Search Results (98)

Non-specific lipid transfer proteins (nsLTPs) constitute a widely distributed family of plant allergens with substantial clinical relevance, particularly in food allergy. Their marked thermal and proteolytic stability enables them to provoke reactions ranging from mild local symptoms to severe anaphylaxis. This narrative review synthesises current knowledge on nsLTP allergens, focusing on their molecular characteristics, taxonomic distribution, exposure routes, and clinical impact. Major allergenic sources include fruits, nuts and seeds, vegetables and cereals, as well as various pollens. Across these sources, Pru p 3 has emerged as the central and most extensively studied allergen, frequently acting as the primary sensitiser and exhibiting broad cross-reactivity with homologous nsLTPs from diverse plant species. Despite growing evidence, significant knowledge gaps remain regarding sensitisation pathways, environmental modifiers, and phenotype stratification. Continued research is required to improve diagnostic precision and guide the development of targeted therapeutic strategies for patients with nsLTP-mediated allergy. Full article

Background: The pathophysiological mechanism of eosinophilic esophagitis (EoE) is complex and is still being investigated. We believe that there is a group of patients with eosinophilic esophagitis which could be differentiated as having an allergic phenotype who exhibit a sensitization profile (aeroallergens, panallergens, foods and specific IgG4 levels) with significant differences compared to patients with conventional allergic disease without associated eosinophilic esophagitis and healthy controls. Method: We measured the prevalence of sensitization to aeroallergens, foods and panallergens by means of molecular diagnostic techniques (ImmunoCAP^TM ISAC) and determined the levels of specific IgG4 against foods and eosinophilic-derived neurotoxin (EDN) (ImmunoCAP technology) in patients with EoE of an allergic phenotype to study whether there are statistically significant differences with respect to the control groups (patients with different allergic pathologies without EoE and healthy patients without documented allergies). The total number of patients under study was 118, distributed among the different study groups. The case group (Allergic phenotype EoE patients) had 48 subjects. The food and respiratory allergy control groups had 30 subjects each. Finally, we included 10 in the healthy control group. Results: We were able to identify statistically significant differences when comparing levels of food-specific IgG4. Milk, egg, wheat, nuts, soy, cod, and Pru p3/LTP stood out. We did not observe significant differences in relation to sensitization to aeroallergens, foods, or panallergens. We also did not observe differences in EDN levels. Conclusions: We present a study in which statistically significant differences in IgG4 levels were observed in response to different types of food, comparing patients with eosinophilic esophagitis of allergic phenotype (case group) against subjects with allergic pathology without EoE and healthy subjects (control groups). Determining whether the detected foods are clinically relevant or not in these patients would be fundamental to establishing their usefulness as a treatment alternative in our patients. Full article

The highly efficient endodyogeny of tachyzoites is a key process driving acute infection by Toxoplasma gondii. The centrocone is a specialized and critical structure for parasite cell division, but the regulatory mechanisms of centrocone proteins in T. gondii remain poorly understood. In this study, we characterized the centrocone protein SMC_N1, which exhibited periodic expression in tachyzoites, peaking during the synthesis phase. Conditional depletion of SMC_N1 was achieved in the type I RH strain and type II cyst-forming PRU strain using the mAID system combined with CRISPR-Cas9. Depletion of SMC_N1 disrupted IMC assembly, endodyogeny and nuclear division, as well as the stable inheritance of the apicoplast and centrosome, resulting in severe defects in intracellular replication and impaired tachyzoite growth. Collectively, these results indicate that SMC_N1 regulates cell division by coordinating organelle inheritance and cytoskeletal dynamics, ensuring proper replication of T. gondii tachyzoites and provide insights into mechanisms controlling parasite proliferation. Full article

Oocysts of Toxoplasma gondii exhibit remarkable resistance to environmental stressors and most conventional disinfectants. Despite its ability to infect a wide variety of host species, sexual reproduction and oocyst formation occur exclusively within felid definitive hosts. Despite the epidemiological significance of oocyst-mediated transmission, the molecular mechanisms governing oocyst production and sporulation remain incompletely understood. Glutaredoxin, serving as a central regulator of cellular redox homeostasis and multiple vital cellular processes in cells, is a potential regulator for oocyst sporulation. Here, we investigated the role of TGME49_227100 (glutaredoxin 5, Grx5) in the T. gondii Pru strain-a type II strain capable of oocyst formation, with a particular focus on its functions during oocyst formation and sporulation. We found that Grx5-knockout tachyzoites exhibited no defects in growth or virulence. Neither in vitro nor in vivo tachyzoite-to-bradyzoite differentiation was affected compared to wild-type parasites. Notably, Grx5 deletion significantly reduced oocyst production in cats by approximately 70%. Additionally, the collected oocysts showed a 50% decrease in sporulation rate. These results indicate that Grx5 plays a predominant role within feline host and the external environmental stage of sporulation, which of these is likely to provide a crucial molecular target for developing a transmission-blocking vaccine. Full article

High platelet reactivity (HPR) in patients with coronary artery disease receiving P2Y12 inhibitors increases ischemic risk. Chronic kidney disease (CKD) is an established contributor to HPR during clopidogrel therapy. The objective of the study was to assess whether CKD influences platelet reactivity (PR) in patients treated with clopidogrel or ticagrelor. This double-blind, double-dummy study enrolled 106 stable patients more than one year after an acute coronary syndrome, with or without CKD. Participants were matched by age and sex and randomized to clopidogrel or ticagrelor. PR was measured using the VerifyNow™ P2Y12 assay, and HPR was defined as P2Y12 reaction units (PRU) ≥ 208. Median glomerular filtration rates were 80 mL/min/1.73 m² in non-CKD patients and 41 mL/min/1.73 m² in CKD patients (p < 0.01). Ticagrelor produced similarly low PR in both groups (36 vs. 35 PRU; p = 0.61). Clopidogrel resulted in a numerically higher PR in CKD patients (209 vs. 180 PRU; p = 0.07). The magnitude of PR reduction with ticagrelor relative to clopidogrel was greater in CKD patients (p-interaction = 0.09). HPR was markedly more common with clopidogrel, particularly in CKD (difference 37%; adjusted OR 4.42; p = 0.01). In conclusion, CKD significantly impairs clopidogrel responsiveness but does not affect ticagrelor, resulting in a greater relative advantage of ticagrelor in patients with CKD. Full article

Sensitization to non-specific lipid transfer proteins (nsLTPs) is highly prevalent in Mediterranean countries. Pru p 3 from peach is a major allergen responsible for IgE-mediated food allergies. As a panallergen, Pru p 3 shows high sequence homology with nsLTPs from other Rosaceae fruits but also from botanically unrelated sources, including nuts and pollens, leading to extensive cross-reactivity complicating diagnosis and management. Given the worldwide prevalence of peanut and tree nut allergies, this study aimed to investigate sensitization patterns in Pru p 3-sensitized patients with tree nut allergy, using artificial intelligence (AI) to identify predictors of clinical reactivity and severity. Data from Pru p 3–sensitized patients with symptoms to peach and/or nuts were analyzed. Sensitization profiles were modeled using an XGBoost algorithm to explore associations with symptoms and severity. Patients sensitized to Pru p 3 and symptomatic for peach and nuts showed predominant sensitization to peanut and hazelnut, but AI revealed stronger associations between clinical reactivity and sensitization to hazelnut, walnut, and almond. Among patients with nut allergy and peach-asymptomatic, peanut and hazelnut sensitization were most frequent, while peach-symptomatic ones, walnut and almond sensitization predominated. Overall, walnut sensitization emerged as the main predictor of clinical severity and increasing number of sensitizations correlated with higher severity. The XGBoost algorithm identified specific allergen combinations associated with symptoms and severity, highlighting walnut sensitization as the strongest severity predictor. Machine learning approaches represent a promising tool for refining risk stratification and personalizing management in nsLTP-related food allergy. Full article

Allergen sensitization profiles are increasingly affected by environmental and climate changes. This study exemplifies fundamental differences in molecular IgE sensitization profiles in two nearby regions in Argentina with different climatic conditions (La Plata and Bahía Blanca). A cross-sectional study was conducted involving 155 patients with allergic symptoms from La Plata and Bahía Blanca (34.0 ± 11.2 years, female/male: 83/72). Serum samples were analyzed for IgE reactivity using a chip containing 101 micro-arrayed allergen molecules. Statistical analyses were performed to compare allergen-specific IgE levels, sensitization prevalences and reported symptoms. Patients from La Plata—with subtropical weather—showed a higher prevalence of IgE reactivity to house dust mite (HDM) allergens (Der p 23: 74%; Der p 1: 53% and Der p 2: 56%) and more frequently reported asthma (AS) symptoms (40% vs. 24%) than patients from Bahía Blanca. In contrast, patients from Bahía Blanca, with dry and windy weather, exhibited higher sensitization rates to pollen allergens, particularly Phl p 1 (49%) and Ole e 1 (22%) as well as to Alternaria alternata (Alt a 1, 35%) and reported a significantly higher prevalence of skin manifestations (54% vs. 31%) than those from La Plata. Cat allergen Fel d 1 was an equally important sensitizer in both regions (La Plata 30% and Bahía Blanca 37%). Sensitization to class 1 food allergens was rare in both groups (1–8%), including non-specific lipid transfer proteins (peanut Ara h 9 and peach Pru p 3) but IgE sensitizations to genuine peanut allergens were almost absent. Important regional differences in allergen sensitization profiles were observed between two geographically close regions with different climatic conditions. Our findings underscore the relevance of region-specific allergen profiling and highlight the clinical utility of molecular allergy diagnosis for a more precise allergen identification and improved management of allergic diseases. Full article

Background/Objectives: Lipid transfer protein (LTP) syndrome is a leading cause of primary food allergy in Mediterranean countries, often associated with severe reactions. Due to in vitro cross-reactivity among plant foods, clinical manifestations are unpredictable, frequently requiring multiple oral food challenges (OFC) to assess nut tolerance. These procedures increase healthcare burden and patient anxiety. This study evaluated the safety and utility of a mixed-nut oral food challenge (MixNut) in LTP-sensitized patients. Methods: In this prospective observational multi-center study, patients with LTP syndrome were enrolled. Group A included individuals allergic to fruits or vegetables who had avoided nuts; Group B included patients with suspected or confirmed nut allergy. Participants underwent a MixNut challenge comprising 2–4 nuts (≥3 g protein per nut). Results: Nineteen patients (73.7% male; median age 32.5 years) underwent MixNut, testing 52 individual nuts. All challenges were negative. The MixNut approach reduced the number of OFC by 63% (from 52 to 19) and total testing time from 208 to 76 h. Specific IgE levels to LTP allergens (Pru p 3, Cor a 8, Ara h 9) varied widely and did not predict clinical reactivity. Conclusions: MixNut is an efficient diagnostic tool for LTP syndrome, significantly reducing testing time, costs, and patient burden. It facilitates accurate dietary management and prevents unnecessary food avoidance. Further studies should optimize MixNut protocols and identify predictive markers for clinical reactivity. Full article

Typical fertilizer applicators are often restricted in performance due to non-uniformity in distribution, required labor and time intensiveness, high discharge rate, chemical input wastage, and fostering weed proliferation. To address this gap in production agriculture, an automated variable-rate fertilizer applicator was developed for the cotton crop that is based on deep learning-initiated electronic control unit (ECU). The applicator comprises (a) plant recognition unit (PRU) to capture and predict presence (or absence) of cotton plants using the YOLOv7 recognition model deployed on-board Raspberry Pi microprocessor (Wale, UK), and relay decision to a microcontroller; (b) an ECU to control stepper motor of fertilizer metering unit as per received cotton-detection signal from the PRU; and (c) fertilizer metering unit that delivers precisely metered granular fertilizer to the targeted cotton plant when corresponding stepper motor is triggered by the microcontroller. The trials were conducted in the laboratory on a custom testbed using artificial cotton plants, with the camera positioned 0.21 m ahead of the discharge tube and 16 cm above the plants. The system was evaluated at forward speeds ranging from 0.2 to 1.0 km/h under lighting levels of 3000, 5000, and 7000 lux to simulate varying illumination conditions in the field. Precision, recall, F1-score, and mAP of the plant recognition model were determined as 1.00 at 0.669 confidence, 0.97 at 0.000 confidence, 0.87 at 0.151 confidence, and 0.906 at 0.5 confidence, respectively. The mean absolute percent error (MAPE) of 6.15% and 9.1%, and mean absolute deviation (MAD) of 0.81 g/plant and 1.20 g/plant, on application of urea and Diammonium Phosphate (DAP), were observed, respectively. The statistical analysis showed no significant effect of the forward speed of the conveying system on fertilizer application rate (p > 0.05), thereby offering a uniform application throughout, independent of the forward speed. The developed fertilizer applicator enhances precision in site-specific applications, minimizes fertilizer wastage, and reduces labor requirements. Eventually, this fertilizer applicator placed the fertilizer near targeted plants as per the recommended dosage. Full article

Toxoplasma gondii is a globally prevalent parasite capable of establishing lifelong infections, which can have severe consequences in immunocompromised individuals and developing fetuses. GRAs are essential secretory effectors that facilitate nutrient acquisition, modulate host immune responses, and support intracellular survival. In this study, we characterized four putative GRAs (GRA85–88) that co-localize with GRA12 in both tachyzoite and bradyzoite stages. Using CRISPR-Cas9-mediated homologous recombination, we successfully generated knockout strains in both type I RH and type II Pru backgrounds. Phenotypic analysis revealed that GRA85, GRA87, and GRA88 were not individually required for parasite replication, invasion, or virulence. However, deletion of gra86 (PruΔgra86) resulted in a significant reduction in virulence and fewer brain cysts in chronically infected mice, although in vitro growth remained unaffected. Transcriptomic profiling of PruΔgra86 revealed downregulation of bradyzoite–related genes and upregulation of GRAs involved in host interaction. Additionally, in vitro differentiation assays showed impaired bradyzoite development in the absence of GRA86. These findings from murine models and in vitro phenotypic assays highlight GRA86 as a regulator of chronic infection and stage conversion, positioning it as an important player in T. gondii pathogenesis and a promising target for therapeutic intervention. Full article

Background: The peach allergen Pru p 7, a member of the Gibberellin-Regulated Protein (GRP) family, has emerged as a key marker of severe fruit-induced allergies. It is hypothesized to mediate cross-reactivity between fruit allergens and cypress pollen. Given the increasing prevalence of food allergies and the complex patterns of cross-sensitization, the role of Pru p 7 in clinical allergy diagnostics warrants further investigation. Objective: This study aims to characterize the sensitization profile to Pru p 7 in a cohort of patients with suspected fruit allergy and to assess its relationship with cypress pollen allergy, particularly to Cup s 7, a homologous GRP from Cupressus sempervirens. Methods: A retrospective analysis was conducted on 20 patients evaluated at the Allergy Unit of the Fondazione IRCCS Policlinico San Matteo. Specific IgE (sIgE) levels to peach extract, Pru p 7, and Cup a 1 (cypress extract) were assessed using the ImmunoCAP^® system (Thermo Fisher Scientific Inc., Waltham, MA, USA). Statistical associations between sensitizations were evaluated using chi-square tests and Spearman’s correlation. Results: Sensitization to peach extract, Pru p 7, and cypress pollen was detected in 38%, 30%, and 45% of patients, respectively. Significant associations were observed between peach and cypress (χ² = 8.80, p = 0.003), peach and Pru p 7 (χ² = 8.23, p = 0.004), and cypress and Pru p 7 (χ² = 6.55, p = 0.01). Notably, all patients sensitized to Pru p 7 also tested positive for both peach and cypress allergens, supporting the hypothesis of pollen–food cross-reactivity. Conclusions: Pru p 7 is a clinically relevant allergen that may account for severe allergic responses in patients not sensitized to classical peach allergens. Its cross-reactivity with Cupressaceae-derived GRPs, such as Cup s 7, highlights the importance of molecular diagnostics in evaluating food allergies, particularly in regions with significant exposure to cypress pollen. Full article

Abscisic acid (ABA) is a key phytohormone involved in regulating plant growth and responses to environmental stress. As receptors of ABA, pyrabactin resistance 1 (PYR)/PYR1-like (PYL) proteins play a central role in initiating ABA signal transduction. In this study, a total of 30 PopPYL genes were identified and classified into three sub-families (PYL I–III) in the pan-genome of 17 Populus species, through phylogenetic analysis. Among these subfamilies, the PYL I subfamily was the largest, comprising 21 members, whereas PYL III was the smallest, with only four members. To elucidate the evolutionary dynamics of these genes, we conducted synteny and Ka/Ks analyses. Results indicated that most PopPYL genes had undergone purifying selection (Ka/Ks < 1), while a few were subject to positive selection (Ka/Ks > 1). Promoter analysis revealed 258 cis-regulatory elements in the PYL genes of Populus euphratica (EUP) and Populus pruinosa (PRU), including 127 elements responsive to abiotic stress and 33 ABA-related elements. Furthermore, six structural variations (SVs) were detected in PYL_EUP genes and significantly influenced gene expression levels (p < 0.05). To further explore the functional roles of PYL genes, we analyzed tissue-specific expression profiles of 17 PYL_EUP genes under drought stress conditions. PYL6_EUP was predominantly expressed in roots, PYL17_EUP exhibited leaf-specific expression, and PYL1_EUP showed elevated expression in stems. These findings suggest that the drought response of PYL_EUP genes is tissue-specific. Overall, this study highlights the utility of pan-genomics in elucidating gene family evolution and suggests that PYL_EUP genes contribute to the regulation of drought stress responses in EUP, offering valuable genetic resources for functional characterization of PYL genes. Full article

Concomitant sensitisation to non-specific lipid transfer proteins (nsLTPs) from olive pollen (Ole e 7) and peach (Pru p 3) has been observed in the south of Spain. In the search for reasons to explain this observation, we studied a potential causal relationship between Human Leukocyte Antigen (HLA) molecules and nsLTP sensitisation. For this purpose, eighteen Ole e 7-monosensitised (MONOLE) patients, 22 Pru p 3-monosensitised (MONPRU) patients, and 22 bisensitised (BI) patients were genotyped for HLA class II alleles. Complementarily, T-cell epitopes were predicted with the Immune Epitope Database analysis tool to test HLA epitope presentation. Our results showed a significant increase in DRB1*11 and DQB1*03 frequencies in MONPRU patients and DRB1*04 frequency in MONOLE patients. Additionally, T-cell epitope analysis revealed high binding affinity between the predicted Pru p 3 epitopes and DRB1*11 and between the predicted Ole e 7 epitopes and DRB1*04, suggesting that presentation of these epitopes may be favoured and predisposing individuals to sensitisation. Conversely, low DQB1*05 frequency and poor binding ability of predicted epitopes from both nsLTPs postulated this allele as a possible protective factor to sensitisation. Variations in the binding affinity between nsLTP epitopes and HLA molecules may underlie individual susceptibility to nsLTP allergy. Full article

The objective of this study was to evaluate the serum biomarkers implicated in the interaction of platelets and endothelium, as well as the efficacy of antiplatelet therapy in patients with aortic stenosis (AS) and coronary artery disease (CAD). A total of 78 adult patients with CAD on aspirin therapy participated in this study, including 49 consecutive patients with AS and 29 control subjects. The analysis included the following serum biomarkers: thrombomodulin (TM), platelet factor 4 (PF4), P-selectin, and CD40L. The efficacy of antiplatelet treatment was evaluated using the VerifyNow Aspirin test (ASPI test) and P2Y12 assay test (ADP test). Patients with AS exhibited increased serum levels of TM (7.64 ± 3.5 ng/mL vs. 6.28 ± 2.1 ng/mL, p = 0.011) and PF4 (25.16; Q1: 8.3; Q3: 29.6 μg/mL vs. 12.85; Q1: 5.7; Q3: 14.5 μg/mL, p = 0.021) compared to the control group. P-selectin and CD40L levels did not differ between groups. There were no significant differences in platelet aggregation in the ASPI (474.04 ± 66.7 ARU vs. 471.31 ± 56.2 ARU; p = 0.822) or ADP (224.88 ± 46.4 PRU vs. 216.62 ± 29.6 PRU; p = 0.394) tests. Bleeding incidence did not differ significantly between groups. The coexistence of AS in patients with CAD is associated with elevated levels of the aforementioned biomarkers, which are indicative of endothelial damage and platelet activation. However, the efficacy of antiplatelet treatment was independent of the presence of AS. Full article

This work systematically addresses the dual challenges of non-inertial dynamic coupling and kinematic constraint redundancy encountered in dynamic modeling of serial–parallel–serial hybrid robotic mechanisms, and proposes an improved Newton–Euler modeling method with constraint compensation. Taking the Skiing Simulation Platform with 6-DOF as the research mechanism, the inverse kinematic model of the closed-chain mechanism is established through $G_F$ set theory, with explicit analytical expressions derived for the motion parameters of limb mass centers. Introducing a principal inertial coordinate system into the dynamics equations, a recursive algorithm incorporating force/moment coupling terms is developed. Numerical simulations reveal a 9.25% periodic deviation in joint moments using conventional methods. Through analysis of the mechanism’s intrinsic properties, it is identified that the lack of angular momentum conservation constraints on the end-effector in non-inertial frames leads to system controllability degradation. Accordingly, a constraint compensation strategy is proposed: establishing linearly independent differential algebraic equations supplemented with momentum/angular momentum balance equations for the end platform. Co-Simulation results demonstrate that the optimized model reduces the maximum relative error of actuator joint moments to 0.98%, and maintains numerical stability across the entire configuration space. The constraint compensation framework provides a universal solution for dynamics modeling of complex closed-chain mechanisms, validated through applications in flight simulators and automotive driving simulators. Full article