Search Results (34,665)

HIV reservoir size and decay represent distinct dimensions of viral persistence, yet whether they are governed by shared or separable immunological mechanisms during early antiretroviral therapy (ART) remains unclear. In this study, we employed multiparameter flow cytometry and bulk RNA sequencing to analyze longitudinal immune profiles across 21 treatment-naïve people living with HIV before ART initiation and at 1 and 5 months thereafter. Our findings revealed an apparent dissociation between HIV-1 DNA levels and decay rates in peripheral blood, and the two indicators appear to be relatively independent dimensions of viral persistence. Specifically, lower HIV-1 DNA levels were associated with higher frequencies of cytotoxic and adaptive-like natural killer (NK) cell subsets, whereas faster HIV-1 DNA decay was linked to restored HIV-specific CD4+ and CD8+ T-cell responses during treatment. Notably, transcriptomic analyses uncovered divergent gene expression signatures related to B cell-associated immunity and type I interferon pathways, with individuals with higher HIV-1 DNA levels exhibiting elevated expression of immunoglobulin and interferon-stimulated genes, while faster decay correlated with enrichment of antiviral and complement-related genes. Collectively, these findings provide a preliminary characterization of immune correlates of peripheral blood total HIV-1 DNA dynamics in the early phase following ART initiation. This work offers potential immune clues for exploring the viral reservoir and generates testable hypotheses for validation in future large cohorts. Full article

FOXP3 is the master transcriptional regulator of regulatory T cells (Tregs) and is expressed in humans as two main alternatively spliced isoforms: full-length FOXP3 (FOXP3-FL) and the exon 2-deficient variant (FOXP3-Δ2). While the role of these isoforms has been mainly studied in CD4⁺ T cells, their distribution across peripheral blood leukocyte populations and their relationship with immune checkpoint expression remain incompletely defined. In this study, we used a multiparametric flow cytometry panel allowing isoform-specific detection of FOXP3-FL and FOXP3-Δ2, together with PD-1 and CTLA-4, to analyze peripheral blood samples from six healthy donors under basal conditions. Major leukocyte populations, including CD4⁺CD25⁺ and CD4⁺CD25⁻ T cells, CD8⁺ T cells, monocytes, and neutrophils, were evaluated. FOXP3-FL predominated in CD4⁺CD25⁺ T cells, whereas FOXP3-Δ2 was more frequently detected in CD8⁺ T cells, monocytes, and neutrophils. However, the absolute frequencies of these FOXP3-Δ2-positive populations were low, consistent with the overall low levels of FOXP3 expression observed in these cell types. In CD4⁺ T-cell subsets, PD-1 expression was generally higher than CTLA-4, regardless of FOXP3 isoform, and FOXP3-Δ2⁺ cells showed relatively higher PD-1 expression compared to FOXP3-FL⁺ cells. In contrast, checkpoint expression in non-CD4⁺ populations was limited. The observed FOXP3-FL⁺/FOXP3-Δ2⁺ ratios across immune cell populations were consistent with a predominant role of FOXP3-FL in maintaining immune tolerance under basal conditions; whether these patterns are preserved or altered in pathological settings warrants further investigation. These results provide a descriptive overview of FOXP3 isoform distribution and checkpoint expression across peripheral blood immune cell subsets in healthy individuals, which may serve as a reference for future studies in immune-mediated diseases. Full article

Industrial contamination can influence the transfer of toxic and essential elements through soil–plant–animal systems and may pose risks to food safety. This study aimed to determine whether contamination patterns in soil are reflected in forage vegetation and meat products and to evaluate trace-element behavior across interconnected components of the soil–plant–animal system. This study assessed the distribution and transfer of 12 elements (As, Be, Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, V, and Zn) in soil, forage vegetation, and meat products from five industrially affected areas of Central Kazakhstan. Element concentrations were determined by inductively coupled plasma mass spectrometry. Soil contained the highest concentrations of most elements, confirming its role as the primary reservoir of contamination, whereas forage vegetation reflected local pollution patterns. The highest levels of contamination were generally observed in the industrial centers of Temirtau and Zhezkazgan, with Zhezkazgan exhibiting the most distinct element profile. Soil-to-forage transfer was most pronounced for Cd, Cu, Pb, and Zn, with significant positive relationships between soil and forage concentrations (p < 0.001). Meat products generally contained lower element concentrations than soil and forage; however, Cd, Hg, and As exceeded regulatory limits in 23 of 279 samples (8.2%). By integrating environmental and animal-derived matrices within a single framework, this study provides new insight into trace-element transfer pathways and facilitates the identification of priority contaminants, high-risk areas, and livestock products requiring enhanced environmental and food safety monitoring in industrial regions. Full article

Background/Objectives: α-lipoic acid (ALA) shows therapeutic potential but faces poor aqueous solubility (BCS Class II), gastric instability, and low oral bioavailability (~30%). This work investigated the formulation of cyclodextrin (CD) inclusion complexes of ALA to overcome the aforementioned limitations and improve nutraceutical applications. Methods: Phase solubility studies in simulated gastric and intestinal fluids screened for optimal CD, followed by molecular dynamics simulations and MM-PBSA binding free energy calculations. Inclusion complexes of choice were prepared by grinding, spray-drying, and lyophilization, followed by solid-state characterization (DSC/XRPD/FTIR). Further analysis was performed using pH-shift dissolution (USP II), permeability (PermeaPad^®, Caco-2), and (photo)stability according to ICH. Results: Hydroxypropyl-β-cyclodextrin (HPβCD) emerged as the optimal host due to favorable complexation, as confirmed by phase solubility studies and supported by molecular modeling, which revealed a favorable balance between inclusion complex stability and pH-triggered drug release. Formulations based on spray-dried and lyophilized HPβCD–ALA complexes (HPβALA-sd and HPβALA-lyo), in which ALA was fully amorphized, achieved near-complete dissolution within five minutes under biorelevant pH-shift conditions. This performance markedly exceeded that of free ALA (approximately 66% dissolution at pH 7.4) while maintaining moderate permeability (Papp 8–9 × 10⁻⁶ cm/s). Storage stability was enhanced markedly (88–90% ALA retention after 6 months at 40 °C/75% RH vs. 36% for free ALA) while UV stability was not improved through CD-complexation, probably due to interaction of UV-VIS light with the exposed portion of ALA. Conclusions: Even though the permeability of ALA–CD inclusion complexes remained medium (Papp ~ 8–9 × 10⁻⁶ cm/s) and unaffected by complexation, a significantly improved dissolution profile indicates better expected bioavailability compared to pure ALA. Full article

The Wadi Al-Ahsaba watershed is an arid to semi-arid catchment situated in southwestern Saudi Arabia, characterized by intermittent surface flow, high evaporation and low rainfall, and a dam reservoir built for flood control. The work aims to assess hydrological and anthropogenic controls on surface and groundwater quality, pollution status, and human health risks using an integrated approach of hydrogeochemical analysis, multivariable statistics, and water quality and contamination indices. A total of 21 water samples (15 surface water, 6 groundwater) were analyzed for general chemistry, major ions, and trace elements. Hydrogeochemical analysis and principal component analysis (PCA) were implemented to differentiate the geogenic from anthropogenic control on water quality. The pollution status and associated risk were evaluated using water quality index (WQI), contamination degree (Cd), Hazard Quotient (HQ), and Hazard Index (HI). Results suggest limited surface–groundwater interaction, with surface water dominated by Ca–Mg–HCO₃ facies, indicating recent recharge and limited water–rock interaction, whereas groundwater exhibits mixed Ca–Mg–Cl and Ca–Na–Cl–SO₄ types, revealing longer residence time and water–rock interaction. Nitrate (9.5–109 mg/L) and TDS (522–1003 mg/L) exceeded drinking water standards in 90% and 95% of tested samples, respectively, and WQI ranged from 43 to 134, reflecting excellent to poor water. High non-carcinogenic risk from nitrate was observed, especially for infants. The study concluded that the geogenic processes (water–rock interaction, evaporation, and mineral dissolution) control the general chemistry of tested water, while anthropogenic input from wastewater and agriculture input are likely contributors to nitrate contamination. The study contributes to the understanding of arid wadi-dam systems by revealing how limited recharge, hydrological connectivity, and episodic flow control contaminant transport and persistence, underscoring the critical role of integrated hydrological analysis and land use management in safeguarding freshwater resources in arid environments. Full article

Estuaries along South Africa’s coastline are increasingly subjected to anthropogenic pressures that disrupt their biogeochemical function and increase the risk of contamination. This study presents the first seasonal assessment of heavy metal contamination and water quality indices in the Qholora Estuary, Eastern Cape Province. Surface water samples collected during wet and dry seasons were analysed for physicochemical properties and heavy metals (As, Cd, Cu, Fe, Hg, and Pb). Multiple pollution metrics (Pollution Index (PI), Nemerow Pollution Index (NPI), Heavy Metal Evaluation Index (HEI), Heavy Metal Pollution Index (HPI)), ecological risk indices ((Ecological Risk Index (ERI), and Potential Ecological Risk Index (PERI)), and the Water Quality Index (WQI) were applied and supported by Principal Component and Cluster Analyses to identify dominant pollutant, contamination sources and seasonal hydro-geochemical controls. Results reveal strong seasonal contrasts: wet-season conditions showed elevated ionic concentrations and enhanced mobilisation of Cu, Pb, Cd, Hg, and Fe due to storm-driven runoff and sediment resuspension, while dry-season patterns reflected evapo-concentration, prolonged residence times, and pH-mediated metal partitioning. Across indices, heavy metal contamination remained low in the dry season but increased significantly in the wet season, especially for Hg, which posed moderate to considerable ecological risk at most sites, indicating emerging ecological pressure under high-flow conditions. These findings highlight a generally low risk under average conditions but a pronounced seasonally vulnerable estuarine system, underscoring the need for intensified monitoring during periods of increased runoff. The study establishes an important baseline for regional water resource management. Full article

This study focuses on the pollution assessment, potential ecological risks, influencing factors, and migration pathways of trace elements from slag heaps of coal gangue and pyrite to farmland in the mountainous areas of southeast China. Based on the pollution index and correlation analysis of trace elements, Cd, As, Pb, and Zn were identified as characteristic pollutants. In the solid waste, surrounding soil, and farmland soil, the mean concentrations of Cd, As, Pb, and Zn of pyrite slag heaps were generally 9.7–86.7 times higher than those of coal gangue dumps. In contrast, higher levels of As were found in coal gangue surrounding soil, while higher Cd and As concentrations existed in coal gangue-affected farmland soil. Mantel test results revealed significant statistical correlations between characteristic pollutants and environmental factors (geographic location, weather, and climate), particularly for pyrite slag heaps. The potential migration pathways from solid waste to the surroundings (soil and water) and then to farmland soil were finally revealed using partial least squares path modeling. This study demonstrated that the pyrite slag heaps were more heavily polluted than the coal gangue dumps. The pyrite slag heap was more susceptible to environmental factors, which could rapidly transfer trace elements to farmland soil via the surrounding soil and water. Therefore, this study offers a statistical framework to infer plausible trace element migration trends via multi-medium monitoring data. It also delivers comparative analytical references for risk assessment of two distinct types of slag heaps. Full article

Determining HIV-1 tropism provides the prognosis of HIV infection and is required before prescribing maraviroc, an entry inhibitor that blocks the interaction between the viral gp120 and the CCR5 coreceptor. However, existing prediction algorithms have been developed primarily for the globally most prevalent subtypes (B, C, and CRF01_AE) and often show reduced performance for other HIV-1 genetic variants. Sub-subtype A6 and circulating recombinant form CRF63_02A6 dominate the HIV-1 epidemic in Russia and other Former Soviet Union (FSU) countries, yet the reliability of tropism prediction for these viruses remains virtually unexplored. We phenotypically determined the tropism of 25 clinical isolates (11 R5, 1 X4, and 7 dual-tropic R5/X4) using U87.CD4.CCR5 and U87.CD4.CXCR4 cell lines and performed a comparative analysis of eight existing genotypic tools (Geno2pheno, WebPSSM, T-CUP 2.0, the Delobel/Garrido rules, and others) or their modifications on a combined dataset that included Los Alamos National Laboratory (LANL) reference sequences (subtypes A, B, C, CRF01_AE, and CRF02_AG) and our laboratory-derived isolates. Most models achieved high accuracy for globally prevalent subtypes (≈95% for B, C, and CRF01_AE) but showed markedly reduced performance for sub-subtype A6 (best accuracy among existing models, 85%) and CRF63_02A6 (best accuracy, 72%), with a poor balance between sensitivity and specificity. To address this problem, we developed HIV-V3Augur, an ensemble stacking model based on the Random Forest and Support Vector Machine (SVM) machine learning algorithms, trained on Pseudo Amino Acid Composition (PseAAC) and Relative Synonymous Codon Usage (RSCU) features with 10-fold stratified cross-validation. HIV-V3Augur achieved an accuracy of 77%, sensitivity of 79%, and specificity of 79% on sub-subtype A6, and on CRF63_02A6 it reached an accuracy of 95%, sensitivity of 87%, and specificity of 100%. Cross-validation demonstrated that HIV-V3Augur represents a balanced genotypic tropism prediction tool for understudied HIV-1 variants circulating in the FSU region. HIV-V3Augur can be used locally through a graphical user interface. Full article

Lead (Pb²⁺) and cadmium (Cd²⁺) are among the most hazardous heavy metal pollutants in wastewater owing to their high toxicity, environmental persistence, and detrimental impacts on human health and aquatic ecosystems. In this study, a novel ternary magnetic composite, SnFe₂O₄/activated carbon/polypyrrole (SnFe₂O₄/AC/PPy), was effectively synthesized and tested as an effective adsorbent in the removal of Pb²⁺ and Cd²⁺ from aqueous water. The composite was prepared by depositing spinel SnFe₂O₄ nanoparticles on activated carbon, followed by in situ polymerization of polypyrrole to enhance surface functionality and adsorption affinity. The successful fabrication of the porous SnFe₂O₄/AC/PPy hybrid composite was confirmed through FTIR, XRD, SEM–EDS, BET, XPS, and VSM characterization. The composite demonstrated a relatively high surface area (352.3 m²/g) and adequate magnetic responsiveness (12.33 emu/g), ensuring facile magnetic separation following wastewater treatment. Batch adsorption experiments showed great removal efficiency of 95.02 and 92.48% for Pb²⁺ and Cd²⁺ ions, respectively, at optimum conditions. The adsorption equilibrium data followed the Langmuir isotherm model with maximum adsorption capacities of 187.07 mg/g for Pb²⁺ and 96.45 mg/g for Cd²⁺ ions, which were attributed to monolayer adsorption on homogenous active sites. The kinetic and isothermal model indicated that the adsorption process was controlled by the combination of physical and chemical interactions. Thermodynamic parameters showed negative Gibbs free energy and enthalpy changes (ΔH° = −49.74 kJ/mol for Pb²⁺ and −38.82 kJ/mol for Cd²⁺ ions), confirming the spontaneous and exothermic nature of adsorption. Furthermore, the increasingly negative ΔG° values at lower temperatures indicated that the adsorption was thermodynamically more favorable under cooler conditions. According to the regeneration studies, the composite maintained a high removal efficiency after five consecutive cycles. In general, SnFe₂O₄/AC/PPy composite has good potential as a stable, reusable, and high-performance adsorbent to treat heavy metal wastewater. Full article

Background/Objectives: The “Land of Fires” (LF) in the Campania Region has attracted considerable attention due to massive environmental contamination deriving from decades of illegal disposal, burial, and burning of urban, industrial, and toxic waste. Cadmium (Cd) has been repeatedly proven to affect male reproductive function by a plethora of endocrine and non-endocrine mechanisms. The scientific literature is almost devoid of large studies addressing semen quality in this area, particularly by directly correlating seminal parameters to objectively measured pollutant burden in biological samples. Therefore, the aim of the current study was to comprehensively evaluate semen quality of males of reproductive age living in the LF, by correlating seminal parameters to cumulative local male reproductive tract Cd burden objectively quantified in whole semen samples. Methods: The current single-center, observational, cross-sectional study evaluated semen quality in 493 males aged 14–50 (29.07 ± 7.17) years living in three LF municipalities. Moreover, the association of semen quality with whole semen Cd (sCd) levels measured by inductively coupled plasma mass spectrometry (ICP-MS) was addressed in a subgroup of participants; semen samples suitable for semen Cd measurements were available from 383/493 (77.7%) participants of the total cohort, and all analyses involving semen Cd were performed within the measured subset. Results: In the total cohort, seminal parameters were as follows: semen pH 8.32 ± 0.3, semen volume 3.13 ± 1.67 mL, sperm concentration 37.58 ± 30.18 × 10⁶/mL, total count 111.2 ± 104 × 10⁶/ejaculate, total motility 56.83 ± 16.09%, progressive motility 50.22 ± 16.63%, in situ motility 6.72 ± 3.43%, immotile spermatozoa 43.07 ± 15.88%, normal morphology 7.97 ± 4.02%, and viability 64.75 ± 15.34%. Prevalence of normozoospermia and pathological seminal parameters was as follows: normozoospermia 66.5% (328/493), pathological seminal parameters 33.5% (165/493), specifically, oligozoospermia 14% (69/493), cryptozoospermia 0.8% (4/493), azoospermia 2.2% (11/493), asthenozoospermia 3% (15/493), teratozoospermia 0.6% (3/493), oligo-astheno-teratozoospermia 6.1% (30/493), necrozoospermia 5.7% (28/493), and different combined seminal parameters alterations 7.1% (35/493). Whole semen Cd was below (undetectable) or above (detectable) the limit of detection (LoD) (0.2 μg/L) in 66.6% (255/383) and 33.4% (128/383) whole semen samples, respectively. In samples with detectable sCd, sCd level was below or above the median value (0.76 μg/L; min–max 0.1–5.95 μg/L) in 23.4% (30/128) and 76.6% (98/128) whole semen samples, respectively. Participants with detectable sCd levels had a significantly reduced sperm total count (93.28 ± 84.88 × 10⁶/ejaculate vs. 113.2 ± 101.5 × 10⁶/ejaculate; p = 0.037), and normal morphology (7.29 ± 3.71% vs. 8.23 ± 3.91%; p = 0.034), and a significantly lower prevalence of normozoospermia (60.2% vs. 72.2%; p = 0.02) and significantly higher prevalence of pathological seminal parameters (39.8% vs. 27.8%; p = 0.02), specifically, a significantly higher prevalence of oligozoospermia (21.1% vs. 12.6%; p = 0.036) than those with undetectable sCd levels. Whole semen Cd levels were significantly higher in participants with pathological seminal parameters (1.08 ± 0.84 μg/L vs. 0.93 ± 0.74 μg/L; p = 0.037) than those with normozoospermia. Participants with sCd levels above the median value (N = 98) had a significantly reduced sperm concentration (29.12 ± 24.84 × 10⁶/mL vs. 43.62 ± 29.55 × 10⁶/mL; p = 0.015) and displayed a trend towards reduced sperm normal morphology (6.92 ± 3.38% vs. 8.55 ± 4.49%; p = 0.057) than those with sCd levels below the median value (N = 30). Moreover, participants with sCd levels above the median value (N = 98) had a significantly reduced sperm concentration (29.12 ± 24.84 × 10⁶/mL vs. 35.3 ± 26.29 × 10⁶/mL; p = 0.03), total count (85.77 ± 80.52 × 10⁶/ejaculate vs. 113.2 ± 101.5 × 10⁶/ejaculate; p = 0.008) and normal morphology (6.92 ± 3.38% vs. 8.23 ± 3.91%; p = 0.006), and a significantly lower prevalence of normozoospermia (57.1% vs. 72.2%; p = 0.008) and significantly higher prevalence of pathological seminal (42.9% vs. 27.8%; p = 0.008), specifically, a significantly higher prevalence of oligozoospermia (23.5% vs. 12.6%; p = 0.014) than those with undetectable sCd levels. Conclusions: The results of the current study demonstrate an association between the environmental Cd exposure and the impairment of seminal parameters, with a significantly poorer semen quality in participants with detectable sCd, and, more markedly, in those with sCd level above the median value, compared to participants with undetectable sCd, although subgroups comparisons highlighted a homogeneous profile in major confounders including age, BMI, and smoking habits among subgroups of participants with different sCd burden. Full article

This study aimed to obtain the lipid fraction from cocoa bean husks by applying ethyl acetate as an extraction solvent in an ultrasound-assisted extraction process. The effects of temperature (T), time (t), and solvent:husk ratio (R) on the lipid fraction (LF) yield were evaluated. The removal of minor compounds (phytosterols and tocopherols) and total phenolics was evaluated under selected conditions, as well as the value of conjugated dienes (CDs). Extraction with n-hexane was performed for comparative purposes. The prediction of the solubility of the main compounds identified in the solvents used was conducted. The influence of the variables on LF removal was T > t > R, which provided the highest result (13.54 ± 0.47 wt%) at the highest levels adopted (70 °C, 60 min, 12 g/mL), a value 23% higher than that obtained using n-hexane. Under these conditions, there is also greater recovery of minor compounds from the peels, especially β-sitosterol, which was quantified at 43 to 50% of the concentration of these compounds. The use of ethyl acetate provided greater removal of minor compounds and total phenolics, resulting in lower primary lipid oxidation products (CD value). The relationship between these properties was evidenced by the Pearson correlation matrix, especially for stigmasterol, campesterol, total phenolics, and total minor compounds. The thermodynamic modeling reveals regimes ranging from full miscibility of liquid solutes to limited solubility of phytosterols and gallic acid; however, the contrast with experimental data indicates that real extraction is limited by kinetic barriers and plant matrix effects. The solvent extractor did not influence the fatty acid profile of the LF obtained, consisting mainly of saturated fatty acids (palmitic and stearic). Full article

Background: Managing neuropathic pain (NP) is particularly challenging in the context of opioid use, and the mechanisms behind chronic pain remain unclear. Objective: This study evaluated the impact of turmeric bioactive compounds on brain regions including frontal cortex (FC), hippocampus (HPC), and hypothalamus (HPT) in the spinal nerve ligation (SNL) in a rat model of NP. Methods: Twenty-four SD rats were assigned to four groups (N = 6 per group), namely sham+vehicle (Sham-V), SNL+vehicle (SNL-V), SNL + 100 mg/kg curcumin (SNL+100CUR), and SNL + 50 mg/kg bisdemethoxycurcumin (SNL+50BDMC), treated daily for four weeks via oral gavage. Gene expression levels related to neuroinflammation, oxidative stress, and mitochondrial homeostasis were measured using qRT-PCR. Protein-level or functional mitochondrial assays were not performed due to limited sample availability. Results: In the FC, SNL decreased the expression level of NRF1 and OPA1, but only OPA1 was increased by BDMC. In the HPC, SNL increased CD11b, NRF2, and MFN1; BDMC decreased CD11b and increased IBA1, NRF1, TFAM, PGC1α and Complex I; and CUR increased NRF1, TFAM, DRP1 and Complex I levels. In the HPT, SNL decreased GFAP and MFN1, with CUR and BDMC further decreasing GFAP but not affecting MFN1. Additionally, CUR and BDMC decreased the expression of several key markers of neuroimmune signaling and mitochondrial homeostasis, including IBA1, CD11b, NFkB, NRF1/2, DRP1, OPA1, PGC1α, TFAM, and PINK1. Conclusions: CUR and BDMC induced region-specific transcriptional remodeling of mitochondrial homeostasis across FC, HPC, and HPT in SNL rats, with somewhat limited effects in the FC, mixed effects in the HPC, and broader downregulation in the HPT. Full article

Background and Clinical Significance: Cardiac and mediastinal angiosarcomas are rare, aggressive malignancies that often present with nonspecific symptoms and pose significant diagnostic challenges. Tumor heterogeneity and necrosis may lead to false-negative biopsy results; Case Presentation: We report a 64-year-old man who initially presented with cardiac tamponade of unclear etiology. Despite an extensive workup, the patient remained asymptomatic for five months before re-presenting with dyspnea and a large mediastinal mass compressing the right heart, along with a lytic rib lesion. Initial ultrasound-guided biopsy of the rib lesion demonstrated a benign vascular proliferation consistent with Masson tumor (intravascular papillary endothelial hyperplasia), which was discordant with aggressive imaging findings. Further evaluation with positron emission tomography–computed tomography (PET-CT) revealed peripheral metabolic activity, and cardiac magnetic resonance imaging (MRI) demonstrated a heterogeneous mass with central necrosis and peripheral enhancement. A repeat CT-guided biopsy targeting the metabolically active region confirmed angiosarcoma, with immunohistochemical staining demonstrating diffuse positivity for ERG, CD31, and CD34. The patient was treated with palliative radiation and paclitaxel-based chemotherapy but experienced rapid clinical decline and transitioned to comfort-focused care; Conclusions: This case highlights the importance of correlating imaging with pathology and emphasizes the risk of sampling error in necrotic tumors. PET-guided biopsy targeting viable tumor regions is essential in cases with discordant findings. Full article

CD19-directed chimeric antigen receptor (CAR) T-cell therapy has fundamentally transformed the treatment landscape for relapsed and refractory B-cell malignancies, yet antigen escape remains a persistent therapeutic challenge that limits long-term remission durability. While antigen loss is typically considered a somatic event acquired during tumor evolution under therapeutic selective pressure, germline CD19 polymorphisms could theoretically influence CAR-binding kinetics, alter epitope presentation, and modulate therapeutic outcomes in ways that remain largely not characterized. Unfortunately, Middle Eastern populations are underrepresented in pharmacogenomic databases and CAR-T clinical trials, creating a knowledge gap that may perpetuate global health disparities in access to precision immunotherapy. We analyzed publicly available whole-exome sequencing data from 1196 individuals of Arab origin to comprehensively characterize CD19 variants with potential relevance to CAR T-cell immunotherapy. The L174V (rs2904880) variant stood out, and showed the Valine/Valine (V/V) genotype frequency was 65.3%, corresponding to a V174 allelic frequency of 76.6%, while the minor allele, L174, has a frequency of 23.4%. The missense mutation (c.520C > G) responsible for this variant results in a leucine-to-valine (L174V) substitution at position 174 of the CD19 protein, relative to the reference genome. The cohort genotypes (CC, CG, and GG) exhibited a significant deviation from Hardy–Weinberg equilibrium (p < 0.00001). While this deviation is consistent with the high consanguinity rates (25–60%) amongst Arab populations, it remains not fully explained, and may be attributed to population structure, relatedness, or technical factors. We further emphasize that our computational analysis cannot establish any direct clinical or functional impact due to this variant, and therefore we refrain from suggesting any specific actions at the current time. In light of these findings, we hypothesize that the distinctive genetic architecture of consanguineous populations should not be viewed as a confounding variable. Instead, it presents a unique opportunity to investigate the clinical relevance of germline variation in the context of precision oncology, particularly at therapy-relevant loci, pending functional validation. Full article

Background: The interaction between human immunoglobulin G (IgG)1 Fc and the Fc gamma receptor (FcγR) IIIa/CD16a elicits protective immune responses. Antibody N-glycosylation stabilizes the FcγR-binding interface and is thus essential for interaction with wildtype IgG1 Fc. Furthermore, the N-glycan introduces substantial compositional and functional heterogeneity, with distinct glycoforms providing different affinities and discrete responses in vivo. Accordingly, various engineering endeavors to improve antibody binding strive to boost the therapeutic efficacy of monoclonal antibodies but do not directly address compositional heterogeneity. Objective: Here, we describe a previously unexplored approach to engineer IgG1 Fc. We eliminated carbohydrate heterogeneity by removing the N-glycan but stabilizing the FcγR-binding interface with disulfide bonds. Conclusions: These newly generated Fc domains served as a starting point for protein engineering through yeast surface display to enhance receptor-binding affinity. We recovered Fc variants from this approach that demonstrated FcγRIIIa binding affinities comparable to the starting sequence and thus serve as a proof-of-principle for this strategy. Full article