Search Results (782)

Nitrogen metabolism in the human body is in a strictly balanced state, which is disturbed in pathologies, including breast cancer. The state of nitrogen balance can be judged by the content of urea and the amount of branched-chain amino acids (BCAAs) (Val, Leu, and Ile), glutamine (Gln), and glutamate (Glu). The study involved 1438 people, including patients with breast cancer (n = 543), fibroadenomas (n = 597), and healthy controls (n = 298). Saliva samples were collected from all patients before treatment, and urea levels were determined in all 1438 samples. Salivary levels of BCAAs, Gln, and Glu were determined in 116 patients with breast cancer, 24 with fibroadenomas, and 25 healthy volunteers. An increase in the concentration of urea in saliva was shown in breast cancer, most pronounced in luminal molecular biological subtypes: luminal A 10.46 [7.69; 12.62] mmol/L (p < 0.0001), luminal B HER2-negative 9.52 [6.72; 12.52] mmol/L (p = 0.0198), and luminal B HER2-positive 8.26 [5.27; 12.07] mmol/L. The Gln/Glu ratio increased in the saliva of the control group (5.43 [3.30; 10.5]) compared with breast cancer (2.22 [0.84; 5.40], p = 0.0094) and fibroadenomas (1.94 [0.89; 6.05], p = 0.0184). For luminal B HER2-positive and TNBC, the Gln/Glu ratio increased sharply to 8.23 [3.24; 10.9] (p = 0.0327) and 11.2 [4.28; 15.2] (p < 0.0001) compared with healthy controls. Thus, an increased Gln/Glu ratio in saliva may characterize a more aggressive subtype of breast cancer. Full article

Natural tyrosinase inhibitors are an important group of compounds with cosmetical and medicinal applications. With the aim of finding new types of natural tyrosinase inhibitors from ornamental and medicinal plants, Lycopodium japonicum was selected and studied. As a result, fifteen structurally diverse secondary metabolites 1–15 were isolated and identified. Their chemical structures were identified by analysis of their spectral data and compared with those reported in the literature. In the tyrosinase inhibitory bioassay, five phytochemicals, 4, 12, 13, 14, and 15, exhibited significant inhibitory effects, with half-maximal inhibitory concentration (IC₅₀) values ranging from 1.46 to 6.82 mM. Additionally, molecular docking studies disclosed that Lys376, Lys379, Gln307, and other amino acid residues played key roles in the potential binding interactions between the active compounds and the tyrosinase. These findings suggest that the species L. japonicum is a warehouse of natural tyrosinase inhibitors. Full article

This study established an efficient in vitro regeneration system using stem nodes from root collar suckers as explants. Subsequently, regenerated shoots were used to establish an in vitro medicinal production protocol that achieved ginkgolic acid production. The self-developed Ginkgo biloba medium (GBM), first reported in this study, was pivotal to system establishment. The plantlet propagation system showed that the bases of stem nodes dipped in GBM with 2 mg·L⁻¹ 6-benzyladenine (BA) and 0.2 mg·L⁻¹ 1-naphthaleneacetic acid (NAA) achieved near-complete axillary bud induction (99.56%). Adventitious shoot induction reached 82.22% (3.5 shoots/explant) using GBM with 0.2 mg·L⁻¹ BA, 0.02 mg·L⁻¹ kinetin (Kin) and 0.2 g·L⁻¹ proline (Pro). Maximum adventitious shoot elongation (92.22%, average 3.35 cm) was observed on GBM containing 0.1 mg·L⁻¹ zeatin (ZT) and 0.01 mg·L⁻¹ BA. After 3-week preculture with 15 mg·L⁻¹ phloroglucinol (PG), treatment with 0.6 mg·L⁻¹ indole-3-butyric acid (IBA) and 0.2% activated carbon (AC) yielded 96.67% rooting (6.19 roots/explant) and 85% acclimatization survival. For medicinal resource production, bud cluster induction at 94.44% (20.89 buds/explant) on GBM with 1 mg·L⁻¹ BA, 0.03 mg·L⁻¹ Kin, and 0.2 g·L⁻¹ Pro. Leaf organs in GBM with 0.3 mg·L⁻¹ BA, 0.01 mg·L⁻¹ Kin, 0.01 mg·L⁻¹ IBA, 0.3 g·L⁻¹ Pro, and 0.01 mg·L⁻¹ glutamine (Gln) accumulated 20.64 g fresh weight and 41.910 mg·g⁻¹ DW ginkgolic acids, representing a 4.93-fold increase over mother plants. This system enables large-scale Ginkgo biloba L. propagation and provides an in vitro strategy for producing medicinal compounds in endangered plants. Full article

Background: Dilated cardiomyopathy (DCM) and long QT syndrome (LQTS) are genetically heterogeneous cardiac disorders that contribute significantly to morbidity and sudden cardiac death. Although they are typically considered distinct entities, co-occurrence within families has been increasingly recognized, complicating diagnosis and genetic counseling. Identifying shared genetic determinants may provide insights into overlapping disease mechanisms. Methods: We investigated a multi-generational family in which several members presented with features of both DCM and LQTS. Exome sequencing was performed to identify potential disease-causing variants, and candidate findings were validated by Sanger sequencing. In silico prediction tools and evolutionary conservation analysis were used to assess the pathogenic potential of the identified variant. Results: We identified a novel heterozygous missense variant in the CRYAB gene, c.368G>A (p.Arg123Gln). This variant is located in a highly conserved region critical for protein function and was consistently predicted to be deleterious across multiple computational algorithms. Segregation analysis demonstrated co-occurrence of the variant with disease phenotypes in affected family members. Clinically, several carriers exhibited overlapping features of both DCM and prolonged QT interval, suggesting a dual cardiac phenotype associated with this mutation. Conclusions: Our findings expand the phenotypic spectrum associated with CRYAB mutations, linking them to a combined presentation of dilated cardiomyopathy and long QT syndrome. This underscores the importance of including CRYAB in comprehensive gene panels for inherited cardiac disorders and highlights the need for integrated clinical and genetic evaluation in families presenting with complex cardiac phenotypes. Full article

In this study, nanoparticles prepared by the heat-induced self-assembly of bovine serum albumin-dextran conjugates (BSA-DX) were utilized as an effective delivery system for the immunoregulatory peptide Gln-Leu-Asn-Trp-Asp (QLNWD) from Meretrix meretrix. The effects of dry-heating duration on the fabrication and characteristics of glycoprotein nanoparticles (GBA NPs) were investigated. Stable GBA NPs (110.84 nm) were obtained after 9 h of dry-heating. Depending on the addition sequence of QLNWD, the QLNWD-loaded nanoparticles were categorized into two types: pre-loading and post-loading. The two strategies were evaluated based on physicochemical characterization, colloidal stability, and RAW264.7 macrophage uptake. Results showed that upon QLNWD incorporation, both pre-loading NPs and post-loading NPs exhibited spherical morphology, with particle sizes decreasing to 105.51 nm and 94.27 nm, respectively. The encapsulation efficiency of pre-loading NPs for QLNWD was higher (87.74%), and the co-localization ability between post-loading NPs and QLNWD was stronger (Pearson’s correlation coefficient = 0.95). In vitro simulated gastrointestinal digestion experiments showed that QLNWD bioaccessibility increased to 47.5% and 42.7% for pre-loaded and post-loaded NPs, respectively. Compared to free QLNWD, NP encapsulation significantly enhanced the uptake of QLNWD by macrophages. Thus, GBA NPs, particularly those prepared by the pre-loading method, are considered promising delivery systems for marine bioactive peptides. Full article

Despite significant advances in understanding the genetics of Parkinson’s disease (PD) and Parkinsonism, the diagnostic yield remains low. Pathogenic variants of GBA1, which encodes the lysosomal enzyme β-glucocerebrosidase and causes recessive Gaucher dis-ease, are recognized as the most important genetic risk factor for PD in heterozygous carriers. This study focuses on the functional genomics of rare genetic variations in other lysosomal hydrolytic enzymes genes in patient-derived fibroblasts. We examined 49 early-onset PD patients using whole exome sequencing and in silico panel analysis based on a curated PD gene list. Two patients were found to carry the p.Asp313Tyr variant in the X-linked GLA gene (encoding GALA, typically associated with Fabry disease), and one patient carried the p.Arg419Gln variant in GLB1 (encoding β-Gal, linked to the recessive GM1 gangliosidosis and mucopolysaccharidosis type IVB). The in silico study of both variants supports a potentially damaging impact on the encoded protein function and structural destabilization. Additional candidate variants were found related to lysosomes, Golgi apparatus and neurodegeneration, suggesting a multifactorial contribution to the disease. However, none of these variants met diagnostic standards. Functional assays showed a significant decrease in GALA expression and partial retention of the enzyme in the trans-Golgi network in fibroblasts with GLA:p.Asp313Tyr, while altered Golgi morphology was observed in fibroblasts with GLB1:p.Arg419Gln. Moreover, all patients exhibited abnormalities in lysosomal morphology, altered lysosomal pH, and impaired autophagic flux. Our findings suggest that rare, heterozygous variants in lysosomal-related genes, even when individually insufficient for monogenic disease, can converge to impair lysosomal homeostasis and autophagic flux in EOPD. The underlying genetic and cellular heterogeneity among patients emphasizes the importance of combining genetic and functional approaches to better understand the mechanisms behind the EOPD, which could enhance both diagnosis and future treatments. Full article

In vitro ovule culture serves as an experimental platform for exploring the growth and development processes of cotton fibers. However, over the decades, research on the in vitro ovule culture of upland cotton has remained underdeveloped. In this study, ovules collected 2 days post-anthesis (2 DPA) from the upland cotton genetic standard line TM-1 were used to investigate the effects of carbon sources (glucose, fructose, sucrose), kinetin (KT), and glutamine (Gln) on ovule growth and observed fiber development in vitro. The results showed that the ovules grew more favorably on a medium supplemented with 0.05 M glucose, 0.02 M fructose, and their degradation products as carbon sources. Regarding the role of KT, it has a slight inhibitory effect on the development of cotton fiber in vitro at a lower concentration (0.1 mg/L). However, as the concentration increased (0.5 mg/L), its effect shifted to promotion. Additionally, Gln demonstrated the ability to enhance the characteristics of fiber fluffiness. In this study, the optimized formula for the in vitro ovule culture of upland cotton was established. This method provides an improved technical system for the in vitro ovule culture of upland cotton, holding great potential for fiber function genomics and seed bioengineering in cotton. Full article

Background: Extra digits on the hands and/or feet are a frequent condition known as polydactyly. Twelve nonsyndromic polydactyly genes have been identified, including KIAA0825. Methods: Four consanguineous Pakistani families that segregate nonsyndromic postaxial polydactyly (PAP) with an autosomal recessive mode of inheritance were clinically and genetically evaluated. Exome sequencing or genotyping of polymorphic microsatellite markers followed by Sanger sequencing were used to identify the variants underlying the PAP etiology. Results: Three novel KIAA0825 variants were identified that segregate with PAP: a nonsense variant c.2319G>A; p.(Trp773*) in two families; a missense variant c.970G>T; p.(Val324Phe) in one family; and a four amino acids in-frame deletion c.2743_2754del; p.(Gln915_Val918del) in one family. The nonsense variant segregated in families with PAP type B (PAPB), while the missense and the in-frame deletion variants segregated with PAP type A and B. Conclusions: The findings of this study expanded the clinical and genetic spectrum of PAP due to KIAA0825 variants including the first KIAA0825 variant specific to PAPB. Full article

Background/Objectives: This study identifies novel dihydroorotate dehydrogenase (DHODH) inhibitors exhibiting potent broad-spectrum antiviral agents, particularly against influenza A virus (A/PR/8/34(H1N1)) and SARS-CoV-2. Methods: Structure-based virtual screening of 1.6 million compounds (ChemDiv and TargetMol databases) yielded 10 candidates, with compounds 6, 9, and 10 demonstrating significant anti-influenza activity (IC₅₀ = 4.85 ± 0.58, 7.35 ± 1.65, and 1.75 ± 0.28 μM, respectively). Building on these, molecular hybridization principles and scaffold hopping principles were applied to design and synthesize six novel compounds (11–16) through cyclization, coupling, and carboxylate deprotection. Prior to subsequent biological assays, the molecular structures of each compound were elucidated by NMR spectroscopy and MS. Their antiviral activities were subsequently assessed against both influenza virus and SARS-CoV-2. The compound 11, demonstrating the most potent antiviral activity, was further subjected to surface plasmon resonance (SPR) analysis to assess its binding affinity for human DHODH. Results: Compound 11 emerged as the most potent DHODH inhibitor (K_D = 6.06 μM), exhibiting superior broad-spectrum antiviral activities (IC₅₀ = 0.85 ± 0.05 μM, A/PR/8/34(H1N1); IC₅₀ = 3.60 ± 0.67 μM, SARS-CoV-2) to the reported DHODH inhibitor (Teriflunomide, IC₅₀ = 35.02 ± 3.33 μM, A/PR/8/34(H1N1); IC₅₀ = 26.06 ± 4.32 μM, SARS-CoV-2). Mechanistic evaluations via 100 ns MD simulations and QM/MM calculations revealed stable binding interactions, particularly hydrogen bonds with GLN47 and ARG136, while alanine scanning mutagenesis confirmed these residues’ critical roles in binding stability. Conclusions: This work identifies compound 11 as a potent broad-spectrum antiviral compound, offering a promising strategy for broad-spectrum antiviral therapy against RNA viruses by depleting pyrimidine pools essential for viral replication. Full article

Background/Objectives: Sanfilippo Syndrome type B or Mucopolysaccharidosis type IIIB (MPS IIIB, OMIM 252920) is a lysosomal storage disease caused by deficiency of alpha-N-acetylglucosaminidase (NAGLU, E.C. 3.2.1.50) due to pathogenic variants in the NAGLU gene (17q21.2). The disease is characterized by progressive neurological manifestations, marked by cognitive decline, with relatively mild somatic involvement. We aim to present relevant information on a cluster of MPS IIIB identified in Ecuador, particularly regarding their clinical, biochemical, genetic, demographic, and ancestry characteristics. Methods: We present a characterization of a clinical, biochemical, genetic and demographic cluster of MPS IIIB patients in Ecuador, located in four main regions: Manabí, Guayas, Los Ríos, and Santo Domingo de los Tsáchilas. The patients included were diagnosed due to increased levels of urinary glycosaminoglycans (uGAG), plus deficient activity of NAGLU, and/or identification of biallelic pathogenic mutations in the NAGLU gene. Patients’ charts were reviewed for biochemical findings, medical history, clinical manifestations and assessments. Results: We present the results of clinical, biochemical, genetic and demographic characterization of a cluster in Ecuador with 24 patients identified with Sanfilippo syndrome type IIIB, resulting in an estimated incidence of 1.5/100,000. The mean age at diagnosis was 8.8 years, with symptom onset at 4.5 years on average. All patients exhibited elevated levels of uGAG and undetectable NAGLU activity, and all of them presented the c.1487T>C (p.Leu496Pro) variant in the NAGLU gene in homozygosis, indicating a possible founder effect, with the exception of one heterozygous one (p.Leu496Pro/p.Arg482Gln). A positive correlation between age of diagnosis and the concentration of one isoform of heparan sulfate (HS-OS) was found (p < 0.05). Clinical findings included neuropsychomotor developmental delay (75%), neurological regression (65%), hepatomegaly (55%), growth deficiency (50%), coarse facies (45%) and hernia (40%). Male patients presented earlier onset of symptoms. Maternal ancestry was successfully determined for 21 of the 24 patients. The majority were of Native American ancestry (71.4%), followed by European (19%), African (4.8%), and Asian (4.8%) lineages. Haplogroup A was the most prevalent (42.9%), followed by haplogroups D (19%), C, U, and H (each 9.5%), and R and L2 (each 4.8%). Conclusions: Ancestry can indicate a possible mechanism to explain the heterogeneous symptomatic presentation. These findings highlight the need for further research on genetic and environmental influences on disease severity in this population. Full article

Background/Objectives: Neurodevelopmental disorders (NDDs) are genetically heterogeneous conditions with a complex molecular etiology involving numerous genes. Biallelic pathogenic variants in INTS1 cause a rare autosomal recessive NDD characterized by congenital cataracts, growth retardation, facial dysmorphism, and global developmental delay. To date, the clinical description of this disorder has been based solely on individual case reports, and its phenotypic spectrum remains incompletely defined. Methods: A 9-year-old female proband was evaluated for developmental delay, multiple congenital anomalies, and distinctive craniofacial features. Whole-exome sequencing (WES) was performed, followed by Sanger validation and segregation analysis. Variant pathogenicity was assessed using in silico prediction tools and 3D protein structural modeling. Results: Whole-exome sequencing identified two novel compound heterozygous missense variants in INTS1, c.1145G>A (p.Arg382Gln) and c.1195G>A (p.Gly399Ser), both located in exon 9. Segregation analysis showed that c.1145G>A was inherited from the father and c.1195G>A from the mother, and both variants are extremely rare in population databases. Conclusions: We report a patient carrying novel biallelic INTS1 variants, whose clinical presentation differs from previously reported cases, including those with milder phenotypes characterized by preserved speech development and absence of intellectual disability. This observation broadens the clinical spectrum of INTS1-related disease and underscores its phenotypic heterogeneity. Full article

Background: Amino acid biomarkers have a crucial influence on our understanding of brain injury mechanisms, and their plasma concentrations may indicate neurological damage and recovery patterns. Pediatric mild traumatic brain injury (mTBI) assessment particularly benefits from such molecular indicators, as clinical presentations can be subtle and variable. However, current diagnostic and prognostic tools lack reliable biochemical markers that can track the temporal evolution of injuries and recovery. Methods: We conducted a prospective longitudinal cohort study involving 36 pediatric mTBI patients and 44 controls to characterize the temporal evolution of key amino acids and their derived indices. Blood samples were collected at 3, 6, 12, and 24 h and at 7, 14, and 28 days post-injury, with amino acids quantified using high-performance liquid chromatography. Results: Our analysis revealed significant temporal changes in glutamate, glutamine, and glycine concentrations, with glutamate peaking at day 7 before declining, while glutamine showed steady increases throughout. The GLN/GLU ratio demonstrated an early excitatory imbalance followed by astrocytic compensation, and the GLX ratio indicated progressive recovery. Conclusions: These patterns represent continuous neurochemical processes involving excitotoxicity and glial regulation, suggesting potential utility as biomarkers for mTBI diagnosis and monitoring. While further validation using larger cohorts is needed, these findings provide compelling evidence of the efficacy of using amino acid profiles to track pediatric mTBI progression and recovery. Full article

The study assesses the relationship between thalamic proton-MR spectroscopy (¹H-MRS) metabolites and thalamic ¹¹C-ER176 translocator-protein positron emission tomography (TSPO-PET) standardized uptake value ratios (SUVR) to advance our understanding of thalamic involvement in multiple sclerosis (MS)-associated neurodegeneration and disability. In this prospective cross-sectional study, patients with MS (pwMS) and controls underwent 3T-MRI, ¹H-MRS, and ¹¹C-ER176-PET targeting the thalamus. MRI-derived thalamic volume was normalized by intracranial volume. ¹H-MRS metabolites—N-acetylaspartate (NAA), glutamate (Glu), glutamine (Gln), total choline (tCho), and myo-inositol (mIns)—were normalized to total creatine (tCr). Clinical disability was evaluated using MS-specific tests of Expanded Disability Status Scale-EDSS and MS-functional composite-MSFC (including Paced Auditory Serial Addition Test-PASAT). Compared to controls (n = 30), pwMS (n = 21) exhibited smaller thalamic volume, higher thalamic ¹H-MRS mIns/tCr (putative gliosis marker), and higher thalamic ¹¹C-ER176-PET SUVR (glial density marker). In pwMS, higher thalamic mIns/tCr (r = −0.67) and tCho/tCr (r = −0.52) correlated with smaller thalamic volume. In pwMS, higher thalamic mIns/tCr correlated with higher thalamic ¹¹C-ER176-PET SUVR (r = 0.48) and decreased cognitive function (PASAT, rho = −0.48). In controls, decreased thalamic NAA/tCr correlated with increased thalamic ¹¹C-ER176-PET SUVR (r = −0.41). Thalamus, a core central nervous system relay, is affected early in MS disease course. Glial-mediated innate immune activation in the thalamus, evaluated by increased ¹H-MRS mIns/tCr and ¹¹C-ER176-PET SUVR, is associated with loss of thalamic volume and increased disability in pwMS. The multimodal imaging approach with ¹H-MRS mIns/tCr and ¹¹C-ER176-PET SUVR emerges as potential glial biomarkers, to better understand disease mechanisms and evaluate therapeutic interventions targeting glial activity in MS. Full article

The global gold extraction industry has been reported to use cyanide-based recovery processes, which pose environmental effects on water resources. The study examined Coptodon zillii liver rhodanese from a gold mining-impacted reservoir with a specific focus on the enzyme’s critical function in cyanide detoxification. Rhodanese was purified using successive chromatographic techniques with 5.4 U/mg specific activity and 3.1-fold purification. The molecular weight of the native enzyme was 36 kDa, and the subunits were 17 kDa, indicative of a dimeric structure. Optimal enzymatic activity was recorded at pH 8.0 and 50 °C. The effect of metal ions was significantly varied: the activity was inhibited by BaCl₂, CaCl₂, NaCl, and MgCl₂, and KCl enhanced performance. The kinetic determinations showed Michaelis-Menten kinetics with a Km of 20.0 mM for sodium thiosulfate and 25.0 mM for potassium cyanide. The enzyme’s minimal activity was identified toward 2-mercaptoethanol, ammonium persulfate, and ammonium sulfate, but with evidence of preference for thiosulfate utilization under the substrate specificity tests. The major interactions between the enzyme and the substrate were revealed by the molecular docking experiments. These showed Glu159, Gln161, and Arg173 formed important hydrogen bonds with thiosulfate, while Arg156 and Val172 were also involved. Other substrates are bound to Gln121 and Trp139 residues with much lower binding energy than thiosulfate. The findings increase our understanding of biochemical adaptation process knowledge in anthropogenically stressed environments, showing strategies of ecological resilience. The characterized enzymatic features showed potent cyanide detoxification potential, and the possible applications are in bioremediation strategies for mining-impacted aquatic ecosystems. Full article

Glutamine (Gln) supplementation during the weaning period can alleviate stress in piglets. However, free Gln has poor stability and low absorption in the small intestine. Glycyl-glutamine (Gly-Gln), a stable dipeptide form of Gln, has been evaluated as a potential alternative in pig nutrition. This study investigated the effects of Gly-Gln at 0, 0.125%, 0.25%, 0.375%, and 0.50%, as well as a Gly + Gln positive control, on growth performance, intestinal morphology, immunity, antioxidant status, and nutrient apparent digestibility in weaned piglets. The results showed that dietary supplementation with 0.25%, 0.375%, or 0.50% Gly-Gln significantly increased average daily gain, average daily feed intake, and final weight (p < 0.05). Linear and quadratic effects (p < 0.05) were observed for growth performance indicators, suggesting that moderate supplementation levels yielded optimal benefits. Dietary Gly-Gln supplementation with 0.25%, 0.375%, or 0.50% Gly-Gln significantly increased serum immunoglobulin (IgG, IgA, and IgM), insulin, insulin growth factor 1, growth hormone, and T4 and T3 contents, and decreased IFN-γ and IL-1β contents (p < 0.05). Diets supplemented with 0.25, 0.375, or 0.50% Gly-Gln increased total antioxidant capacity and superoxide dismutase content in serum and liver, and decreased MDA content (p < 0.05). Compared with the negative control group, dietary supplementation of 0.25%, 0.375%, or 0.50% Gly-Gln significantly increased the mRNA expression of ZO-1, Occludin, and Claudin-1 in the jejunum (p < 0.05). Furthermore, crude protein digestibility was significantly improved in piglets receiving 0.375% and 0.5% Gly-Gln (p < 0.05), with a significant linear relationship between Gly-Gln level and digestibility. In conclusion, 0.25% is the minimum effective dose of Gly-Gln for improving weaning outcomes. Gly-Gln is more effective than equivalent doses of free glycine and glutamine in enhancing growth performance, gut barrier integrity, and nutrient utilization in weaned piglets. Full article