Ethanol Levels Research Articles

Selection of indigenous Saccharomyces cerevisiae strains with good oenological and aroma characteristics for winemaking in Ningxia China

Ningxia is one of the well-known wine producing regions in China. However, the oenological and aroma characteristics of indigenous yeasts remains hidden. The fermentative and oenological properties including stress resistance, hydrogen sulfide, foam production levels; killer phenotype, and flocculation of 89 Ningxia indigenous Saccharomyces cerevisiae isolates and ten commercial yeasts were evaluated. The fermentative and oenological properties of the tested strains varied significantly. They could resist 500 g/L glucose, 300 mg/L SO2, 14% (v/v) ethanol and pH 2.8, and produce more esters. They also produce low levels of ethanol and could conduct fermentations vigorously and at a high rate. Cabernet Sauvignon wines made with NXU 21–24 showed the high intensity of tropical fruit, dry fruit, temperate fruit, and spicy flavor. The floral flavor in NXU 21–102 fermented wine is very intense. The indigenous S. cerevisiae strains of NXU 21–102 and NXU 21–24 exhibited potential use as starter cultures in wine production.

Molecular targets of PXR-dependent ethanol-induced hepatotoxicity in female mice

The pregnane X receptor (PXR, NR1I2), a xenobiotic-sensing nuclear receptor signaling potentiates ethanol (EtOH)-induced hepatotoxicity in male mice, however, how PXR signaling modulates EtOH-induced hepatotoxicity in female mice is unknown. Wild type (WT) and Pxr-null mice received 5 % EtOH-containing diets or paired-fed control diets for 8 weeks followed by assessment of liver injury, EtOH elimination rates, histology, and changes in gene and protein expression; microarray and bioinformatic analyses were also employed to identify PXR targets in chronic EtOH-induced hepatotoxicity. In WT females, EtOH ingestion significantly increased serum ethanol and alanine aminotransferase (ALT) levels, hepatic Pxr mRNA, constitutive androstane receptor activation, Cyp2b10 mRNA and protein, oxidative stress, endoplasmic stress (phospho-elF2α) and pro-apoptotic (Bax) protein expression. Unexpectedly, EtOH-fed female Pxr-null mice displayed increased EtOH elimination and elevated levels of hepatic acetaldehyde detoxifying aldehyde dehydrogenase 1a1 (Aldh1a1) mRNA and protein, EtOH-metabolizing alcohol dehydrogenase 1 (ADH1), and lipid suppressing microsomal triglyceride transport protein (MTP) protein, aldo–keto reductase 1b7 (Akr1b7) and Cyp2a5 mRNA, but suppressed CYP2B10 protein levels, with evidence of protection against chronic EtOH-induced oxidative stress and hepatotoxicity. While liver injury was not different between the two WT sexes, female sex may suppress EtOH-induced macrovesicular steatosis in the liver. Several genes and pathways important in retinol and steroid hormone biosynthesis, chemical carcinogenesis, and arachidonic acid metabolism were upregulated by EtOH in a PXR-dependent manner in both sexes. Together, these data establish that female Pxr-null mice are resistant to chronic EtOH-induced hepatotoxicity and unravel the PXR-dependent and −independent mechanisms that contribute to EtOH-induced hepatotoxicity.

Main Composition and Visual Appearance of Milk Kefir Beverages Obtained from Four Consecutive 24- and 48-h Batch Subcultures

Nowadays, there has been a significant rise in the consumption of kefir, a functional beverage touted for its perceived health benefits. To offer a high-quality beverage to consumers, it is imperative to scrutinize and fine-tune the fermentation process. This study seeks to investigate the impact of fermentation time and the number of subcultures on the physicochemical, microbiological, and volatile composition, as well as the visual appearance, of kefir beverages obtained from four consecutive 24- or 48-h batch subcultures. All fermented beverages exhibited low lactose, ethanol and acids levels, with counts of viable probiotic lactic acid bacteria and yeast exceeding 106 colony forming units/mL. The four kefir beverages from the 48-h batch subcultures notably showed the lowest total concentrations of volatile compounds, likely due to overfermentation and over-acidification of the beverages. This caused the separation of the whey and curd, along with the formation of large gas bubbles, negatively affecting the visual appearance of the products. These findings emphasize the importance of fine-tuning the fermentation process to ensure the production of high-quality kefir beverages that align with consumer preferences. The four beverages from the 24-h batch subcultures exhibited high microbiological and physicochemical stability during storage at 4 °C for 28 days.

Natural variation in yeast reveals multiple paths for acquiring higher stress resistance

BackgroundOrganisms frequently experience environmental stresses that occur in predictable patterns and combinations. For wild Saccharomyces cerevisiae yeast growing in natural environments, cells may experience high osmotic stress when they first enter broken fruit, followed by high ethanol levels during fermentation, and then finally high levels of oxidative stress resulting from respiration of ethanol. Yeast have adapted to these patterns by evolving sophisticated “cross protection” mechanisms, where mild ‘primary’ doses of one stress can enhance tolerance to severe doses of a different ‘secondary’ stress. For example, in many yeast strains, mild osmotic or mild ethanol stresses cross protect against severe oxidative stress, which likely reflects an anticipatory response important for high fitness in nature.ResultsDuring the course of genetic mapping studies aimed at understanding the mechanisms underlying natural variation in ethanol-induced cross protection against H2O2, we found that a key H2O2 scavenging enzyme, cytosolic catalase T (Ctt1p), was absolutely essential for cross protection in a wild oak strain. This suggested the absence of other compensatory mechanisms for acquiring H2O2 resistance in that strain background under those conditions. In this study, we found surprising heterogeneity across diverse yeast strains in whether CTT1 function was fully necessary for acquired H2O2 resistance. Some strains exhibited partial dispensability of CTT1 when ethanol and/or salt were used as mild stressors, suggesting that compensatory peroxidases may play a role in acquired stress resistance in certain genetic backgrounds. We leveraged global transcriptional responses to ethanol and salt stresses in strains with different levels of CTT1 dispensability, allowing us to identify possible regulators of these alternative peroxidases and acquired stress resistance in general.ConclusionsUltimately, this study highlights how superficially similar traits can have different underlying molecular foundations and provides a framework for understanding the diversity and regulation of stress defense mechanisms.

Ethanol distillation performance test of porang flour processing residue in SME scale distillator

Abstract One of the porang processing steps involves the production of porang flour with glucomannan concentrations that meet market requirements. To increase glucomannan levels, impurities are separated. The common solvent used is ethanol. In the extraction procedure, ethanol is not only used once but reused in another extraction procedure by redistilled to increase the concentration of ethanol. This study’s objective is to determine and analyze the effect of input volume variations (5,7.5 and 10 liters) at heating temperatures (85°C, 90°C, and 95°C) on ethanol product volume and concentration. In the distillation process 5; 7.5 and 10 L, ethanol levels respectively in the range of 58.41-71.87%, 61.81-65.87, and 63.76-65.93%. Based on the analysis, the constant value (k) in the change of ethanol distillation level during the distillation process predicted at 5L volume were 0.0149, 0.0160, and 0.0182, for 7.5L volume were 0.0098, 0.0134, and 0.0153, 0.0071, 0.0083, and 0.0102 minutes−1 at 10 L sample volume for each distillation temperature. The optimal values for a distillation device evaluated on a Small Medium Enterprises (SME) scale by varying the volume and temperature of the substance in the distillation vessel were determined to be 5 liters of volume and 95°C, respectively.

Analysis of ethanol content in commercially available Hand sanitizers in Oman using Gas Chromatography- A comparative study

The demand and usage of alcohol-based hand sanitizers have immensely increased worldwide in recent times as a protective measure against COVID-19 and other infectious diseases. To ensure that the public and health workers get hand sanitizers with an alcohol percentage within the label stated range, quality control of these products must be carried out by the governmental authorities. The study was designed to determine the ethanol content of eight commercially available and commonly used hand sanitizer gels available in the Omani market. Eight hand sanitizer gels were collected from various community pharmacies and commercial stores in Oman. The Gas Chromatography-Flame Ionisation Detection (GC-FID) method was employed to analyse ethanol content as it is one of the easiest, most reliable and most accurate methods for the determination of volatile content. Various physical properties of the samples such as pH, density, and refractive index, were also evaluated for comparison purpose. Sample prices were also compared from a pharmacoeconomic point of view. The majority of the hand sanitizers fell within the WHO recommendation range, with ethanol levels in the range of 66% to 76%, except A-7, which was below the recommended concentration for pathogenic microbial inactivation. The five of the samples tested showed ethanol contents below the labelled claim, whereas two sanitizers, A-1 and A-2, had ethanol contents above the labelled claim. The hand sanitizers differed significantly in price and certain physical properties which are likely to influence consumer preferences. Although the hand sanitizer products met the WHO requirements for alcohol content, the authors recommend that the regulatory authority should do a frequent testing of the samples available in the market to ensure quality and safety of these products.

Optimization of coffee vinegar production from cherry Pulp as by product of arabica coffee processing

Abstract The current high consumption of coffee globally has an impact on the abundance of by-products of coffee processing in the form of cherry pulp which are considered waste and cause environmental problems. The waste from arabica coffee is potential economic value products such as fermented vinegar because of its rich nutrients. The aim of this research was to determine the optimal conditions for acetic acid fermentation and the influence of yeast concentration and nitrogen sources fermented for 60 days. The research was carried out in two stages of fermentation using the Orleans method. Anaerobic alcohol fermentation (20 days) followed by aerobic acetic acid fermentation (40 days). A Randomized Block Factorial Design was used with 2 factors, namely yeast concentration (S15% and S210%) and NPK (N10g, N25g, and N310g), repeated 3 times. Observations on alcoholic fermentation were carried out every 5 days for 20 days regarding ethanol levels. Meanwhile, in acetic acid fermentation, 2.5% vinegar starter was added for each treatment and observations were made every 10 days for 40 days regarding the yield, acetic acid content, pH value and total dissolved solids. The results showed that the highest formation of acetic acid was obtained on day 30 of fermentation. The highest acetic acid content was obtained in the treatment S210%: N25g of 5.27%, pH value of 3.46, TSS value of 0.49 °Bx with a total yield of 42.93%. This is in line with the ethanol content previously formed on day 20 of 11.04% in the same treatment. This value meets the quality standards for fermented vinegar according to SNI 01-4371-1996 by at least 4%.

DoE-empowered development and validation of an environmentally sustainable RP-HPLC method for simultaneous estimation of antihypertensive drugs: AQbD perspective

AbstractThe primary objective of the present inquiry is to formulate a sustainable method employing Reverse Phase-High Performance Liquid Chromatography (RP-HPLC) for determination of Amlodipine (AM) and Irbesartan (IRB) simultaneously, compounds commonly prescribed for hypertension treatment. Existing literature underscores the absence of a comprehensive method in this regard. This research endeavors to align with the tenets of green chemistry by seamlessly integrating Analytical Quality by Design (AQbD) with RP-HPLC, replacing environmentally hazardous chemical modifiers with eco-friendly solvents. Identifying the critical variables as the 70% ethanol level and flow rate, a central composite design is applied for optimization. The separation is achieved utilizing a Phenomenex Luna column (C18, 250 mm × 4.6 mm i.d, 5 μm) with a mobile phase comprising ethanol and 0.1 % o-phosphoric acid in a 70:30 v/v ratio, flowing at 0.8 mL min−1, and detection wavelength of 242 nm. Green assessment methodologies are implemented to gauge the adherence of the proposed RP-HPLC method to eco-friendly principles while ensuring efficiency in chromatographic performance. The current developed method is rapid with retention time of 2.3 and 3.3 min for AM and IRB respectively and having a wide linear range from 55 to 130 μg mL−1, which makes the suitable for the accurate quantification of AM and IRB simultaneously in bulk and tablet dosage form, there by minimize environmental impact by providing a conscientious choice for the routine analysis which is achieved through the amalgamation of AQbD with a sustainable approach.

Effects of soil bulk density on respiratory metabolism and medicinal quality of Rehmannia Glutinosa root

As one of the famous "four huai medicines", Rehmannia glutinosa has a longstanding usage history. This study aimed to determine how variations in soil bulk density impact the respiration metabolism of R. glutinosa root tubers and, consequently, the quality of its medicinal components. We evaluated the enzymatic activities and key metabolic compounds of R. glutinosa across a range of soil bulk densities, which included: pyruvate kinase (PK), pyruvate decarboxylase (PDC), alcohol dehydrogenase (ADH), lactate dehydrogenase (LDH), isocitrate dehydrogenase (IDH), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), and the content of pyruvate, ethanol, lactic acid, succinic acid, malic acid, and quality indicators. High soil bulk density increased the activities of PK, PDC, ADH, and LDH, while decreasing SDH, MDH, and IDH in the roots. This shift in enzymatic activities corresponded to elevated levels of ethanol and lactic acid, and lowered levels of succinic and malic acids. In the additionally, the growth and development of aboveground parts of R. glutinosa were seriously hindered under high bulk weight treatment, and the number of root tubers decreased, root coarser and root length shortened, and the contents of catalpa alcohol and mullein were significantly reduced. Our findings suggest an optimal soil bulk density for R. glutinosa growth to be between 1.2 g/cm3 and 1.4 g/cm3. Based on changes in respiratory metabolism and medicinal quality indicators of R. glutinosa under different soil bulk densities, our findings suggest the optimal soil bulk density for R. glutinosa growth is 1.2 g/cm3 to 1.4 g/cm3. Excessively high soil bulk density will affect the growth and quality formation of R. glutinosa by influencing its metabolism. Recognizing this optimal range is crucial for maximizing the medicinal quality of R. glutinosa, offering valuable insights for cultivation practices.

Comparison of Different Spectral Ranges to Monitor Alcoholic and Acetic Fermentation of Red Grape Must Using FT-NIR Spectroscopy and PLS Regression

AbstractWine vinegar is produced through a two-phase fermentation of grape must: initially, yeast converts grape sugars into ethanol, and subsequently, acetobacteria oxidize ethanol into acetic acid. This process, spanning weeks when conducted by surface fermentation, requires constant monitoring of ethanol and total acidity levels. To enhance the quality and efficiency of process monitoring, vinegar production is shifting to faster, environmentally sustainable methods. Near-infrared (NIR) spectroscopy, recognized for its non-invasiveness and speed, is ideal for online implementation in process control. This study tracked dual fermentation in red grape must over an extended period, monitoring two different batches simultaneously to assess fermentation kinetics and reproducibility. Ethanol content and total acidity were analyzed in fermenting musts throughout the whole fermentation process using both classical laboratory analyses and FT-NIR spectroscopy. Principal Component Analysis (PCA) was used to explore the spectral dataset, then Partial Least Squares (PLS) was used to develop calibration models for predicting ethanol and acidity. The models calculated considering the entire spectral range were compared with those obtained for two narrower zones, where more cost-effective and easily miniaturizable sensors are available on the market. FT-NIR allowed to effectively determine ethanol content and acidity (R2Pred > 0.98), both over the entire range (12,500–4000 cm−1, corresponding to 800–2500 nm) and in the 10,526–6060 cm−1 (950–1650 nm) region. Although less satisfactory, still acceptable results were obtained in the 12,500–9346 cm−1 (800–1070 nm) region (R2Pred > 0.81), confirming the potential for cost-effective devices in real-time fermentation monitoring.

HISTOSTRUCTURAL ALTERATIONS IN THE CEREBRAL ARCHITECTURE ASSOCIATED WITH THE PROGRESSION OF ALCOHOL INTOXICATION: AN EXPERIMENTAL ANALYSIS

The aim of this study is to evaluate the histological changes in the cerebellum of rat brains following chronic alcohol intoxication. Materials and Methods. We employed a classical approach for modeling chronic alcohol intoxication by administering 40% ethanol to rats (n=55) in a dosage of 2 ml per 100 g of body weight daily for three months. The cerebellar structure was then analyzed. Results. This study investigates the impact of chronic alcohol intoxication on the histological structures of the cerebellar cortex. The cerebellum, like the nervous system overall, possesses a significant cellular and functional reserve. However, teratogenic factors, including ethanol, notably influence the activity of cerebellar neurons, increasing it. Ethanol exposure during early pregnancy can lead to prenatal growth retardation, stillbirth, cleft palate, hydrocephaly, and reductions in fetal body size. There is evidence suggesting a correlation between blood ethanol levels and a reduction in the number of Purkinje cells. Chronic alcohol consumption results in cerebellar ataxia, alterations in upper limb movements, decreased reaction speeds, reduced attention concentration, impaired coordination accuracy, and disturbances in postural stability and balance. Conclusion. The cerebellum, especially during development, is particularly vulnerable to the toxic effects of alcohol. Recent research suggests that changes in the neurotransmission of gamma-aminobutyric acid (GABA)-dependent receptors may contribute to cerebellar dysfunction induced by ethanol. Ethanol exposure increases the release of GABA not only in Purkinje cells but also in the interneurons of the molecular layer and granule cells.

Changes in Metabolite Profiles of Chinese Soy Sauce at Different Time Durations of Fermentation Studied by 1H-NMR-Based Metabolomics.

Fermentation parameters, especially the duration, are important in imparting a peculiar taste and flavor to soy sauce. The main purpose of this research was to monitor metabolic changes occurring during the various time intervals of the fermentation process. NMR-based metabolomics was used to monitor the compositional changes in soy sauce during fermentation. The 1H-NMR spectra of the soy sauce samples taken from the fermentation tanks at 0 to 8 months were analyzed using 1H-NMR spectroscopy, and the obtained spectra were analyzed by multivariate statistical analysis. The Principal Component Analysis (PCA) and Partial Least Square Discriminate analysis (PLSDA) revealed the separation of samples fermented for various time durations under identical conditions. Key metabolites shown by corresponding loading plots exhibited variations in amino acids (lysine, threonine, isoleucine, etc.), acetate, glucose, fructose, sucrose, ethanol, glycerol, and others. The levels of ethanol in soy sauce increased with longer fermentation durations, which can be influenced by both natural fermentation and the intentional addition of ethanol as a preservative. The study shows that the variation in metabolite can be very efficiently monitored using 1H-NMR-based metabolomics, thus suggestion to optimize the time duration to get the soy sauce product with the desired taste and flavor.

Optimal trade-off between boosted tolerance and growth fitness during adaptive evolution of yeast to ethanol shocks

BackgroundThe selection of Saccharomyces cerevisiae strains with higher alcohol tolerance can potentially increase the industrial production of ethanol fuel. However, the design of selection protocols to obtain bioethanol yeasts with higher alcohol tolerance poses the challenge of improving industrial strains that are already robust to high ethanol levels. Furthermore, yeasts subjected to mutagenesis and selection, or laboratory evolution, often present adaptation trade-offs wherein higher stress tolerance is attained at the expense of growth and fermentation performance. Although these undesirable side effects are often associated with acute selection regimes, the utility of using harsh ethanol treatments to obtain robust ethanologenic yeasts still has not been fully investigated.ResultsWe conducted an adaptive laboratory evolution by challenging four populations (P1–P4) of the Brazilian bioethanol yeast, Saccharomyces cerevisiae PE-2_H4, through 68–82 cycles of 2-h ethanol shocks (19–30% v/v) and outgrowths. Colonies isolated from the final evolved populations (P1c–P4c) were subjected to whole-genome sequencing, revealing mutations in genes enriched for the cAMP/PKA and trehalose degradation pathways. Fitness analyses of the isolated clones P1c–P3c and reverse-engineered strains demonstrated that mutations were primarily selected for cell viability under ethanol stress, at the cost of decreased growth rates in cultures with or without ethanol. Under this selection regime for stress survival, the population P4 evolved a protective snowflake phenotype resulting from BUD3 disruption. Despite marked adaptation trade-offs, the combination of reverse-engineered mutations cyr1A1474T/usv1Δ conferred 5.46% higher fitness than the parental PE-2_H4 for propagation in 8% (v/v) ethanol, with only a 1.07% fitness cost in a culture medium without alcohol. The cyr1A1474T/usv1Δ strain and evolved P1c displayed robust fermentations of sugarcane molasses using cell recycling and sulfuric acid treatments, mimicking Brazilian bioethanol production.ConclusionsOur study combined genomic, mutational, and fitness analyses to understand the genetic underpinnings of yeast evolution to ethanol shocks. Although fitness analyses revealed that most evolved mutations impose a cost for cell propagation, combination of key mutations cyr1A1474T/usv1Δ endowed yeasts with higher tolerance for growth in the presence of ethanol. Moreover, alleles selected for acute stress survival comprising the P1c genotype conferred stress tolerance and optimal performance under conditions simulating the Brazilian industrial ethanol production.

Valorization of Cashew Apple Waste into a Low-Alcohol, Healthy Drink Using a Co-Culture of Cyberlindnera rhodanensis DK and Lactobacillus pentosus A14-6.

This study investigated the potential of microbial fermentative transforming processes in valorizing the cashew apple by-product into a low-alcohol, health-benefiting beverage. We particularly investigated the use of a non-Saccharomyces yeast, Cyberlindnera rhodanensis DK, as the main targeted microbe. At 30 °C without agitation, C. rhodanensis DK caused changes in key parameters during the fermentation of cashew apple juice (CAJ) in terms of varied pH values and initial sugar concentrations. This result indicated that pure CAJ, with pH adjusted to 6 and with the original 6.85% (w/v) total sugar content, was the most feasible condition, as glucose and fructose were mostly consumed at 12 days of fermentation. A co-culture approach with either Saccharomyces cerevisiae TISTR 5088 or Lactobacillus pentosus A14-6 was investigated to improve both physicochemical and fermentation characteristics. Co-fermentation with S. cerevisiae TISTR 5088 resulted in significantly increased ethanol accumulation to 33.61 ± 0.11 g/L, but diminished bioactive compounds, antioxidant activity, and antidiabetic potential. In contrast, co-fermentation with L. pentosus A14-6 demonstrated excellent outcomes, as it significantly increased sugar consumption and finally remained at only 4.95 g/L compared to C. rhodanensis DK alone, produced lower levels of ethanol at only 19.47 ± 0.06 g/L, and higher total titratable acid (TTA), resulting in a final pH of 3.6. In addition, co-fermentation with this lactic acid bacterium significantly enhanced bioactive compounds and antioxidant activity and also retained potential antidiabetic properties. These findings highlight the feasibility of using tailored microbial fermentation strategies to produce low-alcohol beverages with enhanced health-promoting properties from CAJ; however, product-development processes following health food regulations and sensory evaluation are necessary.

Proposal of a New Predictive Score for the Requirement of Intensive Care Unit Admission Among Ethanol-Intoxicated Patients

ABSTRACT The correct selection of ethanol-intoxicated patients that require ICU admission is crucial for reducing morbidity and mortality. This study included 165 patients intoxicated with ethanol, confirmed by blood analysis. The patients were divided into ICU group = 46 and inward group = 119. We created a new predictive score based on seven parameters of APACHE II score and seven laboratory parameters that were significant in ICU-admitted patients by giving each parameter one point. The new predictive score achieved a sensitivity of 87, a specificity of 93.3, and an area under the curve of 0.967 with a cutoff >6. On the other hand, blood ethanol level (BEL) exhibited a weak correlation with both APACHE II and new predictive scores and a weak correlation with the parameters of the new predictive score. The new predictive score included 14 parameters. This score can be used in predicting whether patients need ICU admission during assessment for ethanol intoxication in emergency rooms, independent of BEL analysis. It could potentially reduce morbidity and mortality.

Early feasibility study with an implantable near-infrared spectroscopy sensor for glucose, ketones, lactate and ethanol.

To evaluate the safety and performance of an implantable near-infrared (NIR) spectroscopy sensor for multi-metabolite monitoring of glucose, ketones, lactate, and ethanol. This is an early feasibility study (GLOW, NCT04782934) including 7 participants (4 with type 1 diabetes (T1D), 3 healthy volunteers) in whom the YANG NIR spectroscopy sensor (Indigo) was implanted for 28 days. Metabolic challenges were used to vary glucose levels (40-400 mg/dL, 2.2-22.2 mmol/L) and/or induce increases in ketones (ketone drink, up to 3.5 mM), lactate (exercise bike, up to 13 mM) and ethanol (4-8 alcoholic beverages, 40-80g). NIR spectra for glucose, ketones, lactate, and ethanol levels analyzed with partial least squares regression were compared with blood values for glucose (Biosen EKF), ketones and lactate (GlucoMen LX Plus), and breath ethanol levels (ACE II Breathalyzer). The effect of potential confounders on glucose measurements (paracetamol, aspartame, acetylsalicylic acid, ibuprofen, sorbitol, caffeine, fructose, vitamin C) was investigated in T1D participants. The implanted YANG sensor was safe and well tolerated and did not cause any infectious or wound healing complications. Six out 7 sensors remained fully operational over the entire study period. Glucose measurements were sufficiently accurate (overall mean absolute (relative) difference MARD of 7.4%, MAD 8.8 mg/dl) without significant impact of confounders. MAD values were 0.12 mM for ketones, 0.16 mM for lactate, and 0.18 mM for ethanol. The first implantable multi-biomarker sensor was shown to be well tolerated and produce accurate measurements of glucose, ketones, lactate, and ethanol. Clinical trial identifier: NCT04782934.

Effect of Starter Culture and Temperature on the Flavor and Sensory Characteristics of Dry-Cured Ham.

This study focused on understanding the effects of yeast and mold on the sensory properties of dry-cured ham aged at 20°C and 25°C. Debaryomyces hansenii isolated from Doenjang and fermented sausages, and Penicillium nalgiovense isolated from fermented sausages were utilized. The CIE a* tended to increase in all treatments as the aging period increased. At 6 weeks of aging, DFD25 showed a significantly higher CIE a* value than other treatments. The shear force tended to increase in all treatments as the aging period increased. At 6 weeks of aging, among the treatments aged at 25°C, DFD25 showed a low tendency to shear force. The PC1 of the electronic nose was 42.872%. At 25°C, the hexane content was higher and levels of ethanol, propan-2-one, 2,4,5-trimethylthiazole, and limonene were lower than that at 20°C. DFD25 showed significantly higher hexane content and significantly lower limonene content than other treatments. The PC1 of the electronic tongue was 84.529%. All treatments, except for the C starter, exhibited higher salt and lower sour levels at 25°C compared to 20°C when the same starter was used. The DFD25 showed the lowest sour taste and a higher tendency of umami than the other treatments. Sensory evaluation revealed that DFD25 had significantly higher scores for texture than C25, whereas no significant differences were observed in other aspects. Therefore, the used starters are considered suitable for aging at 25°C; among them, the DFD starter demonstrates superior qualities and enhanced commercial potential compared to the control.

Recurrent Ethanol Treatment does not Produce a Cumulative Effect on Plasma Leptin Levels in Rats

Background: Hypoleptinemia is a potent orexigenic signal. Chronic ethanol (EtOH) consumption is associated with weight gain in humans, and acute ethanol treatment leads to reductions in leptin concentrations in both fasted humans and rats. The objective of this study was to determine whether short-term recurrent ethanol treatment has a cumulative effect on plasma leptin levels in rats, and whether treatment in the fed or fasted state influences the leptin lowering effect of EtOH. We hypothesized that recurrent EtOH treatment would have a cumulative effect on leptin levels, and that these effects would be more pronounced in the fed state, when leptin levels are highest. Methods: Two separate groups of male Sprague Dawley rats were exposed to either oral or intravenous (IV) treatment with EtOH (2g/kg via oral gavage or 0.067 g/kg/min IV for 30 min; n=7/group) on three consecutive days. Each rat served as its own control and received saline treatment either 7 d prior to or 7 d following exposure to EtOH. The oral group was fasted 12-14 h (overnight) prior to treatment, while the IV group was not fasted. Plasma leptin and EtOH levels were measured each day at baseline and 3 h following EtOH treatment. Data are expressed as mean ± SEM. A two-way repeated measures ANOVA was used to test for differences in plasma leptin and EtOH levels due to treatment (saline vs. EtOH) or day (cumulative effect of treatment). Results: Post-treatment ethanol levels in the oral and IV groups (55 ± 6.9 and 45 ± 6.3 mM, respectively) were higher than saline-treated rats (p>0.001). Baseline leptin levels in the oral treatment (fasted) group were 3.4 ± 0.3 and 3.9 ± 0.2 ng/mL under saline and leptin treated conditions, respectively. There was a significant main effect of treatment but not of day in the oral group (p=0.04 and p=0.38, respectively). In the oral group, average daily change in leptin levels 3 h post treatment in saline and leptin treated rats were -0.18 ± 0.05 and -0.72 ± 0.1 ng/mL, respectively. Baseline leptin levels in the IV group (fed) were 7.8 ± 0.3 and 6.7 ± 0.1 ng/mL, in saline and leptin treated rats respectively. There was no significant main effect of treatment or day in the IV group (p=0.17 and p=0.41, respectively). In the IV group, average daily changes in leptin levels 3 h following treatment in saline and leptin treated rats were -1.7 ± 0.2 and -1.3 ± 0.3 ng/mL, respectively. Conclusion: Although we were able to replicate previous work indicating reduced leptin levels following acute EtOH treatment in fasted rats, our results do not support a cumulative leptin-lowering effect of recurrent EtOH treatment. Furthermore, IV EtOH treatment in the fed state did not have any impact on leptin levels. This suggests that the leptin-lowering effects of EtOH may either require the fasted state or may require ingestion and exposure of the GI system to EtOH. NIH R01 DK131165-01A1. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Melatonin reduces brain injury following inflammation-amplified hypoxia-ischemia in a translational newborn piglet study of neonatal encephalopathy.

There is a need to develop therapies for neonatal encephalopathy (NE) in low- and middle-income countries (LMICs) where the burden of disease is greatest and therapeutic hypothermia (HT) is not effective. We aimed to assess the efficacy of melatonin following inflammation-amplified hypoxia-ischaemia (IA-HI) in the newborn piglet. The IA-HI model accounts for the contribution of infection/inflammation in this setting and HT is not cytoprotective. We hypothesised that intravenous melatonin (5% ethanol, at 20 mg/kg over 2 h at 1 h after HI + 10 mg/kg/12 h between 24 and 60 h) is safe and associated with: (i) reduction in magnetic resonance spectroscopy lactate/N-acetylaspartate (MRS Lac/sNAA); (ii) preservation of phosphorus MRS phosphocreatine/phosphate exchange pool (PCr/Epp); (iii) improved aEEG/EEG recovery and (iv) cytoprotection on immunohistochemistry. Male and female piglets underwent IA-HI by carotid artery occlusion and reduction in FiO2 to 6% at 4 h into Escherichia coli lipopolysaccharide sensitisation (2 μg/kg bolus + 1 μg/kg/h over 12 h). At 1 h after IA-HI, piglets were randomised to HI-saline (n = 12) or melatonin (n = 11). There were no differences in insult severity between groups. Target melatonin levels (15-30 mg/L) were achieved within 3 h and blood ethanol levels were <0.25 g/L. At 60 h, compared to HI-saline, melatonin was associated with a reduction of 0.197 log10 units (95% CrI [-0.366, -0.028], Pr(sup) 98.8%) in basal-ganglia and thalamic Lac/NAA, and 0.257 (95% CrI [-0.676, 0.164], Pr(sup) 89.3%) in white matter Lac/NAA. PCr/Epp was higher in melatonin versus HI-saline (Pr(sup) 97.6%). Melatonin was associated with earlier aEEG/EEG recovery from 19 to 24 h (Pr(sup) 95.4%). Compared to HI-saline, melatonin was associated with increased NeuN+ cell density (Pr(sup) 99.3%) across five of eight regions and reduction in TUNEL-positive cell death (Pr(sup) 89.7%). This study supports the translation of melatonin to early-phase clinical trials. Melatonin is protective following IA-HI where HT is not effective. These data guide the design of future dose-escalation studies in the next phase of the translational pipeline.

Simultaneous determination of methanol, ethanol, and formic acid in human serum by HSGC-MS: Method development, validation, and application

The concurrent assessment of methanol, ethanol, and formic acid is imperative for accurate clinical toxicology diagnosis in cases of methanol poisoning. Here, we reported a very simple and reliable headspace gas chromatography coupled with mass spectrometry (HSGC-MS) method for simultaneous quantitative determination of methanol, ethanol and formic acid in blood samples. The derivatization variables for formic acid were optimized. The method exhibits high sensitivity, good selectivity and effective separation factor for methanol (6.51 min), ethanol (8.29 min), and formic acid (11.41 min). The method demonstrates excellent reproducibility and accuracy. The limit of detection (LOD) and the limit of quantification (LOQ) were determined to be 3.14–5.83 μg/mL and 10.47–19.34 μg/mL, respectively. Finally, this method was successfully used for measuring the levels of methanol, ethanol, and formic acid in the serum of patients involved in actual methanol poisoning incidents.