Search Results (768)

Background and Clinical Significance: Adams–Oliver syndrome type 3 (AOS3) is a rare congenital disorder typically characterised by terminal transverse limb defects and variable involvement of other organ systems. Although pathogenic variants in RBPJ are well established in AOS3, associated neurodevelopmental or psychiatric features have been only sporadically documented. Case Presentation: We describe a male patient first evaluated at the age of 10 years and subsequently re-evaluated at 14 years, with AOS3 presenting terminal limb defects together with autistic-like behaviour, cognitive difficulties, dyslexia, and recurrent depressive symptoms. Whole-exome sequencing (WES) identified a heterozygous pathogenic variant in RBPJ (c.505A>G; p.Lys169Glu), confirming the molecular diagnosis of autosomal dominant AOS3. Additional findings included a heterozygous missense variant in CACNA1A (p.Arg1678Cys), a gene linked to neurological disorders with broad phenotypic variability. Because of elevated homocysteine levels, the patient was also tested for MTHFR variants and was found to be heterozygous for C677T and A1298C. Conclusions: This case illustrates a rare combination of a validated AOS3-associated RBPJ variant, along with additional CACNA1A and MTHFR variants that may influence the patient’s neurocognitive and psychiatric characteristics. The results underscore the importance of comprehensive genetic testing in atypical AOS presentations and highlight the complexity of interpreting overlapping genetic factors. Full article

Pleurotus pulmonarius is a high-value, commercially cultivated edible fungus whose primordium formation is a critical phase for yield and commercial value. To better understand the developmental processes of P. pulmonarius, samples from four key developmental stages were collected and subjected to transcriptome analysis. A total of 6530 DEGs were identified, including 50 transcription factors from 10 families. Among these, the PpZCP11 gene, encoding a Zn₂Cys₆ transcription factor, was found to be specifically highly expressed during the primordium stage. We cloned PpZCP11 gene and confirmed its nuclear localization. The OE-PpZCP11 strains produced abundant primordia, while primordium formation in the RNAi-PpZCP11 strains was severely suppressed. Moreover, RNA-seq and yeast-one-hybrid analysis suggested that PpZCP11 may regulate cell wall synthesis. These findings indicate that the PpZCP11 transcription factor acts as a positive regulator of primordium formation by regulating the expression of cell wall-related genes. This study provides a theoretical reference for elucidating the molecular mechanism underlying primordium formation in P. pulmonarius. Full article

Although calpain-5/CAPN5 is widely expressed in mammals, little is known regarding its functions. Pathogenic mutations of CAPN5 are causal for a devastating autoimmune eye disease, neovascular inflammatory vitreoretinopathy (NIV). To provide insight into both the physiological and pathological roles of CAPN5, it is essential to identify candidate interaction partners and possible substrates. Human SH-SY5Y neuroblastoma cells, transfected with full-length catalytically dead (Cys81Ala) CAPN5-3×FLAG, were used for anti-FLAG co-immunoprecipitation (co-IP) and quantitative proteomics using Sequential Window Acquisition of all THeoretical mass spectra (SWATH-MS). Fifty-one proteins were enriched at least four-fold, p < 0.01, relative to cells transfected with an empty FLAG vector. A high proportion (24/51) of candidate CAPN5 interaction partners are associated with protein quality control, including components of the chaperonin, chaperone, and ubiquitin–proteasome systems. Additional candidate interactors include tubulins, kinases, phosphatases, G proteins, and mitochondrial proteins. CAPN5 interactions for 14 of the candidate proteins were confirmed by co-IP and immunoblotting. Of these 14 proteins, 11 exhibited in vitro calcium-induced proteolysis following co-IP with WT CAPN5-3×FLAG. Impaired calcium-induced proteolysis of co-IP proteins was observed for the pathogenic CAPN5 variants R243L and R289W. Further studies are needed to validate the association of candidate CAPN5 interactors with proteins and complexes suggested by the SWATH-MS and co-IP results, and the possible role of CAPN5 within such complexes. The possible involvement of CAPN5 in protein quality control is relevant to NIV, as defects in protein quality control have been implicated in inherited retinal disorders. Proteomic data are available via ProteomeXchange with identifier PXD068008. Full article

A facile and single-step synthesis of poly(L-Cysteine) (p(L-Cys)) particles through microemulsion polymerization using tetrakis(hydroxymethyl) phosphonium chloride (THPC) as crosslinker is accomplished for the first time. The L-Cys:THPC ratio in p(L-Cys) particles was calculated as 80:20% (by weight) with elemental analyses, and the generation of p(L-Cys) particles was confirmed. SEM imaging revealed a popcorn-like morphology of the p(L-Cys) particles with a 1–20 µm particle size range. The isoelectric point of p(L-Cys) particles was determined at pH 1.15 via zeta potential measurements. The hydrolytic degradation of p(L-Cys) particles was determined as about 85% within 3 h (by weight). The p(L-Cys) particles displayed excellent blood compatibility with a hemolysis % ratio of <2.3% and a blood clotting index of 95% at 1 mg/mL concentration. Moreover, cell compatibility tests up to 50 mg/mL against L929 fibroblast cells exhibited about 90% cell viability for p(L-Cys) particles versus 58% for L-Cys molecule. The antimicrobial efficacy of the L-Cys molecules was notably enhanced in p(L-Cys) particles, exhibiting a 5-fold reduction in minimal bactericidal concentration (MBC) values against E. coli (Gram-negative, ATCC 8739) and a 2-fold reduction against S. aureus (Gram-positive, ATCC 6538). Additionally, the antioxidant capacity of p(L-Cys) particles was retained somewhat, measured as 0.14 ± 0.01 µM versus 2.25 ± 0.03 µM Trolox equivalent/g for L-Cys. Therefore, p(L-Cys) particles are versatile and offer a unique avenue for immense biomedical use. Full article

Background: Metabolic syndrome (MetS) comprises interrelated metabolic abnormalities that collectively confer increased risk of cardiovascular disease and hepatic morbidity. The MAF-5 score is a non-invasive prognostic marker of liver fibrosis and mortality, while Cystatin C (CysC) is a sensitive indicator of renal function that also reflects inflammation, atherosclerosis, and metabolic dysfunction. Although both MetS and CysC have been widely studied, their potential interplay via MAF-5 remains unclear. We aimed to investigate the relationship between MAF-5 scores and CysC levels in MetS patients for the first time. Materials and Methods: Adults (≥18 years) with MetS were included in this study. MAF-5 scores (based on waist circumference, BMI, diabetes status, AST, and platelet count) and CysC levels were recorded. The MAF-5–CysC relationship was assessed via Pearson correlation. Participants were grouped into MAF-5 quartiles, and continuous variables were compared using ANOVA with Bonferroni-corrected pairwise tests. Results: We included 347 MetS patients (54.8% female, median age 54 years). The median MAF-5 score was 1.25, and MAF-5 correlated positively with CysC (r = 0.357, p < 0.001). CysC levels differed significantly across MAF-5 quartiles (Q1 = 0.96, Q2 = 0.99, Q3 = 1.06, Q4 = 1.09; p < 0.001), with Q4 showing higher values than Q1 and Q2. Conclusions: A significant correlation was found between MAF-5 scores and CysC in patients with MetS. CysC levels differed significantly across MAF-5 quartiles, suggesting a potential link between systemic inflammation, liver fibrosis, and CysC. These results highlight shared inflammatory and fibrotic pathways, underlying metabolic dysfunction. Clinically, combined assessment of MAF-5 and CysC may improve risk stratification, identifying patients at higher risk for hepatic fibrosis and adverse outcomes. Full article

Background: Chemotherapy-induced peripheral neuropathy (CIPN) is a major dose-limiting adverse effect of paclitaxel and is characterized by cold and mechanical allodynia. Effective therapeutic strategies for CIPN remain limited. This study evaluated the analgesic potential of Corydalis yanhusuo (CY) and Evodia rutaecarpa (ER), as well as their major alkaloids tetrahydropalmatine (THP) and rutaecarpine, in a mouse model of paclitaxel-induced neuropathic pain. Methods: Neuropathic pain was induced by paclitaxel administration (2 mg/kg, i.p., four injections). CY and ER extracts were orally administered at doses of 100 or 300 mg/kg, either alone or in combination, and cold and mechanical allodynia were assessed from days 0 to 8. The analgesic effects of THP and rutaecarpine were also examined. Gene and protein expression analyses were performed to evaluate the involvement of TRPV1 and TRPM8 signaling pathways, and high-performance liquid chromatography (HPLC) was used to confirm the presence of THP in CY and rutaecarpine in ER. Results: Paclitaxel reliably induced robust cold and mechanical hypersensitivity. Oral administration of CY or ER significantly alleviated allodynia in a dose-dependent manner, with greater efficacy at 300 mg/kg. Combined CY–ER treatment produced stronger anti-allodynic effects than either extract alone. THP and rutaecarpine also exhibited dose-dependent analgesic effects, and their co-administration yielded the most pronounced inhibition of paclitaxel-evoked hypersensitivity. Molecular analyses confirmed the involvement of TRPV1- and TRPM8-related pathways in these analgesic effects. Collectively, these findings indicate that CY, ER, and their representative alkaloids effectively attenuate paclitaxel-induced neuropathic pain and highlight CY–ER-based natural products as promising candidates for managing CIPN through modulation of TRPV1/TRPM8 signaling. Full article

The estimated glomerular filtration rate (eGFR) is a cornerstone of kidney function assessment. Widely used Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations based on serum creatinine (eGFR_cr), cystatin C (eGFR_cysC), or both (eGFR_cr-cysC) are influenced by non-glomerular filtration rate (GFR) factors, and their performance may vary across clinical contexts. We retrospectively analyzed 435 adult patients with simultaneous serum creatinine and cystatin C measurements. eGFR was calculated using CKD-EPI 2021 (creatinine), CKD-EPI 2012 (cystatin C), and CKD-EPI 2021 (combined) equations. Patients were classified into Kidney Disease: Improving Global Outcomes (KDIGO) GFR categories (G1–G5), and discrepancies between equations were identified. 44 patients (10.1%) showed discordant GFR categorization across all three equations and underwent detailed clinical assessment. 16 of the 44 discordant cases had clinically confirmed chronic kidney disease (CKD). The combined equation aligned with the clinical diagnosis in all CKD cases. eGFR_cr overestimated kidney function in 10/16 patients, while eGFR_cysC produced lower values in 8/16, consistent with early CKD but potentially influenced by inflammation or obesity. Reclassification occurred in 9/16 patients when switching from eGFR_cr to eGFR_cr-cysC, including four who shifted from G2 to G3a–G4. A significant difference was observed between eGFR_cr and eGFR_cr-cysC (p < 0.05). The combined CKD-EPI equation demonstrated the best clinical concordance, supporting its broader use when diagnostic accuracy is essential. Full article

Currently, there is no report on prediction models of standardized ileal amino acid digestibilities (SIAADs) in soybean meals (SBMs) for medium-growing yellow-feathered chickens. This study firstly analyzed the chemical compositions of 10 SBMs, then determined their SIAADs in chickens, and finally established and verified prediction models for SBM SIAADs based on their chemical compositions and amino acid (AA) profiles. A total of 276 55 d-old Tianluma roosters were selected and randomly divided by body weight into 11 treatment groups. On d 63, chickens were fed either a nitrogen-free diet (NFD) or one of 10 SBM diets for 5 d. On d 67, ileal chyme samples were collected to determine SIAADs. Data from nine SBM samples and stepwise regressions were employed to build prediction models, while one SBM sample was randomly selected to validate model accuracy. Different SBM sources affected (p ≤ 0.007) SIAADs in medium-growing yellow-feathered chickens. The standardized ileal digestibility (SID) of glutamic acid (Glu) was the highest (93.9%), whereas that of cysteine (Cys) was the lowest (81.7%). Fifteen prediction models (R² = 0.567–0.993, p < 0.03) for the SIDs of methionine (Met), isoleucine (Ile), leucine (Leu), phenylalanine (Phe), lysine (Lys), histidine (His), arginine (Arg), aspartic acid (Asp), serine (Ser), Glu, glycine (Gly), alanine (Ala), Cys, tyrosine (Tyr), and proline (Pro) in SBMs for medium-growing yellow-feathered chickens were effectively established based on chemical compositions and AA profiles. Among them, the prediction model for the SID of Cys showed the best fit (R² = 0.993, p = 0.002), while the model for the SID of Ala had the lowest fit (R² = 0.567, p = 0.019). Except for His and Pro, which exhibited poor predictive accuracy, all other models showed good accuracy. These prediction models thus provide a valuable reference for rapidly estimating the SIDs of key AAs in SBMs for medium-growing yellow-feathered chickens. Full article

This paper presents the design of G-IDSS (Graph-based InnoCyPES Data Storage Service): a novel, distributed data storage service that is built around a P2P network overlay to support handling distributed data. G-IDSS is accessible through a standard command-line interface and is based on a graph database to support the schema-less management of the distributed data stored among peers. The mechanisms to facilitate the execution of complex queries requiring distributed data integration and fusion are also presented and discussed. Besides the design, this work also provides relevant details related to the implementation of G-IDSS, reflecting several use cases that demand data that are distributed across different, even geographically spread, locations. G-IDSS scales to thousands of peers in an overlay, it is able to run distributed queries and can integrate data that are stored in different sources. Full article

Background/Objectives: Krabbe disease (KD) is a hereditary lysosomal disorder whose hallmark is progressive demyelination, with variable involvement of the central nervous system. It is caused by pathogenic variants in the GALC gene that disrupt the function of its gene product, the lysosomal enzyme galactosylceramidase. We analyzed two unrelated cases (one early infantile and one adult) with a clinical suspicion of KD. Methods: We used a combination of biochemical techniques (high-performance liquid chromatography–tandem mass spectrometry), NGS (resequencing gene panels), splicing assays, and molecular modeling to identify and analyze the pathogenicity of the variants underlying the disorder. Results: The two probands were compound heterozygotes for disease-causing variants in the GALC gene, encoding the lysosomal hydrolase galactosylceramidase. Three of the variants were novel and caused aberrant splicing, either by exon skipping (c.908+5G>A and c.1034-1G>C) or by inclusion of a cryptic, deep intronic pseudoexon (c.621+772G>C). The fourth variant was a known missense change (c.956A>G, p.(Tyr319Cys)) with conflicting interpretations of pathogenicity in the databases. Conclusions: We demonstrated the pathogenicity of the three novel splicing variants, all with strong impact on galactosylceramidase function. We also concluded that the c.956A>G missense variant is a hypomorph usually underlying the later-onset, milder phenotypes of KD. Our results stress the importance of integrated approaches combining clinical, biochemical, and genetic testing to obtain a definitive diagnosis of lysosomal diseases. Full article

Healthy soil serves as the fundamental basis for sustainable Panax notoginseng (Burkill) F.H. Chen ex C.Y. Wu & K.M. Feng cultivation in understory systems. Current management practices have raised concerns about potential soil degradation and ecological imbalance. To comprehensively assess the soil health status, this study investigated typical understory P. notoginseng plantations in the subtropical mountain monsoon region of western Yunnan. By analyzing 29 soil physical, chemical, and biological indicators, we constructed a Minimum Data Set (MDS) using Principal Component Analysis to evaluate soil health and identify major constraints. The results showed that the MDS for soil health assessment consisted of 11 key indicators: acid phosphatase, fungal ACE index, organic matter, total nitrogen, sucrase, fungal Simpson index, fine sand, non-capillary porosity, silt content, bulk density, and microbial biomass nitrogen. Using both linear and non-linear scoring functions, the Soil Health Index (SHI) calculated based on the MDS showed a significant positive correlation with the SHI derived from the Total Data Set (TDS) (linear scoring: R² = 0.43, p < 0.001; non-linear scoring: R² = 0.305, p < 0.001). This indicates that the MDS captures a substantial and significant portion of the variation explained by the TDS and can serve as a practical and simplified alternative for soil health evaluation in this cultivation system. Based on the MDS, the SHI values obtained using linear and non-linear scoring functions ranged from 0.53 to 0.72 and 0.48–0.59, with mean values of 0.62 and 0.51, respectively, indicating moderate soil health status in the study area. Significant differences in SHI were observed across planting durations and seasons (p < 0.05), with two-year-old plantations showing notably better soil health indices than three-year-old plantations, particularly during the rainy season. The main constraints identified in understory P. notoginseng plantations included microbial community degradation, nutrient imbalance, and physical structural deterioration. Implementing scientific soil management strategies such as optimized rotation cycles, organic amendment applications, and microbial community regulation can effectively mitigate these soil constraints, enhance soil health, and promote the sustainable development of understory P. notoginseng cultivation. Full article

Glucagon-like peptide-1 receptor agonists and lifestyle interventions effectively treat overt obesity, but the benefits/risks of their combined early intervention during middle age remain unclear. This study investigated whether submaximal-dose liraglutide combined with strength–endurance training improves metabolic and liver health, focusing on hepatic oxidative stress and lipid metabolism. Male Wistar rats (16 months old) received liraglutide (L; 0.186 mg/kg/day, s.c.), training (ladder climbing with weights, 3 times/week), both (L+E) or saline for control middle-aged (C) and young adults (CY; 3–4 months old) for 7 weeks (n = 8/group). Middle-aged rats exhibited age-related changes including higher body and visceral fat, increased hepatic and serum cholesterol, hepatic ALT and glutathione imbalance, and decreased soleus muscle (p < 0.05, vs. CY). Exercise increased hepatic glycogen and oxidative stress markers and downregulated lipogenic genes, consistent with liver adaptation to training. L+E synergistically reduced body and visceral fat, hepatic and serum triglycerides, and the triglyceride–glucose index, while reducing oxidative stress (p < 0.05 vs. E, C) and lipogenic gene expression (p < 0.05 vs. C), without affecting pancreas histopathology and function parameters, muscle mass or exercise load volume. In conclusion, submaximal liraglutide safely synergized with training to enhance metabolic health, improve hepatic redox balance and triglyceride metabolism in middle-aged rats, without mitigating cholesterol rise. Full article

This study described TTN gene variants in Ecuadorian patients with hereditary cardiac diseases, integrating genetic ancestry to improve variant interpretation in an underrepresented population. Sixty patients with confirmed hereditary cardiac conditions were analyzed using the TruSight Cardio NGS panel (Illumina, San Diego, CA, USA), which targets 174 cardiac-associated genes. Bioinformatic analyses and classification were performed in accordance with ACMG/AMP guidelines, and ancestry inference was conducted using 46 Ancestry Informative Markers (AIM-InDels). From 4008 detected TTN variants, 29 variants of interest remained after filtering: 27 classified as variants of uncertain significance (VUS) and two as likely pathogenic. All variants were heterozygous and distributed across exons 3–358, primarily in the A-band region, commonly associated with cardiomyopathies and arrhythmic phenotypes. Two truncating variants (exons 267 and 272) met PVS1 criteria, while several missense variants (p.Ser91Gly, p.Pro12140Ser, p.Arg34653Cys) showed possible modulatory effects on hypertrophic or arrhythmic outcomes. Genetic ancestry revealed a predominant Native American background, followed by European and African components. These findings expand the understanding of TTN-related cardiac disease in Latin America, suggesting that TTN functions as a genetic modifier influencing disease expression. Incorporating ancestry information enhances genomic interpretation and supports precision medicine in diverse populations. Full article

Objective: This study investigated whether rapamycin could modulate the polarisation of CD4+ T cells towards T_H1, T_H2, T_H17, and Treg cells using peripheral blood mononuclear cell (PBMC) obtained from patients with granulomatosis with polyangiitis and microscopic polyangiitis (GPA/MPA). Methods: Twenty patients with GPA/MPA were included in this study. Their stored PBMCs were cultured and stimulated with anti-CD3 and anti-CD28 antibodies for 72 h in the presence or absence of rapamycin (10 nM). The cells were stained for surface markers with anti-CD4-FITC and anti-CD25-APC, followed by intracellular staining using anti-interferon (IFN)-γ-PE, anti-IL-4-PerCP-Cy5, anti-IL17A-APC, and anti-Foxp3-PE. The stained cells were analysed using a flow cytometer. Results: The median age of the 20 GPA/MPA patients (10 men and 10 women) was 65.5 years. Rapamycin treatment significantly modulated the polarisation of CD4+IFN-γ+ T (T_H1) cells compared to no treatment among GPA/MPA patients. In addition, the polarisation of CD4+IFN-γ+ T (T_H1) cells was also significantly reduced in rapamycin-treated PBMC obtained from active patients compared to untreated PBMC from the same patients; however, these alterations were not observed in inactive patients. Conversely, rapamycin treatment did not affect the polarisation of CD4+IL-4+ T (T_H2), CD4+IL-17+ T (T_H17), or CD4+FoxP3+CD25+ T (Treg) cells, regardless of GPA/MPA activity. Conclusions: This study was the first pilot study to demonstrate that rapamycin modulates the polarisation of CD4+ T cells towards CD4+IFN-γ+ T cells in active GPA/MPA. Full article

One of the hallmarks of Osteogenesis Imperfecta (OI) is phenotypic variability among individuals with the same mutation. The aim of our study is to investigate the under-explored role of osteoblast differentiation in OI phenotypic variability by using human and murine OI osteoblasts. This is the first comparative study of osteoblasts from OI patients vs. healthy pediatric controls. We investigated osteoblasts carrying COL1A1 substitutions Gly352Ser and Gly589Ser, each expressed in two unrelated patients differing in phenotypic severity. Osteoblasts from type III OI patients with both mutations deposited significantly less mineral vs. type IV. RNA-Seq showed osteoblasts from type IV OI patients with different mutations had downregulated mitochondrial pathways, while osteoblasts from type III OI patients showed downregulation of extracellular matrix pathways. Puromycin assay demonstrated osteoblast protein synthesis was significantly upregulated in type III vs. type IV OI patients. UPR PERK and BiP were reduced in osteoblasts with Gly352Ser from type III and IV OI patients and in osteoblasts with Gly589Ser from a type III OI patient, while both proteins were increased in Gly589Ser osteoblasts from the type IV patient. Additionally, in a murine comparative study, Col1a1 Gly349Ser, called Brtl Ser, showed a much more severe skeletal phenotype than Brtl Cys. Brtl Ser calvarial osteoblasts had reduced collagen secretion and folding with abnormal dermal collagen fibrils vs. wildtype. Also, Brtl Ser osteoblasts showed condensed actin filaments but a similar mineral deposition as Brtl Cys. Electron microscopy revealed elongated mitochondria with cristae dropout in patient and mutant murine osteoblasts. Our study yielded novel insights highlighting osteoblast differentiation, mineralization, and a potential role of mitochondria in OI pathology and phenotypic variability. Full article