bims-supasi 2024-07-28 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2024–07–28
28 papers selected by
Jonathan Wolf Mueller, University of Birmingham

Expression of Podocalyxin Potentially Decorated With Low-sulfated Keratan Sulfate in Human Testicular Embryonal Carcinoma.
Mutual Inhibition of Antithrombin III and SARS-CoV-2 Cellular Attachment to Syndecans: Implications for COVID-19 Treatment and Vaccination.
Multiple Administration of Dexamethasone Possesses a Deferred Long-Term Effect to Glycosylated Components of Mouse Brain.
Structural analysis and bioavailability study of low-molecular-weight chondroitin sulfate‑iron complexes prepared by photocatalysis-Fenton reaction.
Biocompatible Preparation of Beta-Lactoglobulin/Chondroitin Sulfate Carrier Nanoparticles and Modification of Their Colloidal and Hydropathic Properties by Tween 80.
Steroid sulfatase in mouse liver and Testis: Characterization, ontogeny and localization.
Is there a relationship of cortisol and sex steroid hormones with mental health in middle-aged adults? the FIT-AGEING study.
Physicochemical Properties of Different Sulfated Polysaccharide Components from Laetiporus sulphureus and Their Anti-Proliferative Effects on MDA-MB-231 Breast Cancer Cells.
Can Hyaluronic Acid Combined with Chondroitin Sulfate in Viscosupplementation of Knee Osteoarthritis Improve Pain Symptoms and Mobility?
The Influence of Sulfation Degree of Glycosaminoglycan-Functionalized 3D Collagen I Networks on Cytokine Profiles of In Vitro Macrophage-Fibroblast Cocultures.
Auricular cartilage regeneration using chondroitin sulfate-based hydrogel with mesenchymal stem cells in rabbits.
Rapid cellular uptake of citrate-coated iron oxide nanoparticles unaffected by cell-surface glycosaminoglycans.
Enzymatic Modulation of the Pulmonary Glycocalyx Enhances Susceptibility to Streptococcus pneumoniae.
Recent advance for animal-derived polysaccharides in nanomaterials.
Unraveling the intricacies of glycosaminoglycan biosynthesis: Decoding the molecular symphony in understanding complex polysaccharide assembly.
Polysaccharide base electrospun nanofibrous scaffolds for cartilage tissue engineering: Challenges and opportunities.
Magnetic Polysaccharide Mesenchymal Stem Cells Exosomes Delivery Microcarriers for Synergistic Therapy of Osteoarthritis.
Characterization of a novel sulfated-rhamnoglucuronan isolated from Korean seaweed Ulva pertusa and its efficacy for treatment of inflammatory bowel disease in mice.
Hepatic Gene Expression and Metabolite Profiles of Androstenone and Skatole Relative to Plasma Estrone Sulfate Levels in Boars.
Dermatan Sulfate Affects the Activation of the Necroptotic Effector MLKL in Breast Cancer Cell Lines via the NFκB Pathway and Rac-Mediated Oxidative Stress.
7-Phenylheptanoic Acid-Hydroxypropyl β-Cyclodextrin Complex Slows the Progression of Renal Failure in Adenine-Induced Chronic Kidney Disease Mice.
Expressed Protein Ligation in Flow.
Initiation of fluoxetine in a pediatric patient with Mucopolysaccharidosis IIIA: Early observations.
Bioactivation of cinnamic alcohol in a reconstructed human epidermis model and evaluation of sensitizing potency of the identified metabolites.
Hepatic enzyme induction and its potential effect on thyroid hormone metabolism in the metamorphosing tadpole of Xenopus laevis (African clawed frog).
Fucoidan modulates gut microbiota and immunity in Peyer's patches against inflammatory bowel disease.
Tyrosine phenol-lyase inhibitor quercetin reduces fecal phenol levels in mice.
Occurrence of bisphenol analogues and their conjugated metabolites in foodstuff.

J Histochem Cytochem. 2024 Jul 25. 221554241265162

Expression of Podocalyxin Potentially Decorated With Low-sulfated Keratan Sulfate in Human Testicular Embryonal Carcinoma.

Akifumi Muramoto, Hitomi Hoshino, So Inamura, Masataka Murahashi, Tomoya O Akama, Naoki Terada, Motohiro Kobayashi.

  SummaryWe previously demonstrated that among various histological types of human testicular germinal cell tumors (GCTs), embryonal carcinoma (EC) preferentially expresses low-sulfated keratan sulfate (KS) consisting of repeating N-acetyllactosamine (LacNAc) disaccharide units composed of galactose and 6-O-sulfated N-acetylglucosamine (GlcNAc), which is recognized by the R-10G antibody. Recently, we generated another anti-low-sulfated KS monoclonal antibody, 294-1B1. Immunohistochemical analysis of testicular GCTs (n=83) revealed that the low-sulfated KS recognized by 294-1B1 is also preferentially expressed in EC but minimally in other GCT histological types. Moreover, immunolabeling with R-10G and 294-1B1 antibodies was resistant to peptide-N-glycosidase F digestion, and EC was not stained with the MECA-79 antibody, indicating that low-sulfated KS expressed in EC contains mucin-type core 2 O-glycans carrying GlcNAc-6-O-sulfated oligo-LacNAc. Double immunofluorescence staining showed that R-10G and 294-1B1 antibody signals colocalized with those for podocalyxin (PODXL). Furthermore, western blot analysis of recombinant human PODXL•IgG fusion proteins secreted from low-sulfated KS-expressing human embryonic kidney 293T cells revealed that PODXL functions as a core protein for low-sulfated KS. Taken together, these findings strongly suggest that the PODXL glycoform decorated with low-sulfated KS is preferentially expressed in human testicular EC and may therefore serve as a diagnostic marker for this malignancy.

Keywords:  endo-β-galactosidase; keratanase II; testis

DOI:  https://doi.org/10.1369/00221554241265162
Int J Mol Sci. 2024 Jul 09. pii: 7534. [Epub ahead of print]25(14):

Mutual Inhibition of Antithrombin III and SARS-CoV-2 Cellular Attachment to Syndecans: Implications for COVID-19 Treatment and Vaccination.

Anett Hudák, Dávid Pusztai, Annamária Letoha, Tamás Letoha.

  Antithrombin III (ATIII) is a potent endogenous anticoagulant that binds to heparan sulfate proteoglycans (HSPGs) on endothelial cells' surfaces. Among these HSPGs, syndecans (SDCs) are crucial as transmembrane receptors bridging extracellular ligands with intracellular signaling pathways. Specifically, syndecan-4 (SDC4) has been identified as a key receptor on endothelial cells for transmitting the signaling effects of ATIII. Meanwhile, SDCs have been implicated in facilitating the cellular internalization of SARS-CoV-2. Given the complex interactions between ATIII and SDC4, our study analyzed the impact of ATIII on the virus entry into host cells. While ATIII binds to all SDC isoforms, it shows the strongest affinity for SDC4. SDCs' heparan sulfate chains primarily influence ATIII's SDC attachment, although other parts might also play a role in ATIII's dominant affinity toward SDC4. ATIII significantly reduces SARS-CoV-2's cellular entry into cell lines expressing SDCs, suggesting a competitive inhibition mechanism at the SDC binding sites, particularly SDC4. Conversely, the virus or its spike protein decreases the availability of SDCs on the cell surface, reducing ATIII's cellular attachment and hence contributing to a procoagulant environment characteristic of COVID-19.

Keywords:  SARS-CoV-2; antithrombin III; heparan sulfate proteoglycans; spike protein cellular entry; syndecans

DOI:  https://doi.org/10.3390/ijms25147534
Neurol Int. 2024 Jul 22. 16(4): 790-803

Multiple Administration of Dexamethasone Possesses a Deferred Long-Term Effect to Glycosylated Components of Mouse Brain.

Stanislav D Aladev, Dmitry K Sokolov, Anastasia V Strokotova, Galina M Kazanskaya, Alexander M Volkov, Svetlana V Aidagulova, Elvira V Grigorieva.

  Glucocorticoids are used during glioblastoma treatment to prevent the cerebral edema effect surrounding normal brain tissue. The aim of our study was to investigate the long-term effects of multiple administrations of glucocorticoids onto the glycosylated components (proteoglycans and glycosaminoglycans) of normal brain extracellular matrix and the glucocorticoid receptor (GR, Nr3c1) in an experimental model in vivo. Two-month-old male C57Bl/6 mice (n = 90) were injected intraperitoneally with various doses of dexamethasone (DXM) (1; 2.5 mg/kg) for 10 days. The mRNA levels of the GR, proteoglycans core proteins, and heparan sulfate metabolism-involved genes were determined at the 15th, 30th, 60th, and 90th days by a real-time RT-PCR. The glycosaminoglycans content was studied using dot blot and staining with Alcian blue. A DXM treatment increased total GAG content (2-fold), whereas the content of highly sulfated glycosaminoglycans decreased (1.5-2-fold). The mRNA level of the heparan sulfate metabolism-involved gene Hs3St2 increased 5-fold, the mRNA level of Hs6St2 increased6-7-fold, and the mRNA level of proteoglycan aggrecan increased 2-fold. A correlation analysis revealed an association between the mRNA level of the GR and the mRNA level of 8 of the 14 proteoglycans-coding and 4 of the 13 heparan sulfate metabolism-involved genes supporting GR involvement in the DXM regulation of the expression of these genes. In summary, multiple DXM administrations led to an increase in the total GAG content and reorganized the brain extracellular matrix in terms of its glycosylation pattern.

Keywords:  brain extracellular matrix; dexamethasone; glucocorticoid; glucocorticoid receptor; glycosaminoglycan; glycosylation; heparan sulfate biosynthesis; proteoglycan

DOI:  https://doi.org/10.3390/neurolint16040058
Carbohydr Polym. 2024 Oct 15. pii: S0144-8617(24)00661-1. [Epub ahead of print]342 122435

Structural analysis and bioavailability study of low-molecular-weight chondroitin sulfate‑iron complexes prepared by photocatalysis-Fenton reaction.

Qianqian Du, Haoran Song, Chunhong Yan, Chunqing Ai, Sitong Wu, Shuang Song.

  Increasing studies focus on depolymerization of chondroitin sulfate (CS) to enhance its biological activities. In the present study, low-molecular-weight chondroitin sulfate (LMWCS)‑iron complexes were obtained by photocatalysis-Fenton reaction. After degradation with the optimal condition of 0.25 % (w/v) TiO2, 10 mM FeSO4, and 400 mM H2O2 for 0, 15, and 60 min, the average relative molecular weights of CS were reduced to 4.77, 2.47, and 1.21 kDa, respectively. Electron paramagnetic resonance and free radical capture test identified •OH, •O2-, and h+ in the photocatalysis-Fenton system, among them h+ was the major contributor for CS degradation. The structures of degradation products were analyzed by UV, CD, XRD, SEM-EDS, and NMR, and the results indicated that CS chelated iron with its carboxyl and sulfate groups, leading to changes in conformation and microtopography. Then 10 oligosaccharides were identified in the degradation products using HPLC-MSn and the depolymerization mechanism was proposed. Furthermore, iron release was observed in simulated gastrointestinal digestion of LMWCS‑iron complexes. Notably, the everted gut sac experiment demonstrated that LMWCS‑iron complex possessed 3.75 times higher iron absorption than FeSO4 (p < 0.01) and 12.60 times higher CS absorption than original CS (p < 0.0001). In addition, LMWCS‑iron exhibited stronger in vitro antioxidant activity than CS.

Keywords:  Bioavailability; Chondroitin sulfate; Iron complex; Photocatalysis-Fenton

DOI:  https://doi.org/10.1016/j.carbpol.2024.122435
Polymers (Basel). 2024 Jul 12. pii: 1995. [Epub ahead of print]16(14):

Biocompatible Preparation of Beta-Lactoglobulin/Chondroitin Sulfate Carrier Nanoparticles and Modification of Their Colloidal and Hydropathic Properties by Tween 80.

Ioannis Pispas, Nikolaos Spiliopoulos, Aristeidis Papagiannopoulos.

  The electrostatic complexation of the protein beta-lactoglobulin (β-LG) with the anionic polysaccharide chondroitin sulfate (CS) and the subsequent stabilization by thermal treatment were studied to achieve the well-defined nanoparticles (NPs). The formation of the well-defined NPs was obtained at pH 4 with a hydrodynamic radius from 60 to 80 nm. NP aggregation was observed at pH 1.5 because of the loss of the anionic charge of chondroitin sulfate on the surface of the NPs. After thermal treatment, the NPs exhibited stability against a pH increase to pH 7 while a stronger aggregation at pH 1.5 was observed. Core-shell structures were found at pH 7 after thermal treatment, indicating a possible mechanism of partial disintegration. The addition of Tween 80 (T80) before thermal treatment led to the formation of T80 self-assemblies inside the NPs. This caused an increase in the hydrophobicity of the inner and outer surfaces of the NPs as it was observed by fluorescence spectroscopy. The ζ-potential of the complexes and NPs was about -20 mV while the presence of T80 did not affect it. FTIR spectra verified changes of the secondary structure of β-LG in its complexes with CS and T80. The thermally treated NPs exhibited high surface and overall hydrophobicity and stability in high salinity and biocompatible solutions. The thermally treated NPs showed colloidal and physicochemical stability for 1 month, which were enhanced by the addition of T80. Due to the nature of the precursors and their colloidal properties, the NPs are highly promising for applications as biocompatible drug delivery nanocarriers while T80 acts as an agent to modify their properties.

Keywords:  Tween 80; bovine beta-lactoglobulin; electrostatic complexation; nanoparticles; porcine chondroitin sulfate; thermal treatment

DOI:  https://doi.org/10.3390/polym16141995
Steroids. 2024 Jul 23. pii: S0039-128X(24)00121-1. [Epub ahead of print] 109483

Steroid sulfatase in mouse liver and Testis: Characterization, ontogeny and localization.

Barathi Balasubramonian, Kyle W Selcer.

  Steroid hormones often circulate in the plasma as inactive sulfated forms, such as estrone sulfate and dehydroepiandrosterone sulfate. The enzyme steroid sulfatase (STS) converts these steroids into active forms, mainly estrogens, in peripheral tissues. STS is present in most tissues, but it occurs at higher levels in certain organs, notably liver and placenta. In this study, we examined the tissue distribution of STS in a prominent laboratory model, the house mouse (Mus musculus). Tissues included were heart, liver, small intestine, skeletal muscle, and gonads of both sexes. An 3H-estrone-sulfate conversion assay was used to measure STS activity in tissue homogenates and extracts. STS activities were high for hepatic tissue homogenates of both genders. Testicular STS levels were similar to those of liver, while STS activities of ovary, small intestine, heart, and muscle were considerably lower. The specific STS inhibitors, EMATE and STX-64 virtually eliminated STS activity in hepatic microsomes and cytosols, verifying that the observed enzyme activity was due to STS. Enzyme kinetic assays showed Km values of 8.6 µM for liver and 9.1 µM for testis, using E1S as substrate. Hepatic and testicular STS activities, measured in CHAPS-extracted microsome, were low up to 5 weeks of age and were higher through 56 weeks. Western blotting, with a specific STS antibody, confirmed the presence of STS protein (65 Da) in both liver and testis. Immunofluorescence of tissue sections detected the presence of STS protein in hepatocytes, in testicular Leydig cells and in seminiferous tubules (Leydig cells and developing germ cells). These results suggest that STS may have a significant role in testicular function.

Keywords:  Estrogens; Liver; Mouse; Steroid sulfatase; Sulfated steroids; Testis

DOI:  https://doi.org/10.1016/j.steroids.2024.109483
Endocr Pract. 2024 Jul 18. pii: S1530-891X(24)00607-4. [Epub ahead of print]

Is there a relationship of cortisol and sex steroid hormones with mental health in middle-aged adults? the FIT-AGEING study.

Lourdes Herrera-Quintana, Héctor Vázquez-Lorente, Almudena Carneiro-Barrera, Luis Gutiérrez-Rojas, Manuel J Castillo, Francisco J Amaro-Gahete.

   PURPOSE: Mental health has emerged as a worldwide concern given the rising incidence of anxiety and depression disorders in the last years. Cortisol and sex steroid hormones have been demonstrated to be important regulators of mental health processes in older adults. However, the evidence considering these integrated variables in apparently healthy middle-aged individuals has not been thoroughly addressed. The present study was aimed at investigating the association of plasma cortisol, testosterone, free testosterone, Sex Hormone-Binding Globulin (SHBG) and Dehydroepiandrosterone-sulfate (DHEAS) levels with mental health in middle-aged adults.
METHODS: This cross-sectional study included a cohort of 73 middle-aged adults with 45-65 years of age (53% women). Plasma cortisol, testosterone, SHBG, and DHEAS were assessed using a competitive chemiluminescence immunoassay. Free testosterone was calculated from the total testosterone and SHBG. Self-reported depression severity, generic health-related quality of life, hope, satisfaction with life, and optimism-pessimism were evaluated using the Beck Depression Inventory-II (BDI-II), the 36-item Short-Form Health Survey, the Adult Hope Scale (AHS), the Satisfaction with Life Scale (SWLS), and the Life Orientation Test Revised, respectively - higher total scores of these scales indicating greater levels of these variables -.
RESULTS: Testosterone and free testosterone levels were inversely associated with BDI-II values in men (all P≤0.042). Cortisol levels were positively related with SWLS scores, whereas testosterone, free testosterone, SHBG, and DHEAS levels were negatively correlated with BDI-II values in women (all P≤0.045).
CONCLUSION: In summary, these results suggest that increased levels of steroid hormones - within the normal values - may be associated with better mental health in middle-aged adults.

Keywords:  DHEAS; SHBG; cortisol; depression; quality of life; testosterone

DOI:  https://doi.org/10.1016/j.eprac.2024.07.009
J Fungi (Basel). 2024 Jun 28. pii: 457. [Epub ahead of print]10(7):

Physicochemical Properties of Different Sulfated Polysaccharide Components from Laetiporus sulphureus and Their Anti-Proliferative Effects on MDA-MB-231 Breast Cancer Cells.

Chia-I Jen, Lean-Teik Ng.

  Laetiporus sulphureus is an edible and medicinal mushroom widely used in folk medicine for treating cancer and gastric diseases. This study aimed to investigate the physicochemical properties of different sulfated polysaccharide (SPS) components (F1, F2, and F3) isolated from L. sulphureus and evaluate their activity against MDA-MB-231 breast cancer cell proliferation. Compared with F1 and F3, the results showed that F2 exhibited the most potent anti-proliferative activity on MDA-MB-231 cells, which could be attributed to the sulfate and protein contents, molecular weight, and monosaccharide composition. F2 inhibited breast cancer cell proliferation by blocking the cell cycle at the G0/G1 phase but not triggering cell apoptosis. In addition, F2 also showed selective cytotoxicity on breast cancer cells. It modulated the expression of proteins involved in G0/G1 phase progression, cell cycle checkpoints, DNA replication, and the TGFβ signaling pathway in MDA-MB-231 cells. This study demonstrated that F2, the medium-molecular-weight SPS component of L. sulphureus, possessed the most potent inhibitory effect on MDA-MB-231 cell proliferation by arresting the cell cycle at the G0/G1 phase. The main factors contributing to the differences in the potency of anti-breast cancer activity between F1, F2, and F3 could be the sulfate and protein contents, molecular weight, and monosaccharide composition of SPS.

Keywords:  Laetiporus sulphureus; cell cycle arrest; physicochemical properties; protein microarray; sulfated polysaccharides; triple-negative breast cancer

DOI:  https://doi.org/10.3390/jof10070457
Biomolecules. 2024 Jul 11. pii: 832. [Epub ahead of print]14(7):

Can Hyaluronic Acid Combined with Chondroitin Sulfate in Viscosupplementation of Knee Osteoarthritis Improve Pain Symptoms and Mobility?

Augustin Dima, Magda Dragosloveanu, Andreea Ramona Romila, Alexandru Cristea, Georgiana Marinică, Alexandru-Tiberiu Dănilă, Alexandru Mandici, Daniel Cojocariu, Robert-Alexandru Vlad, Adriana Ciurba, Magdalena Bîrsan.

  The objective of the present study was to assess the effect of intra-articular Hyaluronic acid (HA) and Chondroitin sulfate (CS) supplementation (Hialurom® Hondro (HH)) on pain symptoms and joint mobility. In total, 60 mg/mL sodium hyaluronate and 90 mg/mL CS were administered to 21 patients (17 females and 4 males) respecting the in-force requirements, excluding patients with some specific comorbidities. In addition to the clinical study (where the pain intensity (severity) and joint mobility were assessed), rheological characterization was conducted evaluating the following parameters: elastic modulus (G'), loss modulus (G″) oscillatory frequency (fc) at 0.5 Hz and 2.5 Hz, crossover frequency (fc), relaxation time (λ) where it was noticed that the addition of chondroitin sulfate (CS) to sodium hyaluronate (SH) significantly enhances and improves the viscoelastic properties, particularly at higher shear frequencies. A significant decrease in pain intensity felt by the subjects was found, from 7.48 (according to Wong-Baker scale)-pain close to 8 (the patient is unable to perform most activities), to more reduced values of 5.86-at 6 weeks after injection, 4.81-at 3 months after injection, and 5.24-at 6 months after injection, improvements in symptoms was fast and durable. Data related to the evolution of joint mobility show that at 6 weeks after injection, the mobility of joints increased by 17.8% and at 6 months by 35.61%. No serious adverse events were reported with undesired effects so that they would impose additional measures. Better resistance to enzymatic degradation and free radicals could be expected from the synergic combination of sodium hyaluronate and chondroitin sodium sulfate, this having a special importance for the patients, granting them the ability to perform more ample movements and reducing dependency on attendants, thus increasing quality of life.

Keywords:  chondroitin sulfate; hyaluronic acid intra-articular; infiltrations; knee osteoarthritis; pre-filled syringe; viscosupplementation

DOI:  https://doi.org/10.3390/biom14070832
Gels. 2024 Jul 09. pii: 450. [Epub ahead of print]10(7):

The Influence of Sulfation Degree of Glycosaminoglycan-Functionalized 3D Collagen I Networks on Cytokine Profiles of In Vitro Macrophage-Fibroblast Cocultures.

Franziska Ullm, Alexander Renner, Uwe Freudenberg, Carsten Werner, Tilo Pompe.

  Cell-cell interactions between fibroblasts and immune cells, like macrophages, are influenced by interaction with the surrounding extracellular matrix during wound healing. In vitro hydrogel models that mimic and modulate these interactions, especially of soluble mediators like cytokines, may allow for a more detailed investigation of immunomodulatory processes. In the present study, a biomimetic extracellular matrix model based on fibrillar 3D collagen I networks with a functionalization with heparin or 6-ON-desulfated heparin, as mimics of naturally occurring heparan sulfate, was developed to modulate cytokine binding effects with the hydrogel matrix. The constitution and microstructure of the collagen I network were found to be stable throughout the 7-day culture period. A coculture study of primary human fibroblasts/myofibroblasts and M-CSF-stimulated macrophages was used to show its applicability to simulate processes of progressed wound healing. The quantification of secreted cytokines (IL-8, IL-10, IL-6, FGF-2) in the cell culture supernatant demonstrated the differential impact of glycosaminoglycan functionalization of the collagen I network. Most prominently, IL-6 and FGF-2 were shown to be regulated by the cell culture condition and network constitution, indicating changes in paracrine and autocrine cell-cell communication of the fibroblast-macrophage coculture. From this perspective, we consider our newly established in vitro hydrogel model suitable for mechanistic coculture analyses of primary human cells to unravel the role of extracellular matrix factors in key events of tissue regeneration and beyond.

Keywords:  3D hydrogels; coculture; collagen I networks; glycosaminoglycans; macrophages; wound healing

DOI:  https://doi.org/10.3390/gels10070450
Artif Organs. 2024 Jul 19.

Auricular cartilage regeneration using chondroitin sulfate-based hydrogel with mesenchymal stem cells in rabbits.

Masoud Janipour, Amir Soltaniesmaeili, Seyed Hossein Owji, Zahra Shahhossein, Seyedeh-Sara Hashemi.

   BACKGROUND: Cartilage is an avascular and alymphatic tissue that lacks the intrinsic ability to undergo spontaneous repair and regeneration in the event of significant injury. The efficacy of conventional therapies for invasive cartilage injuries is limited, thereby prompting the emergence of cartilage tissue engineering as a possible alternative. In this study, we fabricated three-dimensional hydrogel films utilizing sodium alginate (SA), gelatin (Gel), and chondroitin sulfate (CS). These films were included with Wharton's jelly mesenchymal stem cells (WJ-MSCs) and intended for cartilage tissue regeneration.
METHODS: The hydrogel film that were prepared underwent evaluation using various techniques including scanning electron microscope (SEM), Fourier transform infrared (FTIR) spectroscopy, assessment of the degree of swelling, degradation analysis, determination of water vapor transmission rate (WVTR), measurement of water contact angle (WCA), evaluation of mechanical strength, and assessment of biocompatibility. The rabbit ear cartilage regeneration by hydrogel films with and without of WJ-MSCs was studied by histopathological investigations during 15, 30, and 60 days.
RESULTS: The hydrogel films containing CS exhibited superior metrics compared to other nanocomposites such as better mechanical strength (12.87 MPa in SA/Gel compared to 15.56 in SA/Gel/CS), stability, hydrophilicity, WVTR (3103.33 g/m2/day in SA/Gel compared to 2646.67 in nanocomposites containing CS), and swelling ratio (6.97 to 12.11% in SA/Gel composite compared to 5.03 to 10.90% in SA/Gel/CS). Histopathological studies showed the presence of chondrocyte cells in the lacunae on the 30th day and the complete restoration of the cartilage tissue on the 60th day following the injury in the group of SA/Gel/CS hydrogel containing WJ-MSCs.
CONCLUSIONS: We successfully fabricated a scaffold composed of alginate, gelatin, and chondroitin sulfate. This scaffold was further enhanced by the incorporation of Wharton's jelly mesenchymal stem cells. Our findings demonstrate that this composite scaffold has remarkable biocompatibility and mechanical characteristics. The present study successfully demonstrated the therapeutic potential of the SA-Gel-CS hydrogel containing WJ-MSCs for cartilage regeneration in rabbits.

Keywords:  Wharton's jelly mesenchymal stem cells; auricular cartilage regeneration; chondroitin sulfate; hydrogel films

DOI:  https://doi.org/10.1111/aor.14807
Nanoscale Adv. 2024 Jul 23. 6(15): 3825-3837

Rapid cellular uptake of citrate-coated iron oxide nanoparticles unaffected by cell-surface glycosaminoglycans.

Lena Kampen, Amani Remmo, Shailey Gale Twamley, Andrea Weller, Anke Stach, Paul Turko, Norbert Löwa, Frank Wiekhorst, Antje Ludwig.

Citrate-coated iron oxide nanoparticles, specifically Synomag®-COOH (SynC), are promising tracers in magnetic particle imaging (MPI) due to their high magnetic moments and rapid cellular uptake. The mechanisms driving efficient SynC uptake remain unclear. Previous observations suggest a role of the extracellular glycocalyx during nanoparticle uptake. Here, we ascertain whether the cell-surface glycosaminoglycans (GAGs) regulate the uptake of SynC. Using transmission electron microscopy (TEM), we visualized SynC uptake by THP-1 cells, a human acute monocytic leukemia cell line. We investigated the interaction of SynC with GAGs in living cells using click-chemistry-based labeling. Upon treating THP-1 cells with chondroitinase or hyaluronidase and with a xylosyltransferase-deficient cell line, we quantified SynC uptake and measured interactions of SynC with cells in real time using magnetic particle spectroscopy (MPS). The THP-1 cell membrane engulfed or formed extensions around SynC, indicating uptake through pinocytosis and phagocytosis. We measured an increased MPS signal of SynC within seconds of cell contact, suggesting an interaction with extracellular components like the glycocalyx. Upon adding SynC to THP-1 cells, we could not observe disruption of fluorescently labeled GAGs or an enhanced intracellular fluorescence, implying that SynC does not accelerate the turnover of GAGs by binding. Lack of chondroitin sulfate, heparan sulfate, and hyaluronic acid did not affect the rapid magnetic behavior increase of SynC upon cell contact. Accordingly, we measured no significant differences in SynC uptake between wild type cells and our GAG-deficient models. These findings suggest that GAGs act as a permeable bandpass for SynC nanoparticles with a minor negative surface charge of -13.8 mV. This finding has significant implications for MPI-based cell tracking because it facilitates efficient tracking of cell types that lack a strong repulsion by cell-surface GAGs. It will be crucial to investigate whether the rapid uptake of SynC is cell-type specific and influenced by different extracellular matrix compositions.

DOI: https://doi.org/10.1039/d4na00277f
Am J Respir Cell Mol Biol. 2024 Jul 23.

Enzymatic Modulation of the Pulmonary Glycocalyx Enhances Susceptibility to Streptococcus pneumoniae.

Cengiz Goekeri, Kerstin A K Linke, Karen Hoffmann, Elena Lopez-Rodriguez, Vladimir Gluhovic, Anne Voß, Sandra Kunder, Andreas Zappe, Sara Timm, Alina Nettesheim, Sebastian M K Schickinger, Christian M Zobel, Kevin Pagel, Achim D Gruber, Matthias Ochs, Martin Witzenrath, Geraldine Nouailles.

  The pulmonary epithelial glycocalyx is rich in glycosaminoglycans such as hyaluronan and heparan sulfate. Despite their presence, the importance of these glycosaminoglycans in bacterial lung infections remains elusive. To address this, we intranasally inoculated mice with Streptococcus pneumoniae in the presence or absence of enzymes targeting pulmonary hyaluronan and heparan sulfate, followed by characterization of subsequent disease pathology, pulmonary inflammation, and lung barrier dysfunction. Enzymatic degradation of hyaluronan and heparan sulfate exacerbated pneumonia in mice, as evidenced by increased disease scores and alveolar neutrophil recruitment. However, targeting epithelial hyaluronan in combination with Streptococcus pneumoniae infection further exacerbated systemic disease, indicated by elevated splenic bacterial load and plasma levels of pro-inflammatory cytokines. In contrast, enzymatic cleavage of heparan sulfate resulted in increased bronchoalveolar bacterial burden, lung damage and pulmonary inflammation in mice infected with Streptococcus pneumoniae. Accordingly, heparinase-treated mice also exhibited disrupted lung barrier integrity as evidenced by higher alveolar edema scores and vascular protein leakage into the airways. This finding was corroborated in a human alveolus-on-a-chip platform, confirming that heparinase treatment also disrupts the human lung barrier during Streptococcus pneumoniae infection. Notably, enzymatic pre-treatment with either hyaluronidase or heparinase also rendered human epithelial cells more sensitive to pneumococcal-induced barrier disruption, as determined by transepithelial electrical resistance measurements, consistent with our findings in murine pneumonia. Taken together, these findings demonstrate the importance of intact hyaluronan and heparan sulfate in limiting pneumococci-induced damage, pulmonary inflammation, and epithelial barrier function and integrity.

Keywords:  CAP, pneumococcal virulence, acute lung injury, glycosaminoglycans, heparan sulfate, hyaluronan

DOI:  https://doi.org/10.1165/rcmb.2024-0003OC
Food Chem. 2024 Jun 24. pii: S0308-8146(24)01858-2. [Epub ahead of print]459 140208

Recent advance for animal-derived polysaccharides in nanomaterials.

Meng-Na Li, Xiang-Ze Jia, Qing-Bo Yao, Feng Zhu, Yan-Yan Huang, Xin-An Zeng.

  Inspired by the structure characteristics of natural products, the size and morphology of particles are carefully controlled using a bottom-up approach to construct nanomaterials with specific spatial unit distribution. Animal polysaccharide nanomaterials, such as chitosan and chondroitin sulfate nanomaterials, exhibit excellent biocompatibility, degradability, customizable surface properties, and novel physical and chemical properties. These nanomaterials hold great potential for development in achieving a sustainable bio-economy. This paper provides a summary of the latest research results on the preparation of nanomaterials from animal polysaccharides. The mechanism for preparing nanomaterials through the bottom-up method from different sources of animal polysaccharides is introduced. Furthermore, this paper discusses the potential hazards posed by industrial applications to the environment and human health, as well as the challenges and future prospects associated with using animal polysaccharides in nanomaterials.

Keywords:  Animal polysaccharides; Bottom-up approach; Mechanism; Nanomaterials

DOI:  https://doi.org/10.1016/j.foodchem.2024.140208
Biotechnol Adv. 2024 Jul 20. pii: S0734-9750(24)00110-1. [Epub ahead of print]75 108416

Unraveling the intricacies of glycosaminoglycan biosynthesis: Decoding the molecular symphony in understanding complex polysaccharide assembly.

Zhi-Yuan Yao, Jin-Song Gong, Jia-Yu Jiang, Chang Su, Wen-Han Zhao, Zheng-Hong Xu, Jin-Song Shi.

  Glycosaminoglycans (GAGs) are extensively utilized in clinical, cosmetic, and healthcare field, as well as in the treatment of thrombosis, osteoarthritis, rheumatism, and cancer. The biological production of GAGs is a strategy that has garnered significant attention due to its numerous advantages over traditional preparation methods. In this review, we embark on a journey to decode the intricate molecular symphony that orchestrates the biosynthesis of glycosaminoglycans. By unraveling the complex interplay of related enzymes and thorough excavation of the intricate metabolic cascades involved, GAGs chain aggregation and transportation, which efficiently and controllably modulate GAGs sulfation patterns involved in biosynthetic pathway, we endeavor to offer a thorough comprehension of how these remarkable GAGs are intricately assembled and pushes the boundaries of our understanding in GAGs biosynthesis.

Keywords:  Glycosaminoglycans; Sulfated modification; Synthesis mechanism; Synthetic biology

DOI:  https://doi.org/10.1016/j.biotechadv.2024.108416
Int J Biol Macromol. 2024 Jul 20. pii: S0141-8130(24)04859-1. [Epub ahead of print] 134054

Polysaccharide base electrospun nanofibrous scaffolds for cartilage tissue engineering: Challenges and opportunities.

Atefeh Arash, Fatemeh Dehgan, Soheila Zamanlui Benisi, Milad Jafari-Nodoushan, Mohamad Pezeshki-Modaress.

  Polysaccharides, known as naturally abundant macromolecular materials which can be easily modified chemically, have always attracted scientists' interest due to their outstanding properties in tissue engineering. Moreover, their intrinsic similarity to cartilage ECM components, biocompatibility, and non-harsh processing conditions make polysaccharides an excellent option for cartilage tissue engineering. Imitating the natural ECM structure to form a fibrous scaffold at the nanometer scale in order to recreate the optimal environment for cartilage regeneration has always been attractive for researchers in the past few years. However, there are some challenges for polysaccharides electrospun nanofibers preparation, such as poor solubility (Alginate, cellulose, chitin), high viscosity (alginate, chitosan, and Hyaluronic acid), high surface tension, etc. Several methods are reported in the literature for facing polysaccharide electrospinning issues, such as using carrier polymers, modification of polysaccharides, and using different solvent systems. In this review, considering the importance of polysaccharide-based electrospun nanofibers in cartilage tissue engineering applications, the main achievements in the past few years, and challenges for their electrospinning process are discussed. After careful investigation of reported studies in the last few years, alginate, chitosan, hyaluronic acid, chondroitin sulfate, and cellulose were chosen as the main polysaccharide base electrospun nanofibers used for cartilage regeneration.

Keywords:  Cartilage tissue engineering; Electrospinning; Nanofibrous scaffold; Polysaccharides

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.134054
ACS Nano. 2024 Jul 22.

Magnetic Polysaccharide Mesenchymal Stem Cells Exosomes Delivery Microcarriers for Synergistic Therapy of Osteoarthritis.

Lei Yang, Wenzhao Li, Yuanjin Zhao, Luoran Shang.

  Osteoarthritis (OA) is a prevalent degenerative disease that afflicts more than 250 million people worldwide, impairing their mobility and quality of life. However, conventional drug therapy is palliative. Exosomes (Exo), although with the potential to fundamentally repair cartilage, face challenges in their efficient enrichment and delivery. In this study, we developed magnetic polysaccharide hydrogel particles as microcarriers for synergistic therapy of OA. The microcarriers were composed of modified natural polysaccharides, hyaluronic acid (HAMA), and chondroitin sulfate (CSMA), and were generated from microfluidic electrospray in combination with a cryogelation process. Magnetic nanoparticles with spiny structures capable of capturing stem cell Exo were encapsulated within the microcarriers together with an anti-inflammatory drug diclofenac sodium (DS). The released DS and Exo from the microcarriers had a synergistic effect in alleviating the OA symptoms and promoting cartilage repair. The in vitro and in vivo results demonstrated the excellent performance of the microcarrier for OA treatment. We believe this work has potential for Exo therapy of OA and other related diseases.

Keywords:  exosome; microcarrier; microfluidics; osteoarthritis; stem cell

DOI:  https://doi.org/10.1021/acsnano.4c01406
Carbohydr Polym. 2024 Oct 15. pii: S0144-8617(24)00599-X. [Epub ahead of print]342 122373

Characterization of a novel sulfated-rhamnoglucuronan isolated from Korean seaweed Ulva pertusa and its efficacy for treatment of inflammatory bowel disease in mice.

Seung-U Son, Hyung Joo Suh, Kwang-Soon Shin.

  This study aimed to isolate Ulva pertusa polysaccharide (UPP), which elicits anti-inflammatory bowel disease (IBD) effects, from the Korea seaweed U. pertusa and identify its structure. Firstly, UPP was isolated from U. pertusa using hydrothermal extraction and ethanol precipitation. UPP is a novel polysaccharide that exhibits unique structural features such as 3-sulfated rhamnose, glucuronic acid, iduronic acid, and 3-sulfated xylose, which are repeated in 1,4-glycosidic bonds. Prophylactic oral administration of UPP in mice with dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) suppressed the levels of inflammatory cytokines and MAPK- and NF-κB-related factors in the serum and colon tissue. Tight junction (TJ)-related factors such as occludin, claudin-1, and mucin were effectively augmented by UPP in the colon tissue. In addition, UPP administration prevented the DSS treatment-led cecal short chain fatty acid imbalance, and this effect was most evident for propionic acid. In conclusion, UPP isolated from the Korean U. pertusa demonstrates potent anti-IBD activity. Characterization of this ulvan revealed its unique structure. Moreover, its efficacy may be associated with its anti-inflammatory effects and regulation of gut microbiota and TJ proteins. Thus, this study provides new insights into the biological effects of UPP in IBD.

Keywords:  Dextran sulfate sodium; Inflammatory bowel disease; Sulfated polysaccharide; Sulfated-rhamnoglucuronan; Ulva pertusa; Ulvan

DOI:  https://doi.org/10.1016/j.carbpol.2024.122373
Biomolecules. 2024 Jul 15. pii: 850. [Epub ahead of print]14(7):

Hepatic Gene Expression and Metabolite Profiles of Androstenone and Skatole Relative to Plasma Estrone Sulfate Levels in Boars.

Christine Bone, E James Squires.

  Testicular steroids can alter the activity and expression of enzymes within the liver and may influence the metabolism of skatole and androstenone, which are responsible for boar taint. Plasma levels of estrone sulfate (E1S) are indicative of the steroidogenic capacity of the boar and are variable between animals of similar live weights at slaughter. This study aimed to characterize the relationship between steroidogenic capacity and the metabolism of boar taint compounds by relating plasma E1S levels at slaughter weight to the expression levels of genes regulating the metabolism of androstenone and skatole, along with their respective metabolite profiles. RT-qPCR was used to evaluate gene expression in the liver. Hepatocytes were also isolated and treated with androstenone or skatole, with metabolite levels in the incubation media quantified by high-performance liquid chromatography. Plasma E1S levels ranged from 2.2-108.5 ng/mL and were positively correlated with overall skatole metabolism (p = 0.038), the production of metabolites 3-methyloxindole (p = 0.026) and 3-hydroxy-3-methyloxindole (p = 0.036), and expression levels of key genes involved in skatole metabolism, specifically CYP2C33 (p = 0.0042), CYP2C49 (p = 0.022), and CYB5R1 (p = 0.017). There was no association between androstenone metabolism and plasma E1S concentrations; however, there was evidence of possible co-regulation amongst genes involved in the metabolism of androstenone, skatole, and estrogens. These findings indicate that steroidogenic capacity is related to the rate of skatole, but not androstenone metabolism, in slaughter-weight boars.

Keywords:  androstenone; boar taint; estrone sulfate; metabolism; skatole; steroidogenic capacity

DOI:  https://doi.org/10.3390/biom14070850
Biomolecules. 2024 Jul 10. pii: 829. [Epub ahead of print]14(7):

Dermatan Sulfate Affects the Activation of the Necroptotic Effector MLKL in Breast Cancer Cell Lines via the NFκB Pathway and Rac-Mediated Oxidative Stress.

Grzegorz Wisowski, Adam Pudełko, Monika Paul-Samojedny, Katarzyna Komosińska-Vassev, Ewa M Koźma.

  Dermatan sulfate (DS) is a glycosaminoglycan characterized by having a variable structure and wide distribution in animal tissues. We previously demonstrated that some structural variants of DS were able to rapidly induce moderate necroptosis in luminal breast cancer cells when used at a high concentration. We have now investigated the mechanisms underlying the DS-mediated activation of the necroptotic executor MLKL using immunofluorescence, Western blotting and pharmacological inhibition. The two main processes, by which DS influences the phosphorylation of MLKL, are the activation of NFκB, which demonstrates a suppressive impact, and the induction of oxidative stress, which has a stimulatory effect. Moreover, the triggering of the redox imbalance by DS occurs via the modulatory influence of this glycosaminoglycan on the rearrangement of the actin cytoskeleton, requiring alterations in the activity of small Rho GTP-ase Rac1. All of these processes that were elicited by DS in luminal breast cancer cells showed a dependence on the structure of this glycan and the type of cancer cells. Furthermore, our results suggest that a major mechanism that is involved in the stimulation of necroptosis in luminal breast cancer cells by high doses of DS is mediated via the effect of this glycan on the activity of adhesion molecules.

Keywords:  MLKL; NFκB transcription factor; Rac GTPase; breast cancer; cell death; dermatan sulfate; oxidative stress

DOI:  https://doi.org/10.3390/biom14070829
Toxins (Basel). 2024 Jul 12. pii: 316. [Epub ahead of print]16(7):

7-Phenylheptanoic Acid-Hydroxypropyl β-Cyclodextrin Complex Slows the Progression of Renal Failure in Adenine-Induced Chronic Kidney Disease Mice.

Kindness Lomotey Commey, Airi Enaka, Ryota Nakamura, Asami Yamamoto, Kenji Tsukigawa, Koji Nishi, Masaki Otagiri, Keishi Yamasaki.

  The characteristic accumulation of circulating uremic toxins, such as indoxyl sulfate (IS), in chronic kidney disease (CKD) further exacerbates the disease progression. The gut microbiota, particularly gut bacterial-specific enzymes, represents a selective and attractive target for suppressing uremic toxin production and slowing the progression of renal failure. This study investigates the role of 4-phenylbutyrate (PB) and structurally related compounds, which are speculated to possess renoprotective properties in suppressing IS production and slowing or reversing renal failure in CKD. In vitro enzyme kinetic studies showed that 7-phenylheptanoic acid (PH), a PB homologue, suppresses the tryptophan indole lyase (TIL)-catalyzed decomposition of tryptophan to indole, the precursor of IS. A hydroxypropyl β-cyclodextrin (HPβCD) inclusion complex formulation of PH was prepared to enhance its biopharmaceutical properties and to facilitate in vivo evaluation. Prophylactic oral administration of the PH-HPβCD complex formulation reduced circulating IS and attenuated the deterioration of renal function and tubulointerstitial fibrosis in adenine-induced CKD mice. Additionally, treatment of moderately advanced adenine-induced CKD mice with the formulation ameliorated renal failure, although tissue fibrosis was not improved. These findings suggest that PH-HPβCD can slow the progression of renal failure and may have implications for preventing or managing CKD, particularly in early-stage disease.

Keywords:  7-phenylheptanoic acid; CKD; adenine-induced CKD mice; hydroxypropyl β-cyclodextrin; inclusion complex; indoxyl sulfate; tryptophan indole lyase

DOI:  https://doi.org/10.3390/toxins16070316
J Am Chem Soc. 2024 Jul 25.

Expressed Protein Ligation in Flow.

Lucas Kambanis, Anthony Ayoub, Max J Bedding, Peter H G Egelund, Joshua W C Maxwell, Charlotte Franck, Lucien Lambrechts, Paige M E Hawkins, Timothy S Chisholm, Joel P Mackay, Emma Sierecki, Yann Gambin, Sameer S Kulkarni, Richard J Payne.

The development of a flow chemistry platform for the generation of modified protein targets via expressed protein ligation (EPL) is described. The flow EPL platform enables efficient ligation reactions with high recoveries of target protein products and superior reaction rates compared to corresponding batch processes. The utility of the flow EPL technology was first demonstrated through the semisynthesis of the tick-derived chemokine-binding protein ACA-01 containing two tyrosine sulfate modifications. Full-length, sulfated ACA-01 could be efficiently assembled by ligating a recombinantly expressed C-terminal protein fragment and a synthetic sulfopeptide thioester in flow. Following folding, the semisynthetic sulfoprotein was shown to exhibit potent binding to a variety of pro-inflammatory chemokines. In a second modified protein target, we employed an in-line flow EPL-photodesulfurization strategy to generate both unmodified and phosphorylated forms of human β-synuclein by fusing a recombinant protein thioester, generated through cleavage of an intein fusion protein, and a synthetic (phospho)peptide. The semisynthetic proteins were assembled in 90 min in flow, a significant improvement over corresponding batch protein assembly, and enabled access to tens of milligrams of high purity material. Flow EPL has the potential to serve as a robust technology to streamline access to homogeneously modified proteins for a variety of applications in both academia, as well as in the pharmaceutical and biotechnology sector.

DOI: https://doi.org/10.1021/jacs.4c07462
Mol Genet Metab Rep. 2024 Sep;40 101113

Initiation of fluoxetine in a pediatric patient with Mucopolysaccharidosis IIIA: Early observations.

Lindsay Torrice, Elizabeth Jalazo.

  Fluoxetine has been identified as a potential treatment for mucopolysaccharidosis IIIA (MPS IIIA), a debilitating and progressive lysosomal storage disorder for which no treatments are approved. In the MPS IIIA mouse model, fluoxetine decreases the accumulation of glycosaminoglycans and aggregated autophagic substrates, reducing inflammation, and slowing cognitive deterioration. 1 We treated a single patient, 6 years old, under off-label prescription of fluoxetine, a selective serotonin reuptake inhibitor (SSRI). The primary endpoint was safety. Secondary exploratory assessments included urine quantitative heparan sulfate. Fluoxetine was well-tolerated in this patient and the patient continued treatment following the 12-month monitoring period. The patient experienced an increase in daytime somnolence which resolved with rescheduling fluoxetine administration to bedtime. Quantitative heparan sulfate levels remained elevated during treatment. Parents reported improved sleep latency time and less nighttime waking. These findings support general tolerability and further study of fluoxetine as a potential therapy for MPS IIIA.

Keywords:  Fluoxetine; Glycosaminoglycan; MPS IIIA; Mucopolysaccharidosis; Pediatric; Sanfilippo syndrome

DOI:  https://doi.org/10.1016/j.ymgmr.2024.101113
Front Toxicol. 2024 ;6 1398852

Bioactivation of cinnamic alcohol in a reconstructed human epidermis model and evaluation of sensitizing potency of the identified metabolites.

Lorena Ndreu, Josefine Carlsson, David J Ponting, Ida B Niklasson, E Johanna L Stéen, Lukas McHugh, Niamh M O'Boyle, Kristina Luthman, Ann-Therese Karlberg, Isabella Karlsson.

   Background: Cinnamic alcohol is a natural compound, widely used in fragrances, which can cause allergic contact dermatitis. Cinnamic alcohol lacks intrinsic reactivity and autoxidation or metabolic activation is necessary for it to act as a sensitizer.
Methods: Bioactivation of cinnamic alcohol was explored using human liver microsomes, human liver S9 and SkinEthic™ Reconstructed Human Epidermis. A targeted multiple reaction monitoring mass spectrometry method was employed to study and quantify cinnamic alcohol along with eight potential phase I or phase II metabolites. The reconstructed human epidermis model, treated with cinnamic alcohol, was also analyzed with a non-targeted high-resolution mass spectrometry method to identify metabolites not included in the targeted method.
Results: Two metabolites identified with the targeted method, namely, pOH-cinnamic alcohol and pOH-cinnamic aldehyde, have not previously been identified in a metabolic in vitro system. Their reactivity toward biologically relevant nucleophiles was investigated and compared to their sensitizing potency in vivo in the murine local lymph node assay (LLNA). According to the LLNA, the pOH-cinnamic alcohol is non-sensitizing and pOH-cinnamic aldehyde is a moderate sensitizer. This makes pOH-cinnamic aldehyde less sensitizing than cinnamic aldehyde, which has been found to be a strong sensitizer in the LLNA. This difference in sensitizing potency was supported by the reactivity experiments. Cinnamic sulfate, previously proposed as a potential reactive metabolite of cinnamic alcohol, was not detected in any of the incubations. In addition, experiments examining the reactivity of cinnamic sulfate toward a model peptide revealed no evidence of adduct formation. The only additional metabolite that could be identified with the non-targeted method was a dioxolan derivative. Whether or not this metabolite, or one of its precursors, could contribute to the sensitizing potency of cinnamic alcohol would need further investigation.
Discussion: Cinnamic alcohol is one of the most common fragrance allergens and as it is more effective to patch test with the actual sensitizer than with the prohapten itself, it is important to identify metabolites with sensitizing potency. Further, improved knowledge of metabolic transformations occurring in the skin can improve prediction models for safety assessment of skin products.

Keywords:  biotransformation; cinnamic alcohol; cinnamic sulfate; local lymph node assay (LLNA); mass spectrometry; pOH-cinnamic alcohol; pOH-cinnamic aldehyde; reconstructed human epidermis

DOI:  https://doi.org/10.3389/ftox.2024.1398852
J Appl Toxicol. 2024 Jul 22.

Hepatic enzyme induction and its potential effect on thyroid hormone metabolism in the metamorphosing tadpole of Xenopus laevis (African clawed frog).

Kohei Wada, Takafumi Yamaguchi, Hitoshi Tanaka, Takuo Fujisawa.

  Hepatic enzyme induction, an inherent defense system against xenobiotics, is known to simultaneously affect endocrine system functions in mammals under specific conditions, particularly thyroid hormone (TH) regulation. While this phenomenon has been studied extensively, the pathway leading to this indirect thyroid effect in mammals has unclear applicability to amphibians, despite the importance of amphibian species in assessing thyroid-disruptive chemicals. Here, we investigated the effects of three well-known mammalian enzyme inducers-β-naphthoflavone (BNF), pregnenolone carbonitrile (PCN), and sodium phenobarbital (NaPB)-on the gene expression of phase-I and phase-II metabolizing enzymes in Xenopus laevis tadpoles. Waterborne exposure to BNF and PCN significantly induced the expression of both phase-I (cytochrome P450, CYP) and phase-II enzymes (UDP-glucuronosyltransferase, UGT and sulfotransferase, SULT), but in different patterns, while NaPB exposure induced CYP2B expression without affecting phase-II enzymes in tadpoles, in contrast to mammals. Furthermore, an ex vivo hepatic enzyme activity assay confirmed that BNF treatment significantly increased phase-II metabolic activity (glucuronidation and sulfation) toward TH. These results suggest the potential for certain mammalian enzyme inducers to influence TH clearance in X. laevis tadpoles. Our findings provide insights into the profiles of xenosensing activity and enzyme induction in amphibians, which can facilitate a better understanding of the mechanisms of indirect effects on the thyroid system via hepatic enzyme induction in nonmammalian species.

Keywords:  Xenopus laevis; endocrine disruption; hepatic enzyme induction; pregnenolone carbonitrile; sodium phenobarbital; thyroid hormone; xenosensor; β‐naphthoflavone

DOI:  https://doi.org/10.1002/jat.4672
Carbohydr Polym. 2024 Oct 15. pii: S0144-8617(24)00647-7. [Epub ahead of print]342 122421

Fucoidan modulates gut microbiota and immunity in Peyer's patches against inflammatory bowel disease.

Jia-Kang Li, Suresh Veeraperumal, Jude Juventus Aweya, Yang Liu, Kit-Leong Cheong.

  Although fucoidan has potential use as an anti-inflammatory agent, the specific mechanisms by which it influences signaling and immunomodulatory pathways between gut microbiota and Peyer's patches remain unclear. Therefore, the aim of this study was to investigate the therapeutic potential of fucoidan in a dextran sulfate sodium (DSS)-induced mouse model of inflammatory bowel disease (IBD) by examining the effects on gut microbiota and the underlying anti-inflammatory mechanisms. Purified fucoidan, which upon characterization revealed structural fragments comprising →3)-β-D-Galp-(1→, →4)-α-L-Fucp-(1→, and →3)-α-L-Fucp-(1→ residues with a sulfation at position C2 was used. Treatment of the mice with fucoidan significantly alleviated the symptoms of IBD and restored the diversity of gut microbiota by enhancing the abundance of Bacteroidetes and reducing the proportion of Firmicutes. The administration of fucoidan also elevated levels of short-chain fatty acids while reducing the levels of pro-inflammatory cytokines, including interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ. Most importantly, fucoidan attenuated the expression of integrin α4β7/MAdCAM-1 and CCL25/CCR9, which are involved in homing intestinal lymphocytes within Peyer's patches. These findings indicate that fucoidan is a promising gut microbiota modulator and an anti-inflammatory agent for IBD.

Keywords:  Fucoidan; Gut microbiota; Inflammatory bowel disease; Peyer's patch

DOI:  https://doi.org/10.1016/j.carbpol.2024.122421
PNAS Nexus. 2024 Jul;3(7): pgae265

Tyrosine phenol-lyase inhibitor quercetin reduces fecal phenol levels in mice.

Takuma Kobayashi, Shiori Oishi, Misaki Matsui, Kodai Hara, Hiroshi Hashimoto, Kenji Watanabe, Yasukiyo Yoshioka, Noriyuki Miyoshi.

  Tyrosine phenol-lyase (TPL), which is expressed in intestinal bacteria, catalyzes the formation of phenol from the substrate L-Tyr. Bacterial metabolite phenol and the sulfate conjugate (phenyl sulfate) are known as a type of uremic toxins, some of which exert cytotoxicity. Therefore, pathologically elevated phenol and phenyl sulfate levels are strongly implicated in the etiology and outcome of uremia. In this study, we explored the inhibitory effects of dietary polyphenols on TPL-catalyzed phenol production using a TPL activity assay. Quercetin, one of the most popular polyphenols, exhibited the strongest inhibitory activity (Ki = 19.9 µM). Quercetin competitively inhibited TPL, and its activity was stronger than that of a known TPL inhibitor (Tyr analog; 2-aza-Tyr, Ki = 42.0 µM). Additionally, quercetin significantly inhibited phenol production in TPL-expressing bacterial cultures (Morganella morganii and Citrobacter koseri) and Tyr-rich (5%) diet-fed C57BL/6J mouse feces. Our findings suggest that quercetin is the most promising polyphenol for reducing phenol levels. Because quercetin has a low gastrointestinal absorption rate, TPL inhibition in the intestinal tract by quercetin may be an effective strategy for treating uremia.

Keywords:  intestinal bacteria; phenol; quercetin; tyrosine phenol-lyase inhibitor; uremic toxin

DOI:  https://doi.org/10.1093/pnasnexus/pgae265
Sci Total Environ. 2024 Jul 20. pii: S0048-9697(24)05072-1. [Epub ahead of print]948 174922

Occurrence of bisphenol analogues and their conjugated metabolites in foodstuff.

Jianli Qu, Ruyue Guo, Lin Liu, Fangfang Ren, Hangbiao Jin.

  Bisphenol analogues (BPs) are prevalent in diverse foodstuff samples worldwide. However, the occurrence of conjugated bisphenol A (BPA) and bisphenol S (BPS) metabolites in foodstuff remains poorly understood. This study analyzed eight BPs, and four conjugated BPA and BPS metabolites, in three animal-derived foodstuff and five plant-derived foodstuff samples from China. Results showed that fish foodstuff (9.7 ng/g ww) contained the highest mean concentration of BPA, followed by rice (5.1 ng/g ww) and beans foodstuff (3.6 ng/g ww). BPA-sulfate had higher mean concentrations than BPA-glucuronide in different foodstuff categories, except that in eggs foodstuff (p < 0.05). Compared with other foodstuff items, fish (3.4 ng/g ww) and vegetable (1.6 ng/g ww) foodstuff samples exhibited comparatively higher mean concentrations of BPS. Mean concentrations of BPS-sulfate were consistently higher than BPS-glucuronide in vegetables, meats, and fish foodstuff (p < 0.05). BPA contributed the major total dietary intake (DI) of BPs, with the mean DI of 435 ng/kg bw/day for women and 374 ng/kg bw/day for men, respectively. To our knowledge, this study is the first to investigate the occurrence of conjugated BPA and BPS metabolites in foodstuff, which enhances our comprehension of the origins of these conjugated metabolites in the human body.

Keywords:  BPA; BPA-S; BPS-S; Dietary intake; Foodstuff

DOI:  https://doi.org/10.1016/j.scitotenv.2024.174922