AIM:This study aimed to investigate the effects of glucosamine, a glucose analogue mainly transported via glucose transporter (GLUT) 2, on the progression of cholangiocarcinoma (CCA) cells. METHODS:Differential expressions of GLUTs in CCA and normal bile ducts were analyzed using a public transcriptomic dataset. The effects of glucosamine on CCA cell viability and proliferation were examined using an MTT assay and flow cytometry. Molecular mechanisms were investigated using Western blots and lectin blots. RESULTS:GLUT2 expression was significantly decreased in CCA compared with the normal bile duct tissues. Glucosamine significantly reduced CCA cell viability in a dose-dependent manner (p < .05), while flow cytometry showed increased G1 cell cycle arrest and apoptosis in glucosamine-treated cells (p < .05). Glucosamine significantly suppressed high-mannose type N-glycosylation in CCA cells, as shown in concanavalin A lectin blots and the Western blots of glycoprotein 130 (p < .05), leading to a reduced epidermal growth factor receptor (EGFR) expression and STAT3 phosphorylation. Cell viability and high-mannose type N-glycosylation could be partially rescued by high-glucose supplementation, confirming that disrupting high-mannose type N-glycosylation and EGFR/STAT3 signaling are part of the underlying mechanisms. CONCLUSION:Glucosamine exerted anti-cancer effects on CCA cells, suggesting its potential for further study as a repurposing drug for CCA.

Trimethylamine N-oxide (TMAO), a gut microbiota-derived metabolite, has been implicated in atherosclerosis and adverse cardiovascular outcomes, but prospective data on lower extremity arterial disease (LEAD) in Asian populations remain limited. We conducted a single-center prospective cohort study of 155 Thai patients (mean age 63.5 ± 8.8 years; 64.5% male) with angiographically confirmed coronary artery disease and without LEAD at baseline. Plasma TMAO levels were measured by nuclear magnetic resonance spectroscopy and categorized into quartiles. Incident LEAD was assessed after 3 years, and all-cause mortality after 5 years. Multivariable logistic regression was used to evaluate incident LEAD, whereas Cox proportional hazards models were used to assess mortality. Higher TMAO quartiles were associated with progressively greater odds of incident LEAD (P for trend < 0.001). Compared with the lowest quartile, participants in the third and fourth quartiles had adjusted odds ratios of 5.30 (95% CI 1.32–20.48; P = 0.018) and 14.0 (95% CI 3.38–57.64; P < 0.001), respectively. The median follow-up duration, estimated using the reverse Kaplan–Meier method, was 59.1 months (interquartile range 58.3–60.3 months). Higher TMAO levels were also associated with increased 5-year all-cause mortality. In fully adjusted Cox models, the highest TMAO quartile was independently associated with mortality (hazard ratio 4.17, 95% CI 1.11–15.76; P = 0.035), and Kaplan–Meier analysis demonstrated progressively lower survival across increasing quartiles (log-rank P = 0.005). These findings support elevated plasma TMAO as an independent predictor of incident LEAD and long-term mortality in an Asian population. Trial Registration: This was an observational, non-interventional cohort study. The study was registered in the Thai Clinical Trials Registry to enhance transparency of reporting. Thai Clinical Trials Registry (TCTR20250613001).

DNA topoisomerase IIα (Topo IIα) is essential for maintaining genomic stability during DNA replication and mitosis and is highly expressed in cancer cells, making it a promising target for anticancer therapy. In this study, bis-thiourea derivatives were investigated for their Topo IIα inhibitory activity and anticancer potential using in silico and in vitro studies. Molecular modeling demonstrated that compound 8 exhibited favorable binding affinity and stability within the ATPase domain of Topo IIα. Biochemical assays revealed that compound 8 inhibited Topo IIα activity and showed potent cytotoxicity against several cancer cell lines, particularly A549 cells. Mechanistic studies showed that compound 8 inhibited A549 cell migration and invasion by upregulating E-cadherin while downregulating the mesenchymal markers N-cadherin and vimentin, as well as the EMT-associated transcription factor Slug. Furthermore, compound 8 induced G1-phase arrest by downregulating cyclins D1 and E2 while upregulating p21. These results suggest that compound 8 represents a promising lead for Topo IIα-targeted cancer therapy.

Tyrosinase, a key enzyme in melanin synthesis, serves as a primary target for developing depigmenting agents. The search for novel tyrosinase inhibitors is needed due to the adverse effects of current inhibitors. This study evaluated 16 bis-thiourea derivatives using in vitro and in silico methods, identifying compound 4, with chlorine substituents, as the most potent inhibitor. Compound 4 outperformed kojic acid in inhibiting mushroom tyrosinase activity and interacted with catalytic copper ions and active site residues, as revealed by molecular docking and copper-chelating assay. Molecular dynamics simulation and MM/PBSA-based free energy calculations confirmed the greater stability and binding affinity of the compound 4-tyrosinase complex in an aqueous environment compared to kojic acid-tyrosinase complex. Melanin assay revealed that compound 4 significantly suppressed melanin production in B16F10 melanoma cells, showing stronger anti-melanogenic activity than kojic acid. Drug-likeness predictions confirmed its compliance with Lipinski's rule of five, supporting bis-thiourea derivatives as promising tyrosinase inhibitors.

Artocarpus lakoocha agglutinin (ALA), which specifically targets the Gal/GalNAc components of complex glycans, was isolated from the seeds of Artocarpus lakoocha. This study is the first to explore the role of ALA in identifying aberrant glycans, designated ALA-binding glycans (ALAG), and its implications in cholangiocarcinoma (CCA). ALA-histochemistry was used to evaluate ALAG expression in liver fluke-induced CCA tissues from hamsters (n = 60). Elevated ALAG expression was observed in hyperplastic ducts and significantly increased in CCA tissues, while normal biliary epithelium and hepatocytes showed no expression. Similar results were found in patient CCA tissues (n = 68), where higher ALAG levels correlated with shorter survival rates, indicating the involvement of ALAG in CCA development and progression. Furthermore, ALA treatment inhibited cell viability in CCA cell lines, as demonstrated by MTT and colony formation assays, and Ki-67 expression. ALA treatment also decreased cell migration and invasion, as shown by Transwell assays. Gelatin zymography suggested that these effects might be associated with reduced MMP-9 activity. Overall, these findings may position ALAG as a potential marker for poor prognosis in CCA, while ALA may serve as a novel lectin for both detection and therapeutic applications in CCA.

Monosodium glutamate (MSG) is widely used as a food additive and flavor enhancer, but concerns have emerged regarding its potential effects on kidney function and vitamin B6 metabolism. This study investigated the impact of varying levels of dietary MSG intake on vitamin B6 status in a rat model. Male Wistar rats were divided into four groups and administered MSG in their drinking water at concentrations of 0, 0.5, 1.5, and 3.0 g % for 12 weeks. Three isoforms of vitamin B6, pyridoxal (PL), pyridoxal 5'-phosphate (PLP), and pyridoxic acid (PA), were measured in plasma, liver, kidney, and urine using HPLC-MS. Kidney metabolites were analyzed by 1H nuclear magnetic resonance spectroscopy to assess MSG-related metabolic changes. We found that plasma PL and PLP levels were elevated in all MSG-treated groups compared with controls, regardless of dose. This correlated with increased hepatic PLP levels and a decreased PLP catabolic ratio (PAr). In contrast to the liver, PLP levels in the kidney were significantly lower compared to controls. Metabolomics profiling of kidney tissue revealed elevated levels of various amino acids and metabolic intermediates in the MSG-treated rats. In conclusion, MSG consumption cannot induce vitamin B6 deficiency systematically, such as in the liver and plasma, as long as dietary vitamin B6 is adequate. However, depletion of PLP occurred locally in the kidney. Because MSG consumption may affect renal vitamin B6 status, there is a need for further studies to elucidate the underlying mechanisms and assess the potential health implications of habitual MSG intake.

Cholangiocarcinoma (CCA) is a highly aggressive liver cancer associated with a poor prognosis. Cancer stem cells (CSCs) play critical roles in CCA by driving tumor initiation, metastasis, recurrence, and treatment resistance. CSCs possess unique properties, including self-renewal, quiescence, and metabolic plasticity, enabling them to evade conventional treatments such as chemotherapy and radiation. Several key CCA stem cell markers are involved in interactions with immune cells within the tumor microenvironment (TME) and in metabolic adaptations that support CSC survival and resistance through complex mechanisms. Despite advancements in chemotherapy and targeted, CSCs remain a major challenge in CCA treatment. In this review, we summarize new insights, updated information, emerging therapeutic strategies, and future directions for CSC research in CCA.

glycoPATH integrates RNAseq transcriptomic and LC-MS/MS glycomic data, providing a platform to identify genes implicated in diseases involving N -glycan biosynthesis and enabling the development of targeted therapeutics for these pathways.

Alteration of glycosylation in cancer cells leads to the expression of tumor-associated glycans, which can be used as biomarkers for diagnosis and prognostic prediction of diseases. In this study, we used nano-LC-QToF to identify serum N-glycan biomarkers for the detection of brain tumors. We observed an increase in sialylated N-glycans and a decrease in fucosylated N-glycans in the serum of patients with glioblastoma (GBM) and meningioma (MG) compared to healthy individuals. In GBM, a combination of increased serum sialylated N-glycan (6_4_0_2 compound) and decreased fucosylated N-glycan (4_4_1_0 compound) was identified as the most appropriate panel, with an area under the curve (AUC) of 0.8660, 78.95% sensitivity, 84.21% specificity, and 82.89% accuracy. For MG, a combination of decreased 6_6_2_0 and 5_5_2_0 compounds and increased 4_4_1_1 compound achieved an AUC of 0.9260, 82.35% sensitivity, 78.57% specificity, and 80.26% accuracy for diagnosis of MG. Additionally, an increase in 5_5_1_0 and 4_3_0_0 compounds combined with a decrease in 7_7_4_3 was associated with high-grade MG (WHO grades II-III). In conclusion, we identified serum N-glycan profiles associated with brain tumors, highlighting their potential as biomarkers for the diagnosis and prognosis of these diseases.

Introduction:Kidney injury molecule-1 (KIM-1), encoded by the Hepatitis A Virus Cellular Receptor 1 (HAVCR1) gene, plays a crucial role in kidney injury progression. Although serum and urinary KIM-1 levels are established biomarkers for kidney damage, the relationship between KIM-1 levels, HAVCR1 gene polymorphism, and chronic kidney disease (CKD) stages remains unclear. This study aimed to investigate KIM-1 as a potential biomarker for CKD progression in the Thai population and explore its association with genetic polymorphisms in the HAVCR1 gene. Methods:A total of 250 patients with CKD were recruited from Khon Kaen, Thailand. Serum and urinary KIM-1 levels were measured using an indirect enzyme-linked immunosorbent assay. Single-nucleotide polymorphism (SNP) genotyping was conducted using the TaqMan assay to assess the associations between KIM-1 levels, SNPs, and CKD progression. Statistical analyses were conducted to assess the correlations between estimated glomerular filtration rate (eGFR), KIM-1 levels, and SNPs. Results:Serum and urinary KIM-1 levels showed a significant negative correlation with eGFR, indicating higher KIM-1 levels in patients with more advanced CKD. However, the rs6555820 SNP in the HAVCR1 gene did not show a significant association with KIM-1 levels or eGFR. Interestingly, a significant association between rs6555820 and gender was observed, implying a potential gender-dependent genetic impact. Conclusion:Serum and urinary KIM-1 levels have been found to be associated with CKD stages and eGFR, suggesting their potential as biomarkers for assessing CKD severity. However, no direct associations were observed between the SNP rs6555820 and KIM-1 levels or eGFR. Further research is required to elucidate the genetic mechanisms underlying CKD progression.

Aberrant glycosylation in meningioma (MG) suggests the potential use of glycobiomarkers for diagnosis, monitoring, and prognosis. Using lectin-based glycomics, Dolichos biflorus agglutinin (DBA) is an effective tool for detecting the glyco-biomarker DBA-binding glycan (DBAG) in patient sera. Using an in-house enzyme-linked lectin assay, the level of serum DBAG in MG patients (44.10 AU/mL) was significantly higher than that of healthy controls (10.21 AU/mL), with 86.4% sensitivity and 90.0% specificity. The MG patients with WHO grade II-III (77.35 AU/mL) had higher serum DBAG levels than those with grade I (35.92 AU/mL). Lectin histochemistry staining of MG tissues showed that tumor cells were positive for DBAG in 149 of 150 cases, indicating the tumor origin of serum DBAG. Functional analyses using MG cell lines, IOMM-Lee and HKBMM, suggested the involvement of DBAG in MG progression. The serum DBAG levels in other brain tumors (BT) were also measured. Data showed that serum DBAG levels were higher in malignant BTs (67.44 AU/mL) compared to benign BTs (50.43 AU/mL). In conclusion, DBA can be used to detect a serum glycobiomarker for MG and brain tumors. DBAG was involved in MG cell viability and migration, suggesting its potential as a target for future MG therapy.

Soyasaponin-I (SS-I), a sialyltransferase inhibitor naturally found in soybeans, has antioxidant, anticarcinogenic, and hepatoprotective properties. In this study, we explored the possibility to use SS-I as an antimelanogenic agent for the treatment of skin hyperpigmentation disorders. When melanoma cell lines, MNT-1 and B16F10, were treated with SS-I, significant suppression of both α-2,3 and α-2,6 sialylations was observed by using lectin fluorescence staining with sialic acid-binding lectins-Sambucus nigra agglutinin (SNA) and Maackia amurensis lectin-II (MAL-II). SS-I significantly attenuated the α-MSH-induced melanogenesis of MNT-1 and B16F10 cells without a cytotoxic effect. SS-I could activate ERK and suppress the PKA/CREB signaling pathways of melanoma cells. Moreover, SS-I treatment caused significant downregulation of the expression of melanosome-related proteins; tyrosinase-related protein 1 (TRP1), TRP2, and premelanosome protein (PMEL) and the melanogenic-related transcription factor microphthalmia-associated transcription factor (MITF). Consequently, the expression of tyrosinase-the key enzyme regulating melanin production-was significantly suppressed after SS-I treatment. These results suggest the role of sialylation in melanogenesis and the possibility of using SS-I as an alternative antimelanogenic agent. In conclusion, we have demonstrated the antimelanogenic effect of SS-I, an active compound produced in soybeans. SS-I can be an antimelanogenic agent in cosmetic products for the treatment of hyperpigmentation disorders.

O‐GlcNAcylation is an important biological process in regulating the function of many nucleocytoplasmic proteins in cells. Enhancement of O‐GlcNAcylation was associated with cancer development and progression. Here, we demonstrated the involvement of O‐GlcNAcylation in melanoma metastasis. Using the data from GEO database, we found that O‐GlcNAcylation and its related enzymes, including glutamine fructose‐6‐phosphate amidotransferase (GFAT), O‐GlcNAc transferase (OGT), and O‐GlcNAcase (OGA); were elevated in metastatic melanoma compared with primary tumors and normal tissues. Functional analyses in melanoma cell lines‐‐MNT‐1, SK‐MEL‐28, and A‐375 showed that suppression of O‐GlcNAcylation by siRNA against OGT significantly reduces the migration and invasion abilities of the cells. Phosphorylation of Akt and NFkB was drastically suppressed after knockdown of OGT, suggesting the role of O‐GlcNAcylation in regulating the Akt‐NFkB signaling pathway. In addition, we found that the NFkB target genes, such as ZEB‐2 and MCT‐1, were significantly upregulated in metastatic tumors compared with primary tumors. MCT‐1 expression in melanoma tissues was also correlated with O‐GlcNAcylation level. Taken together, we have demonstrated in this study the possible role of O‐GlcNAcylation in controlling melanoma metastasis via upregulating MCT‐1 expression through activation of Akt‐NFkB signaling pathway.

Meningiomas are the most common primary intracranial tumors, and improved diagnostic and therapeutic strategies are needed. Meningioma (MG) exhibits abnormal glycosylation, which can be used for diagnosis, prognosis, and treatment strategies. In this study, we use Ulex europaeus agglutinin-I (UEA-I) to develop an in-house enzyme-linked lectin assay for detecting meningioma (MG)-associated glycan in patient serum. The level of serum UEA-I binding glycan (UEAG) was significantly higher in MG and other brain tumors compared with healthy controls (HC). Moreover, we found that patients with a higher grade of MG (WHO Grade II) have a higher level of serum UEAG compared to those with Grade I. The functional analysis in MG cell lines showed that UEA-I can inhibit the migration and invasion of MG cells, with no effect on cell viability, suggesting the role of UEAG in MG progression. In conclusion, we have demonstrated the potential of UEAG as a serum glycobiomarker for diagnosis, and it may also serve as a target for MG treatment.

Previous studies demonstrated that diabetes and hyperglycemia promote cholangiocarcinoma (CCA) progression, in vitro and in vivo. However, the predictive abilities of blood glucose levels for CCA prognosis remain unclear. This retrospective cohort analysis included 85 patients with histologically confirmed CCA at Srinagarind Hospital, Khon Kaen University, between 1998 and 2000, comprised 57 males and 28 females with a median age of 56 ± 13 years. The glucose-lymphocyte ratio (GLR) was calculated from preoperative fasting blood glucose and absolute lymphocyte counts, and the cutoff was determined using the receiver operating characteristic curve. Survival analysis revealed that higher GLR was significantly associated with shorter overall survival (P < 0.05). A higher GLR was negatively correlated with total protein, globulin, and lymphocyte counts (P < 0.05). Univariate analysis revealed that a high GLR was associated with male sex and a survival time < 6 months (P < 0.05). The multivariable Cox proportional hazard model showed that a higher GLR was an independent prognostic factor for CCA (adjusted hazard ratio: 1.887; 95% confidence interval: 1.101-3.237), alongside carbohydrate antigen 19-9, staging, neutrophil count, and histological grading. Collectively, these findings indicate that the GLR is a significant prognostic biomarker for CCA, underscoring the importance of metabolic and nutritional status in CCA management.

Native Butea monosperma agglutinin (nBMA), is a lectin isolated from the seeds of the Butea monosperma plant, which binds specifically to galactose, N-acetylgalactosamine, and lactose. This study developed a recombinant β-chain of BMA (rBMA) expressed in Escherichia coli. The rBMA exists in a monomeric form, retains native structure and sugar-binding capacity without exhibiting hemagglutination activity. The binding activity of rBMA was evaluated through lectin-cytofluorescent staining of CCA cell lines. Similar to nBMA, rBMA exhibited a positive signal to CCA cell lines but displayed a strong signal in different cell lines. Sodium periodate treatment abolished rBMA binding in CCA tissues and serum dot blots, confirming carbohydrate-dependent interactions. The neutralizing activity for sugar binding specificity indicated that rBMA binds to the complex glycosylated glycans rather than mono- and di-saccharides. Elevated levels of rBMA binding glycans in serum dot blots were found to differentiate CCA patients from healthy individuals, achieving a diagnostic sensitivity of 92.9%, specificity of 36%, and overall accuracy of 74%. High levels of serum rBMA-binding glycans were associated with poorer survival in CCA patients, and directly correlated with serum alkaline phosphatase levels. No correlation was found with carcinoembryonic antigen and CA19-9 levels. These findings position serum rBMA-binding glycans as potential biomarkers reflecting CCA progression. The monomeric nature and retained glycan specificity of rBMA, coupled with its absence of hemagglutination activity, make it a superior candidate to nBMA for diagnostic applications and a promising platform for targeted therapeutic development for CCA.

Purpose:Melanoma arises from the malignant transformation of melanocytes, a serious health problem in high UV-exposure countries. Ineffective treatments and metastasis have led to poor prognosis and high mortality among melanoma patients. Several underlying mechanisms are suspected. The splicing-error in many genes has been frequently reported in melanoma, therefore, targeting splicing regulator Serine/Arginine protein kinases (SRPKs) is promising. Methods:SRPKs expression in the TCGA dataset was analyzed by GEPIA. A375 and MNT-1 were comparatively treated by SRPK inhibitors, SRPIN340 and SPHINX31. Effects on viability and growth were measured by MTT and hanging drop assay. Apoptotic death was examined by flow cytometry and western blotting. Invasive ability was determined by transwell assay. Invasive-associated genes, proteins, and enzymes were tracked by RT-PCR, western blotting, immunofluorescence, and gelatin zymography. Results:SRPIN340 exhibited higher inhibitory effects on the viability and growth of melanoma cells than SPHINX31. Apoptotic induction was found with downregulated Bcl-2 and upregulated cytochrome c, especially in A375 cells. For metastatic inhibition in A375, lower numbers of invaded cells were counted. Downregulated vimentin mRNA and transcription factors-snail, as well as altered vimentin protein expression and localization were marked. Remarkably, the activities of MMP2 and MMP9 were suppressed. Conclusion:SRPK inhibitors potentially suppressed melanoma cell survivability and metastasis through the triggering of apoptotic proteins and dysregulating vimentin. These collected data serve as a basis for utilizing new alternative therapeutic strategies by targeting splicing regulator SRPK, for melanoma treatment.