Topic: 26. Sickle cell disease Background: Townes and Berkeley mouse models of sickle cell disease (SCD) are commonly used to study disease pathophysiology and to develop novel therapies. Despite similar pathophysiology to SCD patients, there are distinctive differences. For example, Townes HbSS mice have higher ATP and lower 2,3-diphosphyglycerate (2,3-DPG) levels compared to HbAA mice. This contrasts with SCD patients, who show high 2,3-DPG but low ATP, suggesting potential differences between human and murine red blood cell (RBC) metabolism. Pyruvate kinase (PK), a key ATP-generating enzyme in glycolysis, is a target for novel SCD therapies. While PK activators have been shown to increase hemoglobin in clinical trials, such treatments do not always lead to the same hematological change in mouse models, further suggesting that PK features may differ between SCD mouse models and human patients. A detailed characterization of PK and related important cellular metabolites in murine SCD models and SCD patients is therefore important. Aims: To study PK features in murine and human SCD RBCs. Methods: Wild-type (C57BL/6J), Townes and Berkeley mice (non-sickling and sickling) were studied. Human blood was obtained from healthy controls (HbAA) and untreated SCD patients (HbSS). Hematological parameters were measured using the Cell-Dyn Sapphire (Abbott Diagnostics). PK and hexokinase (HK) activity, and PK thermostability (53°C) were measured on purified RBCs. Levels of ATP, 2,3-DPG, and reduced glutathione (GSH) were measured by LC-MS/MS and spectrophotometry, respectively. Results: Samples from 12 wild-type mice, 13 Townes mice (6 HbAA, 7 HbSS), 6 Berkeley mice (3 non-sickling, 3 sickling) and 10 human subjects (6 HbAA, 4 HbSS) were included. High reticulocyte counts compared to control mice were observed in both Townes and Berkeley SCD mice (median 59% and 48% compared to 7% and 5%, respectively). This was accompanied by an increased activity of PK in both SCD models (Figure 1A). When normalizing PK activity to that of HK, another age-dependent enzyme, no difference was found between sickling and non-sickling mice for both mouse models (Figure 1B). However, PK/HK ratio was lower in the Berkeley mice compared to Townes mice (p<0.01 for both non-sickling and sickling mice). ATP/2,3-DPG ratio was higher in Townes and Berkeley sickling mice compared to their control mice (Figure 1C; p<0.01). This contrasts with humans where healthy controls had higher ATP/2,3-DPG ratio than SCD patients (p<0.05). Surprisingly, mouse RBC PK was considerably less stable than its human counterpart, as reflected by a strikingly decreased thermostability (Figure 1D). GSH levels were significantly increased in both Townes and Berkeley sickling mice compared to non-sickling mice, which is in contrast with previous findings that SCD patients have lower GSH levels. Summary/Conclusion: PK/HK ratios are similar between sickling and non-sickling mice in both Berkeley and Townes model strains. In both models, however, there are significant changes in energy level (ATP), 2,3-DPG (important for oxygen affinity), and antioxidant level (GSH) when compared sickling to non-sickling mice. Our results further reveal important distinctions between mouse models and humans: the lack of PK/HK ratio difference between sickling and non-sickling mice, and overall lower PK thermostability in mice. Sickling mice also have higher ATP/2,3-DPG ratio and GSH levels, which is very different from SCD patients. These differences should be considered when studying PK activators in SCD mouse models and extrapolating results to a clinical response.Keywords: Transgenic mice, Pyruvate kinase, Sickle cell disease
Abstract Tea is one of the most popular beverages in the world, especially in Asian societies. Green, oolong and black tea are three main types of tea products. Catechin is the principal polyphenol compound in all tea products including four major subgroup compounds, epigallocatechin gallate (EGCG), epicatehin (EC), epigallocatechin (EGC) and epicatechin gallate (ECG). Green tea contains highest amount of catechin compared to oolong and black tea since fermentation process can significantly reduce the amount of catechin in tea product, which polyphenol oxidase can convert catechins to theaflavins and thearubigins during fermentation process. Therefore, green tea catechin is becoming more and more attractive to nutritionists since it can provide several health benefits to human body. Cholesterol lowering effect is one of the health benefits been studied and proposed over decade. There are well documented evidences that suggested green tea catechin, in particular EGCG has the potential to lower blood cholesterol concentrations. Since the pool bioavailability and absorption ability of catechin, researchers believed that green tea catechin may significantly inhibit lipids absorption in intestine. Mechanisms are including inhibition of pancreatic lipase activity, lipids hydrolysis, and emulsification in intestine and precipitation of micellar cholesterol. In vitro studies, animal studies as well as most of human RCT, consistent results been observed that dietary intake of green tea beverages or extracts could significantly lower circulating cholesterol concentration, in particular lower LDL-C and total cholesterol level. However, in 2010, European Food Safety Authority (EFSA) denied the health claim of cholesterol lowering benefits of dietary intake of green tea or green tea catechins. In this presentation, current scientific evidences and EFSA judgment will be reviewed and discussed.
Abstract Short‐chain fatty acids (SCFAs) are the main metabolites of the intestinal flora and play an important role in the interaction between the intestinal flora and host metabolism. Therefore, reliable methods are needed to accurately measure SCFAs concentrations. SCFAs are commonly analyzed by gas chromatography–mass spectrometry (GC‐MS), which requires lengthy sample treatments and a long run time. This study aimed to develop a fast GC method with formic acid pretreatment for SCFAs quantification in the plasma of rat. Baseline chromatographic resolution was achieved for three SCFAs (acetic, propionic, and butyric) within an analysis time of 10.5 min. The method exhibited good recovery for a wide range of concentrations with a low limit of detection for each compound. The relative standard deviations (RSDs) of all targeted compounds showed good intra‐ and interday precision (<10%). We used our method to measure SCFAs levels in plasma samples from rats fed with a high fructose diet (HFD) to test the accuracy of the developed method. It was shown that SCFAs are indeed affected negatively by a HFD (60% fructose). This method was successfully employed to accurately determine SCFAs in the rat plasma with minimum sample preparation. Results showed potential damage of HFD, which produced lower SCFAs. Practical Application Increasingly, microbiota and gut health research are being conducted by many food scientists to elucidate the relationships among the factors of food components, particularly the nondigestible carbohydrates, food processing conditions, and potential health impact. This research provides a useful, rapid, and accurate method that can save time in the analysis of short‐chain fatty acids, which are commonly analyzed in gut health research.
The present study aimed to develop the utility of a nomogram based on clinical and radiomics as a tool for predicting post-acute pancreatitis diabetes mellitus (PPDM-A).
Traditional edible barbecue products use with lemon juice not only make the barbecue more delicious but also reduce the risk of PAHs in the barbecue products. One of the major economics crops in Taiwan, the waste from citrus fruits was very tremendous mass. However, the peelings of citrus fruits are rich in essential oil, especially, the limonene is the major. Whether the anti-carcinogenesis activities of terpene, such as limonene, in citrus fruits essential oil extraction. This study to demonstrate the PAHs content in fish skin increased markedly after being roasted at 210℃ for 20 minutes and greater mutagenicity risk of roasted fish skin was observed by Ame's test. The reduction of mutagenicity risk of roasted fish skin, which the antimutagenic abilities of substances in descending order were limonene > cold pressure oil > lemon >grapefruit. The antimutagenicity rate and ability of the three extracts were limonene: 18–23%; cold-pressed lemon oil: 18–22%; and steam distilled lemon essential oil: 8–16%. The obvious anti- mutagenicity effects against the PAHs mutagenicity of roasted fish skins can be found in citrus fruits essential oil extraction.
Microfluidic mixers, a pivotal application of microfluidic technology, are primarily utilized for the rapid amalgamation of diverse samples within microscale devices. Given the intricacy of their design processes and the substantial expertise required from designers, the intelligent automation of microfluidic mixer design has garnered significant attention. This paper discusses an approach that integrates artificial neural networks (ANNs) with reinforcement learning techniques to automate the dimensional parameter design of microfluidic mixers. In this study, we selected two typical microfluidic mixer structures for testing and trained two neural network models, both highly precise and cost-efficient, as alternatives to traditional, time-consuming finite-element simulations using up to 10,000 sets of COMSOL simulation data. By defining effective state evaluation functions for the reinforcement learning agents, we utilized the trained agents to successfully validate the automated design of dimensional parameters for these mixer structures. The tests demonstrated that the first mixer model could be automatically optimized in just 0.129 s, and the second in 0.169 s, significantly reducing the time compared to manual design. The simulation results validated the potential of reinforcement learning techniques in the automated design of microfluidic mixers, offering a new solution in this field.
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