development of generic oral cavity products by facilitating the utility of model-integrated evidence to support decision making in generic drug development and regulatory approval. Full article

Background/Objectives: Non-pancreatic hyperlipasemia (NPHL) is associated with high in-hospital mortality, with sepsis being one of the most common etiologies. The prognostic value of hematological parameter-derived inflammatory scores has not been extensively studied in NPHL to date. Methods: The prognostic value of eight inflammatory scores for in-hospital mortality was assessed in a total of 545 NPHL patients from two hospitalized patient cohorts (COVID-19 [n = 144] and non-COVID-19 [n = 401], the latter stratified as bacterial sepsis [n = 111] and absence of systemic infection [n = 290]). We assessed the neutrophil-to-lymphocyte ratio (NLR), derived NLR (dNLR), neutrophil-to-lymphocyte and platelet ratio (N/(LP)), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), aggregate index of systemic inflammation (AISI), systemic inflammation index (SII), and systemic inflammation response index (SIRI), comparing their prognostic value among etiological groups. Results: Patients with bacterial sepsis were older, had more comorbidities, and experienced worse outcomes, including longer hospitalization (median: 15, 7, and 11 days; p < 0.001), higher ICU admission rates (75.7%, 33.8%, and 47.9%, p < 0.001), and increased mortality (45.0%, 13.8%, and 38.2%, p < 0.001), compared to those without systemic infection or with COVID-19-induced NPHL. Overall, NLR, dNLR, and N/(LP) were the most accurate predictors of in-hospital mortality at admission (AUROC: non-infection: 0.747; 0.737; 0.772; COVID-19: 0.810; 0.789; 0.773, respectively). The accuracy of NLR decreased in bacterial sepsis, and only N/(LP) and PLR remained associated with in-hospital mortality (AUROC: 0.653 and 0.616, respectively). Conclusions: The prognostic performance of hematological parameter-derived inflammatory scores in NPHL is etiology-dependent. NLR is the most accurate prognostic tool for mortality in the absence of bacterial sepsis, while N/(LP) is the best score in sepsis-induced NPHL. Full article

Placenta accreta spectrum (PAS) and placenta previa (PP) are severe obstetric disorders associated with high maternal and perinatal morbidity. Early diagnosis of both conditions remains challenging, particularly in cases with subtle imaging findings. This study was aimed to evaluate the diagnostic value of first-trimester maternal serum levels of pregnancy-associated plasma protein-A (PAPP-A) and free beta subunit of human chorionic gonadotropin (β-hCG) in predicting PAS and PP. In this retrospective case–control study, a total of 100 pregnant women were included: 36 with PAS, 32 with PP, and 32 healthy controls. Serum levels were measured at 11–13⁶ weeks of gestation. Both biomarkers were significantly altered in pathological groups compared to controls: PAPP-A was lower in PP (3.04 [1.42–4.52] IU/L) and PAS (3.63 [2.51–5.39] IU/L) vs. controls (5.34 [3.72–8.41] IU/L; p < 0.001), while β-hCG was higher in PP (45.4 [40.1–54.9] IU/L) and PAS (51.4 [32.3–74.8] IU/L) vs. controls (33.5 [22.7–54.1] IU/L; p = 0.044 and p < 0.001, respectively). ROC analysis demonstrated that combined biomarker modeling improved diagnostic accuracy over single-marker use, with AUCs reaching 0.85 (sensitivity 85.2%, specificity 72%) for PAS and 0.88 (sensitivity 100%, specificity 72%) for PP. These findings support the integration of biochemical screening into first-trimester risk assessment protocols. Incorporating maternal serum biomarkers may enhance early identification of high-risk pregnancies, allow timely referral to specialized care, and reduce adverse outcomes. Further prospective studies are warranted to validate the utility of this dual-marker approach across diverse populations and clinical settings. Full article

Ruxolitinib, a clinically approved JAK1/2 inhibitor used in the treatment of hematologic malignancies and inflammatory conditions, has been shown to interfere with the function of cytotoxic T lymphocytes (CTLs). Previous studies supported the involvement of the multidrug resistance transporter P-glycoprotein (Pgp/ABCB1) in CTL biology; however, the nature of its regulation remains unclear. To address this, we investigated the impact of ruxolitinib on Pgp expression and function in human CD8⁺ T cells. We demonstrate that CD8⁺ T lymphocytes express Pgp dynamically at both the mRNA and protein levels across naïve, short-term, and long-term activation states. Ruxolitinib increased the calcein accumulation in human Pgp-overexpressing NIH-3T3 cells and in CTLs and directly modulated Pgp function by increasing its basal ATPase activity in a concentration-dependent manner (10–100 μM), similar to the effect of the known Pgp substrate/modulator verapamil. Although measurable ATPase stimulation and transport inhibition were observed at supratherapeutic concentrations of ruxolitinib, its Pgp-mediated efflux may also occur at therapeutically relevant concentrations. In contrast, at therapeutically relevant plasma concentrations (1–3 μM), ruxolitinib significantly stabilized the mRNA expression of Pgp during early T-cell receptor (TCR) activation and inhibited the TCR-induced upregulation of Pgp, CD8, and PD-1 surface markers, suggesting its interference with activation-associated differentiation. At these same concentrations, ruxolitinib also impaired CCL19-directed transmigration of CTLs across human umbilical vein endothelial cell (HUVEC) monolayers, indicating disruption of lymphoid homing cues. Collectively, these findings demonstrate that ruxolitinib modulates Pgp at both the transcriptional and functional levels, with distinct concentration dependence. The ability of ruxolitinib to alter CTL activation and migration at clinically relevant plasma concentrations highlights the need for careful evaluation of JAK inhibitor–mediated immunomodulation and its implications for vaccination, transplantation, and T cell-based immunotherapies. Full article

Objectives: Ex vivo nasal mucosa models provide physiologically relevant platforms for evaluating nasal drug permeability; however, their application is often limited by high experimental variability and the absence of standardized methodologies. This study aimed to improve experimental design by addressing two major limitations: the confounding effects of mucosal thickness and the questionable reliability of fluorescein sodium (Flu-Na) as an integrity marker for chemically induced mucosal injury. Methods: Permeability experiments were conducted using porcine nasal tissues mounted in Franz diffusion cells, with melatonin and Flu-Na as model compounds. Tissues of varying thickness were collected from both intra- and inter-individual sources, and a numerical simulation-based method was employed to normalize apparent permeability coefficients (Papp) to a standardized mucosal thickness of 0.80 mm. The effects of thickness normalization and chemically induced damage were systematically evaluated. Results: Thickness normalization substantially reduced variability in melatonin Papp, particularly within same-animal comparisons, thereby improving statistical power and data reliability. In contrast, Flu-Na exhibited inconsistent correlations across different pigs and failed to reflect the expected increase in permeability following isopropyl alcohol (IPA)-induced epithelial damage. These results suggest that the relationship between epithelial injury and paracellular transport may be non-linear and not universally applicable under ex vivo conditions, limiting the suitability of Flu-Na as a standalone marker of mucosal integrity. Conclusions: The findings highlight the importance of integrating mucosal thickness correction into standardized experimental protocols and call for a critical reassessment of Flu-Na in nasal drug delivery research. Full article

Background/Objectives: The aim of this study is to develop a hybrid artificial intelligence (AI) approach to improve the accuracy, efficiency, and reliability of Down Syndrome (DS) risk prediction during first trimester prenatal screening. The proposed method transforms one-dimensional (1D) patient data—including features such as nuchal translucency (NT), human chorionic gonadotropin (hCG), and pregnancy-associated plasma protein A (PAPP-A)—into two-dimensional (2D) Aztec barcode images, enabling advanced feature extraction using transformer-based deep learning models. Methods: The dataset consists of 958 anonymous patient records. Each record includes four first trimester screening markers, hCG, PAPP-A, and NT, expressed as multiples of the median. The DS risk outcome was categorized into three classes: high, medium, and low. Three transformer architectures—DeiT3, MaxViT, and Swin—are employed to extract high-level features from the generated barcodes. The extracted features are combined into a unified set, and dimensionality reduction is performed using two feature selection techniques: minimum Redundancy Maximum Relevance (mRMR) and RelieF. Intersecting features from both selectors are retained to form a compact and informative feature subset. The final features are classified using machine learning algorithms, including Bagged Trees and Naive Bayes. Results: The proposed approach achieved up to 100% classification accuracy using the Naive Bayes classifier with 1250 features selected by RelieF and 527 intersecting features from mRMR. By selecting a smaller but more informative subset of features, the system significantly reduced hardware and processing demands while maintaining strong predictive performance. Conclusions: The results suggest that the proposed hybrid AI method offers a promising and resource-efficient solution for DS risk assessment in first trimester screening. However, further comparative studies are recommended to validate its performance in broader clinical contexts. Full article

Oral administration of dipyridamole (DIP) with acetylsalicylic acid (ACA) is recommended in thromboembolic conditions or for the treatment of myocardial infarction and stroke. The present study presents an alternative dosage form of these two active ingredients, consisting of a honey core and a dark chocolate coating. The composition masks the bitter taste, is palatable and ensures compliance of a wide range of patients, mainly pediatric. For the simultaneous quantitative determination of the analytes, a Diode Array Detector/Fluorescence Detector (HPLC-DAD/FLD) method was used with a C18 column (250 mm × 4.6 mm, 5 μm) and an isocratic two-phase system (A: H₂O 0.2% formic acid—B: Acetonitrile-H₂O 90:10 v/v) 65:35 v/v. The method was validated according to ICH guidelines (r² > 0.999, RSD < 2.3%, % Recovery > 95.4%), and a stability study of the two active ingredients as well as salicylic acid (SAL), which is a hydrolysis product of ACA, was followed. Finally, a digestion protocol (oral cavity–stomach–intestine) for edible materials was applied to determine the release rate of ACA, DIP and SAL in the gastrointestinal tract, while an in vitro permeability study (P_app) was subsequently performed in Franz cells. The results show satisfactory behavior of ACA and DIP and provide a trigger for further studies of the formulation. Full article

Background: The 2023 ESC Heart Failure (HF) Guidelines recommend the rapid up-titration of guideline-directed medical therapy (GDMT) for all patients after HF hospitalisation. Real-world data on the implementation of a rapid up-titration programme (RTP) are scarce. Methods: We aimed to summarise the primary experiences of a six-week RTP in a multicentre observational study of five cardiology centres, evaluating the GDMT applied and the target doses (TDs) achieved during the RTP. The safety of RTP in relation to exceeding the “safety indicators” used in the STRONG-HF trial and any serious adverse events were observed. Changes in the left ventricular ejection fraction (LVEF) after RTP were evaluated. Results: Among the 90 consecutive patients (age: 56 [49–63] years, HFrEF: 96%, NT-proBNP at discharge: 1390 [735–2835] pg/mL; continuous variables are presented as median and interquartile ranges, while categorical variables are shown as absolute numbers and percentages, respectively), a remarkable proportion of patients received GDMT at hospital discharge; however, target doses were rarely achieved (RASi: 100%, TD RASi: 11%; βB: 97%, TD βB: 6%; MRA: 99%, TD MRA: 82%; SGLT2i: 98%, TD SGLT2i: 98%; triple therapy [TT: RASi + βB + MRA]: 96%, TD TT: 2%, quadruple therapy [QT: RASi + βB + MRA]: 94%, TD QT: 2%). After the six-week RTP, 100% of the total cohort (TC) were receiving RASi; 99–99–99% were receiving βB, MRA, and SGLT2i medications; and altogether, 98–98% were on TT and QT. In total, 78–78% of the patients received ≥50% of the TDs of TT and QT, while 51–51% of the TC were on TDs of TT and QT. During the RTP, no serious adverse events were observed. Between two and four months after the RTP, 51% of HFrEF patients evolved to the HFimpEF category. Conclusions: The present multicentre, observational study confirms that RTP is feasible and safe in real-world clinical practice, leading to a remarkably large proportion of patients receiving GDMT by the end of the six-week RTP, resulting in a significant increase in LVEF. Full article

A Lipophilic Prodrug Charge Masking (LPCM) strategy involves masking of hydrophilic peptide charges with alkoxycarbonyl groups, which are cleaved by esterases after intestinal absorption. This study investigates the LPCM strategy’s applicability to oxytocin (OT), a peptide with well-defined biological activity. A series of OT prodrugs with varying alkoxycarbonyl chain lengths (2 to 12 carbon atoms) were synthesized, and their permeability was assessed using parallel artificial membrane permeability assay (PAMPA) and Caco-2 cell culture models. The PAMPA results indicated that OT demonstrated poor permeability (P_app = 2.2 × 10⁻⁶ cm/s), while its prodrugs Hoc-OT, Oct-OT, and Dec-OT were characterized by significantly better permeability, with Dec-OT achieving a four-fold increase over OT. The prodrug with a 12-carbon chain (Dod-OT) exhibited poor permeability; however, its high mass retention suggests strong membrane affinity. Further evaluation, using the Caco-2 cell model, revealed a 1.8-fold higher P_app of Oct-OT compared to OT, indicating possible higher oral availability. Conversely, Hoc-OT exhibited lower permeability than OT. Our findings indicate that the LPCM strategy can effectively boost the oral bioavailability of certain peptides, paving the way for their transformation into bioavailable drugs. Full article

This paper presents a novel approach for screening women in their first trimester of pregnancy to identify those at high risk of having a child with Down syndrome (DS), using machine learning algorithms. Various machine learning models, including statistical, linear, and ensemble models, were trained using a pseudo-anonymized dataset of 90,532 screening patients. This dataset, containing less than 1% positive cases, was obtained from Cruces University Hospital, a public health hospital (Osakidetza) in Baracaldo, Basque Country, Spain. The models incorporate a set of input variables, including demographic variables such as maternal age, weight, ethnicity, smoking status, and diabetes status, as well as laboratory variables like nuchal translucency (NT), pregnancy-associated plasma protein-A (PAPP-A), and beta-human chorionic gonadotropin hormone (B-HCG) levels. The trained classification algorithms achieved ROC-AUC values between 0.970 and 0.982, with sensitivity and specificity of 0.94. The results indicate that machine learning techniques can effectively predict Down syndrome risk in first-trimester screening programs. Full article

The impact of short-term feeding of three distinct diets containing insect meals on the intestinal microbiota of African catfish hybrid (Clarias gariepinus × Heterobranchus longifilis) juveniles was examined. The animals received experimental diets containing 30% insect meals derived from black soldier-fly larvae (BSL) (Hermetia illucens), yellow mealworm (Tenebrio molitor) or blue bottle-fly larvae (Calliphora vicina) for 18 days. The relative abundance of the Bacillaceae, the Planococcaceae and other bacteria significantly increased (p < 0.05) in the intestinal microbiota of the BSL group and also in the pooled group of the three catfish groups that received insect meals. Several strains of the Bacillales cultured from BSL feed had higher (p < 0.05) abundance in the intestinal microbiota of the BSL group compared to those of the control group. Among these Bacillales strains, a single fosB antibiotic resistance gene was identified. In the gut resistomes of both the BSL and the control catfish groups, the tetA(P), tetB(P) and lnu(C) antibiotic-resistance determinants were detected, while fosB was detected only in the BSL group. Overall, the study showed that a short-term shift to diets containing insect meals can induce significant (q < 0.05) changes in the gut microbiota of the African catfish without the development of reduced α-diversity and without the overgrowth of bacteria pathogenic to fish. Full article

This study explores alternative methods to combat bacterial infections like fire blight caused by Erwinia amylovora (Ea) using bacteriophages as potential antimicrobial agents. Two lytic phages, Ea PF 7 and Ea PF 9, were isolated from apple samples and classified as Loessnervirus-like based on their genomes. Both phages showed strong efficacy, lysing 95% of the tested 37 Ea strains. They inhibited bacterial growth for up to 10 h, even at low infection rates. The phages had a short latent period of 10 min and produced high burst sizes of 108 and 125 phage particles per infected cell. Stability tests revealed that both phages were stable at moderate temperatures (37–45 °C) and within a pH range of 4–10. However, their viability decreased at higher temperatures and extreme pH levels. Both phages exhibited notable desiccation tolerance and moderate resistance to UV-B radiation during UV testing. The phages were exposed to carefully controlled irradiation, considering factors like lamp type, radiation intensity, exposure time, and object distance. This method introduces a complex approach to research, ensuring repeatable and comparable results. These findings suggest that Ea PF 7 and Ea PF 9 hold promise as antimicrobial agents for therapeutic and biotechnological applications, potentially helping to combat antibiotic resistance in the future. Full article