Liquid Chromatography-tandem Mass Spectrometry Research Articles

C-Cbl induced podocin ubiquitination contributes to the podocytes injury in diabetic nephropathy.

The ubiquitination function in diabetic nephropathy (DN) has attracted much attention, but there is a lack of information on its ubiquitylome profile. To examine the differences in protein content and ubiquitination in the kidney between db/db mice and db/m mice, we deployed liquid chromatography-mass spectrometry (LC-MS/MS) to conduct analysis. We determined 145 sites in 86 upregulated modified proteins and 66 sites in 49 downregulated modified proteins at the ubiquitinated level. Moreover, 347 sites among the 319 modified proteins were present only in the db/db mouse kidneys, while 213 sites among the 199 modified proteins were present only in the db/m mouse kidneys. The subcellular localization study indicated that the cytoplasm had the highest proportion of ubiquitinated proteins (31.87%), followed by the nucleus (30.24%) and the plasma membrane (20.33%). The enrichment analysis revealed that the ubiquitinated proteins are mostly linked to tight junctions, oxidative phosphorylation, and thermogenesis. Podocin, as a typical protein of slit diaphragm, whose loss is a crucial cause of proteinuria in DN. Consistent with the results of ubiquitination omics, the K261R mutant of podocin induced the weakest ubiquitination compared with the K301R and K370R mutants. As an E3 ligase, c-Cbl binds to podocin, and the regulation of c-Cbl can impact the ubiquitination of podocin. In conclusion, in DN, podocin ubiquitination contributes to podocyte injury, and K261R is the most significant site. c-Cbl participates in podocin ubiquitination and may be a direct target for preserving the integrity of the slit diaphragm structure, hence reducing proteinuria in DN.

Synergistic corrosion inhibition of rubber seed extract with KI on cold rolled steel in sulfuric acid solution

BackgroundPlant-based inhibitor has emerged an important topic owing to the combing merits of eco-friendly, low cost, rich natural resources as well as easy preparation. Rubber seed is the agricultural waste, and is mainly composed of oil and fatty acids those contain many polar oxygen atoms, and so it can be extracted to prepare the potential corrosion inhibitor. MethodsRubber seed extract (RSE) is prepared using reflux extraction method with the extracting solvent of ethanol aqueous solution. The synergistic inhibition effect of RSE and potassium iodide (KI) on the corrosion of cold rolled steel (CRS) in 0.50 M H2SO4 solution is firstly studied by weight loss, potentiodynamic polarization (PDP) curves and electrochemical impedance spectroscopy (EIS) methods. CRS surface was thoroughly characterized using metallographic microscope, scanning electron microscope (SEM), atomic force microscope (AFM), X-ray photoelectron spectroscopy (XPS) and contact angle measurements. FTIR, UV–vis and liquid chromatography mass spectrometry (LC-MS) compositional analysis of RSE were used to analyze the efficient components in RSE. Significant findingsThe results indicate that individual RSE or KI cannot function as a successful inhibitor to prevent CRS corrosion, but the mixture of RSE/KI performs better at inhibiting, with a maximal inhibition efficacy of up to 96.4% of 200 mg L−1 RSE/100 mg L−1 KI. The synergistic coefficient of the mixture is larger than 1, indicating that a true synergism. RSE/KI can drastically retard both cathodic and anodic processes, and interfacial capacitance falls as the charge transfer resistance of CRS/solution rises. It exhibits remarkable inhibition properties when the capacitor arc grows. There is only a small corrosion response in the inhibited media, as evidenced by the variations in electrical conductivity and ultraviolet spectrum before and after immersion. The hydrophilicity and surface roughness of CRS surface is dropped as a result of the introduction of RSE/KI. A series of metallurgical microscopy, AFM, SEM or contact angle tests indicate a synergistic inhibition effect is more pronounced for RSE/KI. XPS results clearly demonstrate that elements including Fe, S, N, and I are bonded to the surface of CRS. The primary ingredients of oleic acid, polyphenols, phenylpropanoids can also produce a strong synergism with KI.

Proximate Composition and Phytochemical Analysis of Malaysian Liberica sp. Coffee Bean and Its Pulp

Arabica, Robusta, and Liberica are the three main coffee species cultivated globally. Liberica coffee is a minor species, accounting for less than 1% of global cultivation. Due to favorable climatic conditions in Malaysia, Liberica coffee dominates coffee production, accounting for 73%, while Robusta makes up the remaining 27%. Nevertheless, the substantial coffee production resulted in approximately 15 million tons of discarded skin and pulp, contributing to environmental pollution. This study was conducted due to insufficient information and research on the proximate composition and phytochemical compounds of the coffee bean and pulp from Liberica sp. This study aims to determine the proximate composition of coffee beans and pulp extracts from Liberica sp. and to identify the phytochemical composition using liquid chromatography-mass spectrometry (LC-MS) analysis. The nutritional values of carbohydrates, protein, crude fiber, crude fat, and ash were obtained using proximate analysis. Coffee beans exhibited the highest value for crude protein (11.96%) and crude fiber (11.83%), whereas coffee pulp has the highest significant value for moisture content (68.81%) and ash (7.31%). LC-MS analysis shows emmotin A and deoxymiroestrol were the major phytochemical compounds. These findings contribute to understanding the nutritional value and phytochemical compounds of coffee beans and pulp from Liberica sp. that may contribute to sustainable waste management and other applications in the food and beverage industry.

Enhancing the efficiency of polypyrrole-dodecylbenzene sulfonic acid in-tube solid-phase microextraction coating for analysis of nitrogen-containing pesticides in water environments

Background: With the growing use of nitrogen-containing pesticides in agriculture, their residues in the environment have raised significant public health concerns.Objective: This study aimed to develop a novel PPy-DBSA/IT-SPME coating to enhance the detection efficiency of polar nitrogen-containing pesticides in water matrices.Methods: The preparation conditions were optimized, including pyrrole concentration at 7.0 mol/L, DBSA concentration at 0.014 mol/L, oxidant concentration at 0.35 mol/L, and a coating cycle repeated 10 times. High-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was employed for detection.Results: The novel coating effectively adsorbed 19 different categories of polar nitrogen-containing pesticides, including sulfonylureas, triazolopyrimidines, diphenyl ether herbicides, benzoylurea insecticides, and phenylurea herbicides, with unadsorbed rates below 10%. The analytical method achieved an average recovery rate of 61.92% to 115.21%, with an RSD below 5.0%. Detection and quantification limits ranged from 0.012 to 0.524 μg/L and 0.127 to 5.243 μg/L, respectively.Conclusion: The developed method is green, efficient, simple, and cost-effective. It offers an environmentally friendly and user-friendly approach for the detection of polar nitrogen-containing pesticide residues, demonstrating good recovery and precision. This method holds potential for wide application in environmental monitoring and food safety.

A comparative study of commercially available, minimally invasive, sampling methods on Early Neolithic humeri analysed via palaeoproteomics

Due to methodological advances in the archaeological sciences, an increasing number of archaeological specimens undergo destructive sampling. However, the preservation of cultural heritage is a primary concern. This leads to a dilemma between accessing sample material and obtaining sufficient information for a meaningful analytical outcome. Ideally, sampling a specimen would preserve the object for further macro, micro, and molecular analyses. For palaeoproteomics, a number of minimally invasive sampling approaches have been proposed, representing different benefits and limitations. There have been studies comparing a selection of these protocols, however, these have focused on specimens from a homogenous preservation environment using Zooarchaeology by Mass Spectrometry (ZooMS). Here we expand on earlier work by extending the comparison to specimens from two highly different preservation environments through both ZooMS and liquid-chromatography tandem mass spectrometry (LC-MS/MS). We compare five sampling approaches and seven extraction protocols in total, on 10 Bos sp. humeri from the Early Neolithic site of La Draga, Spain, utilising MALDI-ToF MS and LC-MS/MS to generate proteomic output, while assessing protocol invasiveness using microscopy and 3D imaging. Five humeri originate from Sector A, which is mostly related to dry, terrestrial preservation conditions, while the other five humeri stem from Sector B, which is characterised by its phreatic/aquatic preservation conditions. We show that there is a significant difference in protein recovery and taxonomic specificity between the sampling techniques applied, as well as between burial conditions. Additionally, various surface modifications were observed depending on the specific sampling technique applied. It is therefore essential to assess protein preservation for each sedimentological context within an archaeological site before performing extensive sampling, as protein preservation can be highly inter- and intra-site-specific.

Quantification of 25-hydroxyvitamin D3 in dried blood spots as compared to plasma among Indian adults

Background Vitamin D may play an important role in later-life physical and cognitive health. Vitamin D status is standardly assessed in serum and plasma; however, collection, transport, and storage costs make large epidemiologic studies challenging. We assessed the agreement of 25-hydroxyvitamin D3 (25(OH)D3) quantification from dried blood spots (DBS) as compared to standard plasma assessment among older Indian adults. Methods A total of 58 adults over 45 years of age who resided in Pune, India were enrolled in the study from July 2020 to June 2021. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to assess 25(OH)D3 concentrations in paired plasma and DBS samples. Results Plasma and DBS 25(OH)D3 concentrations were highly correlated (Pearson’s correlation = 0.976). The median 25(OH)D3 concentration of the study population assessed by plasma was 14.6 ng/mL (Q1=12.0, Q3= 18.1) while the median concentration assessed in DBS was 12.8 ng/mL (Q1=11.0, Q3= 16.6). 25(OH)D3 concentrations measured from DBS were on average 6% (95% CI: 2-13%) lower than concentrations assessed by plasma across the observed 25(OH)D3 distribution. Conclusions We found good agreement between 25(OH)D3 quantification between DBS and plasma and our findings indicate that DBS can be used in epidemiologic studies of vitamin D among Indian adults.

Nonthermal Plasma-Stimulated C-N Coupling from CH4 and N2 Depends on the Presence of Surface CHx and Plasma-Phase CN Species.

Formation of C-N containing compounds from plasma-catalytic coupling of CH4 and N2 over various transition metals (Ni, Pd, Cu, Ag, and Au) is investigated using a multimodal spectroscopic approach, combining polarization-modulation infrared reflection-absorption spectroscopy (PM-IRAS) and optical emission spectroscopy (OES). Through sequential experiments utilizing CH4 and N2 nonthermal plasmas, we minimize plasma-phase reactions and identify key intermediates for C-N coupling on metal surfaces. Results show that simultaneous CH4 and N2 exposure with plasma stimulation produces surface C-N species. However, N2-CH4 sequential exposure does not lead to C-N species formation, while CH4-N2 sequential exposure reveals the presence of CHx surface species and CN radical species as key precursors to C-N species formation. From further analysis using X-ray photoelectron spectroscopy and liquid chromatography-mass spectrometry, the influence of exposure conditions on the degree of nitrogen incorporation and the nature of C-N species formed were revealed. The work highlights the importance of surface chemistry and exposure conditions in surface C-N coupling with plasma stimulation.

Lipoprotein glomerulopathy with markedly increased arterial stiffness successfully treated with a combination of fenofibrate and losartan: a case report

BackgroundLipoprotein glomerulopathy (LPG) is a apolipoprotein E (ApoE)-related glomerular disease and has been associated with type III hyperlipidemia. Without appropriate treatment, chronic kidney disease (CKD) caused by LPG progresses, and approximately half of the patients develop end-stage kidney disease within 1–27 years of disease onset. However, few studies have highlighted the clinical course of cardiovascular diseases (CVDs) in patients with LPG. Herein, we report the first case of LPG in which the CVD risk was assessed using arterial stiffness.Case presentationA 32-year-old Japanese man was referred to our hospital due to persistent proteinuria. Kidney biopsy showed markedly dilated capillary lumens containing pale-stained thrombi, which stained positively with Oil Red O. Electron microscopy revealed the presence of thrombi in the capillary lumen with low electron density and vacuoles of various sizes in part of the thrombi. Toluidine blue and Sudan IV stains were used to stain the thin sections of Epon-embedded tissue samples for electron microscopy. Sudan IV-positive droplets were observed in the capillary lumens, vascular walls, and cytoplasm of tubular cells. Increased serum ApoE concentration was observed. Liquid chromatography-tandem mass spectrometry of laser-microdissected glomeruli from paraffin sections revealed an increase in ApoE. Direct deoxyribonucleic acid sequencing of ApoE revealed a heterozygous ApoE Sendai mutation (Arg145Pro). The patient was finally diagnosed with LPG with heterozygosity for ApoE-Sendai mutation (Arg145Pro). Notably, at the time of diagnosis, he had markedly increased arterial stiffness for his age. Arterial stiffness was measured using brachial-ankle pulse wave velocity (baPWV), which was equivalent to that of a 56-year-old man. After three months of treatment with fenofibrate and losartan, a significant reduction in proteinuria was achieved along with an improvement in baPWV. Furthermore, these effects were maintained despite the lack of decrease in serum ApoE levels.ConclusionHerein, we report the case of a patient with LPG with markedly increased arterial stiffness at the time of diagnosis, in whom combination therapy with fenofibrate and losartan successfully improved proteinuria and arterial stiffness. To the best of our knowledge, this is the first case report of LPG in which CVD risk was assessed using arterial stiffness.

Effect of the storage process on quality characteristics of hemp-enriched "tsoureki" a rich-dough baked Greek product.

The effects of the storage process on the quality characteristics of a hemp-enriched "tsoureki" (a rich-dough baked Greek product, rich-dough baked product [RDBP]) were investigated. The wheat flour was substituted with defatted hemp flour at selected ratios (0%-50% hemp:wheat flour). The baked products were stored at 25°C and at specific time intervals (0, 1, 4, 7, 10, and 14 days), and their properties were determined, including moisture content, water activity, structure, texture, color, total phenolic content (TPC), and antioxidant activity. Moreover, analyses of phenolic compounds were performed using quadrupole time of flight liquid chromatography-mass spectroscopy, identifying 14 compounds. Both the first-order kinetic model and modified Avrami equation, including the hemp-to-wheat ratio effect in the rate constant, well described the changes in the quality characteristics. The results showed that storage time and hemp incorporation significantly affected the quality of the product. Water activity decreased from 0.901 to 0.859, whereas moisture content decreased from 30.52%-32.33% (0 days) to 26.97%-27.02% w.b. (14 days) with storage time for all hemp additions. Hardness was greatly affected by hemp flour addition and approached 14.72 and 17.85 N after 14 days of storage for 30% and 50% substitutions, respectively. Springiness and cohesiveness decreased with hemp addition and storage time. The color difference of the hemp-enriched products compared to the control sample increased during storage. TPC increased due to the addition of hemp flour, whereas 14 compounds were identified. Based on property correlation, the hemp-enriched RDBP-tsoureki held its high-quality characteristics for 7 days of storage and contained a significant amount of bioactive compounds. PRACTICAL APPLICATION: Industrially produced, defatted hemp is a promising byproduct that can be used to nutritionally enhance baked goods. Modeling results can be used for the prediction of the properties that define product storage ability and also that the hemp-enriched, rich dough-baked Greek "tsoureki" could be produced while maintaining total phenolic content and antioxidant activity during 7 days of storage. These findings are expected to be used in the future in baked goods industry applications to produce goods with an improved nutritional profile.

Reproducible mass spectrometry data processing and compound annotation in MZmine 3.

Untargeted mass spectrometry (MS) experiments produce complex, multidimensional data that are practically impossible to investigate manually. For this reason, computational pipelines are needed to extract relevant information from raw spectral data and convert it into a more comprehensible format. Depending on the sample type and/or goal of the study, a variety of MS platforms can be used for such analysis. MZmine is an open-source software for the processing of raw spectral data generated by different MS platforms. Examples include liquid chromatography-MS, gas chromatography-MS and MS-imaging. These data might typically be associated with various applications including metabolomics and lipidomics. Moreover, the third version of the software, described herein, supports the processing of ion mobility spectrometry (IMS) data. The present protocol provides three distinct procedures to perform feature detection and annotation of untargeted MS data produced by different instrumental setups: liquid chromatography-(IMS-)MS, gas chromatography-MS and (IMS-)MS imaging. For training purposes, example datasets are provided together with configuration batch files (i.e., list of processing steps and parameters) to allow new users to easily replicate the described workflows. Depending on the number of data files and available computing resources, we anticipate this to take between 2 and 24 h for new MZmine users and nonexperts. Within each procedure, we provide a detailed description for all processing parameters together with instructions/recommendations for their optimization. The main generated outputs are represented by aligned feature tables and fragmentation spectra lists that can be used by other third-party tools for further downstream analysis.

Integrated metabolomic and transcriptomic analysis of triterpenoid accumulation in the roots of Codonopsis pilosula var. modesta (Nannf.) L.T.Shen at different altitudes.

Codonopsis Radix is a beneficial traditional Chinese medicine, and triterpenoid are the major bioactive constituents. Codonopsis pilosula var. modesta (Nannf.) L.T.Shen (CPM) is a precious variety of Codonopsis Radix, which is distributed at high mountain areas. The environment plays an important role in the synthesis and metabolism of active ingredients in medicinal plants, but there is no report elaborating on the effect of altitude on terpenoid metabolites accumulation in CPM. This study aims to analyse the effects of altitude on triterpenoid biosynthetic pathways and secondary metabolite accumulation in CPM. The untargeted metabolomics based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and 10 triterpenoids based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) method were analysed at the low-altitude (1480 m) and high-altitude (2300 m) CPM fresh roots. The transcriptome based on high-throughput sequencing technology were combined to analyse the different altitude CPM triterpenoid biosynthetic pathways. A total of 17,351 differentially expressed genes (DEGs) and 55 differentially accumulated metabolites (DAMs) were detected from the different altitude CPM, and there are significant differences in the content of the 10 triterpenoids. The results of transcriptome study showed that CPM could significantly up-regulate the gene expression levels of seven key enzymes in the triterpenoid biosynthetic pathway. The CPM at high altitude is more likely to accumulate triterpenes than those at low altitude, which was related to the up-regulation of the gene expression levels of seven key enzymes. These results expand our understanding of how altitude affects plant metabolite biosynthesis.

Determination of 13 potential anti-obesity agents in hair by dispersive liquid-liquid microextraction-assisted UHPLC-MS/MS

As the adulteration of dietary supplements with synthetic drugs remains a prevalent issue, the inclusion of anti-obesity agents may pose health risks, potentially leading to central nervous system or cardiovascular diseases. However, surveillance studies on the use of anti-obesity agents by the Chinese population are limited. This study aims to establish an efficient and rapid hair pretreatment method using dispersive liquid-liquid microextraction (DLLME) combined with high-speed grinding and develop a sensitive and accurate analytical method employing ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) for detecting 13 potential anti-obesity agents in hair samples. Herein, hair samples were washed sequentially with 0.1% sodium dodecyl sulfate (SDS), water and acetone, and then ground at high speed using 1 mL of an extraction solution (internal standard solution-n-butanol-1.2 mol/L Na2HPO4, pH10.0, 100:400:500, v/v/v for procaterol; internal standard solution-ethyl acetate-1.2 mol/L Na2HPO4, pH8.0, 100:300:600, v/v/v for other 12 anti-obesity agents) while simultaneously performing DLLME. The developed method successfully detected 13 anti-obesity agents within 11 min, including bambuterol, clenbuterol, ractopamine, clorprenaline, formoterol, salbutamol, terbutaline, procaterol, phentermine, bupropion, sibutramine, desmethyl sibutramine, and N,N-didesmethyl sibutramine, which improved the screening efficiency. The calibration curves exhibited good linearity of 0.025–5 ng/mg, achieving correlation coefficients of r ≥ 0.99. The lower limits of quantification (LLOQs) for the analytes were 0.025 ng/mg, demonstrating acceptable levels of accuracy and precision. Recovery rates ranged between 73.30% and 107.47% across the three concentrations of 0.075, 0.375, and 3.75 ng/mg. The validated method was successfully applied to 369 real cases and detected six analytes, including bambuterol, salbutamol, terbutaline, sibutramine, desmethyl sibutramine, and N,N-didesmethyl sibutramine. This method offers several advantages, including simple pretreatment, high extraction efficiency, rapid extraction, solvent economy, and pollution mitigation, making it highly suitable for large-scale surveillance of usage of added anti-obesity agents.

The mediating role of kynurenine pathway metabolites on the relationship between inflammation and muscle mass in oldest-old men.

Tryptophan (TRP) metabolites along the kynurenine (KYN) pathway (KP) have been found to influence muscle. Pro-inflammatory cytokines are known to stimulate the degradation of TRP down the KP. Given that both inflammation and KP metabolites have been connected with loss of muscle, we assessed the potential mediating role of KP metabolites on inflammation and muscle mass in older men. 505 men (85.0±4.2yrs) from the Osteoporotic Fractures in Men cohort study with measured D3-creatine dilution (D3Cr) muscle mass, KP metabolites, and inflammation markers (C-reactive protein (CRP), alpha-1-acid glycoprotein (AGP) and a subsample (n=305) with interleukin (IL-6, IL-1β, IL-17A) and tumor necrosis factor-α (TNF-α)) were included in the analysis. KP metabolites and inflammatory markers were measured using liquid chromatography-tandem mass spectrometry and immunoassays, respectively. 23-92% of the inverse relationship between inflammatory markers and D3Cr muscle mass was mediated by KP metabolites (indirect effect p<0.05). 3-hydroxyanthranilic acid (3-HAA), quinolinic acid (QA), TRP, xanthurenic acid (XA), KYN/TRP, 3-hydroxykynurenine (3-HK)/3-HAA, QA/3-HAA, and nicotinamide (NAM)/QA mediated the AGP relationship. 3-HAA, QA, KYN/TRP, 3-HK/XA, HKr ratio, 3-HK/3-HAA, QA/3-HAA, and NAM/QA mediated the CRP. KYN/TRP, 3-HK/XA, and NAM/QA explained the relationship for IL-6 and 3-HK/XA and QA/3-HAA for TNF-α. No mediation effect was observed for the other cytokines (indirect effect p>0.05). KP metabolites, particularly higher ratios of KYN/TRP, 3-HK/XA, 3-HK/3-HAA, QA/3-HAA and a lower ratio of NAM/QA, mediated the relationship between inflammation and low muscle mass. Our preliminary cross-sectional data suggest that interventions to alter D3Cr muscle mass may focus on KP metabolites rather than inflammation per se.

Gut microbiota induced abnormal amino acids and their correlation with diabetic retinopathy.

To explore the correlation of gut microbiota and the metabolites with the progression of diabetic retinopathy (DR) and provide a novel strategy to elucidate the pathological mechanism of DR. The fecal samples from 32 type 2 diabetes patients with proliferative retinopathy (PDR), 23 with non-proliferative retinopathy (NPDR), 27 without retinopathy (DM), and 29 from the sex-, age- and BMI- matched healthy controls (29 HC) were analyzed by 16S rDNA gene sequencing. Sixty fecal samples from PDR, DM, and HC groups were assayed by untargeted metabolomics. Fecal metabolites were measured using liquid chromatography-mass spectrometry (LC-MS) analysis. Associations between gut microbiota and fecal metabolites were analyzed. A cluster of 2 microbiome and 12 metabolites accompanied with the severity of DR, and the close correlation of the disease progression with PDR-related microbiome and metabolites were found. To be specific, the structure of gut microbiota differed in four groups. Diversity and richness of gut microbiota were significantly lower in PDR and NPDR groups, than those in DM and HC groups. A cluster of microbiome enriched in PDR group, including Pseudomonas, Ruminococcaceae-UCG-002, Ruminococcaceae-UCG-005, Christensenellaceae-R-7, was observed. Functional analysis showed that the glucose and nicotinate degradations were significantly higher in PDR group than those in HC group. Arginine, serine, ornithine, and arachidonic acid were significantly enriched in PDR group, while proline was enriched in HC group. Functional analysis illustrated that arginine biosynthesis, lysine degradation, histidine catabolism, central carbon catabolism in cancer, D-arginine and D-ornithine catabolism were elevated in PDR group. Correlation analysis revealed that Ruminococcaceae-UCG-002 and Christensenellaceae-R-7 were positively associated with L-arginine, ornithine levels in fecal samples. This study elaborates the different microbiota structure in the gut from four groups. The relative abundance of Ruminococcaceae-UCG-002 and Parabacteroides are associated with the severity of DR. Amino acid and fatty acid catabolism is especially disordered in PDR group. This may help provide a novel diagnostic parameter for DR, especially PDR.

Metabolomics reveals the implication of acetoacetate and ketogenic diet therapy in radioiodine-refractory differentiated thyroid carcinoma.

Patients with radioiodine-refractory (RAIR) differentiated thyroid carcinoma (DTC; RAIR-DTC) have a poor prognosis. The aim of this study was to provide new insights and possibilities for the diagnosis and treatment of RAIR-DTC. The metabolomics of 24 RAIR-DTC and 18 non-radioiodine-refractory (NonRAIR) DTC patients samples were analyzed by liquid chromatograph-mass spectrometry. Cellular radioiodine uptake was detected with γ counter. Sodium iodide symporter (NIS) expression and thyroid stimulating hormone receptor (TSHR) were measured by Western blot analysis. CCK8 and colony formation assays were used to measure cellular proliferation. Scratch and transwell assays were performed to assess cell migration and invasion. Annexin V/PI staining was used to detect cell apoptosis. Cell growth in vivo was evaluated by a tumor xenograft model. The acetoacetate (AcAc) level was measured by ELISA. Pathological changes, Ki67, NIS, and TSHR expression were investigated by immunohistochemistry. The metabolite profiles of RAIR could be distinguished from those of NonRAIR, with AcAc significantly lower in RAIR. The significantly different metabolic pathway was ketone body metabolism. AcAc increased NIS and TSHR expression and improved radioiodine uptake. AcAc inhibited cell proliferation, migration, and invasion, and as well promoted cell apoptosis. Ketogenic diet (KD) elevated AcAc levels and significantly suppressed tumor growth, as well as improved NIS and TSHR expression. Significant metabolic differences were observed between RAIR and NonRAIR, and ketone body metabolism might play an important role in RAIR-DTC. AcAc improved cellular iodine uptake and had antitumor effects for thyroid carcinoma. KD might be a new therapeutic strategy for RAIR-DTC.

Metabolomic prediction of severe maternal and newborn complications in preeclampsia

IntroductionPreeclampsia (PreE) remains a major source of maternal and newborn complications. Prenatal prediction of these complications could significantly improve pregnancy management.ObjectivesUsing metabolomic analysis we investigated the prenatal prediction of maternal and newborn complications in early and late PreE and investigated the pathogenesis of such complications.MethodsSerum samples from 76 cases of PreE (36 early-onset and 40 late-onset), and 40 unaffected controls were collected. Direct Injection Liquid Chromatography–Mass Spectrometry combined with Nuclear Magnetic Resonance (NMR) spectroscopy was performed. Logistic regression analysis was used to generate models for prediction of adverse maternal and neonatal outcomes in patients with PreE. Metabolite set enrichment analysis (MSEA) was used to identify the most dysregulated metabolites and pathways in PreE.ResultsForty-three metabolites were significantly altered (p < 0.05) in PreE cases with maternal complications and 162 metabolites were altered in PreE cases with newborn adverse outcomes. The top metabolite prediction model achieved an area under the receiver operating characteristic curve (AUC) = 0.806 (0.660–0.952) for predicting adverse maternal outcomes in early-onset PreE, while the AUC for late-onset PreE was 0.843 (0.712–0.974). For the prediction of adverse newborn outcomes, regression models achieved an AUC = 0.828 (0.674–0.982) in early-onset PreE and 0.911 (0.828–0.994) in late-onset PreE. Profound alterations of lipid metabolism were associated with adverse outcomes.ConclusionPrenatal metabolomic markers achieved robust prediction, superior to conventional markers for the prediction of adverse maternal and newborn outcomes in patients with PreE. We report for the first-time the prediction and metabolomic basis of adverse maternal and newborn outcomes in patients with PreE.

Bio-electrochemical degradation of carbamazepine (CBZ): A comprehensive study on effectiveness, degradation pathway, and toxicological assessment

Recent attention on the detrimental effects of pharmaceutically active compounds (PhACs) in natural water has spurred researchers to develop advanced wastewater treatment methods. Carbamazepine (CBZ), a widely recognized anticonvulsant, has often been a primary focus in numerous studies due to its prevalence and resistance to breaking down. This study aims to explore the effectiveness of a bio-electrochemical system in breaking down CBZ in polluted water and to assess the potential harmful effects of the treated wastewater. The results revealed bio-electro degradation process demonstrated a collaborative effect, achieving the highest CBZ degradation compared to electrodegradation and biodegradation techniques. Notably, a maximum CBZ degradation efficiency of 92.01% was attained using the bio-electrochemical system under specific conditions: Initial CBZ concentration of 60 mg/L, pH level at 7, 0.5% (v/v) inoculum dose, and an applied potential of 10 mV. The degradation pathway established by identifying intermediate products via High-Performance Liquid Chromatography-Mass Spectrometry, revealed the complete breakdown of CBZ without any toxic intermediates or end products. This finding was further validated through in vitro and in vivo toxicity assays, confirming the absence of harmful remnants after the degradation process.

Comparative proteomic analysis of dental pulp from supernumerary and normal permanent teeth

ObjectivesTo obtain and compare the protein profiles of supernumerary and normal permanent dental pulp tissues.Materials and methodsDental pulp tissues were obtained from supernumerary and normal permanent teeth. Proteins were extracted and analyzed by liquid chromatography-tandem mass spectrometry (LC/MS-MS). Protein identification and quantification from MS data was performed with MaxQuant. Statistical analysis was conducted using Metaboanalyst to identify differentially expressed proteins (DEPs) (P-value < 0.05, fold-change > 2). Gene Ontology enrichment analyses were performed with gProfiler.ResultsA total of 3,534 proteins were found in normal dental pulp tissue and 1,093 in supernumerary dental pulp tissue, with 174 DEPs between the two groups. This analysis revealed similar functional characteristics in terms of cellular component organization, cell differentiation, developmental process, and response to stimulus, alongside exclusive functions unique to normal permanent dental pulp tissues such as healing, vascular development and cell death. Upon examination of DEPs, these proteins were associated with the processes of wound healing and apoptosis.ConclusionsThis study provides a comprehensive understanding of the protein profile of dental pulp tissue, including the first such profiling of supernumerary permanent dental pulp. There are functional differences between the proteomic profiles of supernumerary and normal permanent dental pulp tissue, despite certain biological similarities between the two groups. Differences in protein expression were identified, and the identified DEPs were linked to the healing and apoptosis processes.Clinical relevanceThis discovery enhances our knowledge of supernumerary and normal permanent pulp tissue, and serves as a valuable reference for future studies on supernumerary teeth.

Association Between Vitamin D and Asymptomatic Hyperuricaemia Among Adults––An Observational Study

Introduction: The relationship between Vitamin D levels and asymptomatic hyperuricaemia is gaining attention due to its potential implications for public health. This study investigates this association, focussing on the interplay between Vitamin D, serum uric acid and parathyroid hormone (PTH) levels in adults. Materials and Methods: This cross-sectional observational study, conducted from April to June 2023, involved 100 adult participants (65 males, 35 females) from the general population. Participants were selected based on specific inclusion and exclusion criteria. Data collection included a comprehensive questionnaire, physical examinations and blood sample analysis. Vitamin D levels were measured using liquid chromatography-tandem mass spectrometry, uric acid levels through an enzymatic colorimetric test, and PTH levels via a chemiluminescent immunoassay. Statistical analysis was performed using SPSS version 25.0, IBM Corp, Chicago, IL, USA. Results: The study found an inverse correlation between Vitamin D and serum uric acid levels. Participants with lower Vitamin D levels had higher serum uric acid and PTH levels. The mean serum Vitamin D level was 32.17 ng/mL, and the mean serum uric acid level was 9.99 mg/dL. The study categorised Vitamin D levels as deficient, insufficient and sufficient, observing significant variations in PTH and uric acid levels across these categories. Conclusion: The study suggests a significant relationship between Vitamin D levels and serum uric acid concentrations in adults, with implications for the management of asymptomatic hyperuricaemia and related metabolic conditions. Despite limitations, such as the cross-sectional design and small sample size, this study highlights the potential significance of Vitamin D levels in the management of asymptomatic hyperuricaemia and related metabolic disorders.

A panel of four plasma amino acids is a promising biomarker for newly diagnosed bladder cancer

BackgroundMetastasis and recurrence are the main causes of death in post-operative bladder cancer (BC), emphasizing the importance of exploring early-stage diagnostic markers. Serum biomarkers constitute a promising diagnostic approach for asymptomatic stage cancer as they are non-invasive, have high accuracy and low cost. AimsTo correlate concentrations of plasma amino acids with BC progression to assess their utility as an early-stage diagnostic. MethodsNewly diagnosed BC patients (n=95) and normal controls (n=96) were recruited during the period from 1 December 2018 to 30 December 2020. General and food frequency questionnaires established their basic information and dietary intake data. Venous blood samples were collected from fasting subjects and used to detect levels of plasma amino acids by liquid chromatography-mass spectrometry. Verification was performed on the GSE13507 transcriptome gene expression matrix of BC from Gene Expression Omnibus (GEO) database. ResultsEleven amino acids have been identified as altered in the plasma of newly diagnosed BC patients compared to controls (P < 0.05). Adjusted by gender, education, smoking and other factors, plasma ornithine level (OR = 0.256, 95% CI: 0.104 ∼ 0.630) is a protective factor for BC, plasma levels of methionine (OR = 3.460, 95% CI: 1.384 ∼ 8.651), arginine (OR = 3.851, 95% CI: 1.542 ∼ 9.616), and glutamate (OR = 3.813, 95% CI: 1.543 ∼ 9.419) are all risk factors for BC. ROC analysis demonstrated that the combination of plasma ornithine, methionine, arginine and glutamate could accurately diagnose BC (AUC = 0.84, 95% CI: 0.747 ∼ 0.833). In addition, the mRNA level of arginase 1 was decreased (P < 0.05), while the inducible nitric oxide synthase was increased significantly, which may be linked with the disturbance of arginine metabolism in BC patients. Further analysis of GEO database confirmed the role of arginine metabolism. ConclusionA biomarker panel containing four amino acids may provide a feasible strategy for the early diagnosis of BC. However, further validation is required through prospective studies.