Search Results (74)

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

The sustainable production of plant bioactive compounds is increasingly important as natural habitats decline. This study investigates whether in vitro cell cultures of Eryngium planum, Lychnis flos-cuculi, and Kickxia elatine can serve as alternative sources of biologically active biomass with antimicrobial and anti-Acanthamoeba properties. Callus cultures were established under optimized and controlled conditions, and metabolomic profiling was completed using UPLC-HRMS/MS. In silico analysis, using a molecular docking approach, was applied to understand the interaction between target compounds and Acanthamoeba profilin and identify possible targets for antimicrobial properties. Untargeted metabolomic analysis confirmed the presence of valuable compounds in the callus cultures of the studied species. Biological activity was assessed through anti-Acanthamoeba and antimicrobial assays. Lychnis flos-cuculi and Kickxia elatine callus extracts showed significant inhibitory effects on Acanthamoeba trophozoites, with 87.5% and 80.1% inhibition at 10 mg/mL, respectively. In contrast, E. planum extract stimulated amoebic growth. The anti-Acanthamoeba activity correlated with the presence of ferulic acid and p-coumaric acid in L. flos-cuculi extract, and acteoside in K. elatine extract. Antibacterial testing revealed moderate activity of E. planum and K. elatine extracts against Staphylococcus spp., while Gram-negative bacteria and fungi were largely resistant. These findings highlight the potential of in vitro cultures—particularly those from L. flos-cuculi and K. elatine—as promising, sustainable sources of anti-Acanthamoeba and antimicrobial agents, warranting further investigation into their pharmacologically active constituents. Full article

Axon guidance, a fundamental process in neural circuit formation, is intricately regulated by Fibroblast Growth Factors (FGFs) and their receptors (FGFRs) through dynamic cytoskeletal remodeling. FGF signaling, mediated by heparan sulfate proteoglycans or Klotho co-factors, activates key downstream pathways: PI3K-Akt, JAK-STAT, PLCγ, and RAS-MAPK. These pathways orchestrate actin filament dynamics, microtubule stability, and the organization of intermediate filaments. These pathways converge on Rho GTPases, cofilin, profilin, and tau to balance the cytoskeletal assembly−disassembly cycles, enabling growth cone navigation. Unresolved questions, such as the mechanisms underlying FGF-mediated growth cone steering, highlight critical future research directions. This review integrates structural, molecular, and functional insights into how FGF-FGFR interactions regulate axon pathfinding, emphasizing the crosstalk between signaling cascades and cytoskeletal plasticity. Elucidating these mechanisms not only advances our understanding of neural development but also opens therapeutic avenues for neuro-developmental disorders, nerve injury, and neurodegenerative diseases by targeting FGF-driven cytoskeletal dynamics. Full article

Background: The formin family proteins play an important role in guiding the assembly and nucleation of linear actin and can promote the formation of actin filaments independently of the Arp2/3 complex. As a key protein that regulates the cytoskeleton and cell morphological structure, the formin gene family has been widely studied in plants such as Arabidopsis thaliana and rice. Methods: In this study, we conducted comprehensive analyses, including phylogenetic tree construction, conserved motif identification, co-expression network analysis, and transcriptome data mining. Results: A total of 18 MtFH gene family members were identified, and the distribution of these genes on chromosomes was not uniform. The phylogenetic tree divided the FH proteins of the four species into two major subgroups (Clade I and Clade II). Notably, Medicago truncatula and soybean exhibited closer phylogenetic relationships. The analysis of cis-acting elements revealed the potential regulatory role of the MtFH gene in light response, hormone response, and stress response. GO enrichment analysis again demonstrated the importance of FH for reactions such as actin nucleation. Expression profiling revealed that MtFH genes displayed significant transcriptional responsiveness to cold, drought, and salt stress conditions. And there was a temporal complementary relationship between the expression of some genes under stress. The protein interaction network indicated an interaction relationship between MtFH protein and profilin, etc. In addition, 22 miRNAs were screened as potential regulators of the MtFH gene at the post-transcriptional level. Conclusions: In general, this study provides a basis for deepening the understanding of the physiological function of the MtFH gene and provides a reference gene for stress resistance breeding in agricultural production. Full article

Oral Allergy Syndrome (OAS) is an allergic reaction that occurs upon contact of the mouth and throat with food, leading to symptoms primarily affecting the oral mucosa. In patients with allergic rhinitis, OAS may develop due to cross-reactivity between the pollen allergens responsible for allergic rhinitis, and specific plant-derived foods. This particular type of OAS is known as Pollen Food Allergy Syndrome (PFAS). The difference in prevalence of PFAS across different regions of the world is attributed to various factors, including environmental exposure and dietary habits. Southern Europe’s temperate climate favors the blooming of many allergenic plants, making respiratory allergies and PFAS significant public health concerns. There is a regional variation in pollen in Southern Europe, contributing to differences in the presence of panallergens—such as profilins, pathogenesis-related class 10 (PR-10) proteins and lipid transfer proteins (LTPs)—which mediate PFAS. In order to examine the epidemiology, pathogenesis, and diagnostic approaches of OAS and PFAS, focusing on their prevalence and impact in Southern European adults, a narrative review was performed. Data from Portugal, Spain, France, Italy, Albania, Greece, and Türkiye were retrieved. The main outcome of this review was that the frequency of PFAS varies across studies, not only between countries but also within the same country, due to vegetation variability across regions as well as methodological differences and the year of study. However, despite these differences, PFAS emerges as a common issue in Southern Europe, underscoring the need for effective diagnosis and management. Full article

This study investigates the impact of organic and conventional farming on the allergenic and bioactive properties of wheat. The primary aim was to assess the immunometric parameters and polyphenolic composition in four varieties of winter and four varieties of spring wheat cultivated under both farming systems. Immunometric assays focused on gluten content, the allergenic QQQPP peptide, and the panallergenic profilin Tri a 12. While gluten levels (15–20 g/kg) showed no clear dependence on farming type, organic wheat exhibited a mild yet significant increase in QQQPP-dependent immunoreactivity in five samples (>20 µg/g). However, all organic wheat samples demonstrated a notable reduction in profilin content (<0.6 µg/g), suggesting that the type of wheat cultivation could influence allergenic risk for individuals with wheat-related allergies. Polyphenolic profiling revealed that kaempferol, p-coumaric acid, and gallic acid were the predominant compounds, with organic wheat displaying slightly higher polyphenol levels on average. Despite these differences, the variations were insufficient to determine a superior cultivation method. These findings highlight the potential allergenic and nutritional implications of organic versus conventional wheat farming. Full article