The spoilage of beer significantly threatens the quality and safety of products in the beverage industry. Certain natural foods contain beneficial bioactive components that are considered to be safer than chemical additives. This study aimed to identify extracts of natural foods as potential alternatives to chemical preservatives for controlling beer spoilage microorganisms. Among the two extraction methods applied to 176 natural foods, the extracts of clove alone effectively inhibited the growth of representative beer spoilage microorganisms, specifically Levilactobacillus brevis, Sporolactobacillus vineae, Pectinatus frisingensis, and Saccharomyces cerevisiae var. diastaticus. High-performance liquid chromatography and half-maximal inhibitory concentration analysis revealed that gallic acid and eugenol in cloves were active compounds with antimicrobial properties in vitro that were of a similar order of magnitude to those of commercial preservatives (potassium sorbate and sodium benzoate) under the tested conditions. Scanning electron microscopy observations and a fluorescence leakage assay using 3,3'-dipropylthiadicarbocyanine iodide indicated morphological alterations and membrane perturbation in treated microorganisms, except for the limited effect of gallic acid on S. cerevisiae var. diastaticus. These findings provide insight into the potential role of natural food-derived antimicrobials in controlling beer spoilage microorganisms, pending further validation in real beverage systems.

Multiple sclerosis (MS) is a chronic autoimmune-mediated demyelinating disease of the CNS, characterized by neuroinflammatory, axonal degeneration, and pronounced sexual dimorphism. Experimental data implicate dysregulated 5-HT levels in MS. However, the effects of clinical parameters and disease-modifying-therapies (DMTs) on peripheral 5-HT concentrations remain underexplored. This study aimed to quantify peripheral levels of tryptophan (Trp), 5-HT, and 5-hydroxyindoleacetic acid (5-HIAA) in patients with relapsing–remitting MS (RRMS) and to assess the effects of BMI, DMT duration, and specific DMT regimens. In this cross-sectional analysis, 226 participants were enrolled and stratified into four groups: healthy men (HM; n = 29), healthy women (HW; n = 84), men with RRMS (MMS; n = 29), and women with RRMS (WMS; n = 84). Serum concentrations of Trp, 5-HT, and 5-HIAA were measured using reverse-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Nonparametric statistical tests were applied. Peripheral 5-HT levels were significantly reduced in underweight WMS (BMI < 18 kg/m 2 ; p < 0.05), WMS with DMT duration over 4 years ( p < 0.01), and WMS receiving interferon beta-1a ( p < 0.01) compared to HW. No significant intergroup differences in Trp or 5-HIAA were observed across all stratifications. These findings reveal a novel association between reduced peripheral 5-HT and specific clinical-therapeutic factors in WMS, extending recent MS research on sex-specific vulnerabilities, serotonergic dysregulation in neuroinflammation, and psychiatric comorbidity. By highlighting the influence of low BMI, prolonged DMT exposure, and interferon beta-1a on 5-HT homeostasis, this study underscores the need for multidisciplinary management integrating neurological and psychiatric care in WMS and suggests avenues for precision interventions targeting serotonergic pathways to reduce disease burden.

This study aimed to investigate the tryptophan and tyrosine-derived metabolites in traditional fermented foods. For that purpose, kynurenine, kynurenic acid, indole-3-acetic acid, indole-3-propionic acid, phenol, and p-cresol levels were analyzed using High-performance liquid chromatography (HPLC). Tryptophan metabolite levels ranged from 62.4 to 718.5µg/100g for kynurenine, 1.1 to 340.0µg/100g for kynurenic acid, 0.0 to 36.6µg/100g for indole-3-acetic acid, and 0.0 to 72.5µg/100g for indole-3-propionic acid. Tyrosine metabolite levels ranged from 4.4 to 115.7µg/100g for phenol and 0.0 to 7.2µg/100g for p-cresol. Traditional, fermented, and aged products such as tarhana appear to contain high levels of various metabolites of tryptophan metabolism. Phenol levels were also found to be high in tarhana products and aged dairy products. p-cresol levels remained generally low, with significantly high levels found only in aged cheddar cheese. These results suggest that microbial fermentation and long-term maturation may be associated with differences in the biochemical composition of the fermented foods. It is of great importance to evaluate fermented products in terms of metabolites with neuroactive and inflammatory potential.

Abstract A simple, rapid, and environmentally friendly reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous determination of ketoprofen and famotidine in combined dosage forms. Chromatographic separation was achieved on a Roc C18 column (250 × 4.6 mm, 5 μm) using an isocratic mobile phase of methanol:water:acetonitrile (35:35:30, v/v/v) adjusted to pH 3.1 with formic acid, at a flow rate of 2 mL/min and detection at 264 nm. Retention times were 2.86 min for ketoprofen and 8.56 min for famotidine. The method was validated following ICH Q2(R2) guidelines, demonstrating excellent linearity (R 2 = 0.9996 for ketoprofen, R 2 = 0.9993 for famotidine), accuracy (98.35–101.26%), precision (%RSD < 1%), and sensitivity (LOD: 18.3 and 25 ng/mL; LOQ: 61.8 and 84 ng/mL, respectively). Robustness testing confirmed reliability under varied conditions. Importantly, the method minimizes solvent usage and employs short run times, aligning with green analytical chemistry principles. The proposed RP-HPLC method is precise, accurate, and suitable for routine quality control of ketoprofen–famotidine formulations, offering a sustainable alternative for pharmaceutical analysis.

Long noncoding RNAs (lncRNAs) constitute a substantial portion of the transcriptome and outnumber protein-coding transcripts in humans. lncRNA molecules are bioactive and control cellular and systemic functions by directly or indirectly regulating gene expression at the transcriptional or posttranscriptional levels. Here, we established the use of lncRNA as a modality to treat disease by reengineering three lncRNAs-GAPLINC, MIST, and DRAIR-to treat acute inflammation in mice and human macrophages. For each lncRNA, we established an in vitro transcription synthesis and high-performance liquid chromatography purification workflow and optimized the lncRNA isoforms, 5' cap, 3' poly(A) tail, and chemical base modifications to improve specificity and performance. We also optimized the in vivo delivery of lncRNA with a lipid nanoparticle system that did not induce confounding effects. Using this pipeline and delivery platform, we demonstrated that each lncRNA reduced lipopolysaccharide (LPS)-induced inflammation by specifically regulating distinct subsets of cytokines in cultured mouse macrophages and in mice. GAPLINC and DRAIR reduced the transcription of IL-1β and IL-6, respectively, whereas MIST attenuated TNFα production posttranscriptionally. In addition, we showed that reengineered GAPLINC reduced LPS-induced inflammation in human monocytes, suggesting the clinical potential of this approach. Our engineering approach and findings establish a previously unidentified nucleic acid modality and demonstrate an effective way to reengineer regulatory lncRNAs to treat disease.

Ectoine is a valuable active compound produced by halophilic bacteria with broad applications in the pharmaceutical and cosmetic industries. The increasing industrial demand underscores the need for enhanced production capacity and drives the discovery of effective and efficient ectoine-producing bacteria strains. Although Indonesia possesses numerous high-salinity habitats, such as traditional salt ponds, the potential for novel ectoine-producing strains remains underexplored. Therefore, this study aims to isolate ectoine-producing bacteria from traditional salt ponds in Buleleng Regency and optimize the yield of potential isolates. Halophilic bacteria were initially cultivated from saltwater and soil samples on MM63 medium. The ectoine-producing capability of these isolates was then evaluated using a batch culture approach incorporating an osmotic shock step. Ectoine quantification was performed through reversed-phase High-Performance Liquid Chromatography using a mobile phase of water/acetonitrile (95/5). To maximize production, a series of optimization experiments determined the ideal media composition, initial pH, incubation temperature, and incubation time. The results showed that seven ectoine-producing halophilic bacteria were successfully isolated, with one of the best isolates being Halomonas huangheensis LES5 AG4. The application of osmotic shock increased ectoine production by up to 3.6-fold, specifically from 94 mg/L to 335 mg/L. Furthermore, ectoine production was significantly increased to 646 mg/L after optimizing several key factors, including ammonium chloride (0.47% w/v), glycerol (1.42% w/v), NaCl (11.67% w/v), Initial pH (7.1), incubation temperature (30 °C), and incubation time (16 hours). These results show that Halomonas huangheensis LES5 AG4 holds significant potential for development as an industrial-scale ectoine producer.

Current therapeutic practices have limited effectiveness at targeting acute pain locally. This is in part due to the lack of understanding of the neuroinflammatory pathways active within pulpitis. Increased activity of inadoleamine 2,3 dioxygenase (IDO) was linked to hypersensitivity and pain in several settings. IDO is the rate-limiting enzyme of the kynurenine pathway, but changes in IDO's activity and expression in pulpitis have not been investigated. Tooth samples with a diagnosis of symptomatic irreversible pulpitis (SIP) were collected, and levels and activity of IDO and other markers of the kynurenine (KYN) pathway were examined using high-performance liquid chromatography (HPLC), quantitative polymerase chain reaction (PCR), and immunocytochemistry (ICC). Subsequently, odontoblast-like cells (OLCs) derived from dental pulp stem cells were stimulated in vitro with bacterial lipopolysaccharide and a synthetic proxy for DNA cytosine-phosphate-guanine (CpG) oligonucleotide 2006 (ODN 2006). The effects of these compounds on KYN pathway marker levels and transcription were then analyzed using quantitative PCR and HPLC mass spectrometry. IDO activity was significantly raised in SIP compared with healthy controls. ICC fluorescent imaging showed that IDO and KYN were colocalized with the markers of macrophages and neuronal fibers. Quantitative PCR data of SIP indicated that increased IDO may direct the KYN pathway toward the generation of the neuroinflammatory catabolite quinolinic acid, a metabolite that is opposite to the production of neuroprotective kynurenic acid. Administration of OLCs with ODN 2006 in vitro induced changes similar to those developed in SIP induced by bacterial infection. To our knowledge, this article is the first to demonstrate links between IDO activity, neuroinflammation, and algogenic pathways in SIP and localize this process to neuronal fibers. These observations highlight the potential role of the kynurenine pathway in SIP, but further investigation is required to determine if these changes are mirrored in the levels of neuroexcitatory metabolites in vivo.

Objectives: Reversed-phase high-performance liquid chromatography was used to produce a simple, accurate, and precise method for the estimation of Magnolol (MO) in self-nano-emulsifying drug delivery system. Method: Analysis of this method was carried out using reverse phase Nucleodur C-18 column. For this study Mobile phase used was acetonitrile: H2O (90:10% v/v), the flow rate was 1.0 mL/min, chromatogram of MO was determined at wavelength 290 nm, and sharp peak was obtained at retention time of 3.496 min. Validation of the developed method was done according to International Conference on HarmonizationQ2 (R1) guidelines. Results: MO showed linearity at 2–10 μg/mL with R2 of 0.9983. The % mean recovery was determined to be 95.01%, suggesting that the approach was accurate, and from the relative standard deviation, which was found to be varying from 0.53% to 1.96% for both intra-day and inter-day precision, that is, clearly <2%, indicates that the method was precise. It was observed that the detection and quantification limits, that is, limit of quantification and limit of detection, were, respectively, 0.50 and 1.52 μg/mL. Finally, for the developed method, Robustness study was performed by doing small variations in pH, flow rate, ratio of mobile phase, and the result showed that the developed method was robust, as the percentage relative standard deviation and % recovery were under a specific limit. Conclusion: This indicates that the method that was developed was accurate, linear, précised and robust. Furthermore, it can be used to estimate MO in Pharmaceutical formulations.

Can we integrate plasma protein binding data into IATA using new approach methodologies? A case study on bisphenol analogs.

Abstract Introduction: Vancomycin and Amikacin are drugs with renal excretion primarily and are used widely in critically ill children. Fall in urine output is one of the first events in case of kidney injury in PICU setting. It is not clear how early AKI (eAKI),defined as only fall in urine output by KDIGO 2012 AKI definition before rise in serum creatinine, impacts levels of these drugs.We conducted pilot study to determine drug levels of Vancomycin and Amikacin in critically children with eAKI and 48 hours after modification. Methods: Children on Vancomycin or Amikacin were monitored for hourly urine output . Serum creatinine and Amikacin or Vancomycin levels were measured at 24 hours and again at diagnosis of eAKI.Serum Vancomycin and Amikacin levels were estimated in-house as a research tool by high performance liquid chromatography (HPLC) done on Agilent Infinity II system. Results: Incidence of eAKI was 15.7% in children on Vancomycin and these children also had AKI by serum creatinine criterion. Among children on Amikacin, incidence of eAKI was 12.5% and none developed AKI by serum creatinine criterion. In children on Vancomycin, mean drug levels when eAKI was diagnosed was 22.8 ±4.36 mg/L and creatinine was 1.03 ±0.31 mg/dl, both significantly higher than baseline ( P < 0.0001).Post drug-dose modification trough level was 15.48 ±1.64 mg/L still significantly high ( P < 0.0001) but mean serum creatinine reduced to 0.66 ±0.10 mg/dl. Drug levels and creatinine at D7 were comparable.Mean Amikacin levels in children with eAKI were 59.9 ±13.79mg/L, significantly higher than baseline ( P < 0.0001). 48 h after dose reduction, trough Amikacin levels remained significantly elevated but there was no significant difference in serum creatinine from baseline levels in these children with eAKI.Amikacin level at day 7 also significantly higher despite dose modification. Conclusion: Children on Vancomycin with eAKI had simultaneous rise in serum creatinine. Drug levels remained 48h after drug dose modification and became comparable only at day 7. In children on Amikacin,with eAKI, serum creatinine showed no significant rise but drug levels in these children remained significantly high compared to baseline at 48h after dose modification as well at day 7.

Synthetic glucocorticoids, such as dexamethasone, are widely used in therapy; however, their administration at high doses may be associated with effects on the central nervous system, particularly on neurotransmitter systems, yet the molecular mechanisms underlying these phenomena remain poorly understood. In this study, we investigated the effects of a single intraperitoneal administration of dexamethasone (8mg/kg) on the metabolism of key monoamines and the expression of their metabolic enzymes in various rat brain regions (striatum, hippocampus, and prefrontal cortex) using high-performance liquid chromatography and real-time quantitative reverse transcription polymerase chain reaction. We found that dexamethasone exerts a pronounced, region-specific impact on neurotransmitter systems. In the striatum, dexamethasone increased dopamine and serotonin levels while simultaneously reducing their catabolism, which was associated with decreased messenger ribonucleic acid expression of monoamine oxidase A, monoamine oxidase B, tryptophan hydroxylase and increased expression of tyrosine hydroxylase. In the hippocampus, dexamethasone elevated serotonin levels and reduced its turnover despite an increase in monoamine oxidase A messenger ribonucleic acid expression, suggesting the potential involvement of post-transcriptional regulation or alternative metabolic pathways. In the prefrontal cortex, dexamethasone induced a reduction in norepinephrine levels, accompanied by a decrease in monoamine oxidase A and catechol-O-methyltransferase messenger ribonucleic acid expression. This study highlights the importance of considering region-specific cerebral effects of glucocorticoids for the development of personalized therapeutic and neuroprotective strategies, including the potential use of dexamethasone in conditions such as Parkinson's disease due to its ability to elevate striatal dopamine levels.

Nicotine salts-formed through the reaction of nicotine with organic acids-are widely used in e-liquids due to their enhanced sensory properties. Consequently, there is a growing need for a simple, robust, accurate, and rapid analytical method capable of simultaneously determining multiple nicotine salts in e-cigarette liquids and aerosols. In this study, we developed a novel high-performance liquid chromatography with diode array detection (HPLC-DAD) method for the simultaneous quantification of nicotine and six organic acids (benzoic acid, D,L-tartaric acid, malic acid, citric acid, lactic acid, and levulinic acid) in e-liquids and aerosols. Chromatographic and extraction parameters were fully optimized to ensure analytical reliability. Under optimal conditions, the method exhibited excellent linearity over the range of 0.005-2.5mg/mL (R2 > 0.9999), low limits of detection (0.1-3.0μg/mL), and satisfactory recoveries (96.6-105.9%). The proposed method was successfully applied to quantify nicotine and the six acids in 60 commercial e-liquids, yielding results consistent with GC-FID and LC-MS/MS analyses. Among the tested products, three nicotine salts-nicotine benzoate, nicotine lactate, and nicotine levulinate-were identified, with nicotine benzoate being the most prevalent (found in 25 e-liquids as the sole salt). Finally, analysis of aerosol emissions revealed the differential release behavior of nicotine salts from e-liquids, explaining the selective use of particular salts in commercial formulations.

Gluten proteins are key components in wheat flour that determine the formation of dough and the quality of flour-based products. Upon hydration and mixing, gluten proteins undergo complex structural transformations to form a gluten network, exhibiting a hierarchical multi-scale structure spanning molecular, aggregate, and network scales. Due to the extreme complexity of gluten proteins, accurately characterizing their multi-scale structures remains challenging, requiring the combined application of multiple techniques, which are still relatively limited and thus warrant further exploration. Therefore, this review presents the principles, operational details, and result presentations of current techniques at different structural scales, including electrophoresis, high-performance liquid chromatography, proteomics, Fourier transform infrared spectroscopy, and Fourier transform Raman spectroscopy at the molecular scale; size-exclusion chromatography, asymmetrical flow field-flow fractionation, dynamic light scattering, multi-angle light scattering, differential refractive index, and ultraviolet absorbance at the aggregate scale; and confocal laser scanning microscopy, scanning electron microscopy, confocal Raman microscopy, and two-photon excitation microscopy at the network scale, among others. It further compares the advantages and disadvantages of similar techniques, facilitating their scenario-based selective utilization. Finally, it outlines the ongoing challenges and future perspectives for the development and application of techniques for the multi-scale structural characterization of gluten proteins.

Background: Red/blue- light-emitting diode (LED)-assisted withering provides a controllable spectral input to steer tea quality, yet metabolite-level evidence linking spectrum composition to quantitative taste phenotypes in white tea remains insufficient. Methods: Fresh leaves were withered under supplemental red/blue LEDs—S0, S1, S2, S3, S4, and S5—and the resulting white teas were evaluated by quantitative descriptive analysis (QDA), untargeted metabolomics, weighted gene co-expression network analysis (WGCNA), and high-performance liquid chromatography (HPLC) quantification of caffeine, gallic acid, and eight catechin monomers. Results: Red/blue-mixed spectrum enhanced the overall sensory quality relative to the incandescent lamp; S3 maximized sweetness and freshness, whereas S4 minimized bitterness and astringency and achieved the highest overall score. Untargeted metabolomics showed the largest deviation for S0 vs. S4. A total of 18 common metabolites were identified between the S0 and light-supplemented withering treatments, dominated by saccharides and related derivatives. WGCNA linked a saccharide-centered module to higher sweetness/freshness/smoothness and a lipid-oxylipin-centered module to stronger bitterness/astringency. HPLC independently confirmed that S4 contained the lowest catechins and caffeine, supporting its reduced bitter/astringent attributes. Conclusions: Overall, the application of mixed red-blue spectra offered a promising approach to enhancing the palatability of white tea by coordinately intensifying saccharide metabolism while simultaneously suppressing key bitter and astringent components. Our study provided a scientific basis for standardizing white tea processing to enhance sensory quality.

Background: Traditionally, wild jujube (Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou) has been used to nourish the heart, calm the spirit, and arrest spontaneous sweating. Modern research confirms its broad pharmacological activities, including antioxidant, anti-inflammatory, neuroprotective, and cognitive-enhancing effects. This study aims to isolate and characterize the structure of jujube polysaccharides and evaluate their protective effects against oxidative stress damage in neural stem cells (NSCs). Methods: We successfully isolated and purified a novel pectin polysaccharide (ZJP-2) from wild jujube. Its structure was characterized in detail using high-performance liquid chromatography coupled with multi-angle laser light scattering and refractive index detection (HPLC-MALS-RI), high-performance anion exchange chromatography (HPAEC), gas chromatography–mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) spectroscopy. Results: Structural analysis revealed that ZJP-2 is a pectin heteropolysaccharide with a molecular weight of approximately 67.93 kDa. Its monosaccharide composition primarily includes galac-turonic acid (GalA), arabinose (Ara), rhamnose (Rha), galactose (Gal), and glucose (Glc). The backbone consists of α-GalA and rhamnose-galacturonic acid-I (RG-I) domains linked by (1→4)-glycosidic bonds. NMR spectroscopy further confirmed its glycosidic bond types. In activity assessment, our study demonstrated that ZJP-2 significantly alleviated DMNQ-induced oxidative stress damage in C17.2 neural stem cells. Its protective effect was achieved by reducing intracellular reactive oxygen species (ROS) levels and upregulating the mRNA expression of antioxidant genes associated with the signaling axis (p < 0.05). Moreover, ZJP-2 suppressed DMNQ-induced overexpression of Nestin and NeuN (p < 0.05), contributing to the maintenance of NSCs’ undifferentiated state and functional homeostasis. Conclusions: In conclusion, ZJP-2 possesses distinct structural characteristics and significant neuroprotective potential, supporting its development as a natural functional food or dietary supplement for preventing oxidative stress-related neural damage.

In-vitro refolding of biotherapeutic inclusion bodies has long been recognized as a bottleneck in protein production in host systems such as Escherichia coli. Low throughput, costly reagents, and offline analysis often plague refolding development efforts. Refolding optimization typically employs statistical approaches such as Design of Experiments (DoE). While DOE offers advantage over univariate one-factor-at-a-time analysis, but it requires large subset sampling, which is cost-inefficient and labour-intensive. This paper demonstrates a knowledge-based refolding optimization, contrasted to the typical DoE-based protocol for proinsulin. The reaction is monitored and segmented into two parts (segment 1: 0-2 h and segment 2:2-6 h) based on the Fourier transform infrared (FTIR), Oxidation Reduction Potential (ORP) and Reverse Phase- High Performance Liquid Chromatography (RP-HPLC) analysis. The data is fed to a multi-objective optimization (MOO) method that utilize XGBoost, coupled with an NSGA-II optimizer. Based on the Pareto front, a linear correlation between parameters was observed in segments 1 and 2. An ensemble coupled non-dominated sorting genetic algorithm II (NSGA-II) was developed to optimize the reaction conditions beforehand. The proposed optimizer was then compared with the traditional DoE-based optimization. The developed optimization framework increased the yield to 65% ± 1.78% compared to 54% ± 2.62% in the traditional DoE-based approach (relatively 20% higher). The approach could combine screening and optimization analysis in a single step, dramatically reducing the overall experimental efforts by ∼50%.

Phase-separation mode in HPLC: Theoretical prediction and experimental evaluation of elution times.

Comparative HPLC profiling, antioxidant, enzyme inhibitory activities, and in silico molecular analysis of the aerial parts of Epimedium pubigerum.

Sex-specific associations between second-trimester maternal cortisol and neonatal outcomes in island and mainland populations.

This study aimed to assess the physicochemical stability and compatibility of butorphanol tartrate, nicardipine hydrochloride, urapidil, and tirofiban hydrochloride to support safe infusion practices in the ICU. Compatibility systems involving four-, three-, and two-drug combinations were prepared in 0.9% sodium chloride injection under simulated clinical conditions at room temperature in the absence of light. Evaluations included appearance, pH, and insoluble particle counts at 0, 2, 4, 8, 12, and 24hours. Quantitative analysis of drug concentrations was performed using high performance liquid chromatography (HPLC). All tested combinations retained physical stability within 24hours, showing no measurable variation in appearance, pH, or particle. Nicardipine concentrations declined significantly, reaching 74.35% of baseline in the nicardipine-urapidil-tirofiban mixture at 24hours, and decreasing to 76.29% at 12hours and 71.46% at 24hours in the nicardipine-tirofiban combination. The concentrations of active ingredients in the remaining mixtures remained stable, consistently exceeding 90% of initial values. Butorphanol demonstrated compatibility with the other agents under the tested conditions. In clinical application, infusion of nicardipine combined with tirofiban should be restricted to less than 8hours, and nicardipine combined with urapidil and tirofiban should not exceed 12hours. Other mixtures can be administered for up to 24hours with flexibility in infusion duration.