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Nutrients Jul 2023Combining exercise with fasting is known to boost fat mass-loss, but detailed analysis on the consequential mobilization of visceral and subcutaneous WAT-derived fatty...
Combining exercise with fasting is known to boost fat mass-loss, but detailed analysis on the consequential mobilization of visceral and subcutaneous WAT-derived fatty acids has not been performed. In this study, a subset of fasted male rats (66 h) was submitted to daily bouts of mild exercise. Subsequently, by using gas chromatography-flame ionization detection, the content of 22 fatty acids (FA) in visceral (v) versus subcutaneous (sc) white adipose tissue (WAT) depots was compared to those found in response to the separate events. Findings were related to those obtained in serum and liver samples, the latter taking up FA to increase gluconeogenesis and ketogenesis. Each separate intervention reduced scWAT FA content, associated with increased levels of adipose triglyceride lipase (ATGL) protein despite unaltered AMP-activated protein kinase (AMPK) Thr172 phosphorylation, known to induce ATGL expression. The mobility of FAs from vWAT during fasting was absent with the exception of the MUFA 16:1 n-7 and only induced by combining fasting with exercise which was accompanied with reduced hormone sensitive lipase (HSL) Ser563 and increased Ser565 phosphorylation, whereas ATGL protein levels were elevated during fasting in association with the persistently increased phosphorylation of AMPK at Thr172 both during fasting and in response to the combined intervention. As expected, liver FA content increased during fasting, and was not further affected by exercise, despite additional FA release from vWAT in this condition, underlining increased hepatic FA metabolism. Both fasting and its combination with exercise showed preferential hepatic metabolism of the prominent saturated FAs C:16 and C:18 compared to the unsaturated FAs 18:1 n-9 and 18:2 n-6:1. In conclusion, depot-specific differences in WAT fatty acid molecule release during fasting, irrelevant to their degree of saturation or chain length, are mitigated when combined with exercise, to provide fuel to surrounding organs such as the liver which is correlated with increased ATGL/ HSL ratios, involving AMPK only in vWAT.
Topics: Rats; Male; Animals; Sterol Esterase; Fatty Acids; AMP-Activated Protein Kinases; Lipase; Lipolysis; Obesity; Fasting; Adipose Tissue
PubMed: 37513513
DOI: 10.3390/nu15143095 -
Molecules (Basel, Switzerland) Jul 2023sp. D01, capable of growing in tributyrin medium, was isolated from the gut microbiota of yellow mealworm. By using in silico analyses, we discovered a hypothesized...
sp. D01, capable of growing in tributyrin medium, was isolated from the gut microbiota of yellow mealworm. By using in silico analyses, we discovered a hypothesized esterase encoding gene in the D01 bacterium, and its encoded protein, EstD04, was classified as a bacterial hormone-sensitive lipase (bHSL) of the type IV lipase family. The study revealed that the recombinant EstD04-His(6x) protein exhibited esterase activity and broad substrate specificity, as it was capable of hydrolyzing -nitrophenyl derivatives with different acyl chain lengths. By using the most favorable substrate -nitrophenyl butyrate (C), we defined the optimal temperature and pH value for EstD04 esterase activity as 40 °C and pH 8, respectively, with a catalytic efficiency (/) of 6.17 × 10 mM s at 40 °C. EstD04 demonstrated high stability between pH 8 and 10, and thus, it might be capably used as an alkaline esterase in industrial applications. The addition of Mg and NH, as well as DMSO, could stimulate EstD04 enzyme activity. Based on bioinformatic motif analyses and tertiary structural simulation, we determined EstD04 to be a typical bHSL protein with highly conserved motifs, including a triad catalytic center (Ser, Glu, and His), two cap regions, hinge sites, and an oxyanion hole, which are important for the type IV enzyme activity. Moreover, the sequence analysis suggested that the two unique discrete cap regions of EstD04 may contribute to its alkali mesophilic nature, allowing EstD04 to exhibit extremely distinct physiological properties from its evolutionarily closest esterase.
Topics: Animals; Esterases; Tenebrio; Amino Acid Sequence; Pseudomonas; Gastrointestinal Microbiome; Sterol Esterase; Bacteria; Substrate Specificity; Hydrogen-Ion Concentration; Cloning, Molecular; Enzyme Stability
PubMed: 37513282
DOI: 10.3390/molecules28145410 -
Mikrochimica Acta Jul 2023This work provides a microfluidic-based biosensor to determine total cholesterol in serum based on integrating the reaction/detection zone of a microfluidic chip of a...
This work provides a microfluidic-based biosensor to determine total cholesterol in serum based on integrating the reaction/detection zone of a microfluidic chip of a magnetically retained enzyme microreactor (MREµR) coupled with the remote fluorometric detection through a bifurcated fiber-optic bundle (BFOB) connected with a conventional spectrofluorometer. The method is based on developing the enzymatic hydrolysis and oxidation of cholesterol at microscale size using both enzymes (cholesterol esterase (ChE) and cholesterol oxidase (ChOx)) immobilized on magnetic nanoparticles (MNPs). The biocatalyst reactions were followed by monitoring the fluorescence decreasing by the naphtofluorescein (NF) oxidation in the presence of the previous HO formed. This microfluidic biosensor supposes the physical integration of a minimal MREµR as a bioactive enzyme area and the focused BFOB connected with the spectrofluorometer detector. The MREµR was formed by a 1 mm length of magnetic retained 2:1 ChE-MNP/ChOx-MNP mixture. The dynamic range of the calibration graph was 0.005-10 mmol L, expressed as total cholesterol concentration with a detection limit of 1.1 µmol L (r = 0.9999, s = 0.03, n = 10, r = 3). The precision expressed as the relative standard deviation (RSD%) was between 1.3 and 2.1%. The microfluidic-based biosensors showed a sampling frequency estimated at 30 h. The method was applied to determine cholesterol in serum samples with recovery values between 94.8 and 102%. The results of the cholesterol determination in serum were also tested by correlation with those obtained using the other two previous methods.
Topics: Microfluidics; Hydrogen Peroxide; Enzymes, Immobilized; Cholesterol; Cholesterol Oxidase; Biosensing Techniques; Sterol Esterase
PubMed: 37464062
DOI: 10.1007/s00604-023-05894-w -
International Journal of Molecular... Aug 2023Obesity is associated with high risk of mortality globally because obesity is associated with development of diseases such as diabetes, dyslipidemia, fatty liver...
Obesity is associated with high risk of mortality globally because obesity is associated with development of diseases such as diabetes, dyslipidemia, fatty liver disease, hypertension, and cancer. The present study aimed to identify the mechanism of action related to the anti‑obesity activity of root (PLR) based on its effects on lipid droplet accumulation. The inhibitory activity on lipid accumulation was analyzed through Oil‑Red O staining, and the changes in levels of lipid accumulation‑related proteins were analyzed using Western blot analysis. And the contents of triacylglycerol and free glycerol were analyzed using an ELISA Kit. PLR significantly inhibited the accumulation of lipid droplets and triacylglycerol in differentiating 3T3‑L1 cells. PLR increased phosphorylated‑hormone sensitive lipase (HSL), HSL and adipose triglyceride lipase (ATGL) and decreases perilipin‑1 in differentiating and fully differentiated 3T3‑L1 cells. Furthermore, treatment of fully differentiated 3T3‑L1 cells with PLR resulted in increased free glycerol levels. PLR treatment increased levels of peroxisome proliferator‑activated receptor‑gamma coactivator‑1 alpha (PGC‑1α), PR domain containing 16 (PRDM16) and uncoupling protein 1 (UCP‑1) in both differentiating and fully differentiated 3T3‑L1 cells. However, the PLR‑mediated increase in lipolytic, such as ATGL and HSL, and thermogenic factors, such as PGC‑1a and UCP‑1, were decreased by inhibition of AMP‑activated protein kinase (AMPK) with Compound C. Taken together, these results suggest that PLR exerted anti‑obesity effects by regulating lipolytic and thermogenic factors via AMPK activation. Therefore, the present study provided evidence that PLR is a potential natural agent for the development of drugs to control obesity.
Topics: Mice; Animals; Humans; Lipolysis; AMP-Activated Protein Kinases; Paeonia; 3T3-L1 Cells; Glycerol; Lipase; Sterol Esterase; Triglycerides; Obesity; Thermogenesis
PubMed: 37326061
DOI: 10.3892/ijmm.2023.5268