Secondary Consolidation Characteristics of Organic Soil Modified by Biological Enzymes Based on Gibson Model

In order to study the secondary consolidation characteristics of organic soil modified by enzyme, the secondary consolidation tests of organic soil modified by enzyme were carried out. Firstly, the consolidation coefficient Cv, secondary consolidation coefficient Ca and compression index Cc of modified organic soil under different levels of loads and different enzyme contents were obtained according to the analysis of experimental data. Then, the parameters of Gibson rheological model were fitted respectively according to the experimental results. Finally, the relationship between rheological model parameters, Ca/Cc, secondary consolidation coefficient Ca and loading was analyzed under different enzyme contents. The results show that: (1) The rheological model parameters of organic soil modified by enzyme are negatively correlated with the enzyme content, and positively correlated with the load value. When the enzyme content is 0.01%, some parameters reach the maximum value; (2) The secondary consolidation coefficient is related to the load. The secondary consolidation coefficient of samples with different enzyme contents shows a certain rule under all levels of load. With the increasing of load, the final secondary consolidation coefficient Ca approaches a stable value; (3) The Ca/Cc values of the samples of the modified organic soil with different enzyme contents are between 0.042 and 0.1 under different loads. The results show that the model is suitable for describing the secondary consolidation creep characteristics of organic soil, and can be used to guide the long-term deformation prediction of organic soil foundation.

Optimization of Lipase Catalyzed Biofuel Synthesis From Waste Filter Coffee Using Response Surface Methodology

This study was executed that optimize an environmentally friendly lipase (Resinase® HT with the activity of 135,56 U/ml) catalyzed transesterification process from novel feedstock waste filter coffee oil for biofuel production. Response surface methodology (RSM) with the central composite design was performed to investigate the effect of experimental factors (enzyme content, oil/methanol molar ratio, reaction temperature) on the fatty acid methyl ester (FAME) conversion and also investigated to resolve the optimum design points. After the experimental studies the lowest FAME conversion was found as 71% when the enzyme content was 15%(%gr w/w), oil/methanol molar ratio was 0.17 and the reaction temperature value was 45 °C. According to the design response, 70.83% was observed in the same conditions. The highest FAME conversion of 97% was found when the enzyme content was 5%(%gr w/w), oil/methanol molar ratio was 0.25 and the reaction temperature value was 35 °C. The experimental run gave the FAME conversion of 96.80% at the same reaction conditions. The model fitted with the experimental values with R2 = 0.98. Also, classical soxhlet extraction and Dyer method oil yields were compared. 24 % and 20 % oil removed from waste coffee grounds in traditional soxhlet and Dyer method respectively. The extraction process took 30 minutes with the soxhlet method and 45 minutes with the Dyer method. The results are promising for the application of lipase catalyst for environmentally friendly and sustainable biodiesel production from waste coffee oils all over the world.

Critical Involvement of Calcium-Dependent Cytosolic Phospholipase A2α in Aortic Valve Interstitial Cell Calcification

The involvement of calcium-dependent cytosolic phospholipase A2α (cPLA2α) in aortic valve calcification is not exhaustively elucidated. Here, cPLA2α expression in aortic valve interstitial cell (AVIC) pro-calcific cultures simulating either metastatic or dystrophic calcification was estimated by qPCR, Western blotting, and counting of cPLA2α-immunoreactive cells, with parallel ultrastructural examination of AVIC calcific degeneration. These evaluations also involved pro-calcific AVIC cultures treated with cPLA2α inhibitor dexamethasone. cPLA2α over-expression resulted for both types of pro-calcific AVIC cultures. Compared to controls, enzyme content was found to increase by up to 300% and 186% in metastatic and dystrophic calcification-like cultures, respectively. Increases in mRNA amounts were also observed, although they were not as striking as those in enzyme content. Moreover, cPLA2α increases were time-dependent and strictly associated with mineralization progression. Conversely, drastically lower levels of enzyme content resulted for the pro-calcific AVIC cultures supplemented with dexamethasone. In particular, cPLA2α amounts were found to decrease by almost 88% and 48% in metastatic and dystrophic calcification-like cultures, respectively, with mRNA amounts showing a similar trend. Interestingly, these drastic decreases in cPLA2α amounts were paralleled by drastic decreases in mineralization degrees, as revealed ultrastructurally. In conclusion, cPLA2α may be regarded as a crucial co-factor contributing to AVIC mineralization in vitro, thus being an attractive potential target for designing novel therapeutic strategies aimed to counteract onset or progression of calcific aortic valve diseases.

PL - 030 The effects of ACE gene polymorphisms on ACE content before and after High-Intensity Interval Exercise

Objective Angiotensin Converting Enzyme (ACE) is expressed in human skeletal muscle. The ACE I/D polymorphism (rs4341) has been associated with athletic performance in some studies. Studies suggested that the ACE I/D gene polymorphism is associated with ACE enzyme content in serum, however, the effect of ACE I/D on ACE protein content in human skeletal muscle in unclear. Angiotensin-converting enzyme 2 (ACE2) is a new component of the renin-angiotensin system (RAS), which is counter-regulatory to the ACE enzyme. The polymorphisms in the ACE2 gene (rs1978124 and rs2285666) have been reported to be associated with hypertension, however, their effects on ACE content in the blood and in skeletal muscle have yet to be explored. Utilising the Gene SMART cohort (n=81), we investigated whether the ACE I/D gene polymorphism (rs4341) and two ACE2 gene polymorphisms (rs1978124 and rs2285666) were associated with ACE enzyme content in the blood and skeletal muscle at baseline, and following a single session of High-Intensity Interval Exercise (HIIE). Methods ACE and ACE2 gene polymorphisms were determined using the TaqMan SNP assay (Applied Biosystems, Foster City, California, United States) by Mastercycler® ep realplex2 (Eppendorf, Hamburg, Germany), and QuantStudio™ 7 Flex Real-Time PCR System (Applied Biosystems, Foster City, California, United States). For quantitation of ACE content in the plasma, Abcam Human ELISA Kit (ab119577 –ACE (CD143)) was used (Abcam, Cambridge, United Kingdom). Western blots were used to measure ACE content in skeletal muscle. We used robust linear models adjusted for age to test the effect of the ACE I/D polymorphism on outcomes at baseline, using the MASS package in the R statistical software. p-values were adjusted for multiple comparisons using the Benjamini and Hochberg method, and all reported p-values are adjusted p-values. An adjusted p value < 0.005 was considered significant. Results We found that the ACE I/D gene polymorphism was associated with ACE content in the blood (p<0.005) at baseline, but not the ACE protein content in skeletal muscle at baseline. The ACE2 polymorphisms (rs1978124 and rs2285666) were not associated with ACE enzyme content in the blood or in skeletal muscle at baseline. A single session of HIIE tended (0.005 < p < 0.05) to increase blood ACE content immediately post exercise, while skeletal muscle ACE protein content was lower 3 hours post a single session of HIIE (p<0.005). However, those changes were not related to ACE I/D or ACE2 polymorphisms. Conclusions The ACE I/D gene polymorphism influences ACE enzyme content in the blood but not the ACE protein content of human skeletal muscle. ACE I/D gene polymorphism does not influence the changes of ACE content after a single session of HIIE. ACE2 gene polymorphisms seem to have no effect on ACE content in the blood and skeletal muscle, before or after a session of HIIE.

ACE I/D gene variant predicts ACE enzyme content in blood but not the ACE, UCP2, and UCP3 protein content in human skeletal muscle in the Gene SMART study

Angiotensin-converting enzyme (ACE) is expressed in human skeletal muscle. The ACE I/D polymorphism has been associated with athletic performance in some studies. Studies have suggested that the ACE I/D gene variant is associated with ACE enzyme content in serum, and there is an interaction between ACE and uncoupling proteins 2 and 3 (UCP2 and UCP3). However, no studies have explored the effect of ACE I/D on ACE, UCP2, and UCP3 protein content in human skeletal muscle. Utilizing the Gene SMART cohort ( n = 81), we investigated whether the ACE I/D gene variant is associated with ACE enzyme content in blood and ACE, UCP2, and UCP3 protein content in skeletal muscle at baseline and following a session of high-intensity interval exercise (HIIE). Using a stringent and robust statistical analyses, we found that the ACE I/D gene variant was associated with ACE enzyme content in blood ( P < 0.005) at baseline but not the ACE, UCP2, and UCP3 protein content in muscle at baseline. A single session of HIIE tended (0.005 < P < 0.05) to increase blood ACE content immediately postexercise, whereas muscle ACE protein content was lower 3 h after a single session of HIIE ( P < 0.005). Muscle UCP3 protein content decreased immediately after a single session of HIIE ( P < 0.005) and remained low 3 h postexercise. However, those changes in the muscle were not genotype dependent. In conclusion, The ACE I/D gene variant predicts ACE enzyme content in blood but not the ACE, UCP2, and UCP3 protein content of human skeletal muscle. NEW & NOTEWORTHY This paper describes the association between ACE I/D gene variant and ACE protein content in blood and ACE, UCP2, and UCP3 protein content in skeletal muscle at baseline and after exercise in a large cohort of healthy males. Our data suggest that ACE I/D is a strong predictor of blood ACE content but not muscle ACE content.

Ternary covalent conjugate (antibody–gold nanoparticle–peroxidase) for signal enhancement in enzyme immunoassay

A synthesis of a new horseradish peroxidase (HRP) conjugate and antibody covalently bound with gold nanoparticles (GNPs) with a high enzyme content was first developed.

Skeletal muscle and beyond: the role of exercise as a mediator of systemic mitochondrial biogenesis

It has been known for more than 4 decades that exercise causes increases in skeletal muscle mitochondrial enzyme content and activity (i.e., mitochondrial biogenesis). Increasing evidence now suggests that exercise can induce mitochondrial biogenesis in a wide range of tissues not normally associated with the metabolic demands of exercise. Perturbations in mitochondrial content and (or) function have been linked to a wide variety of diseases, in multiple tissues, and exercise may serve as a potent approach by which to prevent and (or) treat these pathologies. In this context, the purpose of this review is to highlight the effects of exercise, and the underlying mechanisms therein, on the induction of mitochondrial biogenesis in skeletal muscle, adipose tissue, liver, brain, and kidney.