bims-supasi 2022-05-29 papers

bims-supasi

Biomed News

on Sulfation pathways and signalling

Issue of 2022–05–29
twenty papers selected by
Jonathan Wolf Mueller, University of Birmingham

Chondroitin sulfate enhances the barrier function of basement membrane assembled by heparan sulfate.
Chondroitin Sulfate Proteoglycan 4,6 sulfation regulates sympathetic nerve regeneration after myocardial infarction.
Pleiotrophin Interaction with Synthetic Glycosaminoglycan Mimetics.
Hemocompatibility of Galactomannan and Galactoglucomannan Sulfates in In Vitro Experiments.
mTORC1 is a key regulator that mediates OGD- and TGFβ1-induced myofibroblast transformation and chondroitin-4-sulfate expression in cardiac fibroblasts.
Online In-Tube Solid-Phase Microextraction Coupled with Liquid Chromatography-Tandem Mass Spectrometry for Automated Analysis of Four Sulfated Steroid Metabolites in Saliva Samples.
Sulfated Phenolic Substances: Preparation and Optimized HPLC Analysis.
Chondroitin sulfate proteoglycans prevent immune cell phenotypic conversion and inflammation resolution via TLR4 in rodent models of spinal cord injury.
Usefulness of Serum Biomarkers of Endothelial Glycocalyx Damage in Prognosis of Decompensated Patients with Heart Failure with Reduced Ejection Fraction.
The Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 Promotes Ovarian Cancer Pathogenesis.
Hair Cortisol and DHEA-S in Foals and Mares as a Retrospective Picture of Feto-Maternal Relationship under Physiological and Pathological Conditions.
Indoxyl sulfate impairs in vitro erythropoiesis by triggering apoptosis and senescence.
The Role of Gut-Derived, Protein-Bound Uremic Toxins in the Cardiovascular Complications of Acute Kidney Injury.
Sulfated Cyclocarya paliurus polysaccharides improve immune function of immunosuppressed mice by modulating intestinal microbiota.
Advances in Research on Antiviral Activities of Sulfated Polysaccharides from Seaweeds.
The Importance of Val386 in Transmembrane Domain 8 for the Activation of OATP1B3 by Epigallocatechin Gallate.
Polymorphic Molecular Signatures in Variable Regions of the Plasmodium falciparum var2csa DBL3x Domain Are Associated with Virulence in Placental Malaria.
Lung Hyaluronasome: Involvement of Low Molecular Weight Ha (Lmw-Ha) in Innate Immunity.
Inhibitory Effect of Sulfated Polysaccharide from Codium edule P.C. Silva Against 2,4-Dinitrofluorobenzene (DNFB)- Induced Allergic Contact Dermatitis on Female BALB/c Mice.
The Cell Wall Polysaccharides Biosynthesis in Seaweeds: A Molecular Perspective.

Development. 2022 May 24. pii: dev.200569. [Epub ahead of print]

Chondroitin sulfate enhances the barrier function of basement membrane assembled by heparan sulfate.

Chenqi Tao, Neoklis Makrides, Jen-Zen Chuang, Yihua Wu, Steven E Brooks, Jeffrey D Esko, Ching-Hwa Sung, Xin Zhang.

  Glycosaminoglycans (GAGs) are ubiquitously expressed polysaccharides attached to proteoglycans. Here we showed that ablation of heparan sulfate (HS) polymerase Ext1 in retinal progenitor cells did not affect initial progression of retinal angiogenesis, but it disrupted the pruning of blood vessels and establishment of arterioles and venules. In the absence of retinal HS, blood vessels were also vulnerable to high oxygen tension in early postnatal stages, which can be rescued by exogenous VEGF, consistent with the role of retinal HS in the fine-tuning of VEGF signaling. Furthermore, we observed that the retinal inner limiting membrane (ILM) was disrupted by deletion of Ext1 in a timing specific manner, suggesting that retinal HS is required for the assembly but not the maintenance of the basement membrane. Lastly, we showed that further deletion of C4st1, a CS sulfation enzyme, did not affect the assembly of the ILM, but when combined with Ext1 deletion, it aggravated the retinal permeability by disrupting the retinal glycocalyx. These results demonstrated an important role of CS and HS in establishing the barrier function of the extracellular matrix.

Keywords:  Angiogenesis; Astrocyte; Basement membrane; Chondroitin sulfate; Glycocalyx; Retina

DOI:  https://doi.org/10.1242/dev.200569
Elife. 2022 May 23. pii: e78387. [Epub ahead of print]11

Chondroitin Sulfate Proteoglycan 4,6 sulfation regulates sympathetic nerve regeneration after myocardial infarction.

Matthew R Blake, Diana C Parrish, Melanie A Staffenson, Shanice Sueda, William R Woodward, Beth A Habecker.

  Sympathetic denervation of the heart following ischemia/reperfusion induced myocardial infarction (MI) is sustained by chondroitin sulfate proteoglycans (CSPGs) in the cardiac scar. Denervation predicts risk of sudden cardiac death in humans. Blocking CSPG signaling restores sympathetic axon outgrowth into the cardiac scar, decreasing arrhythmia susceptibility. Axon growth inhibition by CSPGs is thought to depend on the sulfation status of the glycosaminoglycans (CS-GAGs) attached to the core protein. Tandem sulfation of CS-GAGs at the 4th (4S) and 6th (6S) positions of n-acetyl-galactosamine inhibits outgrowth in several types of neurons within the central nervous system, but it is not known if sulfation is similarly critical during peripheral nerve regeneration. We asked if CSPG sulfation prevented sympathetic axon outgrowth. Neurite outgrowth of dissociated rat sympathetic neurons across purified CSPGs is restored in vitro by reducing 4S with the 4-sulfatase enzyme Arylsulfatase-B (ARSB). Additionally, we co-cultured mouse cardiac scar tissue with mouse sympathetic ganglia ex vivo and found that reducing 4S with ARSB restored axon outgrowth to control levels. We examined levels of the enzymes responsible for adding and removing sulfation to CS-GAGs by western blot to determine if they were altered in the left ventricle after MI. We found that CHST15 (4S dependent 6-sulfotransferase) was upregulated, and ARSB was downregulated after MI. Increased CHST15 combined with decreased ARSB suggests a mechanism for production and maintenance of sulfated CSPGs in the cardiac scar. We altered tandem sulfated 4S,6S CS-GAGs in vivo by transient siRNA knockdown of Chst15 and found that reducing 4S,6S restored Tyrosine Hydroxylase (TH) positive sympathetic nerve fibers in the cardiac scar and reduced arrhythmias using a mouse model of MI. Overall, our results suggest that modulating CSPG-sulfation after MI may be a therapeutic target to promote sympathetic nerve regeneration in the cardiac scar and reduce post-MI cardiac arrhythmias.

Keywords:  mouse; neuroscience; rat; regenerative medicine; stem cells

DOI:  https://doi.org/10.7554/eLife.78387
Pharmaceuticals (Basel). 2022 Apr 19. pii: 496. [Epub ahead of print]15(5):

Pleiotrophin Interaction with Synthetic Glycosaminoglycan Mimetics.

Jonathan R Miles, Xu Wang, Jose L de Paz, Pedro M Nieto.

  Chondroitin sulfate (CS) E is the natural ligand for pleiotrophin (PTN) in the central nervous system (CNS) of the embryo. Some structures of PTN in solution have been solved, but no precise location of the binding site has been reported yet. Using 15N-labelled PTN and HSQC NMR experiments, we studied the interactions with a synthetic CS-E tetrasaccharide corresponding to the minimum binding sequence. The results agree with the data for larger GAG (glycosaminoglycans) sequences and confirm our hypothesis that a synthetic tetrasaccharide is long enough to fully interact with PTN. We hypothesize that the central region of PTN is an intrinsically disordered region (IDR) and could modify its properties upon binding. The second tetrasaccharide has two benzyl groups and shows similar effects on PTN. Finally, the last measured compound aggregated but beforehand, showed a behavior compatible with a slow exchange in the NMR time scale. We propose the same binding site and mode for the tetrasaccharides with and without benzyl groups.

Keywords:  15N-HSQC; NMR; chondroitin sulphate type E; glycosaminoglycan

DOI:  https://doi.org/10.3390/ph15050496
Bull Exp Biol Med. 2022 May 27.

Hemocompatibility of Galactomannan and Galactoglucomannan Sulfates in In Vitro Experiments.

N N Drozd, S A Kuznetsova, Yu N Malyar, A S Kazachenko.

  We identified compounds that do not independently provoke aggregation of human platelets and do not affect hemolysis of human erythrocytes in vitro: lacking anticoagulant activity sulfated galactoglucomannan (polydispersity 1.43; degree of sulfation 0.66) in concentrations ≤0.2 mg/ml; exhibiting anticoagulant activity (in concentrations up to 0.002 mg/ml) sulfated galactoglucomannan (polydispersity 1.5; degree of sulfation 1.81) and galactomannan obtained by sulfation with the sulfamic acid-urea complex (polydispersity 2.75; degree of sulfation 1.25) and galactomannans obtained by sulfation with chlorosulfonic acid in 1,4-dioxane (polydispersity 1.61/22.27; degree of sulfation 1.00/0.74).

Keywords:  blood/plasma coagulation; erythrocyte hemolysis; galactoglucomannan sulfates; galactomannan sulfates; platelet aggregation

DOI:  https://doi.org/10.1007/s10517-022-05501-7
Exp Ther Med. 2022 Jun;23(6): 413

mTORC1 is a key regulator that mediates OGD- and TGFβ1-induced myofibroblast transformation and chondroitin-4-sulfate expression in cardiac fibroblasts.

Chao Li, Zheng Zhang, Yu Peng, Yanying Zhang, Wanrong Kang, Yingdong Li, Yang Hai.

  Ischemia-reperfusion infarct-derived chondroitin sulfate proteoglycans (CSPGs) are important for sustaining denervation of the infarct. Sympathetic denervation within the heart after myocardial infarction (MI) predicts the probability of a higher risk for serious ventricular arrhythmias. Chondroitin-4-sulfate (C4S) is the predominant chondroitin sulfate component in the heart. However, the mechanisms that induce CSPG expression in fibroblasts following MI remain to be elucidated. The present study found that oxygen-glucose deprivation (OGD) and TGFβ1 stimulation induced myofibroblast transformation and C4S synthesis in vitro by using reverse transcription-quantitative PCR, western blotting and immunofluorescence. MTT assay was used to detect cell viability following OGD or OGD + TGF lotreatment. Using the PI3K inhibitor ZSTK474, the Akt inhibitor MK2206, or the mTOR inhibitor AZD8055, it was observed that OGD and TGFβ1 stimulation induced myofibroblast transformation and that C4S synthesis was mTOR-dependent, whereas the upstream canonical PI3K/Akt axis was dispensable by using western blotting and immunofluorescence. siRNA knockdown of Smad3, Raptor, or Rictor, indicated that mTORC1 was critical for promoting OGD- and TGFβ1-induced myofibroblast transformation and C4S synthesis by using western blotting and immunofluorescence. This response, may be mediated via cooperation between canonical Smad3 and mTORC1 signaling. These data suggested that inhibiting myofibroblast transformation may reduce C4S synthesis. Target mTORC1 may provide additional insight into the regeneration of sympathetic nerves and the reduction of fibrosis after MI at the cellular level. These findings may contribute to the understanding of the mechanism by which C4S overproduction in the hearts of patients with MI is associated with myocardial fibrosis.

Keywords:  Smad3; cardiac fibroblasts; chondroitin sulfate proteoglycans; chondroitin-4-sulfate; mTORC1

DOI:  https://doi.org/10.3892/etm.2022.11340
Molecules. 2022 May 18. pii: 3225. [Epub ahead of print]27(10):

Online In-Tube Solid-Phase Microextraction Coupled with Liquid Chromatography-Tandem Mass Spectrometry for Automated Analysis of Four Sulfated Steroid Metabolites in Saliva Samples.

Hiroyuki Kataoka, Daiki Nakayama.

  Accurate measurement of sulfated steroid metabolite concentrations can not only enable the elucidation of the mechanisms regulating steroid metabolism, but also lead to the diagnosis of various related diseases. The present study describes a simple and sensitive method for the simultaneous determination of four sulfated steroid metabolites in saliva, pregnenolone sulfate (PREGS), dehydroepiandrosterone sulfate (DHEAS), cortisol sulfate (CRTS), and 17β-estradiol-3-sulfate (E2S), by online coupling of in-tube solid-phase microextraction (IT-SPME) and stable isotope dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS). These compounds were extracted and concentrated on Supel-Q PLOT capillary tubes by IT-SPME and separated and detected within 6 min by LC-MS/MS using an InertSustain swift C18 column and negative ion mode multiple reaction monitoring systems. These operations were fully automated by an online program. Calibration curves using their stable isotope-labeled internal standards showed good linearity in the range of 0.01-2 ng mL-1 for PREGS, DHEAS, and CRTS and of 0.05-10 ng mL-1 for E2S. The limits of detection (S/N = 3) of PREGS, DHEAS, CRTS, and E2S were 0.59, 0.30, 0.80, and 3.20 pg mL-1, respectively. Moreover, intraday and interday variations were lower than 11.1% (n = 5). The recoveries of these compounds from saliva samples were in the range of 86.6-112.9%. The developed method is highly sensitive and specific and can easily measure sulfated steroid metabolite concentrations in 50 μL saliva samples.

Keywords:  in-tube solid-phase microextraction (IT-SPME); liquid chromatography–tandem mass spectrometry (LC–MS/MS); online automated analysis; saliva; stable isotope dilution; sulfated steroid metabolites

DOI:  https://doi.org/10.3390/molecules27103225
Int J Mol Sci. 2022 May 20. pii: 5743. [Epub ahead of print]23(10):

Sulfated Phenolic Substances: Preparation and Optimized HPLC Analysis.

Lucie Petrásková, Kristýna Káňová, Katerina Brodsky, Anastasiia Hetman, Barbora Petránková, Helena Pelantová, Vladimír Křen, Kateřina Valentová.

  Sulfation is an important reaction in nature, and sulfated phenolic compounds are of interest as standards of mammalian phase II metabolites or pro-drugs. Such standards can be prepared using chemoenzymatic methods with aryl sulfotransferases. The aim of the present work was to obtain a large library of sulfated phenols, phenolic acids, flavonoids, and flavonolignans and optimize their HPLC (high performance liquid chromatography) analysis. Four new sulfates of 2,3,4-trihydroxybenzoic acid, catechol, 4-methylcatechol, and phloroglucinol were prepared and fully characterized using MS (mass spectrometry), 1H, and 13C NMR. The separation was investigated using HPLC with PDA (photodiode-array) detection and a total of 38 standards of phenolics and their sulfates. Different stationary (monolithic C18, C18 Polar, pentafluorophenyl, ZICpHILIC) and mobile phases with or without ammonium acetate buffer were compared. The separation results were strongly dependent on the pH and buffer capacity of the mobile phase. The developed robust HPLC method is suitable for the separation of enzymatic sulfation reaction mixtures of flavonoids, flavonolignans, 2,3-dehydroflavonolignans, phenolic acids, and phenols with PDA detection. Moreover, the method is directly applicable in conjunction with mass detection due to the low flow rate and the absence of phosphate buffer and/or ion-pairing reagents in the mobile phase.

Keywords:  Desulfitobacterium hafniense; HPLC analysis; aryl sulfotransferase; flavonoids; phenolic acid; polyphenols; sulfates

DOI:  https://doi.org/10.3390/ijms23105743
Nat Commun. 2022 May 25. 13(1): 2933

Chondroitin sulfate proteoglycans prevent immune cell phenotypic conversion and inflammation resolution via TLR4 in rodent models of spinal cord injury.

Isaac Francos-Quijorna, Marina Sánchez-Petidier, Emily R Burnside, Smaranda R Badea, Abel Torres-Espin, Lucy Marshall, Fred de Winter, Joost Verhaagen, Victoria Moreno-Manzano, Elizabeth J Bradbury.

Chondroitin sulfate proteoglycans (CSPGs) act as potent inhibitors of axonal growth and neuroplasticity after spinal cord injury (SCI). Here we reveal that CSPGs also play a critical role in preventing inflammation resolution by blocking the conversion of pro-inflammatory immune cells to a pro-repair phenotype in rodent models of SCI. We demonstrate that enzymatic digestion of CSPG glycosaminoglycans enhances immune cell clearance and reduces pro-inflammatory protein and gene expression profiles at key resolution time points. Analysis of phenotypically distinct immune cell clusters revealed CSPG-mediated modulation of macrophage and microglial subtypes which, together with T lymphocyte infiltration and composition changes, suggests a role for CSPGs in modulating both innate and adaptive immune responses after SCI. Mechanistically, CSPG activation of a pro-inflammatory phenotype in pro-repair immune cells was found to be TLR4-dependent, identifying TLR4 signalling as a key driver of CSPG-mediated immune modulation. These findings establish CSPGs as critical mediators of inflammation resolution failure after SCI in rodents, which leads to prolonged inflammatory pathology and irreversible tissue destruction.

DOI: https://doi.org/10.1038/s41467-022-30467-5
Am J Cardiol. 2022 May 20. pii: S0002-9149(22)00435-0. [Epub ahead of print]

Usefulness of Serum Biomarkers of Endothelial Glycocalyx Damage in Prognosis of Decompensated Patients with Heart Failure with Reduced Ejection Fraction.

Yong-Hyun Kim, Takeshi Kitai, Rommel Morales, Kathryn Kiefer, Thanat Chaikijurajai, W H Wilson Tang.

The surface layer of endothelium contains the endothelial glycocalyx (eGC), consisting of proteoglycan polymers. Syndecan-1, heparan sulfate, and hyaluronic acid are major constituents of eGC, and their increasing detection in serum represents active degradation of eGC. Serum was obtained from patients with no heart failure (non-HF) and with HF with reduced ejection fraction (HFrEF) of <40%, either stable chronic HF (CHF) or acute decompensated HF (ADHF). Syndecan-1, heparan sulfate, and hyaluronic acid were measured for comparisons in the groups, adjusting for clinical and laboratory values. In our study cohort, 51 non-HF, 66 ADHF, and 72 patients with CHF were enrolled. Between ADHF and CHF, left ventricular (LV) mass index, LV ejection fraction, and pulmonary capillary wedge pressure did not differ. Patients with ADHF had significantly higher levels of eGC constituents compared with CHF and non-HF. During follow-up, 21 patients with HF died, and the mortality rate was higher in patients with higher serum syndecan-1 or heparan sulfate (log-rank p = 0.007 and 0.016, respectively). In multivariate analysis, a doubling of serum heparan sulfate concentration amounted to a 31.5% increase in all-cause mortality (hazard ratio = 1.315, confidence interval = 1.012-1.709, p = 0.040). In conclusion, serum biomarkers of eGC were elevated in ADHF (but not in CHF) in patients with HFrEF, suggesting the potential roles of eGC degradation and endothelial dysfunction in HF decompensation. Only elevated heparin sulfate was associated with higher all-cause mortality after adjusting for traditional risk variables in patients with HFrEF.

DOI: https://doi.org/10.1016/j.amjcard.2022.04.036
Int J Mol Sci. 2022 May 21. pii: 5793. [Epub ahead of print]23(10):

The Cell Surface Heparan Sulfate Proteoglycan Syndecan-3 Promotes Ovarian Cancer Pathogenesis.

Lara Hillemeyer, Nancy Adriana Espinoza-Sanchez, Burkhard Greve, Nourhan Hassan, Anca Chelariu-Raicu, Ludwig Kiesel, Martin Götte.

  Syndecans are transmembrane heparan sulfate proteoglycans that integrate signaling at the cell surface. By interacting with cytokines, signaling receptors, proteases, and extracellular matrix proteins, syndecans regulate cell proliferation, metastasis, angiogenesis, and inflammation. We analyzed public gene expression datasets to evaluate the dysregulation and potential prognostic impact of Syndecan-3 in ovarian cancer. Moreover, we performed functional in vitro analysis in syndecan-3-siRNA-treated SKOV3 and CAOV3 ovarian cancer cells. In silico analysis of public gene array datasets revealed that syndecan-3 mRNA expression was significantly increased 5.8-fold in ovarian cancer tissues (n = 744) and 3.4-fold in metastases (n = 44) compared with control tissue (n = 46), as independently confirmed in an RNAseq dataset on ovarian serous cystadenocarcinoma tissue (n = 374, controls: n = 133, 3.5-fold increase tumor vs. normal). Syndecan-3 siRNA knockdown impaired 3D spheroid growth and colony formation as stemness-related readouts in SKOV3 and CAOV3 cells. In SKOV3, but not in CAOV3 cells, syndecan-3 depletion reduced cell viability both under basal conditions and under chemotherapy with cisplatin, or cisplatin and paclitaxel. While analysis of the SIOVDB database did not reveal differences in Syndecan-3 expression between patients, sensitive, resistant or refractory to chemotherapy, KM Plotter analysis of 1435 ovarian cancer patients revealed that high syndecan-3 expression was associated with reduced survival in patients treated with taxol and platin. At the molecular level, a reduction in Stat3 activation and changes in the expression of Wnt and notch signaling constituents were observed. Our study suggests that up-regulation of syndecan-3 promotes the pathogenesis of ovarian cancer by modulating stemness-associated pathways.

Keywords:  STAT3; biomarker; cancer stem cells; chemotherapy; heparan sulfate; notch; ovarian carcinoma; prognosis; proteoglycan

DOI:  https://doi.org/10.3390/ijms23105793
Animals (Basel). 2022 May 14. pii: 1266. [Epub ahead of print]12(10):

Hair Cortisol and DHEA-S in Foals and Mares as a Retrospective Picture of Feto-Maternal Relationship under Physiological and Pathological Conditions.

Aliai Lanci, Jole Mariella, Nicola Ellero, Alice Faoro, Tanja Peric, Alberto Prandi, Francesca Freccero, Carolina Castagnetti.

  Equine fetal hair starts to grow at around 270 days of pregnancy, and hair collected at birth reflects hormones of the last third of pregnancy. The study aimed to evaluate cortisol (CORT) and dehydroepiandrosterone-sulfate (DHEA-S) concentrations and their ratio in the trichological matrix of foals and mares in relation to their clinical parameters; the clinical condition of the neonate (study 1); the housing place at parturition (study 2). In study 1, 107 mare-foal pairs were divided into healthy (group H; n = 56) and sick (group S; n = 51) foals, whereas in study 2, group H was divided into hospital (n = 30) and breeding farm (n = 26) parturition. Steroids from hair were measured using a solid-phase microtiter radioimmunoassay. In study 1, hair CORT concentrations measured in foals did not differ between groups and did not appear to be influenced by clinical parameters. A correlation between foal and mare hair CORT concentrations (p = 0.019; r = 0.312, group H; p = 0.006; r = 0.349, group S) and between CORT and DHEA-S concentrations in foals (p = 0.018; r = 0.282, group H; p < 0.001; r = 0.44, group S) and mares (p = 0.006; r = 0.361, group H; p = 0.027; r = 0.271, group S) exists in both groups. Increased hair DHEA-S concentrations (p = 0.033) and decreased CORT/DHEA-S ratio (p < 0.001) appear to be potential biomarkers of chronic stress in the final third of pregnancy, as well as a potential sign of resilience and allostatic load in sick foals, and deserve further attention in the evaluation of prenatal hypothalamus-pituitary-adrenal (HPA) axis activity in the equine species. In study 2, hormone concentrations in the hair of mares hospitalized for attended parturition did not differ from those that were foaled at the breeding farm. This result could be related to a too brief period of hospitalization to cause significant changes in steroid deposition in the mare's hair.

Keywords:  allostasis; biomarkers; hair; hormones; hypothalamus pituitary adrenal axis; mare; neonatal foal; pregnancy; prenatal; ratio

DOI:  https://doi.org/10.3390/ani12101266
Exp Biol Med (Maywood). 2022 May 25. 15353702221097320

Indoxyl sulfate impairs in vitro erythropoiesis by triggering apoptosis and senescence.

Thitinat Duangchan, Manoch Rattanasompattikul, Narong Chitchongyingcharoen, Sumana Mas-Oodi, Moltira Promkan, Nuttawut Rongkiettechakorn, Suksan Korpraphong, Aungkura Supokawej.

  Anemia is a major complication in over 50% of chronic kidney disease (CKD) patients. One of the main causes of anemia in CKD is the reduction of erythropoietin (EPO) synthesis from renal tubular cells. Therefore, first-line treatment of CKD is EPO administration; however, EPO unresponsiveness in several patients is frequently found. More undefined causes of anemia in CKD are under interest, especially uremic toxins, which are a group of solutes accumulated in CKD patients. The highly detectable protein-bound uremic toxin, indoxyl sulfate (IS) was investigated for its effects on in vitro erythropoiesis in this study. CD34+ hematopoietic stem cells were isolated from human umbilical cord blood and differentiated toward erythrocyte lineage for 14 days in various concentrations of IS (12.5, 25, 50, and 100 µg/mL). The effects of IS on cell proliferation, differentiation, apoptosis, and senescence were determined. Cell proliferation was investigated by manual cell counting. Cell surface marker expression was analyzed by flow cytometry. Wright's staining was performed to evaluate cell differentiation capacity. Apoptosis and senescence marker expression was measured using reverse transcription polymerase chain reaction (RT-PCR). TUNEL assay was performed to detect apoptotic DNA fragmentation. Our results demonstrated that IS reduced cell proliferation and impaired erythrocyte differentiation capacity. In addition, this study confirmed the effects of IS on cell apoptosis and senescence during erythropoietic differentiation. Therefore, the promotion of apoptosis and senescence might be one of the possible mechanisms caused by uremic toxin accumulation leading to anemia in CKD patients.

Keywords:  Uremic toxin; anemia; apoptosis; hematopoietic stem cells; indoxyl sulfate; senescence

DOI:  https://doi.org/10.1177/15353702221097320
Toxins (Basel). 2022 May 11. pii: 336. [Epub ahead of print]14(5):

The Role of Gut-Derived, Protein-Bound Uremic Toxins in the Cardiovascular Complications of Acute Kidney Injury.

Pauline Caillard, Youssef Bennis, Isabelle Six, Sandra Bodeau, Saïd Kamel, Gabriel Choukroun, Julien Maizel, Dimitri Titeca-Beauport.

  Acute kidney injury (AKI) is a frequent disease encountered in the hospital, with a higher incidence in intensive care units. Despite progress in renal replacement therapy, AKI is still associated with early and late complications, especially cardiovascular events and mortality. The role of gut-derived protein-bound uremic toxins (PBUTs) in vascular and cardiac dysfunction has been extensively studied during chronic kidney disease (CKD), in particular, that of indoxyl sulfate (IS), para-cresyl sulfate (PCS), and indole-3-acetic acid (IAA), resulting in both experimental and clinical evidence. PBUTs, which accumulate when the excretory function of the kidneys is impaired, have a deleterious effect on and cause damage to cardiovascular tissues. However, the link between PBUTs and the cardiovascular complications of AKI and the pathophysiological mechanisms potentially involved are unclear. This review aims to summarize available data concerning the participation of PBUTs in the early and late cardiovascular complications of AKI.

Keywords:  acute kidney injury; cardiovascular dysfunction; indole-3-acetic acid; indoxyl sulfate; para-cresyl sulfate; uremic toxins

DOI:  https://doi.org/10.3390/toxins14050336
Int J Biol Macromol. 2022 May 18. pii: S0141-8130(22)01084-4. [Epub ahead of print]212 31-42

Sulfated Cyclocarya paliurus polysaccharides improve immune function of immunosuppressed mice by modulating intestinal microbiota.

Yi Han, Yang Zhang, Kehui Ouyang, Lingli Chen, Meng Zhao, Wenjun Wang.

  The study was aimed to investigate the effect of Cyclocarya paliurus polysaccharides (CPP) and the sulfation derivative (S-CPP) on modulate intestinal mucosal immunity and intestinal microbiota in cyclophosphamide-induced mice. The results showed that CPP and S-CPP effectively alleviated intestinal villi injury, enhanced the contents of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in small intestinal tissue and serum, and upregulated IL-1β at gene levels, zonula occludens-1 (ZO-1), Occludin and Claudin-1 at gene and protein levels, thereby promoting the repair of intestinal mechanical barrier and enhancing intestinal mucosal immunity. Moreover, the beneficial modulation of CPP and S-CPP on the overall structure of intestinal microbiota was revealed by performing 16S ribosomal RNA (16S rRNA) sequencing. Sulfated modification could improve the protection of CPP on the intestinal barrier and the regulation of systemic immunity. S-CPP had a stronger potential to reduce the damage of cyclophosphamide (Cy) on immunity and intestinal microbiota.

Keywords:  Cyclocarya paliurus polysaccharides; Immunity; Immunosuppression; Intestinal microbiota; Sulfated modification

DOI:  https://doi.org/10.1016/j.ijbiomac.2022.05.110
Pharmaceuticals (Basel). 2022 May 06. pii: 581. [Epub ahead of print]15(5):

Advances in Research on Antiviral Activities of Sulfated Polysaccharides from Seaweeds.

Qiang Wei, Guoqiang Fu, Ke Wang, Qiong Yang, Jiarui Zhao, Yuan Wang, Kai Ji, Shuliang Song.

  In recent years, various viral diseases have suddenly erupted, resulting in widespread infection and death. A variety of biological activities from marine natural products have gradually attracted the attention of people. Seaweeds have a wide range of sources, huge output, and high economic benefits. This is very promising in the pharmaceutical industry. In particular, sulfated polysaccharides derived from seaweeds, considered a potential source of bioactive compounds for drug development, have shown antiviral activity against a broad spectrum of viruses, mainly including common DNA viruses and RNA viruses. In addition, sulfated polysaccharides can also improve the body's immunity. This review focuses on recent advances in antiviral research on the sulfated polysaccharides from seaweeds, including carrageenan, galactan, fucoidan, alginate, ulvan, p-KG03, naviculan, and calcium spirulan. We hope that this review will provide new ideas for the development of COVID-19 therapeutics and vaccines.

Keywords:  COVID-19; SARS-CoV-2; algae; antiviral; seaweed; sulfated polysaccharides; viruses

DOI:  https://doi.org/10.3390/ph15050581
J Agric Food Chem. 2022 May 22.

The Importance of Val386 in Transmembrane Domain 8 for the Activation of OATP1B3 by Epigallocatechin Gallate.

Zhongmin Wang, Ying Li, Cecilia E Villanueva, Taotao Peng, Wanjun Han, Zheyue Bo, Hongjian Zhang, Bruno Hagenbuch, Chunshan Gui.

  Estrone-3-sulfate (E3S) uptake mediated by organic anion transporting polypeptide 1B3 (OATP1B3) can be activated by epigallocatechin gallate (EGCG). In this study, by using chimeric transporters and site-directed mutagenesis, we found that Val386 in transmembrane domain 8 (TM8) is essential for OATP1B3's activation by EGCG. Kinetic studies showed that the loss of activation of 1B3-TM8 and 1B3-V386F in the presence of EGCG is due to their decreased substrate binding affinity and reduced maximal transport rate. The overall transport efficiencies of OATP1B3, 1B3-TM8, and 1B3-V386F in the absence and presence of EGCG are 8.6 ± 0.7 vs 15.9 ± 1.4 (p < 0.05), 11.2 ± 2.1 vs 2.7 ± 0.3 (p < 0.05), and 10.2 ± 1.0 vs 2.5 ± 0.3 (p < 0.05), respectively. While 1B3-V386F cannot be activated by EGCG, its transport activity for EGCG is also diminished. OATP1B3's activation by EGCG is substrate-dependent as EGCG inhibits OATP1B3-mediated pravastatin uptake. Furthermore, the activation of OATP1B3-mediated E3S uptake by quercetin 3-O-α-l-arabinopyranosyl(1 → 2)-α-l-rhamnopyranoside is not affected by TM8 and V386F. Taken together, the activation of OATP1B3 by small molecules is substrate- and modulator-dependent, and V386 in TM8 plays a critical role in the activation of OATP1B3-mediated E3S uptake by EGCG.

Keywords:  EGCG; OATP; chimera; kinetics; solute carrier; transmembrane domain

DOI:  https://doi.org/10.1021/acs.jafc.2c02692
Pathogens. 2022 Apr 28. pii: 520. [Epub ahead of print]11(5):

Polymorphic Molecular Signatures in Variable Regions of the Plasmodium falciparum var2csa DBL3x Domain Are Associated with Virulence in Placental Malaria.

Eldin Talundzic, Stephen Scott, Simon O Owino, David S Campo, Naomi W Lucchi, Venkatachalam Udhayakumar, Julie M Moore, David S Peterson.

  The Plasmodium falciparum protein VAR2CSA allows infected erythrocytes to accumulate within the placenta, inducing pathology and poor birth outcomes. Multiple exposures to placental malaria (PM) induce partial immunity against VAR2CSA, making it a promising vaccine candidate. However, the extent to which VAR2CSA genetic diversity contributes to immune evasion and virulence remains poorly understood. The deep sequencing of the var2csa DBL3X domain in placental blood from forty-nine primigravid and multigravid women living in malaria-endemic western Kenya revealed numerous unique sequences within individuals in association with chronic PM but not gravidity. Additional analysis unveiled four distinct sequence types that were variably present in mixed proportions amongst the study population. An analysis of the abundance of each of these sequence types revealed that one was inversely related to infant gestational age, another was inversely related to placental parasitemia, and a third was associated with chronic PM. The categorization of women according to the type to which their dominant sequence belonged resulted in the segregation of types as a function of gravidity: two types predominated in multigravidae whereas the other two predominated in primigravidae. The univariate logistic regression analysis of sequence type dominance further revealed that gravidity, maternal age, placental parasitemia, and hemozoin burden (within maternal leukocytes), reported a lack of antimalarial drug use, and infant gestational age and birth weight influenced the odds of membership in one or more of these sequence predominance groups. Cumulatively, these results show that unique var2csa sequences differentially appear in women with different PM exposure histories and segregate to types independently associated with maternal factors, infection parameters, and birth outcomes. The association of some var2csa sequence types with indicators of pathogenesis should motivate vaccine efforts to further identify and target VAR2CSA epitopes associated with maternal morbidity and poor birth outcomes.

Keywords:  VAR2CSA; chondroitin sulfate A; chronic infection; low birth weight; placental malaria; polymorphism; virulence

DOI:  https://doi.org/10.3390/pathogens11050520
Biomolecules. 2022 Apr 30. pii: 658. [Epub ahead of print]12(5):

Lung Hyaluronasome: Involvement of Low Molecular Weight Ha (Lmw-Ha) in Innate Immunity.

Antony Hoarau, Myriam Polette, Christelle Coraux.

  Hyaluronic acid (HA) is a major component of the extracellular matrix. It is synthesized by hyaluronan synthases (HAS) into high-molecular-weight chains (HMW-HA) that exhibit anti-inflammatory and immunomodulatory functions. In damaged, infected, and/or inflamed tissues, HMW-HA are degraded by hyaluronidases (HYAL) or reactive oxygen species (ROS) to give rise to low-molecular-weight HAs (LMW-HAs) that are potent pro-inflammatory molecules. Therefore, the size of HA regulates the balance of anti- or pro-inflammatory functions. The activities of HA depend also on its interactions with hyaladherins. HA synthesis, degradation, and activities through HA/receptors interactions define the hyaluronasome. In this review, a short overview of the role of high and low-molecular-weight HA polymers in the lungs is provided. The involvement of LMW-HA in pulmonary innate immunity via the activation of neutrophils, macrophages, dendritic cells, and epithelial cells is described to highlight LMW-HA as a therapeutic target in inflammatory respiratory diseases. Finally, the possibilities to counter LMW-HA's deleterious effects in the lungs are discussed.

Keywords:  hyaluronic acid; inflammation; innate immunity; lung

DOI:  https://doi.org/10.3390/biom12050658
Adv Pharm Bull. 2022 Mar;12(2): 410-418

Inhibitory Effect of Sulfated Polysaccharide from Codium edule P.C. Silva Against 2,4-Dinitrofluorobenzene (DNFB)- Induced Allergic Contact Dermatitis on Female BALB/c Mice.

Martin Raemond Brondial Mallabo, Mary Jho-Anne T Corpuz, Reginald B Salonga, Ross D Vasquez.

  Purpose: Sulfated polysaccharide from Codium species has been reported for its antiinflammatoryactivities. However, the effect of sulfated polysaccharide from Codium edule on allergic responses has not been studied. The study was conducted to determine the effect ofsulfated polysaccharide (F1) from C. edule on allergic contact dermatitis (ACD) induced by2,4-dinitrofluorobenzene (DNFB) in female BALB/c mice. Methods: F1 was isolated using DEAE sepharose gel chromatography and chemically identifiedby LC-MS analyses. The effects of F1 on changes in ear thickness, allergic responses, andhistology were evaluated. The effects of F1 on the production of inflammatory cytokinesinterferon gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-ɑ) in serum were also quantifiedand compared with standard prednisolone therapy. Results: F1 was identified as a heteropolysaccharide with β-D-galactans and β-L-arabinans units.F1 was non-toxic at 2000 mg/kg. Administration of F1 in DNFB-challenged mice significantlysuppressed the increase in ear thickness, erythema, desquamation, and proliferation ofinflammatory cells. F1 significantly decreased the production of inflammatory markers, IFN-γand TNF-α in a dose-dependent manner when compared to the untreated group (P < 0.05). Conclusion: Results suggest that F1 from C. edule is a bioactive sulfated heteropolysaccharidewith anti-inflammatory activity and might be a valuable candidate molecule for the treatmentof allergic diseases such as ACD.

Keywords:  Allergic contact dermatitis; Codium edule; Cytokines; Inflammation; Seaweed; Sulfated polysaccharide

DOI:  https://doi.org/10.34172/apb.2022.042
Front Plant Sci. 2022 ;13 902823

The Cell Wall Polysaccharides Biosynthesis in Seaweeds: A Molecular Perspective.

Zhanru Shao, Delin Duan.

  Cell wall polysaccharides (CWPS) of seaweeds play crucial roles in mechanical shear resistance, cell-cell adhesion and the interactions with changeable marine environments. They have diverse applications in food, cosmetics, agriculture, pharmaceuticals and therapeutics. The recent boost of multi-omics sequence analysis has rapidly progressed the mining of presumed genes encoding enzymes involved in CWPS biosynthesis pathways. In this review, we summarize the biosynthetic pathways of alginate, fucoidan, agar, carrageenan and ulvan in seaweeds referred to the literatures on published genomes and biochemical characterization of encoded enzymes. Some transcriptomic data were briefly reported to discuss the correlation between gene expression levels and CWPS contents. Mannuronan C-5 epimerase (MC5E) and carbohydrate sulfotransferase (CST) are crucial enzymes for alginate and sulfated CWPS, respectively. Nonetheless, most CWPS-relevant genes were merely investigated by gene mining and phylogenetic analysis. We offer an integrative view of CWPS biosynthesis from a molecular perspective and discuss about the underlying regulation mechanism. However, a clear understanding of the relationship between chemical structure and bioactivities of CWPS is limited, and reverse genetic manipulation and effective gene editing tools need to be developed in future.

Keywords:  biosynthesis; carbohydrate sulfotransferase; cell wall polysaccharide; gene mining; mannuronan C-5 epimerase

DOI:  https://doi.org/10.3389/fpls.2022.902823