bims-supasi 2026-09-27 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2026–09–27
twenty-one papers selected by
Jonathan Wolf Mueller, University of Birmingham

Discovery and characterization of Chondroitinase A with specificity for 4-O-sulfated N-acetylgalactosamine in Chondroitin sulfate/Dermatan sulfate.
Synthesis of the endogenous non-reducing end heparan sulfate disaccharides for newborn screening and diagnosis of mucopolysaccharidosis types I and II.
Laminarin sulfate from Lessonia flavicans as a potential non-animal anticoagulant with minimal hemorrhagic risk: Structural characterization by NMR spectroscopy and activity analysis.
Sulfation Rewrites the Biological Profile of Dietary Flavonoids: Implications for Stability, Redox Signaling, and Inflammation.
Chikungunya Virus E2 N218K Enhances Heparan Sulfate-Dependent Attachment and Viral Internalization While Conferring Partial Attenuation in A129 Mice.
Sulfation of Sugarcane-Bagasse Xylan: Process Optimization, Physicochemical Properties and In Vitro Anticoagulant Activity.
Indoxyl Sulfate, Cardiac Biomarkers, and Left Ventricular Wall Thickening in Cats with Chronic Kidney Disease.
Dysregulation of the Heparanase/TFPI-2/Heparan Sulfate Axis in Recurrent Pregnancy Loss.
One-pot fabrication and in vitro characterization of hydroxyapatite-enriched cryogels for bone tissue engineering.
Alpha-N-acetylglucosaminidase (NAGLU) inactivation causes morphological damage to the ultrastructural appearance of epididymal mononuclear phagocytes (eMPs) and epithelial cells of the epididymis in adult mice.
Determining Carrageenan Sulfate Groups Using Ion Association with Alcian Blue Dye.
The effect and mechanism of TIPE in promoting M2 polarization through DcR3 in the microenvironment of colorectal cancer.
Clinical Characteristics and Thyroid Findings in Children with Isolated Premature Adrenarche.
Sulfated Hyaluronan Drives Cell Adaptation and Matrix Composition in Advanced 3D Breast Cancer Cell Models.
Establishment of Serum Cholesterol Sulfate Reference Intervals and Evaluation of Its Diagnostic and Metabolic Associations in Chinese Adults.
Clinical and biochemical correction of a patient with neurodegenerative Mucopolysaccharidosis IIIA using hematopoietic stem cell gene therapy.
Apolipoprotein E Mimetic Peptide-Mediated Modulation of Lipid Homeostasis.
Agmatine Sulfate Is Associated with Altered Redox Status, Apoptosis-Associated Responses, and AKT/JNK Signalling in Ishikawa Endometrial Cancer Cells.
Functional and Mechanistic Studies of the EST Gene in the Gonads of Chlamys farreri.
Biochemical analysis of catalytic and disease-associated residues in mouse Golgi-resident 3'-phosphoadenosine-5'-phosphate (PAP) phosphatase BPNT2/gPAPP.
Seaweed sulfated polysaccharides regulate the gut microbiota-mucosal barrier-innate immunity axis: structural heterogeneity, immunological evidence, and translational prospects.

J Biol Chem. 2026 Sep 22. pii: S0021-9258(26)02466-X. [Epub ahead of print] 113594

Discovery and characterization of Chondroitinase A with specificity for 4-O-sulfated N-acetylgalactosamine in Chondroitin sulfate/Dermatan sulfate.

Wenshuang Wang, Haixin Yi, Ping'en Zi, Xu Wang, Rouyi Zou, Yu-Zhong Zhang, Fuchuan Li.

  Chondroitin sulfate (CS) and dermatan sulfate (DS) are glycosaminoglycans whose diverse biological functions are largely determined by specific sulfation patterns. The GlcA/IdoA-GalNAc4S (A/iA unit), where GlcA, IdoA, GalNAc and 4S denote D-glucuronic acid, L-iduronic acid, N-acetylgalactosamine and 4-O-sulfation, respectively, is the most prevalent sulfation motif in animal tissues and participates in a broad spectrum of physiological and pathological processes. However, chondroitinase A (CSase A), a lyase capable of selectively degrading CS/DS rich in A/iA units, has not yet been identified, which has limited our understanding of the functional roles of such structures in CS/DS. Here, we report the first discovery of a CSase A from the metagenome of sediments near the Haima cold seep. This enzyme belongs to polysaccharide lyase family 8 and exhibits strict selectivity for A/iA units, yielding disaccharides composed almost exclusively of the 4-O-sulfated disaccharide, with no other sulfated disaccharide variants detectable even under exhaustive digestion. Structural analysis of resistant tetrasaccharides and hexasaccharides, and degradation assays with structure-defined oligosaccharides demonstrate that CSase A exclusively cleaves the β-1,4 glycosidic bond preceding a disaccharide unit containing GalNAc or GalNAc4S, thereby defining its rigorous substrate specificity. Site-directed mutagenesis and activity profiling identified Glu-579, Arg-480, Tyr-427, and His-420 as residues essential for catalysis, supporting a proposed tyrosine-histidine-mediated acid-base mechanism. The discovery of CSase A provides a unique molecular tool for dissecting the functional roles of A/iA units in CS/DS-related biology and disease.

Keywords:  Chondroitin sulfate; Deep sea; Glycosaminoglycan; Lyase; Substrate specificity

DOI:  https://doi.org/10.1016/j.jbc.2026.113594
Carbohydr Res. 2026 Sep 17. pii: S0008-6215(26)00289-2. [Epub ahead of print]570 110100

Synthesis of the endogenous non-reducing end heparan sulfate disaccharides for newborn screening and diagnosis of mucopolysaccharidosis types I and II.

Gurpreet Singh, Sachin Popat Gholap, Sanghyun Lim, Zackary M Herbst, Michael H Gelb.

  There is ongoing newborn screening (NBS) for mucopolysaccharidoses types I and II (MPS I and MPS II) because initiation of treatment early in life leads to better clinical outcomes. NBS starts with measurement of the relevant enzymatic activities in dried blood spots (DBS) on NBS cards. A second-tier test is carried out to eliminate false positives when enzymatic activity is low. In MPS I and MPS II, heparan sulfate accumulates, and previous studies have shown that the non-reducing end disaccharide fragments found endogenously in patient DBS and urine are elevated in patients but not in false positives. The proposed biomarker for MPS I is l-iduronic acid α(1,4)-linked to N-acetyl-d-glucosamine-6-sulfate, and the proposed biomarker for MPS II is l-iduronic acid(2-sulfate) α(1,4)-linked to N-acetyl-d-glucosamine. We now report the total synthesis of these candidate biomarkers for the first time and show by liquid chromatography-tandem mass spectrometry that they are identical to the analytes that accumulate in urine from MPS I and MPS II patients. The biomarkers were also prepared in heavy isotope labeled form so that they can be added to samples as internal standards. These reagents will allow for the first time the absolute moles of biomarkers to be determined in patient samples.

Keywords:  Dermatan sulfate; Glycosaminoglycans; Heparan sulfate; Inborn errors of metabolism; Lysosomal storage disease; Mucopolysaccharidosis-I; Mucopolysaccharidosis-II; Newborn screening; Tandem mass spectrometry

DOI:  https://doi.org/10.1016/j.carres.2026.110100
Carbohydr Polym. 2026 Nov 01. pii: S0144-8617(26)00915-X. [Epub ahead of print]391 125798

Laminarin sulfate from Lessonia flavicans as a potential non-animal anticoagulant with minimal hemorrhagic risk: Structural characterization by NMR spectroscopy and activity analysis.

Minghao Shi, Jiewen Fu, Xiang Shi, Xusheng Lv, Yonghong Liu, Qingxia Yuan, Longyan Zhao.

  Chemical sulfation of well-defined, low-molecular-weight, non-animal polysaccharides represents a potential approach to anticoagulants that potently inhibit the intrinsic tenase complex (iFXase) while causing fewer side effects, notably a low risk of bleeding. From Lessonia flavicans, we isolated a 5.5-kDa laminarin (LFP0) with a (1 → 3)-β-D-glucan core and trace branching. LFP0 was sulfated under anhydrous and short-time conditions to afford four derivatives (LFP0-S1-S4) with degrees of substitution (DS, 0.1-1.5), molecular weights (6.2-10.9 kDa), and sulfate contents (5.8%-46.2%). Physicochemical analysis and 2D NMR confirmed an intact structure with predominant C-6-O-sulfation, then C-2/C-4 sulfation. The sulfated laminarins prolonged APTT in proportion to DS and left PT unaltered. LFP0-S4, with high FIXa affinity, surpassed LMWH in iFXase inhibition by 80.6% but showed only 58.7% of its TT prolongation, indicating higher selectivity for intrinsic-pathway inhibition. In mice, LFP0-S4 exhibited potent antithrombotic activity without obvious bleeding risk, and showed no toxicity at doses up to 2000 mg/kg. These findings position L. flavicans laminarin as a promising structural blueprint for developing intrinsically targeted anticoagulants and warrant further investigation of its pharmacological and toxicological profiles.

Keywords:  Anticoagulant; Intrinsic coagulation pathway; Laminarin; Lessonia flavicans; Sulfated polysaccharide

DOI:  https://doi.org/10.1016/j.carbpol.2026.125798
Food Sci Nutr. 2026 Sep;14(9): e72382

Sulfation Rewrites the Biological Profile of Dietary Flavonoids: Implications for Stability, Redox Signaling, and Inflammation.

Jiří Vrba, Veronika Zapletalová, Jakub Havlásek, Barbora Papoušková, Barbora Petránková, Helena Pelantová, Kateřina Valentová.

  Dietary flavonoids undergo extensive Phase II metabolism, yet the functional consequences of sulfation for their biological activity remain poorly understood. In this study, we compared the physicochemical properties and cellular effects of nine structurally diverse flavonoids and their sulfated derivatives. Flavonoid 3'-O- or 4'-O-sulfates were prepared enzymatically using bacterial aryl sulfotransferase, structurally confirmed, and evaluated as potential metabolites that can be formed by human cytosolic sulfotransferases (SULTs). The results showed that sulfation significantly increased the stability of otherwise labile flavonoids under physiologically relevant conditions, while generally reducing their antiradical capacity and cytotoxicity toward murine RAW264.7 macrophage cells. In contrast to nonsulfated flavonoids, particularly fisetin and quercetin, all tested flavonoid sulfates failed to activate the NRF2 (nuclear factor erythroid 2 p45-related factor 2) pathway in RAW264.7 cells. On the other hand, some of the parent flavonoids as well as flavonoid sulfates partially attenuated lipopolysaccharide-stimulated expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), indicating that sulfated metabolites retain modulatory effects on inflammatory responses. Using recombinant human sulfotransferases, we identified multiple enzymes responsible for the formation of the tested sulfates, confirming their relevance as plausible metabolites. Overall, our findings demonstrate that metabolic sulfation profoundly reshapes flavonoid stability and biological activity, underscoring the importance of considering conjugated metabolites when evaluating the health effects of dietary flavonoids.

Keywords:  NF‐kappaB; NRF2; flavonoid; metabolism; sulfate

DOI:  https://doi.org/10.1002/fsn3.72382
Pathogens. 2026 Sep 19. pii: 993. [Epub ahead of print]15(9):

Chikungunya Virus E2 N218K Enhances Heparan Sulfate-Dependent Attachment and Viral Internalization While Conferring Partial Attenuation in A129 Mice.

Teng Chen, Jie Zhang, Zherui Zhang, Chenglin Deng, Jing Wang, Xin-Can Peng, Yu-Xin Li, Nan Chen, Xing-Yao Huang.

  The E2 glycoprotein of chikungunya virus (CHIKV) binds negatively charged cell-surface heparan sulfate (HS) to facilitate viral attachment and modulate pathogenicity, yet the E2 determinants governing HS-dependent attachment and the effects of altered HS utilization on subsequent infection remain incompletely defined. From a low-passage clinical isolate obtained during the 2010 Guangdong outbreak, we identified a previously unreported E2 substitution, N218K, which introduces a basic residue. In vitro, this mutation enhanced early infection and attachment to mammalian cells and reduced the RNA/PFU ratio; in A129 mice, however, it reduced early serum viral RNA levels, was associated with lower tissue viral RNA burdens at 5 dpi, and delayed mortality, consistent with partial attenuation. Soluble HS competition assays showed that N218K increased HS dependence during viral attachment, whereas DEAE-dextran assays supported a role for charge-dependent interactions in restricting local viral spread. N218K also enhanced viral internalization after normalization of initial attachment, identifying an additional post-attachment phenotype associated with this substitution. Our findings identify N218K as a determinant of HS-dependent attachment and partial attenuation in A129 mice, reveal a "high attachment-low dissemination" phenotype, and further show that this charge-altering E2 substitution enhances post-attachment internalization.

Keywords:  E2 N218K; E2 glycoprotein; attachment; chikungunya virus; heparan sulfate; internalization; pathogenicity

DOI:  https://doi.org/10.3390/pathogens15090993
Polymers (Basel). 2026 Sep 19. pii: 2296. [Epub ahead of print]18(18):

Sulfation of Sugarcane-Bagasse Xylan: Process Optimization, Physicochemical Properties and In Vitro Anticoagulant Activity.

Mingjun Zhang, Xuefeng Wang, Qi Wang, Jianbin Li.

  Xylan sulfates are established anticoagulant polysaccharides, but their preparation requires balancing sulfate substitution and product recovery. Sugarcane-bagasse xylan was sulfated using sulfur trioxide-pyridine complex (SO3·Py), 4-dimethylaminopyridine (DMAP), and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI). An entropy-weighted five-factor Box-Behnken design jointly optimized degree of substitution (DS) and isolated, DS-adjusted yield, assigning respective weights of 0.570 and 0.430. Pareto analysis identified a trade-off between increasing DS and decreasing yield, while accounting for model uncertainty produced little change in the selected optimum. Validation used adjusted reagent-to-xylan mass ratios of 0.2, 3.2, and 2.2 g/g for DMAP, SO3·Py, and EDCI, respectively, at 50.7 °C for 3.8 h. Three independent batches yielded DS = 1.53 ± 0.01, yield = 78.0 ± 0.1%, and a composite score of 0.792 ± 0.008, within the model's 95% prediction interval. Operational water solubility increased from 21.2 to 96.8-130.4 mg/mL after sulfation, alongside changes in aqueous aggregation and rheology. In citrated sheep plasma, the derivatives prolonged activated partial thromboplastin time and thrombin time with increasing concentration. Responses increased across the derivative series, in which DS and the reported apparent molar mass covaried. These results provide a basis for balancing substitution and product recovery and comparing the water solubility and in vitro anticoagulant responses of the resulting derivatives.

Keywords:  bagasse xylan; coagulation time; response surface methodology; sugarcane; sulfation; water solubility

DOI:  https://doi.org/10.3390/polym18182296
Vet Sci. 2026 Sep 19. pii: 989. [Epub ahead of print]13(9):

Indoxyl Sulfate, Cardiac Biomarkers, and Left Ventricular Wall Thickening in Cats with Chronic Kidney Disease.

Murat Akbaba, Idil Bastan, Filiz Bakar-Ates, Osman Safa Terzi.

  Factors associated with left ventricular wall thickening (LVWT) in cats with chronic kidney disease (CKD) remain unclear. This prospective observational study compared renal variables, systolic blood pressure (SBP), echocardiographic measurements, cardiac biomarkers, and uremic toxins in 31 cats: 12 with CKD without LVWT, 8 with CKD and LVWT, and 11 healthy controls. LVWT was defined as IVSd and/or LVPWd ≥ 6 mm in at least two of three examinations. IRIS stage distribution did not differ between the CKD groups (p = 1.000). Urea, creatinine, and phosphorus concentrations were higher in both CKD groups than in controls but were similar between CKD groups. SBP, NT-proBNP, and cardiac troponin I concentrations were highest in the CKD-LVWT group (p < 0.001). Plasma indoxyl sulfate concentrations were significantly higher in the CKD-LVWT group than in both other groups, whereas serum p-cresyl sulfate concentrations did not differ between CKD groups. However, SBP was markedly higher in the CKD-LVWT group and represents a major potential confounder. However, because SBP was also higher in the CKD-LVWT group and reliable multivariable adjustment could not be performed, the independent contribution of indoxyl sulfate could not be determined. The observed association may have been influenced by systemic hypertension, CKD stage, antihypertensive treatment and adherence, and other potential confounders and should not be interpreted as independent or causal.

Keywords:  NT-proBNP; cardiac troponin I; feline chronic kidney disease; indoxyl sulfate; left ventricular wall thickening; p-cresyl sulfate; systolic hypertension

DOI:  https://doi.org/10.3390/vetsci13090989
Int J Mol Sci. 2026 Sep 15. pii: 8210. [Epub ahead of print]27(18):

Dysregulation of the Heparanase/TFPI-2/Heparan Sulfate Axis in Recurrent Pregnancy Loss.

Chen Yanovich, Nadin Sabbah, Keren Asayag, Yonatan Crispel, Haim Cohen, Anat Keren-Politansky, Yona Nadir.

  Recurrent pregnancy loss (RPL) is a clinically heterogeneous reproductive disorder, and in many women the underlying mechanism remains unexplained by standard thrombophilia evaluation. Pregnancy is characterized by a tightly regulated hemostatic balance at the maternal-placental interface. Heparanase, tissue factor pathway inhibitor 2 (TFPI-2), and heparan sulfate (HS) are expressed in placental and hemostatic pathways and may contribute to coagulation-related pregnancy complications. We investigated whether the heparanase/TFPI-2/HS axis is altered in women with RPL. Blood samples were obtained from 69 women with RPL and 47 control women with at least two normal deliveries and no history of pregnancy-related vascular complications. Samples were collected at least three months after pregnancy and in the absence of hormonal therapy. Plasma and white blood cell samples were analyzed using an enzyme-linked immunosorbent assay, Western blotting, real-time polymerase chain reaction, co-immunoprecipitation, and Sanger sequencing. Mechanistic experiments using heparanase-derived peptide 16 and TFPI-2-derived peptide 6 were performed in MCF-7 and JAR cells. We found that women with RPL had increased plasma heparanase levels, heparanase procoagulant activity, HS levels, and heparanase mRNA expression compared with controls. TFPI-2 protein and mRNA expression were also elevated. The heparanase rs4693608 AA genotype was more frequent in women with RPL and, together with rs4364254 TT, was associated with increased heparanase expression and procoagulant activity. Although TFPI-2 levels were higher in RPL, co-immunoprecipitation showed reduced heparanase/TFPI-2 complex formation, suggesting impaired inhibitory regulation. HS modulated heparanase binding to TF and TFPI-2 in a concentration-dependent, bell-shaped manner. Heparanase-derived peptide 16 increased TFPI-2 expression, supporting a regulatory feedback loop. In conclusion, RPL is associated with dysregulation of the heparanase/TFPI-2/HS axis, including increased heparanase expression and activity, altered heparanase/TFPI-2 interaction, and heparanase-related genetic variation. These findings suggest a coagulation-related reproductive mechanism that may contribute to pregnancy loss and warrants validation in larger prospective studies.

Keywords:  heparan sulfate; heparanase; recurrent pregnancy loss; thrombophilia; tissue factor pathway inhibitor 2

DOI:  https://doi.org/10.3390/ijms27188210
Biomater Adv. 2026 Sep 17. pii: S2772-9508(26)00486-3. [Epub ahead of print]190 215186

One-pot fabrication and in vitro characterization of hydroxyapatite-enriched cryogels for bone tissue engineering.

Laura Di Muzio, Vito Cosimo Carriero, Patrizia Paolicelli, Susi Zara, Alessia Ricci, Simone Carradori, Paolo Ballirano, Claudia Sergi, Jacopo Tirillò, Stefania Petralito, Maria Antonietta Casadei.

  Cryogels have emerged as promising biomimetic scaffolds for extracellular matrix emulation in bone tissue engineering (BTE). In this study, we investigated how hydroxyapatite (HA) incorporation influences the physicochemical, mechanical, and biological performance of cryogels based on methacryloyl derivatives of type A gelatin, type B gelatin, and chondroitin sulfate (GelMAA, GelMAB, and CSMA, respectively). HA-enriched cryogels were fabricated through a one-pot cryogelation process in which HA particles were dispersed within the polymer solutions prior to crosslinking, without the use of additional dispersing agents. Scaffolds incorporating either commercial spheroidal HA or ultrasound-synthesized needle-to-plate HA were compared with non-mineralized counterparts as well as with each other, to evaluate the effect of the mineral phase on the resulting cryogels properties. The different polymer matrices exhibited distinct capacities to stabilize and homogeneously incorporate HA. Negatively charged polymers (GelMAB and CSMA) enabled HA incorporation up to 10% w/w, whereas GelMAA efficiently incorporated only 2.5% w/w. However, increasing mineral content did not result in improved mechanical properties or biological performance, suggesting that alterations in cryogel pore architecture and interconnectivity outweighed the benefits of higher mineral loading. Biological evaluation revealed limited cell response in CSMA-based cryogels, while GelMA-based systems supported cell adhesion and viability, with GelMAA yielding marginally more favorable outcomes. Furthermore, HA crystal morphology had only a minor influence on scaffold performance, including osteoinductive potential. Overall, the results indicate that the composition of the polymer matrix plays a more critical role than HA loading or crystal morphology in determining the functional performance of mineralized cryogels for BTE.

Keywords:  Chondroitin sulfate; Cryogels; Gelatin; Hydroxyapatite; Osteogenic differentiation; Tissue engineering

DOI:  https://doi.org/10.1016/j.bioadv.2026.215186
PLoS One. 2026 ;21(9): e0359319

Alpha-N-acetylglucosaminidase (NAGLU) inactivation causes morphological damage to the ultrastructural appearance of epididymal mononuclear phagocytes (eMPs) and epithelial cells of the epididymis in adult mice.

Lorena Carvelli, Louis Hermo, Daniel G Cyr, Mary Gregory, Alexey V Pshezhetsky, Carlos R Morales.

Heparan sulfate (HS) a glycosaminoglycan family of carbohydrates, is covalently attached to core proteins forming heparan sulfate proteoglycans (HSPGs) which are found on the surface of cells. Following endocytosis, HSPGs are degraded within lysosomes by several enzymes, including α-N-acetylglucosaminidase (NAGLU). Absence of NAGLU leads to Mucopolysaccharidosis III type B (Sanfilippo type B), a lysosomal storage disease, which in Naglu-/- mice was suggested to result in infertility. Since endocytosis is a feature of epididymal epithelial and immune cells, a plausible explanation could be excessive accumulation of lysosomes in these cells in 7-month-old mice. EM analysis revealed a massive accumulation of pale lysosomes (LM-IHC/LAMP2 + /cathepsin B+) of varying shapes and sizes in the cytoplasm of principal cells, unlike the small dense spherical lysosomes of wild type mice. Their presence had a profound effect on the integrity of organelles such as the ER, Golgi, mitochondria and nucleus. In addition, the epithelium demonstrated sporadic large, highly vacuolated cells, identified as CD68+ and MOMA+ epididymal mononuclear phagocytes (eMPs), with a cytoplasm dominated by pale lysosomes, some of gigantic size. The plethora of lysosomes overwhelming the cytoplasm suggested an ongoing phagocytic activity to protect the epithelium despite the excessive disruption of their organelles. A distinguishing feature of the content of lysosomes of eMPs was the presence of aggregates of small flattened membranous discs aligned parallel to one another, referred to as zebra bodies. The latter had the appearance of mitochondrial cristae of principal cells and were reactive for a mitochondrial marker, suggesting a phagocytic activity for eMPs of degenerating principal cells. Taken together, our data demonstrates that the accumulation of lysosomes of epithelial cells and eMPs dramatically altered their structural appearance in the absence of NAGLU inducing an inactivation of their critical role in supporting a proper epithelial environment for sperm protection and maturation.

DOI: https://doi.org/10.1371/journal.pone.0359319
Molecules. 2026 Sep 10. pii: 3196. [Epub ahead of print]31(18):

Determining Carrageenan Sulfate Groups Using Ion Association with Alcian Blue Dye.

Alexander Shyichuk, Dorota Ziółkowska, Iryna Schyychuk.

  The purpose of this study is to develop a simple and reliable method to measure sulfate content and determine the purity of carrageenan raw materials. The method uses cationic Alcian Blue dye that binds strongly to anionic carrageenan macromolecules, resulting in an insoluble ion associate. At a specific carrageenan-to-dye ratio, the hydrophobic ion-associated particles form macroscopic flocs and precipitate quickly. The resulting deep sedimentation leaves almost colorless supernatants, indicating the stoichiometric charge ratio. The critical polymer-to-dye ratio has been found to be independent of the dye concentration. The Alcian Blue reagent has been standardized against a synthetic polymer, poly(sodium styrene sulfonate). The method of ion-associate precipitation was used to determine the content of sulfate groups in commercial carrageenans of different types: kappa, iota, and lambda. The results obtained agree well with the IR spectra of the tested carrageenans.

Keywords:  Alcian Blue; carrageenan; ion association; sulfate ester content

DOI:  https://doi.org/10.3390/molecules31183196
Cancer Immunol Immunother. 2026 Sep 04. pii: 233. [Epub ahead of print]75(10):

The effect and mechanism of TIPE in promoting M2 polarization through DcR3 in the microenvironment of colorectal cancer.

Shiying Zhang, Chunlin Shen, Zeyang Lin, Qiang Zhu, Yihan Feng, Liwei Sun, Chengjia Zhang, Haole Zhang, Lumin Wang, Shuyu Chen, Jiyi Huang, Yuhan Ye, Xingfeng Qiu, Guohong Zhuang.

   BACKGROUND: Elevated expression of tumor necrosis factor α-induced protein 8 (TIPE) is observed in colorectal cancer (CRC) and is known to regulate decoy receptor 3 (DcR3) expression while promoting M2 polarization of macrophages. However, the precise role of DcR3 in macrophage M2 polarization remains to be elucidated.
METHODS: We employed bioinformatic analysis alongside in vitro and in vivo experimental approaches to investigate the regulatory mechanism by which TIPE influences macrophage M2 polarization in CRC.
RESULTS: TIPE expression in CRC tissues positively correlates with M2 macrophage markers (CD206, CD163, IL-10). Functionally, TIPE promotes M2 polarization of macrophages by upregulating DcR3. Mechanistically, DcR3 binds to heparan sulfate proteoglycan 2 (HSPG2) in a heparan sulfate (HS)-dependent manner, leading to activation of the protein kinase Bα1 (AKT1) signaling pathway. AKT1 activation not only directly drives M2 polarization but also enhances macrophage chemotaxis and recruitment toward CRC cells by upregulating the chemokine CXCL8.
CONCLUSIONS: This study reveals a novel mechanism through which TIPE regulates M2 macrophage polarization and function via DcR3, offering new perspectives for the diagnosis and therapeutic targeting of CRC.

Keywords:  CRC; DcR3; M2 polarization; TIPE; Tumor microenvironment

DOI:  https://doi.org/10.1007/s00262-026-04531-8
Children (Basel). 2026 Sep 09. pii: 1221. [Epub ahead of print]13(9):

Clinical Characteristics and Thyroid Findings in Children with Isolated Premature Adrenarche.

Sibel Aka, Seymanur Kocyigit, Saygin Abali, Serap Semiz.

  Objectives: Premature adrenarche (PA) is associated with accelerated adrenal maturation, but data regarding thyroid characteristics in children with isolated PA are limited. We aimed to describe the clinical features of children with idiopathic PA and investigate the relationships between thyroid function, thyroid autoimmunity, and adrenal androgen secretion. Methods: This retrospective single-center study included 435 children (378 girls, 57 boys) with idiopathic PA evaluated between 2014 and 2024. Clinical, anthropometric, biochemical, and thyroid data were analyzed. Associations between adrenal and thyroid parameters were examined using correlation and group comparison analyses. Results: Girls accounted for 86.9% of the cohort. Children were generally taller than their genetic target height and frequently had excess weight. Boys exhibited higher serum dehydroepiandrosterone sulfate (DHEAS) concentrations than girls and more often met criteria for exaggerated adrenarche despite less advanced clinical androgenization. Lower birth weight (BW) was associated with higher DHEAS concentrations, and children born with low BW more frequently demonstrated exaggerated adrenarche. BW SDS showed a significant inverse correlation with DHEAS concentrations. In contrast, DHEAS concentrations were unrelated to body mass index (BMI) SDS or bone age advancement. Height SDS increased progressively with increasing BMI category in both sexes, whereas DHEAS concentrations remained comparable across weight groups. Thyroid abnormalities were identified in 8.0% of the cohort. Free thyroxine and thyroid-stimulating hormone concentrations were within the normal range in most children and showed no association with DHEAS concentrations or exaggerated adrenarche status. Although thyroid autoantibody positivity was frequent among the tested subgroup, it was not associated with adrenal androgen concentrations. Conclusions: Idiopathic PA is characterized by accelerated linear growth, frequent excess weight, and an inverse relationship between birth weight and adrenal androgen secretion. Thyroid function was not associated with adrenal androgen production within this PA cohort. The frequent thyroid autoantibody positivity observed in the selected tested subgroup should be interpreted cautiously and cannot be considered PA-specific in the absence of healthy control groups; prospective controlled studies are needed.

Keywords:  adrenal; autoimmunity; nonclassical congenital adrenal hyperplasia; premature adrenarche; thyroid

DOI:  https://doi.org/10.3390/children13091221
Cells. 2026 Sep 17. pii: 1687. [Epub ahead of print]15(18):

Sulfated Hyaluronan Drives Cell Adaptation and Matrix Composition in Advanced 3D Breast Cancer Cell Models.

Christos Koutsakis, Katerina Mineschou, Konstantinos Spanopoulos, Sylvia Mangani, Marco Franchi, Evgenia Karousou, Zoi Piperigkou, Nikos K Karamanos.

  Hyaluronan (HA), a major extracellular matrix (ECM) glycosaminoglycan, plays a key role in breast cancer progression. Although native HA lacks sulfate groups, chemically modified sulfated hyaluronan (sHA) has demonstrated promising antitumor activity. Previous work from our group showed that sHA alters cellular functions and modulates ECM-related gene expression in triple-negative breast cancer (TNBC) cells. The aim of this study was to investigate the effects of low-molecular-weight HA (50 kDa) and its sulfated derivative of the same molecular weight, sHA, in MDA-MB-231 and MCF-7 breast cancer cells using advanced 3D cell culture models. Gene expression analyses focused on ECM components, including HA receptors and matrix metalloproteinases (MMPs) that were evaluated. The 3D cell morphology was examined using scanning electron microscopy (SEM). Spheroid growth and expression profiles linked to ECM remodeling and invasiveness were also assessed. Notably, sHA significantly inhibited 3D spheroid growth, reduced cell spreading in a cell line-dependent manner and influenced the expression of genes correlated with ECM remodeling and HA synthesis. These findings emphasize the importance of 3D models for studying ECM-driven breast cancer progression, further supporting sHA as a potential therapeutic modulator.

Keywords:  3D cell platforms; breast cancer; extracellular matrix; hyaluronan; spheroids; sulfated hyaluronan

DOI:  https://doi.org/10.3390/cells15181687
Diagnostics (Basel). 2026 Sep 12. pii: 2953. [Epub ahead of print]16(18):

Establishment of Serum Cholesterol Sulfate Reference Intervals and Evaluation of Its Diagnostic and Metabolic Associations in Chinese Adults.

Yin Liu, Xucong Ji, Xiaoqian Yu, Hongmei Zhang, Xinhua Dai, Zhiguang Su.

  Background/Objectives: Cholesterol sulfate (CS) is a multifunctional signaling molecule implicated in diverse physiological and pathological processes. Its clinical translation as a biomarker is hindered by the lack of established reference intervals, unclear disease-specific alterations, and undefined relationships with routine laboratory parameters. This study aimed to establish a serum CS reference range for Chinese populations, evaluate its associations with various diseases and clinical laboratory parameters, and define its clinical diagnostic utility. Methods: A total of 642 subjects were enrolled, comprising 372 healthy controls and 270 patients with one of six diseases: Alzheimer's disease, osteoporosis, obesity, type 2 diabetes mellitus (T2DM), non-alcoholic fatty liver disease (NAFLD), or Crohn's disease. Serum CS concentrations were quantified using LC-MS/MS. Reference intervals were established using non-parametric methods. Univariate analyses included Spearman's rank correlation, Mann-Whitney U test, and receiver operating characteristic (ROC) curve analysis; multivariate analysis employed a gamma-family generalized linear model (GLM) with a log-link function, incorporating age, sex, clinical laboratory parameters, and disease status as covariates. The age-by-sex interaction was examined to assess synergistic effects. Incremental diagnostic value of CS was evaluated using the DeLong test and likelihood ratio test. Results: The serum CS reference interval for healthy individuals was 0.55-2.24 mg/L (males: 0.60-2.25 mg/L; females: 0.54-2.20 mg/L), with a significant sex difference. The age-by-sex interaction effect was significant (two-way ANOVA interaction p = 0.005; GLM interaction β = 0.102, p = 0.002). Stratified analysis revealed a significant positive correlation between age and CS in males, but no such association in females. In univariate analysis, CS correlated significantly with total cholesterol (TC), non-HDL-C, hemoglobin A1c (HbA1c), and other biomarkers; CS levels were elevated in Alzheimer's disease and reduced in Crohn's disease. In the multivariate GLM, only TC and male sex emerged as independent significant determinants of CS, whereas HbA1c and NAFLD showed borderline effects; no disease group retained independent significance. After adjustment for TC and sex, the residual area under the curves (AUCs) of CS for all diseases were close to 0.50, and DeLong's test indicated no incremental diagnostic value beyond the baseline model (all p > 0.70). Conclusions: This study is the first to establish a serum CS reference range for the Chinese population. Serum CS levels are independently regulated by TC and sex, with a significant age-by-sex interaction. The associations between CS and disease status were largely mediated by confounding from lipid profiles and age. CS is not suitable as an independent disease biomarker but may serve as a supplementary indicator for assessing lipid metabolism-particularly in male populations, where it may indirectly reflect age-related changes in cholesterol metabolism.

Keywords:  Alzheimer’s disease; Crohn’s disease; LC-MS/MS; cholesterol sulfate; metabolic biomarkers; reference interval

DOI:  https://doi.org/10.3390/diagnostics16182953
Mol Ther. 2026 Sep 25. pii: S1525-0016(26)00803-8. [Epub ahead of print]

Clinical and biochemical correction of a patient with neurodegenerative Mucopolysaccharidosis IIIA using hematopoietic stem cell gene therapy.

Jane L Kinsella, Simon A Jones, Karen F Buckland, Rebecca J Holley, Helena Lee, Wendy Ogden, Jane Potter, Heather J Church, Kathryn L Brammeier, Karen L Tylee, Stuart Ellison, Rachel Searle, Claire Booth, Farzin Farzeneh, Louise Sherwin, Adrian J Thrasher, Stewart Rust, Brian W Bigger, Robert F Wynn.

Autologous haematopoietic stem cell gene therapy (HSC-GT) is an emerging therapy for treatment-refractory neurodegenerative lysosomal storage disorders (LSD). Mucopolysaccharidosis IIIA (MPSIIIA) is a devastating LSD where pathogenic variants in the SGSH gene lead to toxic heparan sulfate accumulation, inflammation and cell death. Children present with developmental delay, neurodegeneration, significant behavioural disturbance, and die in late teens. We report 80-month outcomes for a patient with severe MPSIIIA following treatment with investigational autologous HSC-GT (OTL-201), at 30-months of age. Haematopoietic stem and progenitor cells (HSPC) were collected from peripheral blood following mobilisation with G-CSF and Plerixafor. HSPC were transduced with a lentiviral vector containing the SGSH gene and the CD11b promoter. The product was infused following myeloablative conditioning with busulfan. Product engraftment was rapid, with production of sustained supra-physiological SGSH enzyme measured in leukocytes, plasma and cerebrospinal fluid (CSF), and reductions in substrate heparan sulfate to within the normal range in urine, plasma and CSF. Neurocognitive outcomes on the Bayley Scale of Infant Development (BSID-III) and parental-reported outcomes showed improvements compared to natural history. This demonstrates sustained long-term efficacy and safety of HSC-GT in a patient with severe MPSIIIA, setting the scene for a phase I/II clinical trial in younger patients.

DOI: https://doi.org/10.1016/j.ymthe.2026.09.032
Biomolecules. 2026 Sep 17. pii: 1352. [Epub ahead of print]16(9):

Apolipoprotein E Mimetic Peptide-Mediated Modulation of Lipid Homeostasis.

Dilan R Patel, Cole L Martin, G M Anantharamaiah, Stephen G Aller.

  Lipid homeostasis is essential for maintaining cellular function and metabolic balance in all organisms, and its dysregulation can contribute to hypercholesterolemia, atherosclerosis, and cardiovascular disease. Adverse health outcomes are associated with elevated levels of low-density lipoprotein cholesterol (LDL-C), retention of which causes macrophages to transition into foam cells, leading to widespread inflammation, cardiovascular disease, and atherosclerosis. Statins (HMG-CoA reductase inhibitors) are considered the gold standard for LDL and very-low-density lipoprotein (VLDL) reduction, yet they exhibit side effects like myalgia and insulin resistance. Apolipoprotein E (apo E)-based lipid-binding synthetic mimetic peptides have been shown to clear cholesterol from the blood through canonical hepatic pathways such as HSPG receptors and are currently being studied to reduce plasma cholesterol. This review examines the development, biological properties, and therapeutic potential of the apo E mimetic peptide AEM-28. AEM-28 reduces atherogenic lipoproteins from plasma efficiently in multiple preclinical models as well as Phase I clinical trials in humans, where general tolerance of the treatment was observed. AEM-28 utilizes a unique mechanism of action that involves binding to atherogenic lipoproteins with multiple receptor-binding domains, facilitating enhanced clearance via the highly abundant heparan sulfate proteoglycans on hepatocytes. These attributes support the potential of AEM-28 as a complementary therapeutic or even an alternative for patients requiring LDL apheresis and those with hypercholesterolemia, atherosclerosis, or cardiovascular disease.

Keywords:  LDL; apo E; atherosclerosis; cholesterol; hypercholesterolemia; peptide

DOI:  https://doi.org/10.3390/biom16091352
Life (Basel). 2026 Sep 05. pii: 1483. [Epub ahead of print]16(9):

Agmatine Sulfate Is Associated with Altered Redox Status, Apoptosis-Associated Responses, and AKT/JNK Signalling in Ishikawa Endometrial Cancer Cells.

Neziha Senem Arı, Ayşe Çakır Gündoğdu.

  Agmatine is a naturally occurring polyamine with context-dependent effects on cellular redox regulation, survival, and apoptosis; however, its effects in endometrial cancer cells remain poorly characterized. This exploratory in vitro study investigated cellular responses associated with agmatine sulfate exposure in Ishikawa endometrial adenocarcinoma cells, using human dermal fibroblasts (HDFs) as a non-cancerous, non-tissue-matched comparator. Metabolic activity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay following exposure to 0.5-15 mM agmatine sulfate for 24 and 48 h. Based on the 48-h concentration-response analysis in Ishikawa cells, 4.1 and 6.5 mM were selected for subsequent analyses of intracellular reactive oxygen species (ROS), malondialdehyde (MDA), cleaved caspase-3, and p-AKT/p-JNK immunoreactivity. Agmatine sulfate produced concentration- and time-dependent reductions in MTT-derived metabolic activity in both cell lines. In Ishikawa cells, 4.1 and 6.5 mM agmatine sulfate significantly increased ROS, MDA, and cleaved caspase-3 levels, accompanied by decreased p-AKT and increased p-JNK immunoreactivity. In HDF cells, ROS levels increased at both concentrations, whereas significant increases in MDA, cleaved caspase-3, and p-JNK were observed only at 6.5 mM; p-AKT immunoreactivity remained unchanged. These findings indicate that agmatine sulfate exposure is associated with reduced metabolic activity, increased oxidative stress and apoptosis-associated responses, and changes in AKT/JNK-related signalling in Ishikawa cells. However, the responses observed in HDF cells do not support a conclusion of tumour-selective activity. Further studies using additional endometrial cancer cell lines, tissue-matched non-malignant controls, and targeted experimental approaches are warranted to clarify the biological relationships among these responses.

Keywords:  AKT/JNK signalling; agmatine sulfate; apoptosis; endometrial cancer; oxidative stress

DOI:  https://doi.org/10.3390/life16091483
Biology (Basel). 2026 Sep 08. pii: 1574. [Epub ahead of print]15(18):

Functional and Mechanistic Studies of the EST Gene in the Gonads of Chlamys farreri.

Yuxin Li, Wenzhu Cui, Xiaoling Liu.

  Estrogen sulfotransferase (EST), a member of the sulfotransferase (SULT) family, catalyzes estrogen sulfation and regulates its activity and content. EST has been mainly studied in vertebrates. Our previous analysis of EST expression in kidney, hepatopancreas, mantle, adductor muscle, gill, and gonads of Chlamys farreri (C. farreri) revealed highest expression in the gonads, suggesting an important role in gonadal function. We knocked down EST by RNAi, examined gonadal histology and E2 levels (via ELISA), expressed recombinant EST in prokaryotes, and assessed its activity using a novel MCF-7 proliferation assay, where E2 promotes cell growth and EST inactivation reduces it. After EST knockdown, EST mRNA levels decreased by 67% in the ovary and 63% in the testis, indicating effective silencing. Histological analysis revealed that RNAi accelerated oocyte proliferation and development, enhanced oocyte vitellogenesis and delayed testicular development in the experimental group. Moreover, estradiol (E2) levels in both ovaries and testes were higher in the experimental groups than in the controls (p < 0.05). The recombinant EST protein was expressed as inclusion bodies and, after purification, denaturation, refolding, and ultrafiltration, yielded high-purity, concentrated protein. Enzyme activity assays showed a decreased proliferation rate of MCF-7 cells, indicating that EST possesses enzymatic activity that can inactivate E2. It is worth mentioning that the traditional isotope-labeling method for detecting EST activity is limited by high cost, operational complexity, technical demands, and potential health risks; therefore, this study established a simpler, more economical, and safer method for measuring EST enzymatic activity. In conclusion EST contributes to the maintenance of estrogen balance during gonadal development.

Keywords:  ELISA; EST; RNAi; enzyme activity assay; histological analysis; prokaryotic expression

DOI:  https://doi.org/10.3390/biology15181574
Biosci Biotechnol Biochem. 2026 Sep 24. pii: zbag144. [Epub ahead of print]

Biochemical analysis of catalytic and disease-associated residues in mouse Golgi-resident 3'-phosphoadenosine-5'-phosphate (PAP) phosphatase BPNT2/gPAPP.

Ryota Urushihara, Yasuhisa Maruno, Saki Maruoka, Moe Tsuruta, Ryotaro Matsuzaki, Etsuko Nishimoto, Takamasa Teramoto, Yoshimitsu Kakuta.

  Golgi-resident 3'-phosphoadenosine-5'-phosphate (PAP) phosphatase BPNT2/gPAPP plays an essential role in maintaining sulfation homeostasis, but its biochemical characterization has been limited by the difficulty of in vitro analyses. Here, we established a stable expression and activity assay system using a Trigger factor-fused BPNT2 construct. Using this system, we performed AlphaFold 3-guided mutational analyses. The predicted model and mutational analyses showed that several acidic residues are required for efficient PAP phosphatase activity, likely through their contribution to Mg²⁺ coordination. In addition, T177 was suggested to help maintain an active-site geometry that could accommodate a catalytic water molecule. Conservation analysis and lithium inhibition supported the idea that BPNT2 shares a conserved Mg²⁺-dependent catalytic framework with related PAP phosphatases. Disease-associated variants D175N and T181P showed substantially reduced activity, possibly due to impaired Mg2+ coordination and local structural distortion. These findings provide a biochemical framework for understanding BPNT2 function in Golgi sulfation regulation.

Keywords:  BPNT2; PAP phosphatase; disease-associated mutation; gPAPP; sulfation

DOI:  https://doi.org/10.1093/bbb/zbag144
Front Immunol. 2026 ;17 1920049

Seaweed sulfated polysaccharides regulate the gut microbiota-mucosal barrier-innate immunity axis: structural heterogeneity, immunological evidence, and translational prospects.

Wei Liu, Meijing Zhang, Jianan Huang, Sihan Liu, Ran Chen, Yingqiang Li, Jinlin Liu, Rui Jia.

  Seaweed sulfated polysaccharides, fucoidan, carrageenan, ulvan, and porphyran, are structurally diverse marine glycans that modulate the gut microbiota, mucosal barrier, and innate immunity, yet the literature remains fragmented across classes, models, and processing histories. This review synthesizes evidence on their structure-function relationships, focusing on microbial enzymatic accessibility, selective fermentation, metabolite remodeling, barrier regulation, and immune signaling, while also assessing translational barriers. A systematic literature search prioritized mechanistic studies, structural analyses, animal models, human trials, and high-quality reviews. Our analysis reveals that porphyran provides the clearest microbiota-barrier coupling, fucoidan and carrageenan give the richest, but most heterogeneous, immune readouts, and ulvan is an emerging class with growing intestinal evidence. Molecular weight, sulfation pattern, and degradation history consistently shape microbial utilization and downstream effects across all classes. Disease contexts with the strongest support include inflammatory bowel disease, constipation, barrier injury, and infection-driven intestinal inflammation. Time-dependent immune shifts have been reported, but durable epigenetic memory remains unproven, and extra-intestinal evidence through gut-liver and gut-brain axes is still preliminary. We conclude that these polysaccharides are best understood as structurally defined substrates that bridge the microbiota, barrier, and immunity; future progress toward clinical translation will depend on stricter standardization, integrated mechanistic designs, and robust human data.

Keywords:  gut microbiota; innate immunity; mucosal barrier; porphyran; seaweed sulfated polysaccharides; ulvan

DOI:  https://doi.org/10.3389/fimmu.2026.1920049