AKR1B1 drives hyperglycemia-induced metabolic reprogramming in MASLD-associated hepatocellular carcinoma.

Background & Aims: The mechanism behind the progressive pathological alteration in metabolic dysfunction-associated steatotic liver disease/steatohepatitis (MASLD/MASH)-associated hepatocellular carcinoma (HCC) is poorly understood. In the present study, we investigated the role of the polyol pathway enzyme AKR1B1 in metabolic switching associated with MASLD/MASH and in the progression of HCC. Methods: AKR1B1 expression was estimated in the tissue and plasma of patients with MASLD/MASH, HCC, and HCC with diabetes mellitus. The role of AKR1B1 in metabolic switching in vitro was assessed through media conditioning, lentiviral transfection, and pharmacological probes. A proteomic and metabolomic approach was applied for the in-depth investigation of metabolic pathways. Preclinically, mice were subjected to a high-fructose diet and diethylnitrosamine to investigate the role of AKR1B1 in the hyperglycemia-mediated metabolic switching characteristic of MASLD-HCC. Results: A significant increase in the expression of AKR1B1 was observed in tissue and plasma samples from patients with MASLD/MASH, HCC, and HCC with diabetes mellitus compared to normal samples. Mechanistically, in vitro assays revealed that AKR1B1 modulates the Warburg effect, mitochondrial dynamics, the tricarboxylic acid cycle, and lipogenesis to promote hyperglycemia-mediated MASLD and cancer progression. A pathological increase in the expression of AKR1B1 was observed in experimental MASLD-HCC, and expression was positively correlated with high blood glucose levels. High-fructose diet + diethylnitrosamine-treated animals also exhibited statistically significant elevation of metabolic markers and carcinogenesis markers. AKR1B1 inhibition with epalrestat or NARI-29 inhibited cellular metabolism in in vitro and in vivo models. Conclusions: Pathological AKR1B1 modulates hepatic metabolism to promote MASLD-associated hepatocarcinogenesis. Aldose reductase inhibition modulates the glycolytic pathway to prevent precancerous hepatocyte formation. Impact and implications: This research work highlights AKR1B1 as a druggable target in metabolic dysfunction-associated steatotic liver disease (MASLD) and hepatocellular carcinoma (HCC), which could provide the basis for the development of new chemotherapeutic agents. Moreover, our results indicate the potential of plasma AKR1B1 levels as a prognostic marker and diagnostic test for MASLD and associated HCC. Additionally, a major observation in this study was that AKR1B1 is associated with the promotion of the Warburg effect in HCC.

Investigating the impact of inbuilt cold atmospheric pressure plasma on molecular assemblies of tryptophan enantiomers: in vitro fabrication of self-assembled supramolecular structures.

The advancements in understanding the phenomenon of plasma interactions with matter, coupled with the development of CAPP devices, have resulted in an interdisciplinary research topic of significant importance. This has led to the integration of various fields of science, including plasma physics, chemistry, biomedical sciences, and engineering. The reactive oxygen species and reactive nitrogen species generated from cold atmospheric plasma on interaction with biomolecules like proteins and peptides form various supramolecular structures. CAPP treatment of amino acids, which are the fundamental building blocks of proteins, holds potential in creating self-assembled supramolecular architectures. In this work, we demonstrate the process of self-assembly of aromatic amino acid tryptophan (Trp) enantiomers (l-tryptophan and d-tryptophan) into ordered supramolecular assemblies induced by the reactive species generated by a cold atmospheric pressure helium plasma jet. These enantiomers of tryptophan form organized structures as evidenced by FE-SEM. To assess the impact of CAPP treatment on the observed assemblies, we employed various analytical techniques such as zeta potential, dynamic light scattering and FTIR spectroscopy. Also, photoluminescence and time-resolved lifetime measurements revealed the transfiguration of individual Trp enantiomers. The LC-ESI-QTOF-MS analysis demonstrated that CAPP irradiation led to the incorporation of oxygenated ions into the pure Trp molecule. These studies of the self-assembly of Trp due to ROS and RNS interactions will help us to understand the assembly environment. This knowledge may be utilized to artificially design and synthesize highly ordered functional supramolecular structures using CAPP.

Urinary Metabolomics Identified Metabolic Perturbations Associated with Gutka, a Smokeless Form of Tobacco.

Gutka, a form of smokeless tobacco, is widely used in the Indian subcontinent and in other regions of South Asia. Smokeless tobacco exposure is most likely to increase the incidence of oral cancer in the Indian population, and metabolic changes are a hallmark of cancer. The development of biomarkers for early detection and better prevention measures for smokeless tobacco users at risk of oral cancer can be aided by studying urinary metabolomics and offering insight into altered metabolic profiles. This study aimed to investigate urine metabolic alterations among smokeless tobacco users using targeted LC-ESI-MS/MS metabolomics approaches to better understand the effects of smokeless tobacco on human metabolism. Smokeless tobacco users' specific urinary metabolomics signatures were extracted using univariate, multivariate analysis and machine learning methods. Statistical analysis identified 30 urine metabolites significantly associated with metabolomic alterations in humans who chew smokeless tobacco. Receiver operator characteristic (ROC) curve analysis evidenced the 5 most discriminatory metabolites from each approach that could differentiate between smokeless tobacco users and controls with higher sensitivity and specificity. An analysis of multiple-metabolite machine learning models and single-metabolite ROC curves revealed discriminatory metabolites capable of distinguishing smokeless tobacco users from nonusers more effectively with higher sensitivity and specificity. Furthermore, metabolic pathway analysis depicted several dysregulated pathways in smokeless tobacco users, including arginine biosynthesis, beta-alanine metabolism, TCA cycle, etc. This study devised a novel strategy to identify exposure biomarkers among smokeless tobacco users by combining metabolomics and machine learning algorithms.

Analysis of NSAIDs in Rat Plasma Using 3D-Printed Sorbents by LC-MS/MS: An Approach to Pre-Clinical Pharmacokinetic Studies.

Analytical sample preparation techniques are essential for assessing chemicals in various biological matrices. The development of extraction techniques is a modern trend in the bioanalytical sciences. We fabricated customized filaments using hot-melt extrusion techniques followed by fused filament fabrication-mediated 3D printing technology to rapidly prototype sorbents that extract non-steroidal anti-inflammatory drugs from rat plasma for determining pharmacokinetic profiles. The filament was prototyped as a 3D-printed sorbent for extracting small molecules using AffinisolTM, polyvinyl alcohol, and triethyl citrate. The optimized extraction procedure and parameters influencing the sorbent extraction were systematically investigated by the validated LC-MS/MS method. Furthermore, a bioanalytical method was successfully implemented after oral administration to determine the pharmacokinetic profiles of indomethacin and acetaminophen in rat plasma. The Cmax was found to be 0.33 ± 0.04 µg/mL and 27.27 ± 9.9 µg/mL for indomethacin and acetaminophen, respectively, at the maximum time (Tmax) (h) of 0.5-1 h. The mean area under the curve (AUC0-t) for indomethacin was 0.93 ± 0.17 µg h/mL, and for acetaminophen was 32.33± 10.8 µg h/mL. Owing to their newly customizable size and shape, 3D-printed sorbents have opened new opportunities for extracting small molecules from biological matrices in preclinical studies.

Extrusion 3D printing of minicaplets for evaluating in vitro & in vivo praziquantel delivery capability.

This study aimed to explore extrusion three dimensional (3D) printing technology to develop praziquantel (PZQ)-loaded minicaplets and evaluate their in vitro and in vivo delivery capabilities. PZQ-loaded minicaplets were 3D printed using a fused deposition modelling (FDM) principle-based extrusion 3D printer and were further characterized by different in vitro physicochemical and sophisticated analytical techniques. In addition, the % PZQ entrapment and in vitro PZQ release performance were evaluated using chromatographic techniques. It was in vitro observed that PZQ was fully released in the gastric pH medium within the period of gastric emptying, that is, 120 min, from the PZQ-loaded 3D printed minicaplets. Furthermore, in vivo pharmacokinetic (PK) profiles of PZQ-loaded 3D printed minicaplets were systematically evaluated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The PK profile of the PZQ-loaded 3D printed minicaplets was established using different parameters such as Cmax, Tmax, AUC0-t, AUC0-∞, and oral relative bioavailability (RBA). The Cmax value of pristine PZQ was found at 64.79 ± 13.99 ng/ml, while PZQ-loaded 3D printed minicaplets showed a Cmax of 263.16 ± 47.85 ng/ml. Finally, the PZQ-loaded 3D printed minicaplets showed 9.0-fold improved oral RBA compared with that of pristine PZQ (1.0-fold). Together, these observations potentiate the desired in vitro and improved in vivo delivery capabilities of PZQ from the PZQ-loaded 3D printed minicaplets.

Determination of bisphenols and parabens in cow urine distillate from India: implication of human exposure and risk assessment.

The exposures of a wide range of endocrine-disrupting chemicals to cows have been linked to diseases and are a major concern. In Indian scripts, cow urine is believed to be a treatment for many diseases. Nonetheless, exposure of bisphenols and parabens to cow urine distillates is unknown. Hence, in this study, we determined the concentration of bisphenols and parabens in commercially available cow urine distillate collected from India. The mean concentration of total bisphenols and parabens ranges from the limit of quantification (LOQ) to 149.3 ng/mL and 1479.88 ng/mL, respectively. Predominant bisphenol-F and bisphenol-A were accounted for 88.6% and 6% of total bisphenol concentration, respectively, in marketed cow urine distillate, whereas methyl and propyl parabens were dominant and accounted for 33% and 65%, respectively. The estimated mean daily intake (EDI) of bisphenol for males and females were 45.94 ng/kg-bw/day and 54.29 ng/kg-bw/day, respectively, while for parabens EDI was 455.35ng/kg-bw/day and 538.14 ng/kg-bw/day for males and females, respectively. Hazard quotient, to evaluate the potential risk of exposure, showed no risk in the studied samples. Even though the EDI results from the Monte-Carlo risk assessment analysis did not exceed the acceptable daily intake, their estrogenic actions cannot be ignored in general populations. The estrogenic activities contributed by parabens and bisphenol A measured by estradiol equivalency quotient (EEQ) ranged from 0.00033-42 pg/mL and 2.3 pg/mL, respectively. Our results revealed higher concentrations of bisphenols and parabens in cow urine distillates; hence, special attention should be given to the quality and safety of cow urine distillates. Moreover, strict guidelines should be enforced for the quality of cow urine distillates.