bims-supasi 2024-07-14 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2024–07–14
eightteen papers selected by
Jonathan Wolf Mueller, University of Birmingham

Steroid sulfatase and sulfotransferases in the estrogen and androgen action of gynecological cancers: current status and perspectives.
Chondroitin sulfate glycan sulfation patterns influence histochemical labeling of perineuronal nets: a comparative study of interregional distribution in human and mouse brain.
Structural characteristics of sulfated xylogalactomannan isolated from Caulerpa okamurae and its anticoagulant activity.
F-actin and heparan sulfate proteoglycan distribution in female mosquito salivary glands and ducts.
Real-Time Study of the Specific Interactions of Lactoferrin with Mimicked Heparan Sulfate Meshes Using Ordered Porous Layer Interferometry.
Alterations in the Structure, Composition, and Organization of Galactosaminoglycan-Containing Proteoglycans and Collagen Correspond to the Progressive Stages of Dupuytren's Disease.
Cell-recruited microspheres for OA treatment by dual-modulating inflammatory and chondrocyte metabolism.
Characterising glycosaminoglycans in human breastmilk and their potential role in infant health.
Amphiphilic Heparinoids as Potent Antiviral Agents against SARS-CoV-2.
Impact of Composition and Autoclave Sterilization on the Mechanical and Biological Properties of ECM-Mimicking Cryogels.
Genome-Wide Identification of the Sulfate Transporters Gene Family in Blueberry (Vaccinium spp.) and Its Response to Ericoid Mycorrhizal Fungi.
Adipose Tissue Sex Steroids in Postmenopausal Women with and without Menopausal Hormone Therapy.
Therapeutic effect of San Bi Tang combined with glucosamine sulfate capsules in cold-dampness-type knee osteoarthritis.
Identification of heparin interaction sites on thrombin-activatable fibrinolysis inhibitor that modulate plasmin-mediated activation, thermal stability, and antifibrinolytic potential.
Steroid metabolism and hormonal dynamics in normal and malignant ovaries.
Bioceramic-mediated chondrocyte hypertrophy promotes calcified cartilage formation for rabbit osteochondral defect repair.
Sex- and age-specific reference intervals of 16 steroid metabolites quantified simultaneously by LC-MS/MS in sera from 2458 healthy subjects aged 0 to 77 years.
Uremic toxin indoxyl sulfate induces trained immunity via the AhR-dependent arachidonic acid pathway in end-stage renal disease (ESRD).

Essays Biochem. 2024 Jul 12. pii: EBC20230096. [Epub ahead of print]

Steroid sulfatase and sulfotransferases in the estrogen and androgen action of gynecological cancers: current status and perspectives.

Tea Lanišnik Rižner, Marija Gjorgoska.

  Sulfatase (STS) and sulfotransferases (SULT) have important role in the biosynthesis and action of steroid hormones. STS catalyzes the hydrolysis of estrone-sulfate (E1-S) and dehydroepiandrosterone-sulfate (DHEA-S), while sulfotransferases catalyze the reverse reaction and require 3-phosphoadenosine-5-phosphosulfate as a sulfate donor. These enzymes control the concentration of active estrogens and androgens in peripheral tissues. Aberant expression of STS and SULT genes has been found in both, benign hormone-dependent diseases and hormone-dependent cancers. The aim of this review is to present the current knowledge on the role of STS and SULT in gynecological cancers, endometrial (EC) and ovarian cancer (OC). EC is the most common and OC the most lethal gynecological cancer. These cancers primarily affect postmenopausal women and therefore rely on the local production of steroid hormones from inactive precursors, either DHEA-S or E1-S. Following cellular uptake by organic anion transporting polypeptides (OATP) or organic anion transporters (OAT), STS and SULT regulate the formation of active estrogens and androgens, thus disturbed balance between STS and SULT can contribute to the onset and progression of cancer. The importance of these enzymes in peripheral estrogen biosynthesis has long been recognized, and this review provides new data on the important role of STS and SULT in the formation and action of androgens, their regulation and inhibition, and their potential as prognostic biomarkers.

Keywords:  androgens; estrogens; intracrinology; sulfatase; sulfotransferase

DOI:  https://doi.org/10.1042/EBC20230096
Glycobiology. 2024 Jul 12. pii: cwae049. [Epub ahead of print]

Chondroitin sulfate glycan sulfation patterns influence histochemical labeling of perineuronal nets: a comparative study of interregional distribution in human and mouse brain.

Claudia Belliveau, Stéphanie Théberge, Stefanie Netto, Reza Rahimian, Gohar Fakhfouri, Clémentine Hosdey, Maria Antonietta Davoli, Aarun Hendrickson, Kathryn Hao, Bruno Giros, Gustavo Turecki, Kimberly M Alonge, Naguib Mechawar.

  Perineuronal nets (PNNs) are a condensed subtype of extracellular matrix that form a net-like coverings around certain neurons in the brain. PNNs are primarily composed of chondroitin sulfate (CS) proteoglycans from the lectican family that consist of CS-glycosaminoglycan (CS-GAG) side chains attached to a core protein. CS disaccharides can exist in various isoforms with different sulfation patterns. Literature suggests that CS disaccharide sulfation patterns can influence the function of PNNs as well as their labeling. This study was conducted to characterize such interregional CS disaccharide sulfation pattern differences in adult human (n = 81) and mouse (n = 19) brains. Liquid chromatography tandem mass spectrometry was used to quantify five different CS disaccharide sulfation patterns, which were then compared to immunolabeling of PNNs using Wisteria Floribunda Lectin (WFL) to identify CS-GAGs and anti-aggrecan to identify CS proteoglycans. In healthy brains, significant regional and species-specific differences in CS disaccharide sulfation and single versus double-labeling pattern were identified. A secondary analysis to investigate how early-life stress (ELS) impacts these PNN features discovered that although ELS increases WFL+ PNN density, the CS-GAG sulfation code and single versus double PNN-labeling distributions remained unaffected in both species. These results underscore PNN complexity in traditional research, emphasizing the need to consider their heterogeneity in future experiments.

Keywords:  adverse childhood experiences; chondroitin sulfate; extracellular matrix; human; mouse

DOI:  https://doi.org/10.1093/glycob/cwae049
Int J Biol Macromol. 2024 Jul 08. pii: S0141-8130(24)04548-3. [Epub ahead of print]275(Pt 2): 133743

Structural characteristics of sulfated xylogalactomannan isolated from Caulerpa okamurae and its anticoagulant activity.

Yanlin Zhu, Sitong Wu, Feng Guo, Zhe Dong, Yan Chen, Yin Chen.

  Due to wonderful taste, rich nutrition and biological functions, many marine green algae in the genus Caulerpa have been recently developed as candidates for green caviar. A novel water-soluble sulfated xylogalactomannan CO-0-1 was obtained from the green algae Caulerpa okamurae. CO-0-1 was mainly composed of mannose (Man), galactose (Gal), and xylose (Xyl) at the ratio of 4.4:4.0:1.4 with the molecular weight at 470 kDa and the sulfate content at 12.78 %. The sulfated xylogalactomannan had Man at the backbone with →4)-β-D-Manp-(1→ and →2)-β-D-Manp-(1→ as the main chain and branches at O-3 position. The side chains contained →3)-β-D-Galp-(1→ and minor →2)-β-D-Xylp(1→. The sulfate groups only distributed at the side chains and at O-6 position of →3)-β-D-Galp-(1→ and O-4 position of (1→2)-β-D-Xylp. The anticoagulant activity indicated that CO-0-1 displayed intrinsic anticoagulant and specific anti-thrombin activities. The investigation expanded the utilization and development scene and scope of the green algae Caulerpa okamurae.

Keywords:  Anticoagulant activity; Caulerpa okamurae sulfated polysaccharides; Structure determination

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.133743
J Med Entomol. 2024 Jul 12. pii: tjae060. [Epub ahead of print]

F-actin and heparan sulfate proteoglycan distribution in female mosquito salivary glands and ducts.

Doria Fassbinder Bowers, Kristin Ann Ciano.

  Directly involved in the "suck-and-spit" physiology, female mosquito salivary glands (SGs) primarily imbibe blood for egg development and release anticoagulants to keep blood flowing. Indirectly involved, mosquitoes can uptake arboviruses during blood feeding from a viremic host. This research examined the presence of the filamentous cytoplasmic contractile protein (F-actin) and heparan sulfate proteoglycan (HSPG), in the female mosquito SGs. Immunofluorescent antibody labeling of actin molecules or HSPG combined with anatomy suggests that F-actin forms a network in the SG lobe parenchymal cells attached to intralobar ducts by HSPG. In addition, F-actin twists around intralobar SG ducts in a beaded manner, altogether involved in the expulsion of SG secretions. This arrangement in female Aedes aegypti SGs, suggests that F-actin structures are integrally involved in transmitting infectious agents into hosts.

Keywords:  Sindbis virus; arbovirus; blood feeding; immunohistochemistry

DOI:  https://doi.org/10.1093/jme/tjae060
Anal Chem. 2024 Jul 11.

Real-Time Study of the Specific Interactions of Lactoferrin with Mimicked Heparan Sulfate Meshes Using Ordered Porous Layer Interferometry.

Yu Zhang, Ning Ma, Lu Wang, Liming Liu, Tianze Wang, Hao Liu, Weiping Qian.

Heparan sulfate (HS) meshes within the glycocalyx on cell surfaces have protein recognition ability and have been crucial for gaining insights into vital bioprocesses, such as viral infection, cancer development, and inflammation. The protein recognition ability is determined by the mesh property and compositions of HS, although little attention has been paid to the effect of the mesh property on the recognition. An in-depth specificity study of protein-HS-mesh recognition is essential to illustrate related biological functions. Here, ordered porous layer interferometry is applied to study the interaction behavior between mimicked HS meshes and lactoferrin (LF). Our work aimed at mimicking HS meshes with heparin, a widely used substitute of HS, and analyzing the specific LF-heparin-mesh interaction mechanism by inhibiting the nonspecific interaction in a blended sample. We found that the counterion release-based electrostatic interaction is dominant in the specific LF-heparin-mesh recognition. Furthermore, we detail the contributions of nonspecific and specific interactions to the recognition. We illustrate that the concentrated charge distribution of the proteins appears to be primarily related to this robust, specific recognition.

DOI: https://doi.org/10.1021/acs.analchem.4c01808
Int J Mol Sci. 2024 Jun 29. pii: 7192. [Epub ahead of print]25(13):

Alterations in the Structure, Composition, and Organization of Galactosaminoglycan-Containing Proteoglycans and Collagen Correspond to the Progressive Stages of Dupuytren's Disease.

Luiz Guilherme S Lenzi, João Baptista Gomes Dos Santos, Renan P Cavalheiro, Aline Mendes, Elsa Y Kobayashi, Helena B Nader, Flavio Faloppa.

  Dupuytren's disease (DD) is a prevalent fibroproliferative disorder of the hand, shaped by genetic, epigenetic, and environmental influences. The extracellular matrix (ECM) is a complex assembly of diverse macromolecules. Alterations in the ECM's content, structure and organization can impact both normal physiological functions and pathological conditions. This study explored the content and organization of glycosaminoglycans, proteoglycans, and collagen in the ECM of patients at various stages of DD, assessing their potential as prognostic indicators. This research reveals, for the first time, relevant changes in the complexity of chondroitin/dermatan sulfate structures, specifically an increase of disaccharides containing iduronic acid residues covalently linked to either N-acetylgalactosamine 6-O-sulfated or N-acetylgalactosamine 4-O-sulfated, correlating with the disease's severity. Additionally, we noted an increase in versican expression, a high molecular weight proteoglycan, across stages I to IV, while decorin, a small leucine-rich proteoglycan, significantly diminishes as DD progresses, both confirmed by mRNA analysis and protein detection via confocal microscopy. Coherent anti-Stokes Raman scattering (CARS) microscopy further demonstrated that collagen fibril architecture in DD varies importantly with disease stages. Moreover, the urinary excretion of both hyaluronic and sulfated glycosaminoglycans markedly decreased among DD patients.Our findings indicate that specific proteoglycans with galactosaminoglycan chains and collagen arrangements could serve as biomarkers for DD progression. The reduction in glycosaminoglycan excretion suggests a systemic manifestation of the disease.

Keywords:  chondroitin and dermatan sulfates; collagen; decorin; extracellular matrix; fascia palmar; hyaluronic acid; proteoglycans; sulfated glycosaminoglycans; versican

DOI:  https://doi.org/10.3390/ijms25137192
Mater Today Bio. 2024 Aug;27 101127

Cell-recruited microspheres for OA treatment by dual-modulating inflammatory and chondrocyte metabolism.

Yun Zhou, Xu He, Wen Zhang, Weiguo Zhang, Huan Zhao, Xichao Zhou, Qiaoli Gu, Hao Shen, Huilin Yang, Xingzhi Liu, Lixin Huang, Qin Shi.

  Osteoarthritis (OA) is a degenerative disease potentially exacerbated due to inflammation, cartilage degeneration, and increased friction. Both mesenchymal stem cells (MSCs) and pro-inflammatory macrophages play important roles in OA. A promising approach to treating OA is to modify multi-functional hydrogel microspheres to target the OA microenvironment and structure. Arginyl-glycyl-aspartic acid (RGD) is a peptide widely used in bioengineering owing to its cell adhesion properties, which can recruit BMSCs and macrophages. We developed TLC-R, a microsphere loaded with TGF-β1-containing liposomes. The recruitment effect of TLC-R on macrophages and BMSCs was verified by in vitro experiments, along with its function of promoting chondrogenic differentiation of BMSCs. And we evaluated the effect of TLC-R in balancing OA metabolism in vitro and in vivo. When TLC-R was co-cultured with BMSCs and lipopolysaccharide (LPS)-treated macrophages, it showed the ability to recruit both cells in substantial numbers. As the microspheres degraded, TGF-β1 and chondroitin sulfate (ChS) were released to promote chondrogenic differentiation of the recruited BMSCs, modulate chondrocyte metabolism and inhibit inflammation induced by the macrophages. Furthermore, in vivo analysis showed that TLC-R restored the narrowed space, reduced osteophyte volume, and improved cartilage metabolic homeostasis in OA rats. Altogether, TLC-R provides a comprehensive and novel solution for OA treatment by dual-modulating inflammatory and chondrocyte metabolism.

Keywords:  Chondroitin sulfate; Hydrogel microspheres; Osteoarthritis; RGD; Transforming growth factor-β1

DOI:  https://doi.org/10.1016/j.mtbio.2024.101127
Microb Cell. 2024 ;11 221-234

Characterising glycosaminoglycans in human breastmilk and their potential role in infant health.

Melissa Greenwood, Patricia Murciano-Martínez, Janet Berrington, Sabine L Flitsch, Sean Austin, Christopher Stewart.

  Human breastmilk is composed of many well researched bioactive components crucial for infant nutrition and priming of the neonatal microbiome and immune system. Understanding these components gives us crucial insight to the health and wellbeing of infants. Research surrounding glycosaminoglycans (GAGs) previously focused on those produced endogenously; however, recent efforts have shifted to understanding GAGs in human breastmilk. The structural complexity of GAGs makes detection and analysis complicated therefore, research is time consuming and limited to highly specialised teams experienced in carbohydrate analysis. In breastmilk, GAGs are present in varying quantities in four forms; chondroitin sulphate, heparin/heparan sulphate, dermatan sulphate and hyaluronic acid, and are hypothesised to behave similar to other bioactive components with suspected roles in pathogen defense and proliferation of beneficial gut bacteria. Chondroitin sulphate and heparin, being the most abundant, are expected to have the most impact on infant health. Their decreasing concentration over lactation further indicates their role and potential importance during early life.

Keywords:  breastmilk; chondroitin sulphate; glycosaminoglycan; heparan sulphate; heparin; microbiome

DOI:  https://doi.org/10.15698/mic2024.07.827
J Med Chem. 2024 Jul 12.

Amphiphilic Heparinoids as Potent Antiviral Agents against SARS-CoV-2.

Mohit Chhabra, Chethan D Shanthamurthy, Nanjudaswamy Vijendra Kumar, Sandhya Mardhekar, Sharath S Vishweshwara, Norbert Wimmer, Naphak Modhiran, Daniel Watterson, Alberto A Amarilla, Jonathan S Cha, James R Beckett, James J De Voss, Yasmin Kayal, Israel Vlodavsky, Lauren R Dorsett, Raymond A A Smith, Neha S Gandhi, Raghavendra Kikkeri, Vito Ferro.

Herein, we report the synthesis and biological evaluation of a novel series of heparinoid amphiphiles as inhibitors of heparanase and SARS-CoV-2. By employing a tailor-made synthetic strategy, a library of highly sulfated homo-oligosaccharides bearing d-glucose or a C5-epimer (i.e., l-idose or l-iduronic acid) conjugated with various lipophilic groups was synthesized and investigated for antiviral activity. Sulfated higher oligosaccharides of d-glucose or l-idose with lipophilic aglycones displayed potent anti-SARS-CoV-2 and antiheparanse activity, similar to or better than pixatimod (PG545), and were more potent than their isosteric l-iduronic acid congeners. Lipophilic groups such as cholestanol and C18-aliphatic substitution are more advantageous than functional group appended lipophilic moieties. These findings confirm that fine-tuning of higher oligosaccharides, degree of sulfation, and lipophilic groups can yield compounds with potent anti-SARS-CoV-2 activity.

DOI: https://doi.org/10.1021/acs.jmedchem.4c00487
Polymers (Basel). 2024 Jul 07. pii: 1939. [Epub ahead of print]16(13):

Impact of Composition and Autoclave Sterilization on the Mechanical and Biological Properties of ECM-Mimicking Cryogels.

Laura Di Muzio, Susi Zara, Amelia Cataldi, Claudia Sergi, Vito Cosimo Carriero, Barbara Bigi, Simone Carradori, Jacopo Tirillò, Stefania Petralito, Maria Antonietta Casadei, Patrizia Paolicelli.

  Cryogels represent a valid strategy as scaffolds for tissue engineering. In order to adequately support adhesion and proliferation of anchorage-dependent cells, different polymers need to be combined within the same scaffold trying to mimic the complex features of a natural extracellular matrix (ECM). For this reason, in this work, gelatin (Gel) and chondroitin sulfate (CS), both functionalized with methacrylic groups to produce CSMA and GelMA derivatives, were selected to prepare cryogel networks. Both homopolymer and heteropolymer cryogels were produced, via radical crosslinking reactions carried out at -12 °C for 2 h. All the scaffolds were characterized for their mechanical, swelling and morphological properties, before and after autoclave sterilization. Moreover, they were evaluated for their biocompatibility and ability to support the adhesion of human gingival fibroblasts and tenocytes. GelMA-based homopolymer networks better withstood the autoclave sterilization process, compared to CSMA cryogels. Indeed, GelMA cryogels showed a decrease in stiffness of approximately 30%, whereas CSMA cryogels of approximately 80%. When GelMA and CSMA were blended in the same network, an intermediate outcome was observed. However, the hybrid scaffolds showed a general worsening of the biological performance. Indeed, despite their ability to withstand autoclave sterilization with limited modification of the mechanical and morphological properties, the hybrid cryogels exhibited poor cell adhesion and high LDH leakage. Therefore, not only do network components need to be properly selected, but also their combination and ability to withstand effective sterilization process should be carefully evaluated for the development of efficient scaffolds designed for tissue engineering purposes.

Keywords:  cell adhesion; chondroitin sulfate; cryogels; gelatin; macroporous networks; scaffolds; tissue engineering

DOI:  https://doi.org/10.3390/polym16131939
Int J Mol Sci. 2024 Jun 26. pii: 6980. [Epub ahead of print]25(13):

Genome-Wide Identification of the Sulfate Transporters Gene Family in Blueberry (Vaccinium spp.) and Its Response to Ericoid Mycorrhizal Fungi.

Mei Dong, Jiawei He, Xiaoxuan Tang, Siwen Liu, Jinjie Xing, Xuyang Chen, Li Chen, Yadong Li, Haiyue Sun.

  Sulfur metabolism plays a major role in plant growth and development, environmental adaptation, and material synthesis, and the sulfate transporters are the beginning of sulfur metabolism. We identified 37 potential VcSULTR genes in the blueberry genome, encoding peptides with 534 to 766 amino acids. The genes were grouped into four subfamilies in an evolutionary analysis. The 37 putative VcSULTR proteins ranged in size from 60.03 to 83.87 kDa. These proteins were predicted to be hydrophobic and mostly localize to the plasma membrane. The VcSULTR genes were distributed on 30 chromosomes; VcSULTR3;5b and VcSULTR3;5c were the only tandemly repeated genes. The VcSULTR promoters contained cis-acting elements related to the fungal symbiosis and stress responses. The transcript levels of the VcSULTRs differed among blueberry organs and changed in response to ericoid mycorrhizal fungi and sulfate treatments. A subcellular localization analysis showed that VcSULTR2;1c localized to, and functioned in, the plasma membrane and chloroplast. The virus-induced gene knock-down of VcSULTR2;1c resulted in a significantly decreased endogenous sulfate content, and an up-regulation of genes encoding key enzymes in sulfur metabolism (VcATPS2 and VcSiR1). These findings enhance our understanding of mycorrhizal-fungi-mediated sulfate transport in blueberry, and lay the foundation for further research on blueberry-mycorrhizal symbiosis.

Keywords:  Vaccinium; blueberry; genome-wide identification; mycorrhiza; sulfate transporters

DOI:  https://doi.org/10.3390/ijms25136980
J Clin Endocrinol Metab. 2024 Jul 10. pii: dgae458. [Epub ahead of print]

Adipose Tissue Sex Steroids in Postmenopausal Women with and without Menopausal Hormone Therapy.

Natalia Hetemäki, Alexandra Robciuc, Veera Vihma, Mikko Haanpää, Esa Hämäläinen, Matti J Tikkanen, Tomi S Mikkola, Hanna Savolainen-Peltonen.

   CONTEXT: The decrease in serum estrogens after menopause is associated with a shift from a gynoid to an android adipose tissue (AT) distribution. Menopausal hormone therapy (HT) mitigates this change and accompanying metabolic dysfunction, but its effects on AT sex steroid metabolism have not been characterized.
OBJECTIVE: We studied effects of HT on subcutaneous and visceral AT estrogen and androgen concentrations and metabolism in postmenopausal women.
DESIGN, SETTING, PATIENTS, AND INTERVENTIONS: Serum and subcutaneous and visceral AT from 63 postmenopausal women with (n=50) and without (n=13) per oral HT were analyzed for estrone, estradiol, progesterone, testosterone, androstenedione, dehydroepiandrosterone, and serum estrone sulfate using liquid chromatography-tandem mass spectrometry. Steroid sulfatase activity was measured using radiolabeled precursors. mRNA expression of genes encoding sex steroid-metabolizing enzymes and receptors was performed using real-time reverse transcription quantitative polymerase chain reaction.
RESULTS: HT users had 4- to 7-fold higher concentrations of estrone and estradiol in subcutaneous and visceral AT, and 30% lower testosterone in visceral AT compared to non-users. Estrogen-to-androgen ratios were 4- to 12-fold higher in AT of users compared to non-users of HT. In visceral AT, estrogen-to-androgen ratios increased with HT estradiol dose. AT to serum ratios of estrone and estradiol remained high in HT users.
CONCLUSIONS: Higher local estrogen to androgen ratios and high AT to serum ratios of estrogen concentrations in HT users suggest that HT may significantly influence intracrine sex steroid metabolism in AT, and these local changes could be involved in the preventive effect of HT on menopause-associated abdominal adiposity.

Keywords:  Adipose tissue; menopausal hormone therapy; postmenopausal; sex steroids

DOI:  https://doi.org/10.1210/clinem/dgae458
World J Clin Cases. 2024 Jul 06. 12(19): 3854-3865

Therapeutic effect of San Bi Tang combined with glucosamine sulfate capsules in cold-dampness-type knee osteoarthritis.

Hui-Ying Ni, Yao-Ping Zhang, Xiao-Feng Zhang.

   BACKGROUND: Cold-dampness-type knee osteoarthritis is a common middle-aged and elderly disease, but its pathogenesis is not fully understood, and its clinical treatment has limitations. Glucosamine sulfate capsules are commonly used for treating arthritis, and San Bi Tang is a classic formula of traditional Chinese medicine (TCM) that has the effects of warming yang, dispelling dampness, relaxing muscles, and activating collaterals. This research hypothesized that the combination of modified San Bi Tang and glucosamine sulfate capsules could enhance the clinical efficacy of treating cold-dampness-type knee osteoarthritis through complementary effects.
AIM: To analyze the clinical efficacy of San Bi Tang combined with glucosamine sulfate capsules when treating cold-dampness-type knee osteoarthritis.
METHODS: A total of 110 patients with cold-dampness-type knee osteoarthritis were selected as research subjects and randomly divided into a control group and an experimental group of 55 cases each. The control group received only treatment with glucosamine sulfate capsules, while the experimental group received additional treatment with modified San Bi Tang for a duration of 5 wk. The patients' knee joint functions, liver and kidney function indicators, adverse reactions, and vital signs were evaluated and analyzed using SPSS 26.0 software.
RESULTS: Before treatment, the two groups' genders, ages, and scores were not significantly different, indicating comparability. Both groups' scores improved after treatment, which could indicate pain and knee joint function improvement, but the test group had better scores. The TCM-specific symptoms and the clinical efficacy of the treatment in the test group were higher. Before and after treatment, there were no abnormalities in the patients' liver and kidney function indicators.
CONCLUSION: The combination of modified San Bi Tang and glucosamine sulfate capsules is superior to treatment with sulfated glucosamine alone and has high safety.

Keywords:  Aggravated by cold; Combination therapy; Glucosamine sulfate capsules; Knee osteoarthritis; San Bi Tang

DOI:  https://doi.org/10.12998/wjcc.v12.i19.3854
Res Pract Thromb Haemost. 2024 May;8(4): 102459

Identification of heparin interaction sites on thrombin-activatable fibrinolysis inhibitor that modulate plasmin-mediated activation, thermal stability, and antifibrinolytic potential.

Tanya T Marar, Michael B Boffa.

   Background: Thrombin-activatable fibrinolysis inhibitor (TAFI) is a plasma zymogen that provides a molecular link between coagulation and fibrinolysis. Studies have shown that the presence of glycosaminoglycans accelerates TAFI activation by plasmin and stabilizes activated TAFI (TAFIa).
Objectives: We aimed to define the elements of TAFI structure that allow these effects.
Methods: Based on crystallographic studies and homology to heparin-binding proteins, we performed mutagenesis of surface-exposed charged residues on TAFI that putatively constitute heparin-binding sites. We determined heparin binding, kinetics of activation by plasmin in the presence or absence of heparin, thermal stability, and antifibrinolytic potential of each variant.
Results: Mutagenesis of Lys211 and Lys212 did not impair heparin binding but affected the ability of TAFI to be activated by plasmin. Mutagenesis of Lys306 and His308 did not impair heparin binding, but mutation of His308 had a severe negative effect on TAFI/TAFIa function. Mutation of Arg320 and Lys324 in combination markedly decreased heparin binding but had no effect on heparin-mediated acceleration of TAFI activation by plasmin while somewhat decreasing TAFIa stabilization by heparin. Mutagenesis of Lys327 and Arg330 decreased (but did not eliminate) heparin binding while decreasing the ability of heparin to accelerate plasmin-mediated TAFI activation, stabilize TAFIa, and increase the antifibrinolytic ability of TAFIa. A quadruple mutant of Arg320, Lys324, Lys327, and Arg330 completely lost heparin-binding ability and stabilization of the enzyme by heparin.
Conclusion: Basic residues in the dynamic flap of TAFIa define a functionally relevant heparin-binding site, but additional heparin-binding sites may be present on TAFI.

Keywords:  carboxypeptidase B2; fibrinolysis; glycosaminoglycans; heparin; kinetics; mutagenesis; plasmin

DOI:  https://doi.org/10.1016/j.rpth.2024.102459
Essays Biochem. 2024 Jul 12. pii: EBC20240028. [Epub ahead of print]

Steroid metabolism and hormonal dynamics in normal and malignant ovaries.

Lucy I Beevors, Sudha Sundar, Paul A Foster.

  The ovaries are key steroid hormone production sites in post-pubertal females. However, current research on steroidogenic enzymes, endogenous hormone concentrations and their effects on healthy ovarian function and malignant development is limited. Here, we discuss the importance of steroid enzymes in normal and malignant ovaries, alongside hormone concentrations, receptor expression and action. Key enzymes include STS, 3β-HSD2, HSD17B1, ARK1C3, and aromatase, which influence ovarian steroidal action. Both androgen and oestrogen action, via their facilitating enzyme, drives ovarian follicle activation, development and maturation in healthy ovarian tissue. In ovarian cancer, some data suggest STS and oestrogen receptor α may be linked to aggressive forms, while various oestrogen-responsive factors may be involved in ovarian cancer metastasis. In contrast, androgen receptor expression and action vary across ovarian cancer subtypes. For future studies investigating steroidogenesis and steroidal activity in ovarian cancer, it is necessary to differentiate between disease subtypes for a comprehensive understanding.

Keywords:  Ovarian Cancer; Ovary; Steroid Metabolism; Steroid Sulfatase; androgens; estrogens

DOI:  https://doi.org/10.1042/EBC20240028
Bioact Mater. 2024 Oct;40 306-317

Bioceramic-mediated chondrocyte hypertrophy promotes calcified cartilage formation for rabbit osteochondral defect repair.

Rachel H Koh, Junhee Kim, Jeong-Uk Kim, Seunghyun L Kim, Arun Kumar Rajendran, Seunghun S Lee, Heesoo Lee, Joo Hyun Kim, Ji Hoon Jeong, Yongsung Hwang, Jong Woo Bae, Nathaniel S Hwang.

  Osteochondral tissue is a highly specialized and complex tissue composed of articular cartilage and subchondral bone that are separated by a calcified cartilage interface. Multilayered or gradient scaffolds, often in conjunction with stem cells and growth factors, have been developed to mimic the respective layers for osteochondral defect repair. In this study, we designed a hyaline cartilage-hypertrophic cartilage bilayer graft (RGD/RGDW) with chondrocytes. Previously, we demonstrated that RGD peptide-modified chondroitin sulfate cryogel (RGD group) is chondro-conductive and capable of hyaline cartilage formation. Here, we incorporated whitlockite (WH), a Mg2+-containing calcium phosphate, into RGD cryogel (RGDW group) to induce chondrocyte hypertrophy and form collagen X-rich hypertrophic cartilage. This is the first study to use WH to produce hypertrophic cartilage. Chondrocytes-laden RGDW cryogel exhibited significantly upregulated expression of hypertrophy markers in vitro and formed ectopic hypertrophic cartilage in vivo, which mineralized into calcified cartilage in bone microenvironment. Subsequently, RGD cryogel and RGDW cryogel were combined into bilayer (RGD/RGDW group) and implanted into rabbit osteochondral defect, where RGD layer supports hyaline cartilage regeneration and bioceramic-containing RGDW layer promotes calcified cartilage formation. While the RGD group (monolayer) formed hyaline-like neotissue that extends into the subchondral bone, the RGD/RGDW group (bilayer) regenerated hyaline cartilage tissue confined to its respective layer and promoted osseointegration for integrative defect repair.

Keywords:  Bilayer scaffold; Bioceramic; Chondrocyte hypertrophy; Osteochondral defect repair; Whitlockite

DOI:  https://doi.org/10.1016/j.bioactmat.2024.06.018
Clin Chim Acta. 2024 Jul 06. pii: S0009-8981(24)02104-1. [Epub ahead of print]562 119852

Sex- and age-specific reference intervals of 16 steroid metabolites quantified simultaneously by LC-MS/MS in sera from 2458 healthy subjects aged 0 to 77 years.

Hanne Frederiksen, Trine Holm Johannsen, Stine Ehlern Andersen, Jørgen Holm Petersen, Alexander Siegfried Busch, Marie Lindhardt Ljubicic, Margit Bistrup Fischer, Emmie N Upners, Casper P Hagen, Katharina M Main, Lise Aksglaede, Niels Jørgensen, Line Lund Kårhus, Allan Linneberg, Anna-Maria Andersson, Christa E Flück, Anders Juul.

   BACKGROUND: Reference intervals covering the whole life span for all the metabolites in the steroid hormone biosynthesis quantified by sensitive and robust analytical methods are sparse or not existing.
OBJECTIVE: To develop a state-of-the-art LC-MS/MS method for simultaneous quantification of multiple steroid metabolites and to establish detailed sex- and age-specific reference intervals for 16 steroid metabolites.
MATERIALS AND METHOD: An isotope diluted LC-MS/MS method was developed for simultaneous quantitation of 16 steroid hormones. Serum samples from cross-sectional cohorts of healthy infants, children, adolescents, and adults aged 0.17 months to 77 years (n = 2458) were analysed.
RESULTS: With this novel, specific, and sensitive LC-MS/MS method, it was possible to quantify progesterone, 17-hydroxypregnenolone, 17-hydroxyprogesterone, dehydroepiandrosterone sulfate, androstenedione, testosterone, dihydrotestosterone, 11-deoxycorticosterone, corticosterone, 11-deoxycortisol, cortisol, and cortisone in ≥90 % of the samples, while estrone sulfate, aldosterone and dehydroepiandrosterone were quantified in 77 %, 75 % and 60 % of the samples, respectively. 21-deoxycortisol was only detectable in 2.5 % of samples from healthy subjects. Sex- and age-dependent fluctuations observed in minipuberty, puberty and adulthood including the menopausal transition were modelled. This enabled us to establish valid reference intervals from birth to late adult life for both males and females.
CONCLUSION: Detailed sex- and age-specific reference intervals of multiple, simultaneously quantified steroid metabolites by a novel and specific LC-MS/MS method provides a valuable tool for clinical practice and for future research.

Keywords:  Infant; LC-MS/MS; Puberty; Reference interval; Sex difference; Steroid hormone

DOI:  https://doi.org/10.1016/j.cca.2024.119852
Elife. 2024 Jul 09. pii: RP87316. [Epub ahead of print]12

Uremic toxin indoxyl sulfate induces trained immunity via the AhR-dependent arachidonic acid pathway in end-stage renal disease (ESRD).

Hee Young Kim, Yeon Jun Kang, Dong Hyun Kim, Jiyeon Jang, Su Jeong Lee, Gwanghun Kim, Hee Byung Koh, Ye Eun Ko, Hyun Mu Shin, Hajeong Lee, Tae-Hyun Yoo, Won-Woo Lee.

  Trained immunity is the long-term functional reprogramming of innate immune cells, which results in altered responses toward a secondary challenge. Despite indoxyl sulfate (IS) being a potent stimulus associated with chronic kidney disease (CKD)-related inflammation, its impact on trained immunity has not been explored. Here, we demonstrate that IS induces trained immunity in monocytes via epigenetic and metabolic reprogramming, resulting in augmented cytokine production. Mechanistically, the aryl hydrocarbon receptor (AhR) contributes to IS-trained immunity by enhancing the expression of arachidonic acid (AA) metabolism-related genes such as arachidonate 5-lipoxygenase (ALOX5) and ALOX5 activating protein (ALOX5AP). Inhibition of AhR during IS training suppresses the induction of IS-trained immunity. Monocytes from end-stage renal disease (ESRD) patients have increased ALOX5 expression and after 6 days training, they exhibit enhanced TNF-α and IL-6 production to lipopolysaccharide (LPS). Furthermore, healthy control-derived monocytes trained with uremic sera from ESRD patients exhibit increased production of TNF-α and IL-6. Consistently, IS-trained mice and their splenic myeloid cells had increased production of TNF-α after in vivo and ex vivo LPS stimulation compared to that of control mice. These results provide insight into the role of IS in the induction of trained immunity, which is critical during inflammatory immune responses in CKD patients.

Keywords:  aryl hydrocarbon receptor; chronic kidney disease; epigenetic; human; immunology; indoxyl sulfate; inflammation; metabolic reprogramming; trained immunity

DOI:  https://doi.org/10.7554/eLife.87316