bims-supasi 2025-10-26 papers

Glycobiology. 2025 Oct 22. pii: cwaf068. [Epub ahead of print]

Establishment of a method for measuring the 3-O-sulfated structure in heparan sulfate.

Hideo Mochizuki, Yuya Otsuka, Koji Kimata.

Heparan sulfate is present on the cell surface and within the extracellular matrix of most animal species, and it regulates various physiological processes by binding to a wide variety of functional proteins. The diverse array of sulfation patterns on the heparan sulfate chain enables specific binding to each protein. Of particular interest is the 3-O-sulfated (3S) structure of glucosamine residues, which is considered a key structure in determining binding specificity to functional proteins. However, 3-O-sulfation is a rare modification, which makes it difficult to analyze and hinders elucidation of its physiological function. Establishing a general method for measuring the 3S structure is thus important for continued progress in this research field. We previously developed an HPLC method to separate and quantify 13 components of heparan sulfate, including five 3S components, as a complete heparin lyase digestion product. Application of this method in routine analysis required the development of a standard sample for quantitation that is simple to prepare in large amounts and exhibits good stability. A heparan sulfate standard composed of 13 components in known quantities, designated HS13, was prepared for this purpose. A standard mixture of the 13 components can be obtained by digesting HS13 with a heparin lyase. A compositional analysis of heparan sulfate derived from various rat organs was then conducted to test the newly developed standard. The organ-specific distribution of each 3S component was elucidated, and it was confirmed that the 3S components in biological samples can be quantified by routine HPLC analysis.

Keywords: 3-O-sulfate; glycosaminoglycan; heparan sulfate; organ distribution; quantitative analysis

DOI: https://doi.org/10.1093/glycob/cwaf068

JOR Spine. 2025 Dec;8(4): e70131

Enzymatic Behavior of Condoliase in Porcine Nucleus Pulposus: Ex Vivo and In Vitro Assessment.

Ippei Watanabe, Ko Takeda, Taiichi Shirogane.

Background: Condoliase is a treatment for lumbar disc herniation. This enzyme exerts its medicinal effects by digesting chondroitin sulfate (CS), which is abundant in the nucleus pulposus. However, the behavior of administered condoliase in the nucleus pulposus is not clear. Because the purpose of this study is to understand the mechanism of enzyme action, we evaluated the properties of condoliase in the nucleus pulposus.
Methods: The following were evaluated: (1) The diffusibility of fluorescein labeled condoliase injected into isolated porcine nucleus pulposus. (2) The time dependence of condoliase activity in porcine nucleus pulposus or CS solution. (3) The morphology of the enzyme-treated nucleus pulposus tissue was characterized using scanning electron microscopy.
Results: After injection into the nucleus pulposus, condoliase was difficult to diffuse spontaneously, and the rate of CS-disaccharides production was significantly increased up to a peak at 24 h and decreased thereafter. Not all of the CS in the nucleus pulposus was digested by condoliase. These results suggested that condoliase digested CS locally without causing its spontaneous diffusion within the nucleus pulposus. Moreover, condoliase did not digest the collagen fibers that form the supportive architecture of the nucleus pulposus.
Conclusions: We demonstrated that condoliase is retained in the nucleus pulposus and exerts its pharmacological effects by locally degrading CS without degrading collagen fibers. The results obtained in this study can be useful in predicting the mechanism of the pharmacological action of condoliase in clinical practice.

Keywords: chemonucleolysis; chondroitin sulfate; condoliase; lumbar disc herniation; nucleus pulposus

DOI: https://doi.org/10.1002/jsp2.70131

Int J Pharm X. 2025 Dec;10 100403

Chondroitin sulfate-decorated cupper-benzene dicarboxylate framework as an efficient passive and active targeting nanomedicine for anticancer methotrexate delivery.

Siamak Javanbakht, Reza Mohammadi.

In this research, an advanced drug delivery system was developed by decorating the copper-benzene dicarboxylate framework (Cu(BDC)) with the multifunctional chondroitin sulfate (ChS), termed Cu(BDC)/ChS. This novel system is designed for both active and passive targeting, featuring a pH-sensitive release mechanism that enhances drug effectiveness. Different characterization techniques confirmed the successful synthesis of the Cu(BDC)/ChS nanocomposite. In-vitro experiments evaluating the loading and release of methotrexate (MTX) showed that the release rate was significantly higher at pH 4.5, releasing 70 % over 92 h at 41 °C, in contrast to less than 20 % at pH 7.4 at 37 °C. This pH responsiveness of the Cu(BDC)/ChS promotes drug release in environments alike to tumor tissues. Additionally, cytotoxicity tests revealed that MTX-loaded Cu(BDC)/ChS exhibited considerable cytotoxic effects on MCF-7 cancer cells, with IC50 value of ∼250 μg/mL after 48 h, accompanied by an increase in apoptosis rates. Remarkably, the overexpression of CD44 receptors on cancer cell surfaces underscores the significance of ChS-functionalized systems in promoting selective cancer cell apoptosis, while exhibiting minimal cytotoxicity toward normal HUVEC cells. Overall, the findings indicate that the combination of Cu(BDC) and ChS holds promise for developing effective platforms for anticancer drug delivery.

Keywords: Chondroitin sulfate; Drug delivery; Metal-organic framework; Methotrexate; Receptor

DOI: https://doi.org/10.1016/j.ijpx.2025.100403

Steroids. 2025 Oct 17. pii: S0039-128X(25)00142-4. [Epub ahead of print] 109701

Exploring the relationship between endogenous steroids and nausea and vomiting in pregnancy: A longitudinal prospective study.

Jana Benešová, Martin Hill, Daniela Dlouhá, Kateřina Roberts, Jana Ullmann, Peter Koliba, Marta Velíková, Šárka Kaňková.

Nausea and vomiting in pregnancy (NVP) affect approximately 70 % of women worldwide. It is thought to have an adaptive function in the first trimester, when it protects the mother and the fetus against potential dangers from the diet. Proximate causes of NVP include hormonal changes during pregnancy. This longitudinal prospective study examined associations between various endogenous steroids and NVP. In the first and third trimester, pregnant women (N = 179) completed the Index of Nausea, Vomiting, and Retching questionnaire (92.1 % of women reported at least some symptoms of NVP in the first trimester and 37.4 % in the third trimester) and we analyzed their blood serum concentrations of 91 endogenous steroids. In the first trimester, NVP intensity was significantly positively associated with progesterone metabolites from the C21 5α/β-reduced steroid group (e.g., allopregnanolone sulfate) and with conjugated 5α-androstane-3α,17β-diol and conjugated 5α-androstane-3β,17β-diol. In the third trimester, we found significant negative associations between NVP and progesterone, conjugated testosterone, 7-oxo-DHEA, 5-androstene-3β,16α,17β-triol sulfate, some C21 Δ5 steroids (e.g., pregnenolone sulfate, 17-hydroxypregnenolone sulfate), and C21 5α/β-reduced steroids (such as allopregnanolone sulfate and conjugated pregnanolone). Our findings suggest that sulfated 3α-hydroxy-5α-steroids may contribute to NVP in early pregnancy by affecting brainstem regions involved in the vomiting reflex. In late pregnancy, higher levels of immunomodulatory androstanes and progesterone may reduce NVP severity via immune regulation and smooth muscle relaxation.

Keywords: Allopregnanolone sulfate; Hormones; Immunomodulation; Nausea and vomiting in pregnancy; Neuroactive steroids; Progesterone

DOI: https://doi.org/10.1016/j.steroids.2025.109701

J Clin Transl Endocrinol. 2025 Dec;42 100420

Hormone responses to buserelin in polycystic ovary syndrome and in eumenorrheic women with hyperandrogenism/hyperandrogenemia, and the relationship of these responses with insulin resistance.

Salvatore Benvenga, Michele Russo, Fausto Famà, Gianpiero Forte, Vittorio Unfer.

We compared 37 women with polycystic ovary syndrome (PCOS) with 24 women with eumenorrhea plus hyperandrogenism and/or hyperandrogenemia without ultrasound evidence of PCO morphology (EuHyperA) to assess their hormone responses to a GnRH-agonist (buserelin). Following our recent paper on PCOS and EuHyperA, we selected patients who performed the 2 h-oral glucose tolerance test (OGTT), and stratified them according to presence/absence of insulin resistance (IR), viz. HOMA-index ≥ 2.5. IR impacted on the PCOS group since IR-yes-PCOS women had significantly higher body weight, BMI, total testosterone (TT), free androgen index (FAI), and 17-hydroxyprogesterone (17-OHP), borderline higher delta-4 androstenedione (Δ4-ASD) and ovarian volume, and significantly lower sex hormone-binding globulin (SHBG) vs IR-no-PCOS. IR-no-EuHyperA had significantly higher follicle-stimulating hormone (FSH), borderline higher dehydroepiandrosterone sulfate (DHEAS) and borderline lower 17-OHP vs IR-no-PCOS. IR-yes-EuHyperA had significantly higher DHEAS, borderline lower TT and FAI vs IR-yes-PCOS. The insulin curve was significantly higher in the IR-yes vs IR-no-PCOS, and IR-yes vs IR-no-EuHyperA. Compared to PCOS, EuHyperA had insignificantly lower glucose and insulin responses regardless of IR status. After steroidogenic ovarian stimulation (24 h-buserelin test), IR presence vs IR absence impacted on 4 curves in PCOS (significantly higher TT, borderline higher 17-OHP, significantly lower Δ4-ASD and DHEAS), and only one curve in EuHyperA (significantly higher TT). IR-no-EuHyperA had two curves significantly lower than IR-no-PCOS (Δ4-ASD and TT). Instead, IR-yes-EuHyperA had significantly lower Δ4-ASD, TT and 17-OHP curves, and significantly higher DHEAS curve vs IR-yes-PCOS. In conclusion, of 35 parameters (baseline, OGTT-related, buserelin-related), 28 (80%) were statistically similar in EuHyperA vs PCOS regardless of IR status. However, IR presence impacted on more parameters in PCOS than EuHyperA. Given the known ovary sparing by IR in PCOS, it appears that IR exacerbates androgen production of PCOS women more markedly than EuHyperA women.

Keywords: 17-OH-progesterone; Eumenorrhea; Gonadotropin-releasing hormone-agonist test; Hyperandrogenemia; Hyperandrogenism; Insulin resistance; Polycystic ovary syndrome

DOI: https://doi.org/10.1016/j.jcte.2025.100420