A novel PDMS-based digital magnetofluidic platform for lab-on-a-chip applications.

Superhydrophobic gold film embedded PDMS was employed as a novel platform for digital magnetofluidics. The device performance for lab-on-a-chip applications was investigated by demonstrating four types of reactions. First, acid-based titration was introduced as a simple mixing reaction. Second, colorimetric detection of phosphate based on the molybdenum blue method was represented as a more complicated reaction. The fabricated device was able to determine the amount of phosphate in the concentration range of 10-100 ppm with %RSD of color intensity of less than 5%. Third, colorimetric detection of glucose using glucose oxidase was demonstrated as an enzymatic reaction. A linear range of 1-20 mM for determination of glucose was applied for measuring glucose in beverages with recovery percentages of glucose in the acceptable range of 89.6-106.8%. Finally, multistep analysis of C-reactive protein (CRP) based on immunomagnetic separation was successfully demonstrated on this proposed device. Therefore, the superhydrophobic gold-coated PDMS has shown its ability to be a simple platform for digital magnetofluidics for a variety of applications in the field of lab-on-a-chip technology.

Total flavonoids of Hippophae rhamnoides L. improves type 2 diabetes symptoms in rats through down-regulating of the DAG/PRKCA/MAPK10/p65/TNF-α signalling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Dry mature fruits of Hippophae rhamnoides L. (HRL), Elaeagnaceae, have traditional functions of invigorating spleen and improving spleen insufficiency. Traditional Chinese medicine (TCM) clinics have been proved that HRL is in favor of diabetes treatment. Modern pharmacological studies demonstrated that total flavones of Hippophae rhamnoides (TFH) are the main substance for HRL to develop anti-inflammation and anti-diabetes functions. However, chemical features, active ingredients and anti-diabetes pharmacological mechanism of HRL still remain unclear. AIM OF THE STUDY: Key targets and metabolites in anti-type-II diabetes mellitus (T2DM) of TFH have been explored based on AGE-RAGE signaling pathway in diabetic complications. The anti-T2DM mechanism of TFH has been elaborated from comprehensive perspectives, including target prediction, metabolites, potential metabolic pathways, and so on. MATERIALS AND METHODS: In this study, a qualitative test of chemical composition of HRL was carried out based on ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The anti-T2DM targets and pathways of HRL were predicted through network pharmacological approach. The T2DM rat model was induced by high-fat and high-glucose diet combined with streptozotocin (STZ). The T2DM model was evaluated through fasting blood glucose level, body weight, serum biochemical indicators, insulin levels and homeostatic model assessment of insulin resistance. The key metabolic pathways were screened through the correlation between metabolites and key targets. Finally, the quantitative analysis of key targets and metabolites was verified through experiments. RESULTS: After TFH intervention, the fasting blood-glucose level of T2DM rats induced by high-fat and high-glucose diet combined with streptozotocin (STZ) was downregulated significantly, while body weight, serum liquid level, insulin levels and homeostatic model assessment of insulin resistance (HOMA-IR) were improved. According to ELISA, Western blotting (WB) and reverse transcriptase polymerase chain reaction (RT-PCR), TFH significantly downregulates expression levels of diglyceride (DAG)-activated protein kinase C (PRKCA), mitogen activated protein kinase 10 (MAPK10), human nuclear factor κB subunit p65 (NF-κB p65) and tumor necrosis factor-α (TNF-α) in pancreas of STZ-induced rats. CONCLUSIONS: TFH downregulates expressions of PRKCA, MAPK10 and p65 TNF-α as well as level of the key metabolite DA in the DAG/PRKCA/MAPK10/TNF-α/p65 pathways, improves lipid metabolism disorder, inhibits inflammatory response and thereby relieves symptoms of T2DM.

Antrodia salmonea suppresses epithelial-mesenchymal transition/metastasis and Warburg effects by inhibiting Twist and HIF-1α expression in Twist-overexpressing head and neck squamous cell carcinoma cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Antrodia salmonea (AS), linked to the genus Taiwanofungus, is a medicinal fungus, and exhibits anti-inflammatory, anti-oxidant, and tumor inhibiting properties. AIM OF THE STUDY: In this study, we investigated the metabolic reprogramming and anti-metastasis/epithelial-mesenchymal transition (EMT) effects of AS exposure in Twist-overexpressing head and neck squamous cell carcinoma (HNSCC, OECM-1 and FaDu-Twist) cells. MATERIALS AND METHODS: MTT assay, Western blot, migration/invasion assay, immunofluorescence, glucose uptake assay, lactate assay, oxygen consumption rate (OCR)/Extracellular acidification rate (ECAR) assay, Liquid Chromatography-Electrospray Ionization Tandem Mass Spectrometry (LC-ESI-MS), and qRT-PCR experimental techniques were used to evaluate the therapeutic potential of AS treatment in HNSCC cells. RESULTS: This study showed that AS exhibits anti-EMT and anti-metastatic effects as well as metabolic reprogramming in Twist-overexpressing HNSCC cells. AS exposure inhibited Twist and hypoxia-inducible factor-1α (HIF-1α) protein and/or mRNA expression in Twist-overexpressing OECM-1 and FaDu-Twist cells. AS markedly suppressed EMT by enhancing the expression of E-cadherin; while the N-cadherin was suppressed. Furthermore, glucose uptake and lactate accumulation, together with HIF-1α-regulated glycolysis genes were diminished by AS in OECM-1 cells. AS decreased the ECAR, and enhanced the OCR together with basal respiration, ATP production, maximal respiration, and spare respiratory capacity under normoxia and hypoxia (CoCl2) in OECM-1 cells. There was a marked reduction in the level of glycolytic intermediate's; while TCA cycle metabolites were increased by AS treatment in OECM-1 cells. CONCLUSIONS: We concluded that AS treatment suppresses EMT/metastasis and Warburg effects through Twist and HIF-1α inhibition in Twist-overexpressing HNSCC cells.

Antidiabetic effect of Ardisia elliptica extract and its mechanisms of action in STZ-NA-induced diabetic rat model via 1H-NMR-based metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Ardisia elliptica Thunb. (AE) (Primulaceae) is a medicinal plant found in the Malay Peninsula and has been traditionally used to treat diabetes. However, limited studies to date in providing scientific evidence to support the antidiabetic efficacy of this plant by in-vitro and in-vivo models. AIM OF THE STUDY: To investigate the anti-hyperglycemic potential of AE through in-vitro enzymatic activities and streptozotocin-nicotinamide (STZ-NA) induced diabetic rat models using proton-nuclear magnetic resonance (1H-NMR)-based metabolomics approach. MATERIALS AND METHODS: Anti-α-amylase and anti-α-glucosidase activities of the hydroethanolic extracts of AE were evaluated. The absolute quantification of bioactive constituents, using ultra-high performance liquid chromatography (UHPLC) was performed for the most active extract. Three different dosage levels of the AE extract were orally administered for 4 weeks consecutively in STZ-NA induced diabetic rats. Physical assessments, biochemical analysis, and an untargeted 1H-NMR-based metabolomics analysis of the urine and serum were carried out on the animal model. RESULTS: Type 2 diabetes mellitus (T2DM) rat model was successfully developed based on the clear separation observed between the STZ-NA induced diabetic and normal non-diabetic groups. Discriminating biomarkers included glucose, citrate, succinate, allantoin, hippurate, 2-oxoglutarate, and 3-hydroxybutyrate, as determined through an orthogonal partial least squares-discriminant analysis (OPLS-DA) model. A treatment dosage of 250 mg/kg body weight (BW) of standardized 70% ethanolic AE extract mitigated increase in serum glucose, creatinine, and urea levels, providing treatment levels comparable to that obtained using metformin, with flavonoids primarily contribute to the anti-hyperglycemic activities. Urinary metabolomics disclosed that the following disturbed metabolism pathways: the citrate cycle (TCA cycle), butanoate metabolism, glycolysis and gluconeogenesis, pyruvate metabolism, and synthesis and degradation of ketone bodies, were ameliorated after treatment with the standardized AE extract. CONCLUSIONS: This study demonstrated the first attempt at revealing the therapeutic effect of oral treatment with 250 mg/kg BW of standardized AE extract on chemically induced T2DM rats. The present study provides scientific evidence supporting the ethnomedicinal use of Ardisia elliptica and further advances the understanding of the fundamental molecular mechanisms affected by this herbal antidote.

Akebia saponin D from Dipsacus asper wall. Ex C.B. Clarke ameliorates skeletal muscle insulin resistance through activation of IGF1R/AMPK signaling pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: Dipsacus asper Wall. Ex C.B. Clarke (DA), a perennial herb, is one of the most commonly used herbs in Traditional Chinese Medicine for strengthening muscles and bones and regulating blood vessels. Akebia saponin D (ASD/AVI) is a triterpenoid saponin extracted from the root of DA, which has favorable pharmacological properties such as anti-osteoporosis, anti-apoptosis, liver protection and hypolipidemic. AIM OF THE STUDY: To explore the underlying mechanisms and regulatory role of Akebia saponin D (ASD/AVI) on high-fat diet-induced insulin resistance in skeletal muscle. MATERIALS AND METHODS: C2C12 cells were used to explore the best concentration in the skeletal muscle insulin resistance model in an in vitro experiment. The protective effect of AVI on insulin resistance and the corresponding signaling pathway were detected by glucose content measurement, quantitative PCR, and Western blot. A high-fat diet STZ-induced insulin resistance mice model was used to evaluate the protective function of AVI in vivo. After four weeks of treatment, ITT, OGTT, and treadmill tests were applied to examine insulin sensitivity and their serum and skeletal muscle tissues were collected for further analysis. RESULTS: AVI effectively reduced body weight, blood glucose levels and calorie intake in insulin-resistant mice, and reduced lipid accumulation and in their muscle tissue. AVI also improved glucose uptake and insulin sensitivity in both in vivo and in vitro experiments. Following AVI administration, there was an increase in the expression of the AMPK signaling pathway. Our experiments further confirmed that AVI specifically targets the IGF1R, thereby more effectively regulating the insulin signaling pathway. CONCLUSION: AVI improves type 2 diabetes-induced insulin resistance in skeletal muscle by activating the IGF1R-AMPK signaling pathway, promoting glucose uptake and energy metabolism in IR.

Quantitative saccharide release of hydrothermally treated flours by validated salivary/pancreatic on-surface amylolysis (nanoGIT) and high-performance thin-layer chromatography.

The susceptibility of hydrothermally treated flour products to amylolysis was studied. The human salivary α-amylase and porcine pancreatin enzyme mixture containing α-amylase were used on-surface to investigate the release of glucose, maltose, and maltotriose. On the same adsorbent surface (all-in-one), their high-performance thin-layer chromatography separation and detection via selective chemical derivatization was performed. For the first time, the all-in-one nanoGIT system was studied quantitatively and validated for the simulated static oral and intestinal on-surface amylolysis of ten different hydrothermally treated flours and soluble starch. Differences were detected in the digestibility of refined and whole flours from wheat, spelt, and rye as well as from einkorn, amaranth, emmer, and oat. Amaranth released the lowest amount of saccharides and spelt the highest in both oral and intestinal digestion systems. The results suggest that consumption of whole grain products may be beneficial because of their lower saccharide release, with particular attention to rye.

Integrated Cu-Au stereo microelectrode arrays and microfluidic channels for the electrochemical detection of glucose.

Noble and transition metal nanomaterials are widely used in glucose sensing. However, the fabrication of these sensors still suffers from complex nanomaterial synthesis process and unstable nanomaterial loading on sensing surfaces. Herein, a Cu-Au bimetallic microelectrode array was prepared via local electrochemical deposition and electrochemical reduction without the need for templates and additional nanomaterial preparation processes. Based on the COMSOL computational fluid study, the obtained microelectrode arrays combined with microfluidic channels allow the continuous and rapid detection of glucose. Large number of active sites on the surface of 3D nano-arrays contributes to excellent sensing performance for glucose, with good linear detection ranges in 10 µM to 4 × 102 µM and 4 × 102 µM to 4 × 105 µM, and a low detection limit of 284 nM. The feasibility of sensor in real sample was verified by detecting glucose in beverages with good recoveries ranging from 95.50% to 104.31%.

Synergistic effect of residual sugar on freeze-thaw stability of high internal phase emulsions using glycosylated cod protein as interface stabilizer.

Nowadays, glycosylated protein seems to be one of the most effective stabilizers for preparing freeze-thaw stabile emulsion; nevertheless, few papers mentioned the relationship between the residual free sugars after the glycosylation reaction and the freeze-thaw stability of high internal phase emulsions (HIPEs). Herein, glucose was used to prepare glycosylated cod proteins (GCPs). The synergistic effect was related to the grafting degree of GCP, and the amount of glucose added to prepare freeze-thaw stable HIPEs was reduced from 20% to 4% when the grafting degree of GCP increased from 0% to 31.58% (i.e. 12% GCP). This might be due to fewer ice crystals forming in water phase or less destruction of emulsion droplets by ice crystals. The obtained results in this study will allow developing freeze-thaw stable HIPEs or new frozen ingredients.

A colorimetric assay for H2O2 and glucose based on the morphology transformation of Au/Ag nanocages to nanoboxes.

Herein, we introduced a sensitive colorimetric platform for hydrogen peroxide (H2O2) assay based on gold/silver (Au/Ag) nanocages with porous structure. In the presence of H2O2, the morphology of hollow Au/Ag nanocages was converted to closed nanoboxes, altering their localized surface plasmon resonance (LSPR) peak position and the solution color from light blue to deep blue. The morphology transformation and LSPR peak position of Au/Ag nanocages were proportional to H2O2 concentration at the range of 0.1 to 50 µM. The limit of detection (LOD) was obtained to be 0.02 µM, and the relative standard deviation (RSD, for 0.2, 2.0, and 20 µM) was 2.7, 2.3, and 2.9%, respectively. Moreover, a smartphone-based colorimetric sensor was developed for H2O2 assay at the concentration range of 0.25-4.0 µM, with LOD of 0.2 µM and RSD of 3.2, 2.5, and 2.9% (for 0.5, 1.0, and 3.0 µM, respectively). We exploited the established sensor for glucose assay by measuring the generated H2O2 from the enzymatic reaction between glucose and glucose oxidase. There was a linear relationship between LSPR peak wavelength variations and the amount of glucose from 1.0 to 50 µM, with LOD of 0.4 µM and RSD of 3.2, 3.1, and 3.8% (for 2.0, 10, and 30 µM, respectively). The sensor was successfully applied to determine H2O2 and glucose in food and human serum samples, respectively.

Sargentodoxa cuneata and Patrinia villosa extract inhibits LPS-induced inflammation by shifting macrophages polarization through FAK/PI3K/Akt pathway regulation and glucose metabolism reprogramming.

ETHNOPHARMACOLOGICAL RELEVANCE: Sargentodoxa cuneata and Patrinia villosa (S&P) are two natural herbal medicine widely used for treatment of various inflammatory diseases in Traditional Chinese Medicine, whereas the mode of action needs to be further investigated. AIM OF THE STUDY: This study aimed to explore the anti-inflammatory effects and unravel the involved mechanism of S&P extract. MATERIALS AND METHODS: The components of S&P extract were first detected using the liquid chromatography-tandem mass spectrometry (LC-MS/MS). The effects of S&P extract on the viability and migration ability of macrophages were detected using CCK8, LDH, adhesion and transwell assays. Cytokine release and macrophage phenotype transition were measured using a cytometric bead array and flow cytometry. The potential mechanism was uncovered using an integrative approach combining RNA sequencing and LC-MS/MS-based metabolic analysis. The expression of related proteins was further validated using western blotting. RESULTS: S&P extract inhibited the proliferation and migration of LPS-induced macrophages, changed the morphology of macrophages, and inhibited the production of NO and the expression of iNOS. Furthermore, the extract inhibited tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) production and the expression of the M1 phenotype markers CD11c and CD16/32, whereas it promoted interleukin-10 (IL-10) production and the expression of the M2 phenotype markers CD206 and arginase 1 (Arg1). RNA sequencing analysis demonstrated that the upregulated genes by S&P extract treatment were involved in M2 macrophages: Il10, Ccl17, Ccl22, Cd68. The downregulated genes were involved in M1 macrophages and glycolysis processes: Stat1, Il18, Cd80, Cd86, Nos2, Il6, Pik3ap1, Raf1, Pdhb, etc. Metabolomics results showed that the S&P extract strongly ameliorated lipopolysaccharide (LPS)-induced metabolic disturbances. KEGG analysis indicated that most of these metabolites were involved in glucose metabolism, which is involved in the tumor necrosis factor (TNF), phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt), Glycolysis, and mitogen-activated protein kinase (MAPK) pathways. In vitro experiments further confirmed that the extract significantly inhibited the phosphorylation of focal adhesion kinase (FAK), PI3K and Akt, and the expression of glucose metabolism-related proteins. Adding a FAK inhibitor (defactinib) further inhibited the expression of M1/M2 phenotypic markers and the phosphorylation of FAK, PI3K, and Akt. CONCLUSIONS: S&P extract can induce M2 polarization and shift macrophages from M1 to M2 tissue repair in LPS-induced inflammation by regulating glucose metabolism and the FAK/PI3K/Akt pathway.

Development of a biodegradable microfluidic paper-based device for blood-plasma separation integrated with non-enzymatic electrochemical detection of ascorbic acid.

In the present article, we developed an electrochemical microfluidic paper-based device (EµPAD) for the non-enzymatic detection of Ascorbic Acid (AA) concentration in plasma using whole human blood. We combined LF1 blood plasma separation membrane and Whatman grade 1 filter paper to separate plasma from whole blood through wax printing. A screen-printed electrode (SPE) was modified with spherical-shaped MgFe2O4 nanomaterial (n-MgF) to improve the catalytic properties of SPE. The n-MgF was prepared via hydrothermal method, and its material phase and morphology were confirmed via XRD, FTIR, TEM, SEM, and AFM analysis. The fabricated n-MgF/SPE/EµPAD exhibited detection of AA ranging from 0 to 80 µM. The obtained value of the detection limit, limit of quantification, sensitivity, and response time are 2.44 µM, 8.135 µM, 5.71 × 10-3 mA µM-1 cm-2, and 10 s, respectively. Our developed n-MgF/SPE/EµPAD shows marginal interference with the common analytes present in plasma, such as uric acid, glutamic acid, glucose, urea, lactic acid, and their mixtures. Overall, our low-cost, portable device with its user-friendly design and efficient plasma separation capability offers a practical and effective solution for estimating AA concentration from whole human blood in a single step.

Glucose oxidation induced pH stimuli response controlled release electrochemiluminescence biosensor for ultrasensitive detection of CYFRA 21-1.

Herein, a self on electrochemiluminescence (ECL) biosensor was constructed by pH stimuli response controlled release strategy, in which SiO2-PEI as the carrier, BSA/luminol-Ab2 as the encapsulated substance, gold nanoparticles (Au NPs) as the blocking cap, glucose as the inducer. In addition, CeO2-Au was used as catalyst, which generated more O2•- to increase the ECL signal. Under the action of voltage, the glucose was oxidized to gluconic acid, which induced the pH to decrease accordingly. Therefore, Au NPs were stimulated to fall from the surface of SiO2-PEI, releasing the BSA/luminol-Ab2 to realize self on mode. With such design, the constructed self on ECL biosensor owned an ultrasensitive detection capacity of CYFRA 21-1, showing an excellent linear relationship in the range of 0.001-100000 ng/L and 0.4 fg/mL low limit of detection (LOD). It provided an innovative idea for the biosensor construction to clinical detection of lung cancer.

Qingjie decoction attenuated E.coli-induced diarrhea by regulating energy metabolism and alleviating inflammation based on network analysis and metabolomics.

ETHNOPHARMACOLOGICAL RELEVANCE: Diarrhea is a frequently encountered gastrointestinal complication in clinical practice, and E. coli is one of the main causative agents. Although Qingjie decoction (QJD) has been shown to be highly effective in treating diarrhea by eliminating heat-toxin, the underlying molecular mechanisms and pathways of QJD remain unclear. AIM OF REVIEW: The aim of this research was to explore the effects and fundamental mechanism of QJD on diarrhea induced by E.coli in rats. MATERIALS AND METHODS: Initially, we used UHPLC-MS/MS analysis to identify the chemical composition of QJD. Then, we constructed a visualization network using network pharmacology. Next, we utilized metabolomics to identify differentially expressed metabolites of QJD that are effective in treating diarrhea. RESULTS: The chemical composition of QJD was analyzed using UHPLC-MS/MS, which identified a total of 292 components. Using a network pharmacology approach, 127 bioactive compounds of QJD were screened, targeting 171 potential diarrhea treatment targets. TNF-α, IL-6, IL-1ß, and CAT were identified as important targets through visualizing the PPI network. Enrichment analysis demonstrated significant enrichment in the TNF signaling pathway, IL-17 signaling pathway, and PI3K-Akt signaling pathway. QJD showed beneficial effects, such as increased body weight, decreased fecal water content, and reduced inflammatory cell infiltration in the duodenum and colon, as well as maintaining the structure of the duodenum and colon. Metabolomic analysis revealed 32 differentially expressed metabolites in the control, model and QJD-H groups, including glucose, valine, and cysteine. Functional analysis indicated that differential metabolites were related to energy metabolism, including glucose metabolism, TCA cycle, and amino acid metabolism. CONCLUSION: QJD significantly increased body weight, decreased water content in feces, relieved inflammatory cell infiltration, maintained the structure of duodenum and colon. Combining network analysis and metabolomics, QJD exerted therapeutic effects by inhibiting inflammation and oxidative stress, regulating glucose metabolism, tricarboxylic acid metabolism, and amino acid metabolism.

Failure to replicate the diabetes alleviating effect of a maternal gluten-free diet in non-obese diabetic mice.

Type 1 diabetes (T1D) is an autoimmune disease with an unexplained rising incidence for which environmental factors like gluten may play a role. Previously, we showed that a gluten-free (GF) diet provided strictly in utero reduces the autoimmune diabetes incidence in Non-Obese Diabetic (NOD) mice compared to a gluten-containing standard (STD) diet. The current study was initiated to elucidate possible mechanisms behind the diabetes-alleviating effect of the same diet intervention. NOD mice received either a GF Altromin diet or a STD Altromin diet during pregnancy. Female offspring from both groups were fed a STD diet throughout life and their diabetes incidence was recorded for 200 days. The following parameters were measured in 13-week-old female offspring: insulitis degree, glucose and insulin tolerance, and plasma insulin autoantibody titer. The diet intervention showed no reduction in autoimmune diabetes incidence, insulitis degree, glucose nor insulin tolerance and plasma insulin autoantibody titer. In conclusion, this study could not replicate the previously observed diabetes alleviative effects of a maternal gluten-free diet in NOD mouse offspring and could therefore not further elucidate potential mechanisms.

Effect of fenofibrate and selective PPARα modulator (SPPARMα), pemafibrate on KATP channel activity and insulin secretion.

OBJECTIVE: Insulin secretion is regulated by ATP-sensitive potassium (KATP) channels in pancreatic beta-cells. Peroxisome proliferator-activated receptors (PPAR) α ligands are clinically used to treat dyslipidemia. A PPARα ligand, fenofibrate, and PPARγ ligands troglitazone and 15-deoxy-∆12,14-prostaglandin J2 are known to close KATP channels and induce insulin secretion. The recently developed PPARα ligand, pemafibrate, became a new entry for treating dyslipidemia. Because pemafibrate is reported to improve glucose intolerance in mice treated with a high fat diet and a novel selective PPARα modulator, it may affect KATP channels or insulin secretion. RESULTS: The effect of fenofibrate (100 µM) and pemafibrate (100 µM) on insulin secretion from MIN6 cells was measured by using batch incubation for 10 and 60 min in low (2 mM) and high (10 mM) glucose conditions. The application of fenofibrate for 10 min significantly increased insulin secretion in low glucose conditions. Pemafibrate failed to increase insulin secretion in all of the conditions experimented in this study. The KATP channel activity was measured by using whole-cell patch clamp technique. Although fenofibrate (100 µM) reduced the KATP channel current, the same concentration of pemafibrate had no effect. Both fenofibrate and pemafibrate had no effect on insulin mRNA expression.

Changes in Branched-Chain Amino Acids One Year after Sleeve Gastrectomy in Youth with Obesity and Their Association with Changes in Insulin Resistance.

Adults with obesity have a reduction in branched-chain amino acid (BCAA) levels following metabolic and bariatric surgery (MBS), which is hypothesized to contribute to the metabolic advantages of MBS. We examined this relationship in 62 youth 13-24 years old with severe obesity (47 female) over 12 months. Thirty had sleeve gastrectomy (SG) and 32 were non-surgical controls (NS). We measured fasting insulin, glucose, glycated hemoglobin (HbA1c), isoleucine, leucine, and valine concentrations, and post-prandial insulin and glucose, following a mixed meal tolerance test. Twenty-four-hour food recalls were collected. At baseline, groups did not differ in the intake or the serum levels of BCAAs, HbA1C, HOMA-IR, Matsuda index, insulinogenic index, or oral Disposition index (oDI). Over 12 months, SG vs. NS had greater reductions in serum BCAAs, and SG had significant reductions in BCAA intake. SG vs. NS had greater reductions in HbA1c and HOMA-IR, with increases in the Matsuda index and oDI. In SG, baseline leucine and total BCAA concentrations were negatively correlated with the baseline Matsuda index. Reductions in serum leucine were positively associated with the reductions in HOMA-IR over 12 months. These associations suggest a potential role of BCAA in regulating metabolic health. Reducing dietary intake and serum BCAA concentrations may reduce insulin resistance.

Analytical Performance of the Factory-Calibrated Flash Glucose Monitoring System FreeStyle Libre2TM in Healthy Women.

Continuous glucose monitoring (CGM) is used clinically and for research purposes to capture glycaemic profiles. The accuracy of CGM among healthy populations has not been widely assessed. This study assessed agreement between glucose concentrations obtained from venous plasma and from CGM (FreeStyle Libre2TM, Abbott Diabetes Care, Witney, UK) in healthy women. Glucose concentrations were assessed after fasting and every 15 min after a standardized breakfast over a 4-h lab period. Accuracy of CGM was determined by Bland-Altman plot, 15/15% sensor agreement analysis, Clarke error grid analysis (EGA) and mean absolute relative difference (MARD). In all, 429 valid CGM readings with paired venous plasma glucose (VPG) values were obtained from 29 healthy women. Mean CGM readings were 1.14 mmol/L (95% CI: 0.97 to 1.30 mmol/L, p < 0.001) higher than VPG concentrations. Ratio 95% limits of agreement were from 0.68 to 2.20, and a proportional bias (slope: 0.22) was reported. Additionally, 45% of the CGM readings were within ±0.83 mmol/L (±15 mg/dL) or ±15% of VPG, while 85.3% were within EGA Zones A + B (clinically acceptable). MARD was 27.5% (95% CI: 20.8, 34.2%), with higher MARD values in the hypoglycaemia range and when VPG concentrations were falling. The FreeStyle Libre2TM CGM system tends to overestimate glucose concentrations compared to venous plasma samples in healthy women, especially during hypoglycaemia and during glycaemic swings.

Organic Electrochemical Transistor with MoS2 Nanosheets Modified Gate Electrode for Sensitive Glucose Sensing.

An organic electrochemical transistor (OECT) with MoS2 nanosheets modified on the gate electrode was proposed for glucose sensing. MoS2 nanosheets, which had excellent electrocatalytic performance, a large specific surface area, and more active sites, were prepared by liquid phase ultrasonic exfoliation to modify the gate electrode of OECT, resulting in a large improvement in the sensitivity of the glucose sensor. The detection limit of the device modified with MoS2 nanosheets is down to 100 nM, which is 1~2 orders of magnitude better than that of the device without nanomaterial modification. This result manifests not only a sensitive and selective method for the detection of glucose based on OECT but also an extended application of MoS2 nanosheets for other biomolecule sensing with high sensitivity.

Comparison of Oxidant and Antioxidant Status in the Aqueous Humour of Cataract Patients with and without Type 2 Diabetes Mellitus.

OBJECTIVES: To compare the total oxidant status (TOS), total antioxidant capacity (TAC), and oxidative stress index (OSI) in the aqueous humour of cataract patients with and without type 2 diabetes mellitus (T2DM). STUDY DESIGN: Cross-sectional analytical study. Place and Duration of the Study: Department of Physiology, Army Medical College, National University of Medical Sciences, Rawalpindi, Pakistan in collaboration with Armed Forces Institute of Ophthalmology, from 1st June 2022 to 31st January 2023. METHODOLOGY: A total of 120 individuals were recruited and divided into 2 equal groups. Sample of aqueous humour was collected, and the total oxidant status, and total antioxidant status were measured by spectrophotometry. Blood samples were collected to measure the fasting glucose and glycosylated haemoglobin levels. Results were analysed by SPSS version 24 and p-value ≤0.05 was considered significant. RESULTS: Total oxidative stress and OSI were (55.57±5.46) vs. (33.75±4.28) = p (0.001), (32.79±9.23) vs. (13.28±3.02) = p (0.001) found significantly increased in aqueous humour of diabetic patients while TAC was reduced (1.86±0.41) vs. (2.62 ± 0.45) = p (0.001) compared to the non-diabetics. Fasting blood glucose (109.18±12.32 vs. 91.61±5.71 mg/dl, p=0.001) and glycosylated haemoglobin levels (6.46±0.85 vs.5.43±0.85, p=0.001) were also found significantly high in the diabetic group compared to the non-diabetics. CONCLUSION: Increased levels of glucose and glycosylated haemoglobin increase oxidative stress in the blood of T2DM patients. This raises the total oxidative stress in the aqueous homor while lowering the TAC. This disturbed oxidant-antioxidant status in the aqueous humour of diabetic patients plays an important role in the development of cataract in T2DM patients compared to non-diabetics. KEY WORDS: Cataract, Aqueous humour, Oxidative stress, Type 2 diabetes mellitus.