Metabolic Activity Research Articles

Comparative Analysis of Microbial Diversity and Metabolic Profiles during the Spontaneous Fermentation of Jerusalem Artichoke (Helianthus tuberosus L.) Juice.

Jerusalem artichoke juice is valued for its nutritional content and health benefits. Spontaneous fermentation enhances its flavor, quality, and functional components through microbial metabolic activities. This study used high-throughput sequencing to analyze microbial community changes, and LC-MS and GC-MS to detect secondary metabolites and flavor compounds during fermentation. During natural fermentation, beneficial bacteria like Lactobacillus and Pediococcus increased, promoting lactic acid production and inhibiting harmful bacteria, while environmental bacteria decreased. Similarly, fungi shifted from environmental types like Geosmithia and Alternaria to fermentation-associated Pichia and Penicillium. A total of 1666 secondary metabolites were identified, with 595 upregulated and 497 downregulated. Key metabolic pathways included phenylpropanoid biosynthesis, with significant increases in phenylalanine, tryptophan, and related metabolites. Lipid and nucleotide metabolism also showed significant changes. Flavor compounds, including 134 identified alcohols, esters, acids, and ketones, mostly increased in content after fermentation. Notable increases were seen in Phenylethyl Alcohol, Ethyl Benzenepropanoate, 3-Methylbutyl Butanoate, Ethyl 4-Methylpentanoate, 5-Ethyl-3-Hydroxy-4-Methyl-2(5H)-Furanone, Ethyl Decanoate, Hexanoic Acid, and 1-Octanol. γ-aminobutyric acid (GABA) and other functional components enhanced the health value of the juice. This study provides insights into microbial and metabolic changes during fermentation, aiding in optimizing processes and improving the quality of fermented Jerusalem artichoke juice for functional food development.

Combined PET Radiotracer Approach Reveals Insights into Stromal Cell-Induced Metabolic Changes in Pancreatic Cancer In Vitro and In Vivo.

Background/Objective Pancreatic stellate cells (PSCs) in pancreatic adenocarcinoma (PDAC) are producing extracellular matrix, which promotes the formation of a dense fibrotic microenvironment. This makes PDAC a highly heterogeneous tumor-stroma-driven entity, associated with reduced perfusion, limited oxygen supply, high interstitial fluid pressure, and limited bioavailability of therapeutic agents. Methods In this study, spheroid and tumor xenograft models of human PSCs and PanC-1 cells were characterized radiopharmacologically using a combined positron emission tomography (PET) radiotracer approach. [18F]FDG, [18F]FMISO, and [18F]FAPI-74 were employed to monitor metabolic activity, hypoxic metabolic state, and functional expression of fibroblast activation protein alpha (FAPα), a marker of activated PSCs. Results In vitro, PanC-1 and multi-cellular tumor spheroids demonstrated comparable glucose uptake and hypoxia, whereas FAPα expression was significantly higher in PSC spheroids. In vivo, glucose uptake as well as the transition to hypoxia were comparable in PanC-1 and multi-cellular xenograft models. In mice injected with PSCs, FAPα expression decreased over a period of four weeks post-injection, which was attributed to the successive death of PSCs. In contrast, FAPα expression increased in both PanC-1 and multi-cellular xenograft models over time due to invasion of mouse fibroblasts. Conclusion The presented models are suitable for subsequently characterizing stromal cell-induced metabolic changes in tumors using noninvasive molecular imaging techniques.

Cytotoxicity assessment of eluates from vacuum-forming thermoplastics.

This study aimed to evaluate possible cytotoxic effects of thermoplastic materials commonly used for occlusal splints and orthodontic appliances. Seven thermoplastics were included: three variants of the Essix sheets (C+, Plus, and Tray Rite; Dentsply Sirona), three thermoplastics (Bleach Heavy, Splint, and X-Heavy; Cavex Holland) and Invisalign (Align Technology). Cylindrical specimens (n = 24; 10mm diameter) were incubated in cell culture medium for 24h and 14 days. After incubation, the medium was collected, serially diluted, and dosed to primary human gingival fibroblasts in triplicate. Medium processed like the samples was used as negative control. Cell viability was evaluated by XTT and LDH assay to assess metabolic activity and membrane integrity, respectively. Next, cell cycle was assessed with flow cytometry after exposing HGFs to undiluted extracts. The 24-hour and 14-day extracts did not evoke cytotoxicity after 24-hour incubation. No significant differences in cell viability (one-way ANOVA, p > 0.05 ) in the XTT and LDH assays or in cell cycle distribution between the different materials (two-way ANOVA, p > 0.05 ). The thermoplastics tested in the study showed no evident in-vitro cytotoxic effects. Further investigation should focus on determining which compounds are released from thermoplastic materials and assessing potential toxicity related to exposure to these compounds. Our study adds to the growing body of evidence on the biocompatibility of dental thermoplastics. This can aid clinical decision-making, as thermoplastics are expected to be safe to use in terms of cytotoxicity.

Pancreatic cancer tumor organoids exhibit subtype-specific differences in metabolic profiles

BackgroundPancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease characterized by complex metabolic rewiring that enables growth in changing nutrient availability and oxygen conditions. Transcriptome-based prognostic PDAC tumor subtypes, known as ‘basal-like’ and ‘classical’ subtypes are associated with differences in metabolic gene expression including genes involved in glycolysis. Tumor subtype-specific metabolism phenotypes may provide new targets for treatment development in PDAC, but their functional relevance has not been fully elucidated. We aimed to investigate differences in metabolic profiles and transcriptomes in tumor models derived from patients with basal-like and classical tumors.MethodsPatient-derived organoids (PDOs) were established from tumor biopsies collected from patients with metastatic PDAC, including three PDOs from basal-like and five PDOs from classical tumors. Metabolic analyses included assessment of differences in metabolic activity using Seahorse Glycolysis and Mito Stress tests and 13C-glucose metabolites tracing analysis. In order to investigate the influence of mitochondrial pyruvate transport on metabolic differences, PDOs were treated with the mitochondrial pyruvate carrier 1 (MPC1) inhibitor UK-5099. Prognostic relevance of MPC1 was determined using a tumor tissue microarray (TMA) in resectable, and proteomics profiling in metastatic PDAC datasets. Whole genome and transcriptome sequencing, differential gene expression and gene set enrichment analyses were performed in PDOs.ResultsMetastatic PDAC PDOs showed subtype-specific differences in glycolysis and oxidative phosphorylation (OXPHOS). Basal-like tumor-derived PDOs had a lower baseline extracellular acidification rate, but higher glycolytic reserves and oxygen consumption rate (OCR) than classical tumor-derived PDOs. OCR difference was eliminated following treatment with UK-5099. In the 13C-glucose metabolites tracing experiment, a basal-like tumor PDO showed lower fractions of some M + 2 metabolites but higher sensitivity to UK-5099 mediated reduction in M + 2 metabolites than a classical tumor PDO. Protein level analyses revealed lower MPC1 protein levels in basal-like PDAC cases and association of low MPC1 levels with clinicopathologic parameters of tumor aggressiveness in PDAC. PDO differential gene expression analyses identified additional subtype-specific cellular pathways and potential disease outcome biomarkers.ConclusionsOur findings point to distinct metabolic profiles in PDAC subtypes with basal-like tumor PDOs showing higher OXPHOS and sensitivity to MPC1 inhibition. Subtypes-specific metabolic vulnerabilities may be exploited for selective therapeutic targeting.

Early elevations in arterial pressure: a contributor to rapid depressive symptom emergence in female Zucker rats with metabolic disease?

One of the growing challenges to public health and clinical outcomes is the emergence of cognitive impairments, particularly depressive symptom severity, because of chronic elevations in metabolic disease and cerebrovascular disease risk. To more clearly delineate these relationships and to assess the potential for sexual dimorphism, we used lean (LZR) and obese Zucker rats (OZR) of increasing age to determine relationships between internal carotid artery (ICA) hemodynamics, cerebral vasculopathies, and the emergence of depressive symptoms. Male OZR exhibited progressive elevations in perfusion pressure within the ICA, which were paralleled by endothelial dysfunction, increased cerebral arterial myogenic activation, and reduced cerebral cortex microvessel density. In contrast, female OZR exhibited a greater degree of ICA hypertension than male OZR but maintained normal endothelial function, myogenic activation, and microvessel density to an older age range than did males. Although both male and female OZR exhibited significant and progressive elevations in depressive symptom severity, these were significantly worse in females. Finally, plasma cortisol concentration was elevated higher and at a younger age in female OZR as compared with males, and this difference was maintained to final animal usage at ∼17 wk of age. These results suggest that an increased severity of blood pressure waves may penetrate the cerebral circulation more deeply in female OZR than in males, which may predispose the females to a more severe emergence of depressive symptoms with chronic metabolic disease, whereas males may be more predisposed to more direct cerebral vasculopathies (e.g., stroke, transient ischemic attack).NEW & NOTEWORTHY We provide novel insight that the superior maintenance of cerebrovascular endothelial function in female versus male rats with chronic metabolic disease buffers myogenic activation of cerebral resistance arteries/arterioles despite worsening hypertension. As hypertension development is earlier and more severe in females, potentially due to an elevated stress response, the blunted myogenic activation allows greater arterial pressure wave penetrance into the cerebral microcirculation and is associated with accelerated emergence/severity of depressive symptoms in obese female rats.

The role of lactylation in plasma cells and its impact on rheumatoid arthritis pathogenesis: insights from single-cell RNA sequencing and machine learning.

Rheumatoid arthritis (RA) is a chronic autoimmune disorder characterized by persistent synovitis, systemic inflammation, and autoantibody production. This study aims to explore the role of lactylation in plasma cells and its impact on RA pathogenesis. We utilized single-cell RNA sequencing (scRNA-seq) data and applied bioinformatics and machine learning techniques. A total of 10,163 cells were retained for analysis after quality control. Clustering analysis identified 13 cell clusters, with plasma cells displaying the highest lactylation scores. We performed pathway enrichment analysis to examine metabolic activity, such as oxidative phosphorylation and glycolysis, in highly lactylated plasma cells. Additionally, we employed 134 machine learning algorithms to identify seven core lactylation-promoting genes and constructed a diagnostic model with an average AUC of 0.918. The RA lactylation score (RAlac_score) was significantly elevated in RA patients and positively correlated with immune cell infiltration and immune checkpoint molecule expression. Differential expression analysis between two plasma cell clusters revealed distinct metabolic and immunological profiles, with cluster 2 demonstrating increased immune activity and extracellular matrix interactions. qRT-PCR validation confirmed that NDUFB3, NGLY1, and SLC25A4 are highly expressed in RA. This study highlights the critical role of lactylation in plasma cells for RA pathogenesis and identifies potential biomarkers and therapeutic targets, which may offer insights for future therapeutic strategies.

Evaluating the Impact of Novel Incretin Therapies on Cardiovascular Outcomes in Type 2 Diabetes: An Early Systematic Review.

Background This systematic review is registered with CRD42024507397 protocol number and aims to compare the known data about retatrutide on long-term cardiovascular (CV) protection with tirzepatide, an incretin with recent proven CV benefits. Material and Methods The inclusion criteria were (i) original full-text articles that are randomized control or clinical trials; (ii) published within the last ten years; (iii) published in English; and (iv) conducted on adult human populations. The exclusion criteria were articles deruled on cell cultures or mammals. Studies were selected if they (1) included patients with type 2 diabetes mellitus (DM) and CV risk; (2) patients that received either tirzepatide or retatrutide; and (3) provided sufficient information such as the corresponding 95% confidence intervals or at least a sufficient p-value. Studies were excluded if they were a letter to the editor, expert opinions, case reports, meeting abstracts, or reviews; redundant publications; or needed more precise or complete data. Results The seven included studies were assessed for bias with the Newcastle Ottawa scale, heterogenous, and emphasized the potential CV beneficial effect of type 2 DM (T2DM) therapies (glycemia, glycated A1c hemoglobin, body weight, lipid profile, blood pressure and renal parameter). Discussions Further, longer follow-up studies are necessary to verify the long-term CV protection, standardize the specific aspects of CV risk, and compare with subjects without T2DM for a more integrative interpretation of the CV effects independent of the improvement of metabolic activity.

Heterocyclic phytometabolites formononetin and arbutin prevent in vitro oxidative and alkylation-induced mutagenicity

Phenolic phytometabolites are promising bioactive compounds for management of genomic instability related diseases. Formononetin (FMN) and arbutin (ARB) are found in several plant sources. Our goal was to investigate the safety and efficacy of FMN and ARB using in vitro both standardized and alternative toxicogenetic methods. FMN and ARB were evaluated through the OECD’S guidelines No. 471 (Bacterial Reverse Mutation Test –Salmonella/microsome) and No. 487 (In vitro Mammalian Micronucleus Test – CBMN assay), accordingly to the mentioned recommendations. Also, antimutagenicity of FMN and ARB was assessed in S. Typhimurium strains TA98, TA100 and TA1535, following pre-, co- and post- treatment protocols. Liver human lineages HepG2 and F C3H were assayed for cytotoxicity after exposure to FMN and ARB (24, 48 and 72 h) using in vitro WST-1 test. ARB showed no mutagenicity in the Salmonella/microsome test under both metabolic conditions (in presence or absence of 4 % S9 mix), but FMN was cytotoxic to the TA97 and TA100 strains after metabolic activation. Under this same condition, FMN induced an increase in the mutagenic index of strain TA1535 at two of the highest tested concentrations. Even so, ARB and FMN exhibited protection against the induced alkylation of DNA in multiple action modes. In the antimutagenicity assay, FMN reached the maximum of 80 % of oxidative-provoked mutagenicity reduction in TA98 strain in co-treatment with known mutagen, besides 69 % of reduction in TA100 in the same exposure condition. ARB showed up to reduce induced mutagenicity in strains TA100 and TA1535, reaching percentages from 55 % to 100 % of antimutagenicity in all of the tested exposure models against alkylating agent. In the CBMN assay, no increase in micronuclei formation was observed. The results suggest that FMN and ARB prevent DNA from mutation using multi-targeted antimutagenic roles. Finally, our data suggests that FMN and ARB are not genotoxic and presented encouraging antimutagenicity action in vitro, being promising compounds for use in genomic instability-related diseases therapeutics.

Hormonal regulation of the phenotype into environmentally appropriate pace‐of‐life syndromes

AbstractThe risk of predation is an important driver that tailors life histories in various ways. Using an evolutionary model based on hormonal control, we study how different predation regimes affect adaptive risk‐taking and growth in fish populations. Growth, metabolism and foraging in the modelled fish are regulated by three simplified hormone functions: growth hormone, orexin, and thyroid hormone. A dynamic state‐dependent optimization model finds optimal hormone profiles for adaptive growth strategies in juvenile fish. We consider a gradient from species where behaviour and metabolic activity have large consequences for risk (typically benthic and camouflaged species), to the opposite endpoint where behaviour may modify predation risk to a smaller degree (as in the pelagic). Along this gradient, the model predicts changes in the pace of life from slow to fast, enacted by up‐regulation of the three hormone functions which in turn increase foraging and metabolism and change the priorities of energy reserves versus growth. Under all types of predation risk investigated, growth is faster when food availability is higher. Energy reserves are maintained primarily during periods of poor food availability and are used to accelerate growth during periods when food availability is high. The thyroid hormone function is up‐regulated predominantly when food availability is high and has an important role in trade‐offs balancing energetic gain and survival. At the individual time scale, the hormone system improves organismic flexibility and robustness. Over the phylogenetic time scale, hormone system adaptations have also restricted the phenotypic plasticity of individuals.

Maximum standardized uptake value-to-tumor size ratio in fluorodeoxyglucose F18 positron emission tomography/computed tomography: a simple prognostic parameter for non-small cell lung cancer.

By correcting the effect of tumor size on metabolic activity, the maximum standardized uptake value-to-tumor size (SUVmax:tumor size) ratio on fluorodeoxyglucose F18 positron emission tomography (18F-FDG PET)/computed tomography (CT) scans can be a prognostic parameter of non-small cell lung cancer (NSCLC). The current study evaluates the prognostic value of SUVmax:tumor size ratio on pretreatment 18F-FDG PET/CT scans in patients with NSCLC. Furthermore, the SUVmax:tumor size ratio is compared with other established PET parameters. This study included 108 patients with NSCLC who underwent pretreatment 18F-FDG PET/CT scans and curative lung surgery. The associations between the SUVmax:tumor size ratio and other conventional PET parameters were investigated. The recurrence-free survival according to the SUVmax:tumor size ratio was also analyzed. In addition, the SUVmax:tumor size ratio was compared according to postoperative pathologic findings. In total, 72 (66.7%) of the 108 participants presented with adenocarcinoma (ADC). Nineteen (17.6%) patients experienced recurrence during a median follow-up period of 32.3 months. The median SUV max:tumor size ratio was 2.37 (1.23 for ADCs and 3.90 for other histologic types). The SUVmax:tumor size ratio was associated with SUVmax and mean SUV, as well as metabolic tumor volume and total lesion glycolysis. Patients with an SUVmax:tumor size ratio higher than the median had a worse recurrence outcome than those with an SUVmax:tumor size ratio lower than the median. Participants with ADC who presented with lymphovascular invasion had a higher SUVmax:tumor size ratio than those without. The presence of lymph node metastasis and advanced histologic grade were associated with a high SUVmax:tumor size ratio in patients with ADC. The SUVmax:tumor size ratio on pretreatment 18F-FDG PET/CT scans was associated with aggressive tumor behavior and poor outcome in NSCLCs, particularly ADC. The SUVmax:tumor size ratio on pretreatment 18F-FDG PET/CT scans has a prognostic value in patients with NSCLCs, especially ADC.

B-124 Structure-based Variant Effect Prediction and Design of Functional CYP2C9 Variants by Virtual Evolution

Abstract Background The decryption of massive variants of uncertain significance (VUS) discovered by sequencing poses the main challenge in the post-sequencing era. VUS also resulted in a particular problem in the field of pharmacogenomics, which can be epitomized by drug-metabolizing enzymes, such as CYP2C9. Therefore, an accurate model that incorporates the inner connection between massive protein variants and their function alterations is crucial for clinical annotation of human variome and the virtual evolution of enzyme engineering for biotherapeutics. Methods We developed a model named variant effect recognition network for CYP2C9 (vERnet-P), which learned features from AlphaFold2-predicted protein structures and predicted the enzyme activity of missense single-nucleotide variants in CYP2C9. The two crucial strategies of vERnet-P are the construction of amino acid interaction networks and the application of deep learning. Therefore, vERnet-P leveraged highly accurate protein structures predicted by AlphaFold2, combined with novel techniques for enriching and capturing useful information. Based on the accurate and preemptive prediction of CYP2C9 variants, we performed the saturation mutation prediction on several sites of CYP2C9 to identify new variants with the expected function. Furthermore, the drug metabolic activities of these newly identified variants were investigated with two probe drugs by in vitro metabolic activity assessments. Results An accuracy of up to 93.5% for the classification of activity levels was yielded in the testing dataset. In addition, the tasks for recognizing the high-activity and low-activity variants both achieved 93.5% accuracies, demonstrating that vERnet-P truly learned the features related to enzyme activity, rather than the selection bias to one kind of samples. Strikingly, the AUC values of the ROC and the PR reached 0.971 and 0.966 respectively in the testing cohort for vERnet-P. vERnet-P achieved the strongest agreement with massively parallel assessments relative to the other 8 computational variant effect predictors in the task of predicting CYP2C9 variant enzyme activity. We explored 6 mutant sites in CYP2C9 by using vERnet-P to perform saturation mutation prediction and found 12 novel variants with a high possibility of activity changing. We identified 6 variants with most likely increased activity by using the prediction of wild-type as a cutoff, consequently, 6 variants with most likely decreased activity were selected. According to these 12 discovered variants, 10 were strongly confirmed by the in vitro metabolic activity assays of both probe drugs. Conclusions vERnet-P achieved state-of-the-art performance in predicting the CYP2C9 variant enzyme activity. The preemptive prediction for CYP2C9 variants can efficiently provide the clinical drug dosing guideline. Furthermore, by saturating mutation prediction on several sites, our results indicated the evolutionary direction of CYP2C9 enzyme activity and discovered some brand-new CYP2C9 variants with ultra-strong functional alterations, which have not previously been reported. The results of in vitro metabolic activity assessment were highly consistent with our in silico prediction, indicating the potential of AI models for inferring evolutionary direction and designing novel variants with the expected function.

Early wave reflection of carotid artery is associated with 18F-FDG PET hypometabolism in Alzheimer's brain areas of cognitively normal adults.

Arterial stiffening likely plays a role in Alzheimer disease (AD) pathogenesis. The current study investigated whether inter-individual variations in arterial stiffness and pressure wave parameters were associated with 18F-FDG positron emission tomography (PET) metabolism in AD-associated brain areas throughout adulthood, independently of age and before the onset of any neuropsychological disorders. A prospective, large age-range population of 67 patients (17 young, 16 middle-aged, and 34 older adults; 37 women) underwent a: brain 18F-FDG PET, blood pressure recording, and carotid/femoral pulse wave-based measurements, including the time-to-peak of the reflected backward carotid pulse wave (bT), on the same day. Multivariable and quantitative voxel-to-voxel analyses (P-voxel < 0.005, corrected for cluster volumes) were conducted to assess associations between vascular parameters and 18F-FDG PET metabolism in AD-associated brain areas. In the multivariable analysis, only increased age and decreased bT were independently associated with the decline of metabolic activity in AD-associated brain areas (P < 0.001). In the voxel-to-voxel analysis with age as a covariate, bT was strongly associated with the metabolic activity of 40 clusters in AD-associated brain areas (clusters cumulative volume: 63 cm3; T score max: 5.7). In a large age-range population of adult patients, who are still unaffected by neuropsychological disorders, an early reflected arterial pressure wave, as evidenced by a decreased bT value, is strongly associated with hypometabolic activity of AD-associated brain areas, independently of age.

Nonthermal fluctuations accelerate biomolecular motors

AbstractIntracellular transport is essential for maintaining cellular function. This process is driven by different mechanisms in prokaryotic and eukaryotic cells. In small prokaryotic cells, diffusion is the primary means of transport, while larger eukaryotic cells also rely on active transport by molecular motors such as kinesin and dynein. Recently, it has become evident that, in addition to diffusion based on thermal fluctuations (Brownian motion), which was conventionally considered a diffusion mechanism within living cells, nonthermal fluctuations generated by metabolic activities play a crucial role in intracellular diffusion. Similarly, while molecular motors have been proposed to exploit thermal fluctuations in the environment following the direct observation and manipulation of single molecules, they have also been reported to utilize nonthermal fluctuations in recent years. This review begins with a brief overview of the historical knowledge of diffusive intracellular transport, which has been extended from the thermal fluctuations to the nonthermal fluctuations generated by metabolic activity. It then introduces recent findings on how nonthermal fluctuations accelerate the motion of molecular motors and discusses future perspectives on the general effects of these fluctuations on molecules in living cells.

A-134 Comparative Analysis of ALT and AST Measurements: Impact of P5P Activators on Platform Discrepancies

Abstract Background Vitamin B6 (VB6), notably its active form pyridoxal 5'-phosphate (P5P or PLP), is essential for ALT and AST enzyme activity in amino acid metabolism. Insufficient P5P can yield inaccurately low ALT and AST measurements in individuals with VB6 deficiency. This study assesses VB6 deficiency prevalence and its impact on ALT and AST measurements on Beckman Coulter compared to Roche and Siemens platforms. Methods VB6 levels were analyzed in 996 patients, with &amp;lt; 20 nmol/L considered deficient. ALT and AST levels were compared between Beckman Coulter AU5800 versus Roche cobas c, and Beckman Coulter AU5800 versus Siemens Dimension Vista. Roche and Siemens assays included P5P supplements, while Beckman Coulter did not. 29 ALT specimens (23 VB6 deficient and 6 normal) and 34 AST specimens (28 VB6 deficient and 6 normal) were compared between Beckman Coulter and Roche. Additionally, 16 specimens (4 VB6 deficient and 12 normal) were tested for ALT and AST for the comparison between Beckman Coulter and Siemens. The study then assessed the impact of P5P-containing reagents on test results by comparing the percentage differences in average concentration and their clinical decision implications across different platforms, within the low, normal, and high ranges. Results Out of 996 patients, 356 exhibited VB6 results below 20 nmol/L, indicating a striking 36% prevalence of VB6 deficiency within our patient population. Beckman Coulter and Roche comparison: In VB6 deficient patients, ALT displayed an average concentration difference of 11.5%: Beckman ALT mean 23 (+/-24.6) U/L versus Roche ALT mean 26 (+/-30) U/L. AST demonstrated a 41% difference in average concentration: Beckman AST mean 28.2 (+/-28.9) U/L versus Roche AST 48.9 (+/-49) U/L. In normal patients, ALT displayed an average concentration difference of 2.4%: Beckman ALT mean 21.3 (+/-21.3) U/L versus Roche ALT mean 20.8 (+/-25.7) U/L. AST demonstrated a 23.5% difference in average concentration: Beckman AST mean 25.5. (+/-25.4) U/L versus Roche AST 33.3 (+/-37.5) U/L. Additionally, 13% of ALT and 18% of AST displayed noteworthy effects on clinical decisions, resulting in shifts from low abnormal to normal range or normal range to high abnormal. Beckman Coulter and Siemens comparison: In VB6 deficient patients, ALT displayed an average concentration difference of 36.8%: Beckman ALT mean 18 (+/-15.7) U/L versus Siemens ALT mean 28.5 (+/-20.5) U/L. AST demonstrated a 43.4% difference in average concentration: Beckman AST mean 18.3 (+/-8.1) U/L versus Siemens AST 32.3 (+/-17.2) U/L. In normal patients, ALT displayed an average concentration difference of 32.8%: Beckman ALT mean 22.2 (+/-16.1) U/L versus Siemens ALT mean 33.1 (+/-20.1) U/L. AST demonstrated a 31.3% difference in average concentration: Beckman AST mean 25.4 (+/-12.3) U/L versus Siemens AST 36.1 (+/-19.7) U/L. While 6% of ALT specimens would have altered clinical decisions, the impact was more pronounced for AST, with 50% of results changing to an upper reference point. Conclusions In clinical settings where accurate assessment of enzyme activity is vital for effective patient care, our study emphasizes the crucial necessity for standardizing AST and ALT methodologies to incorporate P5P supplementation.

Effect of time series on the degradation of lignin by Trametes gibbosa: Products and pathways

Lignin is the third most abundant organic resource in nature. The utilization of white-rot fungi for wood degradation effectively circumvents environmental pollution associated with chemical treatments, facilitating the benign decomposition of lignin. Trametes gibbosa is a typical white-rot fungus with rapid growth and strong wood decomposition ability. The lignin content decreased from 23.62 mg/mL to 17.05 mg/mL, which decreased by 27 % in 30 days. The activity of manganese peroxidase increased steadily by 9.44 times. The activities of laccase and lignin peroxidase had the same trend of change and reached peaks of 49.88 U/L and 10.43 U/L on the 25th day, respectively. The change in H2O2 content in vivo was opposite to its trend. For FTIR and GC–MS analysis, the fungi attacked the side chain structure of lignin phenyl propane polymer and benzene ring to crack into low molecular weight aromatic compounds. The side chains of low molecular weight aromatic compounds are oxidized, and long-chain carboxylic acids are formed. Additionally, the absorption peak in the vibration region of the benzene ring skeleton became complex, and the structure of the benzene rings changed. In the beginning, fungal growth was inhibited. Fungal autophagy was aggravated. The metal cation binding proteins of fungi were active, and the genes related to detoxification metabolism were upregulated. The newly produced compounds are related to xenobiotic metabolism. The degradation peak focused on the redox process, and the biological function was enriched in the regulation of macromolecular metabolism, lignin metabolism, and oxidoreductase activity acting on diphenols and related substances as donors. Notably, genes encoding key degradation enzymes, including lcc3, lcc4, phenol-2-monooxygenase, 3-hydroxybenzoate-6-hydroxylase, oxalate decarboxylase, and acetyl-CoA oxidase were significantly upregulated. On the 30th day, the N-glycan biosynthesis pathway was significantly enriched in glycan biosynthesis and metabolism. Weighted correlation network analysis was performed. A total of 1452 genes were clustered in the coral1 module, which were most related to lignin degradation. The genes were significantly enriched in oxidoreductase activity, peptidase activity, cell response to stimulation, signal transduction, lignin metabolism, and phenylpropane metabolism, while the rest were concentrated in glucose metabolism. In this study, the lignin degradation process and products were revealed by T. gibbosa. The molecular mechanism of lignin degradation in different stages was explored. The selection of an efficient utilization time of lignin will help to increase the degradation rate of lignin. This study provides a theoretical basis for the biofuel and biochemical production of lignin. SynopsisTrametes gibbosa degrades lignin in a pollution-free way, improving the utilization of carbon resources in an environmentally friendly spontaneous cycle. The products are the new way towards sustainable development and low-carbon technology.

The complex interplay of Selenoprotein P, NO metabolites, and pancreatic enzymes in chronic pancreatitis and hypothyroidism is partially orchestrated by the SEPP1 gene’s rs7579 polymorphism: focus on gender aspect

Objective To establish the association of the SEPP1 gene’s (rs7579) polymorphism with enzymatic, metabolic and hormonal activity in patients with chronic pancreatitis (CP) and primary hypothyroidism. Materials and methods Eighty CP patients (40 with comorbid hypothyroidism) and 30 healthy controls participated in the case-control study. Pancreatic enzymes, Selenoprotein P, NO metabolites (NO2 -+NO3 -), glucose, total cholesterol (TC), triglycerides (TG) and low/high density lipoprotein cholesterol (LDL-, HDL-C), Atherogenicity Index (AI), thyroid-stimulating hormone (TSH), Thyroxine (T4), glomerular filtration rate (GFR) were studied. SEPP1 (rs7579) genotyping performed by PCR (FlexCycler BU). Results and discussion SEPP1 (rs7579) gene’s A-allele increases the risk of hypothyroidism twice in CP [OR=2.0; OR 95%CI:1.09-3.66; р=0.023]. Pancreatic patients with hypothyroidism and SEPP1 gene’s (rs7579) AA-genotype had 60.0% (рАА=0.013) lower elastase and 13.78% (р=0.003) higher α-amylase as well as TSH by 10.81% (рАА=0.009) and 15.64% (рАА=0.009), especially in women 14.55-45.18% (р&lt;0.001) at a lower value of Selenoprotein P – by 28.65-48.08% (р≤0.048) regardless of the SEPP1 gene (rs7579) variants. Women have 17.43-18.14% (pW≤0.032) higher NO metabolites than men; A-allele women have free T4 value 2.62 times lower (рGG=0.01) than GGwomen and 2.97 times lower (рW=0.011) than men. Comorbid patients with A-allele had elevated TC and LDL-C, by 11.77-26.45% (рАА≤0.01) and 18.06-26.21% (рАА≤0.019), respectively. Women have 54.50% (pW=0.002) higher AI with 39.30% lower (pW=0.034) Selenoprotein P and 21.96-24.56% (pW≤0.033) GFR than men. Conclusion SEPP1 (rs7579) gene polymorphism influences the risk of hypothyroidism in CP patients, changes of pancreatic enzymes, dyslipidaemia particular in AA-genotype carriers, mainly women. There was no dependence of SEPP1 and total NO metabolites values on the SEPP1 gene (rs7579) polymorphism. Bangladesh Journal of Medical Science Vol. 23 No. 04 October’24 Page : 1038-1047

Insights into the functional properties of thioredoxin domain-containing protein 12 (TXNDC12): Antioxidant activity, immunological expression, and wound-healing effect in yellowtail clownfish (Amphiprion clarkii)

Thioredoxin domain-containing protein 12 (TXNDC12) is a member of the thioredoxin-like superfamily that contributes to various thiol-dependent metabolic activities in all living organisms. In this research, the TXNDC12 gene from yellowtail clownfish (Amphiprion clarkii) was structurally characterized using in silico tools, assessed for immunological expression, and evaluated for biological activity using recombinant protein and cellular overexpression. The deduced coding sequence of AcTXNDC12 comprised a 522-bp nucleotide, encoding 173 amino acids with a predicted molecular mass of 19.198 kDa. The AcTXNDC12 protein consists of a66WCGAC70 active motif and a170GDEL173 signature. The highest tissue-specific expression of AcTXNDC12 was observed in the brain tissue, with significant modulation observed in the blood and gill tissues following stimulation of polyinosinic: polycytidylic acid, lipopolysaccharides (LPS), and Vibrio harveyi. In functional assays, recombinant AcTXNDC12 protein (rAcTXNDC12) showed insulin disulfide reduction activity, 2,2′-azino-di-(3-ethylbenzthiazoline sulfonic acid) decolorization antioxidant capacity, and ferric (Fe3+) reducing antioxidant potential. Additionally, a significant reduction in nitric oxide production was observed in AcTXNDC12-overexpressed RAW 264.7 cells upon LPS stimulation. Furthermore, genes associated with the regulation of oxidative stress, including nuclear factor erythroid 2-related factor 2 (Nrf2), catalase (Cat), peroxiredoxin 1 (Prx1), and ribonucleotide reductase catalytic subunit M1 (Rrm1) were significantly upregulated in fathead minnow cells overexpressing AcTXNDC12 in response to H2O2 treatment. The scratch wound healing assay demonstrated tissue regeneration and cell proliferation ability upon AcTXNDC12 overexpression. Altogether, the current study elucidated the antioxidant activity, immunological importance, and wound-healing effect of the AcTXNDC12 gene in yellowtail clownfish, providing valuable insights for advancing the aquaculture of A. clarkii fish.

Effects of Dietary Agavin on the Gut Microbiota of the Nile Tilapia (Oreochromis niloticus) Reared at High Densities.

High-density stress can lead to dysbiotic microbiota, affecting the organism's metabolic, and protective functions. Agavin is a fructan with prebiotic properties that regulate the gut microbiota by promoting the growth of beneficial bacteria. This study evaluated the effect of agavin on the gut microbiota using Next-Generation Sequencing (NGS) and its correlation with the growth parameters. Four groups of fish were fed different diets: a control diet (negative and positive control), without agavin supplementation, and two experimental diets supplemented with agavin at 20gkg-1 and 40gkg-1. Nile tilapias (1.04g ± 0.01g) were fed for 110days. After 90days of feeding, fish were subjected to high-density stress (63kgm-3) for 20days, except for the negative control. NGS detected 1579 different operational taxonomic units in the samples. In the correlation analysis of growth parameters, the families Vibrionaceae and Methyloligillaceae showed a positive correlation with fish growth parameters, these results may serve to know the relation of agavin and microbiota on the growth performance, as well as the metabolic activities of families in tilapia. Furthermore, high-density stress and agavin supplementation modify the gut microbiota in tilapia. At a low-density, supplementation with 20gkg-1 agavin promoted the growth of the potentially beneficial families Sphingomonadaceae, Oxalobacteriaceae, and Chitinophagaceae; at high densities, reduced the abundance of pathogenic families (Vibrionaceae and Aeromonadaceae). These results suggest that, under stress conditions, agavin can stimulate the growth of potentially beneficial bacteria and reduce the growth of potentially pathogenic bacteria, suggesting its potential use as a prebiotic in aquaculture.

Mixed species biofilms act as planktonic cell factories despite isothiazolinone exposure under continuous-flow conditions.

The primary approach to managing biofouling in industrial water systems involves the large-scale use of biocides. It is well-established that biofilms are 'cell factories' that release planktonic cells even when challenged with antimicrobials. The effect of isothiazolinone on the metabolic activity and biomass of mixed Pseudomonas biofilms was monitored in real-time using the CEMS-BioSpec system. The exposure of biofilms to the minimum inhibitory concentration (1.25 mg L-1) of biocide did not impact planktonic cell production (log 7.5 CFU mL-1), while whole-biofilm metabolic activity and biomass accumulation increased. Only the maximum biocide concentration (80 mg L-1) resulted in a change in planktonic cell yields and temporal inhibition of biofilm activity and biomass, a factor that needs due consideration in view of dilution in industrial settings. Interfacing the real-time measurement of metabolic activity and biomass with dosing systems is especially relevant to optimizing the use of biocides in industrial water systems.

A case of neoadjuvant targeted therapy with pralsetinib for locally advanced lung adenocarcinoma with RET fusion mutation

This case report details the successful treatment of a 68-year-old male patient with locally advanced RET-rearranged lung adenocarcinoma using neoadjuvant pralsetinib. The patient initially presented with a suspicious right upper lobe nodule, which was later diagnosed as lung adenocarcinoma following genetic testing that revealed a RET exon 12 fusion. After 2 months of neoadjuvant treatment with pralsetinib, a significant radiological response was observed, with a reduction in tumor size and metabolic activity. Subsequently, the patient underwent video-assisted thoracoscopic right upper lobectomy and mediastinal lymph node dissection. Postoperative pathological analysis revealed a major pathological response, with only 5% residual tumor cells in the primary lesion and no viable tumor cells in the lymph nodes. Postoperative pathological staging of TNM was ypT1aN0M0, stage IA1(AJCC, 8th edition). The patient recovered well after surgery, demonstrating the potential efficacy of neoadjuvant pralsetinib in locally advanced RET-rearranged lung adenocarcinoma. However, further clinical validation is required to establish the role of neoadjuvant targeted therapy and postoperative adjuvant therapy in this patient population.