Search Results (79)

Although mango is not classified among the nine major allergenic foods reported by the Food and Drug Administration (FDA), the increasing global and domestic consumption of mango has been accompanied by a growing number of reported cases of mango allergy. Currently, reports on cross-reactive allergens and cross-reactive linear epitopes in mango are limited. This study employed BLASTp (version 2.11.0+) to predict potential allergens that may cross-react with mango protein allergens and other food protein allergens. Subsequently, cross-reactive allergens were identified using sera from mango-allergic patients. Furthermore, similar sequences of the identified cross-reactive allergens were predicted by BLAST. These similar sequences were then synthesized by the solid-phase peptide synthesis method. Finally, the cross-reactive linear epitopes were determined by assessing their IgE-binding capacity using serum IgE from the same patient cohort. The results demonstrated that the sera from mango-allergic patients exhibited IgE-binding cross-reactivity with those from peanut, wheat, cashew, pistachio, and hazelnut, particularly with IgE-binding cross-reactivity to wheat and hazelnut, which has not been previously reported. The following novel cross-reactive linear epitopes were identified: the AA80–88 sequence of mango chitinase with the AA37–45 sequence of wheat Tri a 27 and the AA15–22 sequence of mango profilin with the AA65–72 sequence of pistachio Pis v 1. Furthermore, multiple cross-reactive epitopes were mapped between mango profilin and peanut Ara h 5, corresponding to the sequences AA31–51/AA31–50, AA50–65/AA52–65, AA76–96/AA76–96, and AA103–117/AA104–117, respectively. Full article

Tomato (Solanum lycopersicum L.) is a key source of bioactive compounds and essential minerals, but it also contains clinically relevant allergens. Despite growing concern about the effects of climate change on crop quality, the impact of heat stress during specific reproductive stages on fruit allergen accumulation remains poorly understood. This study aimed to investigate how the timing of heat stress affects tomato fruit quality, antioxidant traits, and the expression of major pan-allergens. Plants of the cultivar Micro-Tom were exposed to heat stress (40 °C for 8 h) at three flowering stages: pre-anthesis, anthesis, and post-anthesis. Ripe fruits were evaluated for morphological parameters, mineral composition, nutraceutical properties, antioxidant responses, and the expression of profilin and cyclophilin. Heat stress applied at post-anthesis significantly reduced fruit weight and diameter, while earlier treatments had limited morphological effects. Mineral composition was largely unchanged across treatments. In contrast, total phenolic content increased progressively with later stress application, whereas flavonoid content and antioxidant capacity (FRAP) remained relatively stable. Antioxidant enzyme activity showed only minor stage-dependent variation, suggesting a controlled oxidative response. Notably, allergen-related proteins exhibited distinct patterns: profilin accumulation increased progressively under heat stress, while cyclophilin showed a transient peak at anthesis. These findings demonstrate that the timing of reproductive heat stress differentially affects tomato fruit quality and allergen accumulation. This study provides novel insights into the stage-specific modulation of food allergens under heat stress, contributing to a better understanding of crop nutritional and allergenic properties in the context of climate change. Full article

Exposure to airborne pollen allergens is a major trigger of respiratory allergy, whose prevalence continues to rise throughout Europe. In southern Portugal, the Mediterranean climate and extensive vegetation diversity promote high pollen loads, particularly from the Amaranthaceae family. This retrospective observational study aimed to characterise the sensitisation profiles of patients with positive skin prick tests (SPTs) to Chenopodium album and/or Salsola kali, the dominant Amaranthaceae species in the region. Data from 346 patients were analysed, including demographic and clinical characteristics, SPT results, and specific IgE sensitisation to molecular allergens. Of these, 35% were positive for C. album only, 13% for S. kali only, and 51% for both. In molecular testing, 54% of S. kali-sensitised patients were positive to Sal k 1, whereas only 4% of C. album-sensitised patients were positive to Che a 1. Sensitisation to panallergens such as profilins and Ole e 1-like proteins was frequent, suggesting extensive IgE cross-reactivity between these taxa. A significant correlation in wheal size (r = 0.53, p < 0.0001) further supports shared allergenic determinants. Despite higher SPT positivity to C. album, S. kali is likely the predominant sensitising source in this population. These findings highlight the importance of molecular-based diagnostics to distinguish genuine sensitisation from cross-reactivity in Mediterranean settings. Full article

To address cryodamage in Culter alburnus sperm, this study evaluated the effects of trehalose supplementation in a conventional cryomedium (D-15 + 10% ethylene glycol). Six experimental groups were established: fresh sperm (G1), a conventional cryomedium (G2), groups supplemented with 10, 100, or 200 mmol/L trehalose (G3–G5), and a control group with extender only (G6). The group with 100 mmol/L trehalose (G4) was associated with improved post-thaw motility parameters (activation rate, movement time, and lifespan) and higher antioxidant (superoxide dismutase and catalase) and energy metabolism (ATPase, succinate dehydrogenase, lactate dehydrogenase) enzyme activities. Ultrastructural damage in G4 included partial plasma membrane rupture and mitochondrial swelling, while G6 exhibited additional damage features including membrane disintegration, mitochondrial disruption, and flagellar fracture. Proteomic analysis revealed that, compared to G1, G4 exhibited higher abundance of proteins (e.g., Histone H2A, cytochrome c oxidase, profilin) involved in structural integrity and energy homeostasis, whereas G6 showed signatures of oxidative stress and metabolic dysfunction (lower abundance of NADH dehydrogenase and higher abundance of calcium-transporting ATPase and glutathione S-transferase). In conclusion, 100 mmol/L trehalose was associated with improved cryopreservation outcomes, and the proteins identified provide a basis for further investigation. This approach offers a framework for refining germplasm conservation strategies in aquaculture. Full article

Background/Objectives: Diabetic small fiber neuropathy and related sensory and epidermal problems affect up to 70% of all patients with diabetes. Long-term hyperglycemia disrupts cytoskeletal organization and axonal transport; however, molecular changes within human diabetic epidermis remain understudied. Diaph1 and its cytoskeletal ligands, including β-Actin and Profilin, are key regulators of cytoskeletal dynamics and may be associated with diabetes-related alterations in skin structure and innervation. Methods: Sixteen patients with type 2 diabetes, aged 43.3 ± 9.6 years (disease duration 18.9 ± 8.7 years), and twelve non-diabetic controls, aged 43.9 ± 8.9 years, were enrolled in the study. All participants provided informed consent. Skin punch biopsies were obtained under local anesthesia and processed for staining of PGP 9.5, Diaph1, β-Actin, and Profilin. Quantitative image analysis was performed to assess stained area fraction, signal intensity, and intraepidermal nerve fiber density. Statistical comparisons and Spearman’s rank correlation analyses were used to evaluate group differences and associations between staining parameters. Results: Diabetic skin samples exhibited a significant reduction in PGP 9.5-positive intraepidermal nerve fibers, indicating reduced cutaneous innervation. In contrast, Diaph1 and Profilin showed broader and more diffuse epidermal staining, while β-Actin displayed altered staining patterns and intensity. Significant correlations between Diaph1- and β-Actin-related staining measures indicated an association consistent with altered cytoskeletal organization under chronic hyperglycemic conditions. Conclusions: Long-standing type 2 diabetes was associated with reduced PGP 9.5-positive intraepidermal nerve fibers, together with altered epidermal staining patterns of Diaph1, Profilin and β-Actin. These findings indicate coexisting cutaneous denervation and cytoskeletal alterations in diabetic skin. Full article

Background: Papillary Thyroid Cancer (PTC) harboring CCDC6-RET translocation is typically classified as a differentiated epithelial tumor. Although Selpercatinib, a RET-selective drug, was recently approved for use in advanced PTC, the emergence of drug resistance has already been observed. Tumor plasticity, including non-canonical Epithelial–Mesenchymal Transition (EMT) programs, is recognized as a key mechanism underlying drug resistance. The downregulation of the transcription factor Wilms’ Tumor 1 (WT1) in cancer is associated with increased motility, invasiveness, and metastatic potential. Methods: In this study, we developed a selpercatinib-resistant PTC-derived cell line, TPC-1-SelpR. Bioinformatic analyses were conducted to study the promoter of the CCDC6-RET gene and the transcriptomic landscape of PTC from RNAseq data. Subsequent real-time PCR, Western blot, and imaging techniques, such as confocal microscopy (CM) and fluorescence microscopy (FM), were employed to study the effects of WT1 loss-of-function following RNAi silencing. Results: In TPC-1-SelpR, WT1 expression appears downregulated compared to its counterpart, TPC-1. Crucially, WT1 silencing induced a context-dependent modulation of the CCDC6-RET driver: while WT1 silencing reduced CCDC6-RET expression in TPC-1, in TPC-1-SelpR, a post-transcriptional compensation of CCDC6-RET was observed. The gene expression of several factors involved in EMT, such as Twist, Vimentin, Integrin beta-1, and Profilin, was rewired in TPC-1-SelpR^{WT1-knockdown}. Although the Vimentin protein product decreased, CM and FM analyses confirmed a reorganization of residual protein: the subcellular redistribution was more dispersed in TPC-1-SelpR^{WT1-knockdown}. Further upregulation of the stemness factor Sox2 over the differentiation factor Sox17 occurred. These molecular changes were associated with higher cell motility of TPC-1-SelpR^{WT1-knockdown}. Conclusions: Collectively, these findings suggest that WT1 is a critical regulator involved in tumor plasticity, thereby supporting selpercatinib resistance. Full article

Wheat allergy (WA) poses a diagnostic challenge due to its diverse clinical phenotypes—ranging from classic food allergy and wheat-dependent exercise-induced anaphylaxis (WDEIA) to baker’s asthma. An additional diagnostic aspect is serological cross-reactivity with grass pollen. Undoubtedly, the transition from extract-based diagnostics to precise component-based diagnostics (CRDs) facilitates the management of wheat allergy. It has significantly improved the diagnostic accuracy for WDEIA ω-5-gliadin (Tri a 19), although considering new knowledge about wheat proteins, it seems necessary to include them in the diagnostic scheme, especially where Tri a 19 remains negative despite clinical symptoms. Therefore, in this review, we evaluate the clinical utility of new wheat molecules with a high risk of anaphylaxis. We pay particular attention to Tri a 37 (α-purothionin), a thermally stable allergen associated with a 4-fold increase in the risk of severe anaphylaxis, and Tri a 36 (LMW glutenin), which shows higher sensitivity than Tri a 19 in specific pediatric cohorts. In addition, we emphasize the role of Tri a 14 (nsLTP) in distinguishing true wheat sensitization from pollen-related cross-reactivity caused by profilins (Tri a 12) or carbohydrate determinants (CCDs). Beyond diagnostics, the review discusses dynamic changes in sensitization profiles in relation to the allergic march and the phenomenon of spontaneous remission in children. New management strategies are also discussed, including the potential of omalizumab (based on the data from the OUtMATCH study) in facilitating the reintroduction of allergens into the diet. Full article

Tomato (Solanum lycopersicum L.), one of the most widely consumed horticultural crops worldwide, is also a notable source of food allergens, with a higher prevalence of allergic reactions in pollen-sensitized patients. Profilin, a small actin-binding protein, is a major allergen in tomato fruits encoded by two homologous genes, Lyc e1.01 and Lyc e1.02. Although various strategies have been proposed to reduce allergenicity in foods, no prior study has successfully achieved the complete elimination of profilin proteins in tomato using precise genome editing. In this study, we designed a single-guide RNA targeting a conserved region of both genes and employed the CRISPR/Cas9 system to generate loss-of-function mutations. We first evaluated single-guide RNA editing efficiency in tomato protoplasts and then performed Agrobacterium-mediated stable transformation, which yielded 13 transgenic T₀ lines. Genotyping and Western blot analyses confirmed successful editing at both target loci and significantly decreased profilin accumulation in mutant tomato fruits. Notably, two homozygous Cas9-free lines were identified in the T1 generation, and one of these (line 23-15) showed significantly decreased profilin protein levels in the fruit. These findings demonstrate that the CRISPR/Cas9-mediated disruption of allergenic Lyc e1 genes effectively removes profilin from tomato fruits. This strategy provides a promising framework for developing hypoallergenic tomato cultivars that may be extended to other related crop species. Full article

Background/Objectives: Monkeypox is endemic to the African continent and has recently garnered global attention due to reported outbreaks in non-endemic nations. No approved drug is available for non-severe cases, and some isolates gained resistance to approved antivirals. In this study, we employed a drug repositioning strategy to evaluate the efficacy of existing FDA-approved antiviral drugs if repurposed for use against emerging Monkeypox, representing a cost-effective method for identifying novel therapeutic interventions. Methods: Methodology including Egyptian virus strain isolation, propagation and titration followed by in vitro studies, molecular docking and molecular dynamics simulations combined with binding free energy were carried out. Twenty-three FDA-approved drugs, including Abacavir, Acyclovir, Amantadine, Chloroquine, Daclatasvir, Dolutegravir, Entecavir, Favipiravir, Hydroxychloroquine, Lamivudine, Molnupiravir, Nevirapine, Oseltamivir, Penciclovir, Remdesivir, Ribavirin, Sofosbuvir, Tenofovir, Valaciclovir, Valganciclovir, Velpatasvir, Zanamivir, and Zidovudine, were screened for potential anti-monkeypox activity in vitro. In silico studies were carried out against three monkeypox proteins, Thymidylate Kinase, A42R Profilin-Like Protein, and VACV D13, to identify their potential targets. Results: In vitro testing showed that two antiviral drugs are positive. The employed computational methods indicate that remdesivir demonstrated superior binding patterns with elevated scores and stable complexes throughout the simulation. Conclusions: Our findings showed that Remdesivir therapeutic compound is potent against the tested strain of MPXV, and exhibited a robust binding affinity for Thymidylate Kinase, A42R Profilin-Like Protein, and VACV D13 enzymes, and thus may potentially be utilized as antiviral for the treatment of monkeypox virus. Full article

Diabetic peripheral neuropathy (DPN) is a serious consequence of prolonged hyperglycemia and contributes to the morbidity associated with diabetes. Hyperglycemia enhances the non-enzymic glycation of proteins and the accumulation of Advanced Glycation End Products (AGEs). We employed a diabetic mouse model lacking both Diaph1 and RAGE to elucidate the role of RAGE-Diaph1 signaling in the pathogenesis of DPN. We demonstrate that simultaneous deletion of Diaph1 and RAGE did not change the course or the intensity of hyperglycemia-induced weight loss in mice. However, abrogating RAGE-Diaph1 signaling affects actin cytoskeleton remodeling rates in nerve axons by altering the ratio of the actin-regulating molecules cofilin and profilin. Our experimental results suggest that the loss of RAGE-Diaph1 signaling protects neurons from hyperglycemic conditions. We observed a beneficial effect of abolishing RAGE-Diaph1 signaling on the axonal structure of neuropathic nerves. In addition, we observed that abolishing RAGE-Diaph1 signaling improved motor nerve conduction velocity in the sciatic nerves of hyperglycemic mice. Our data indicate that RAGE-Diaph1 signaling is likely enhanced in chronic hyperglycemia, resulting in aberrant actin dynamics in nerve axons. These defective actin dynamics play a key role in the progression of DPN, leading to structural and functional loss in peripheral nerves. Full article

Pearl color is the primary determinant of its value, and the mantle donor tissue (saibo) used in pearl culture plays a critical role in color formation. To elucidate the molecular mechanisms underlying nacre color, we performed comparative transcriptomic analyses of saibo tissues from Sinohyriopsis cumingii displaying three representative phenotypes: white (W), purple (P), and golden (G). A total of 364 differentially expressed genes (DEGs) (102 upregulated and 162 downregulated genes) were identified in W vs. P. A total of 770 DEGs (467 upregulated and 303 downregulated genes) were identified in W vs. G. KEGG pathway analysis revealed significant differences in the expression of genes mainly involved in pigment-based and structural coloration, including amino sugar and nucleotide sugar metabolism (ko00520), cell adhesion molecules (ko04514), tyrosine metabolism (ko00350), ECM-receptor interaction (ko04512), and PI3K-Akt signaling pathway (ko04151). Subsequent integrative analyses across W vs. P and W vs. G groups identified 45 key regulatory genes, which were classified into four functional categories: extracellular matrix protein synthesis and biomineralization (e.g., chit, silkmaxin, bmp2/7, profilin, perlucin2), organic pigment metabolism (e.g., tyr, typ, dbh, bco2, gst5, ldlr, cpox, pks-like 1, pks15), metal ion metabolism and accumulation (e.g., hip-like, fcr1, ferritin 2), and epigenetic regulation (e.g., metK, mbd4/6, mettl24/27, alkbh6). Taken together, our findings reaffirm the complex nature of pearl coloration and reveal that structural coloration, pigment biosynthesis, and epigenetic modulation collectively shape nacre color formation, which paves the way for further functional elucidation of color-related genes and facilitates selective breeding practices in S. cumingii. Full article

A series of six silver(I) complexes, namely bromo(1-benzyl-3-cinnamyl-benzimidazol-2-ylidene)silver (I) (1a), bromo[1-(4-methylbenzyl)-3-cinnamyl-benzimidazol-2-yliden]silver(I) (1b), bromo[1-(3-methoxylbenzyl)-3-cinnamyl-benzimidazol-2-yliden]silver(I) (1c), bromo[1-(3,5-dimethoxy-benzyl)-3-cinnamyl-benzimidazol-2-ylidene]silver(I) (1d), bromo[1-(naphthalen-1-ylmethyl)-3-cinnamyl-benzimidazol-2-ylidene]silver(I) (1e) and bromo[1-(pyren-1-ylmethyl)-3-cinnamyl-benzimidazol-2-yliden]silver(I) (1f), were synthetized and characterized by microanalyses and mass spectrometry and characterized by FT-IR and NMR spectroscopic techniques. The in vitro effects of silver(I) complexes on trophozoites of two Acanthamoeba isolates obtained from patients with keratitis were investigated. The parasites were exposed to concentrations of 10, 100 and 1000 µM for 24, 48 and 72 h. The complexes exhibited potent, dose- and time-dependent activity. Complete inhibition was observed within 24 h at a concentration of 1000 µM. At a concentration of 100 µM, complexes 1c–e exhibited reduced viability to less than 10% within 48 to 72 h. At a concentration of 10 µM, partial inhibition was observed. Preliminary morphological changes included the loss of acanthopodia, rounding, and detachment. These effects were not observed in the presence of the pre-ligands or commercially available silver compounds. Furthermore, molecular docking was utilized to analyze the molecules against Acanthamoeba castellanii CYP51, A. castellanii profilin IA, IB, and II. The highest recorded interactions were identified as −9.85 and −11.26 kcal/mol for 1e and 1f, respectively, when evaluated against the A. castellanii CYP51 structure. Full article

Vertebrate embryonic development relies on tightly regulated signaling pathways that guide morphogenesis, cell fate specification, and tissue organization. Among these, the Wnt signaling pathway plays a central role, orchestrating key developmental events. The non-canonical Wnt pathways, including the Planar Cell Polarity and Wnt/Ca²⁺ branches, are especially critical for regulating cytoskeletal dynamics during gastrulation. Recent studies highlight that these pathways interface with cytoskeletal effectors to control actin remodeling in response to extracellular cues. One such effector is Profilin, a small, evolutionarily conserved actin-binding protein that modulates actin polymerization and cellular architecture. Profilins, particularly Profilin1 and 2, are known to interact with Daam1, a formin protein downstream of PCP signaling, thereby linking Wnt signals to actin cytoskeletal regulation. Emerging evidence suggests that Profilins are active signaling intermediates that contribute to morphogenetic processes. Their context-dependent interactions and differential expression across species also suggest that they play specialized roles in development and disease. This review synthesizes the current understanding of Profilin’s role in non-canonical Wnt signaling, examining its molecular interactions and contributions to cytoskeletal control during development. By integrating data across model systems, we aim to clarify how Profilins function at the intersection of signaling and cytoskeletal dynamics, with implications for both developmental biology and disease pathogenesis. Full article

Neospora caninum, as well as Toxoplasma gondii, secrete proteins that facilitate the invasion of host cells and the regulation of host immune response and metabolism. However, the localization of the secretory proteins in infected animal brains has not been studied in detail. Here, we investigate the brain and intracellular distribution of the secretory proteins in experimentally infected mice and naturally infected calves through histopathology and immunohistochemistry (IHC) to detect surface antigen 1 (NcSAG1), cyclophilin (NcCYP), profilin (NcPF), dense granule protein 6 (NcGRA6), and NcGRA7. These methods revealed that numerous tachyzoites positive for NcSAG1, NcCYP, NcPF, NcGRA6, and NcGRA7 were localized in and around the animals’ necrotic lesions, and NcGRA7 was diffusely observed in the necrotic lesions of the infected mice. Moreover, IHC revealed that NcGRA6 and NcGRA7 were distributed in the cytoplasm of infected neurons around the parasites in the infected mice and calves. This suggests that NcGRA6 and NcGRA7 might be directly related to the alteration of neuronal metabolism and activity, and that NcGRA7 might be related to the formation of necrotic lesions. Full article

Orange allergy is estimated to account for up to 3–4% of food allergies. Major allergens identified in orange (Citrus sinensis) include Cit s 1 (germin-like protein) and Cit s 2 (profilin), while Cit s 3 (non-specific lipid transfer protein, nsLTP) and Cit s 7 (gibberellin-regulated protein) have also been described. The objective of this study was to investigate the presence and IgE-binding capacity of germin-like proteins in citrus fruits other than oranges. We describe five patients with immediate allergic reactions after orange ingestion. All patients underwent skin prick tests (SPT) to aeroallergens and common food allergens, prick-by-prick testing with orange, lemon, and mandarin (pulp, peel, seeds), total IgE, specific IgE (sIgE), anaphylaxis scoring (oFASS), and the Food Allergy Quality of Life Questionnaire (FAQLQ-AF). Protein extracts from peel and pulp of orange, lemon, and mandarin were analyzed by Bradford assay, SDS-PAGE, and IgE immunoblotting using patient sera. Selected bands were identified by peptide mass fingerprinting. A 23 kDa band was recognized by all five patients in orange (pulp and peel), lemon (peel), and mandarin (peel). This band was consistent with Cit s 1, a germin-like protein already annotated in the IUIS allergen database for orange but not for lemon or mandarin. Peptide fingerprinting confirmed the germin-like identity of the 23 kDa bands in all three citrus species. Germin-like proteins of approximately 23 kDa were identified as IgE-binding components in peel extracts of orange, lemon, and mandarin, and in orange pulp. These findings suggest a potential shared allergen across citrus species that may contribute to allergic reactions independent of LTP sensitization. Full article