A simple method for the determination of the cholesterol esterase activity.

Stępień, Agnieszka Ewa; Gonchar, Mykhailo

doi:10.18388/abp.2013_1999

Cited by 12 publications

(8 citation statements)

References 15 publications

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“…DAPI is known to interact tightly with DNA13, 14 by binding mainly at the minor groove,15 specially in the presence of AT sequences 5. 14, 16 On the other hand, several lines of evidence exist for the occurrence of other binding modes, including its intercalation into DNA, especially at GC sites,17, 18 and external binding19.…”

Section: Introductionmentioning

confidence: 99%

“…DAPI is known to interact tightly with DNA [13,14] by binding mainly at the minor groove, [15] specially in the presence of AT sequences. [5,14,16] On the other hand, several lines of evidence exist for the occurrence of other binding modes, including its intercalation into DNA, especially at GC sites, [17,18] and external binding [19] . [ The knowledge of the details of DAPI binding to polynucleotides is important, if one takes into account that this dye continues to find wide application.…”

Section: Introductionmentioning

confidence: 99%

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The Fluorophore 4′,6‐Diamidino‐2‐phenylindole (DAPI) Induces DNA Folding in Long Double‐Stranded DNA

Beccia

Biver

Pardini

et al. 2012

Chemistry An Asian Journal

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DAPI (4',6-diamidino-2-phenylindole) is a widely used fluorescent dye, whose complicated binding features to DNAs and RNAs have been the object of debates and are still not fully understood. In this study, different approaches were employed, including binding equilibrium measurements (spectrofluorometry), melting experiments (spectrophotometry), viscometric measurements, circular dichroism, and T-jump kinetic analyses; all data concur in shedding light on the complex mechanistic aspects of the binding mode of DAPI to natural DNA. Conditions are found that induce the mode of the DAPI/DNA interaction to change from groove binding to intercalation. Moreover, it is observed, for the first time, that DAPI is able to induce the formation of a rather compact polymer-dye adduct under particular conditions. The results suggest that this form is a folded or coiled DNA structure stabilized by DAPI dye bridges.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The Fluorophore 4′,6‐Diamidino‐2‐phenylindole (DAPI) Induces DNA Folding in Long Double‐Stranded DNA

Beccia

Biver

Pardini

et al. 2012

Chemistry An Asian Journal

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“…Two sources of cholesterol esterase were tested to assess their ability to cleave hydroxycholesterol esters, one from Pseudomonas and the other from bovine pancreas. Despite the fact that both enzymes have been reported to have similar activity regarding cholesterol esters [41], a different behavior was observed with oxysterols. The use of Pseudomonas enzyme resulted in a stronger signal for both 25-and 27-HC (data not shown).…”

Section: Preliminary Exploration Of Cholesterol Esterases To Hydrolyzmentioning

confidence: 91%

Determination of total plasma oxysterols by enzymatic hydrolysis, solid phase extraction and liquid chromatography coupled to mass-spectrometry

Mendiara

Domeño

Nerı́n

et al. 2018

Journal of Pharmaceutical and Biomedical Analysis

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The potential use of cholesterol esterases was tested to avoid alkaline hydrolysis for cleavage of plasma esterified oxysterols. The enzymatic hydrolysis was optimized by testing two sources of enzyme-Pseudomonas and bovine pancreas, presence of surfactants, incubation time and amount of enzyme. Free forms of 4β-, 7-, 24-, 25- and 27-hydroxycholesterol (HC) as well 7-ketocholesterol (7-KC) were analyzed by liquid chromatography and mass-spectrometry using the deuterated internal standard, 25-HC(d6). Enzymatic hydrolysis was more effective using the Pseudomonas enzyme and in presence of surfactants. Compared to alkaline hydrolysis, it generated a cleaner chromatographic baseline and better recovery of the internal standard. Oxysterols were assayed with detection limits between 7 and 31 pg/mL. Interassay coefficients of variation were lower than 10% and extraction recovery efficiencies, higher than 90%. The procedure was used to characterize plasma levels of Cyp7b1-deficient rat, where it showed increased plasma levels of 7, 24 and 25-HC. Due to the low volume of sample required, it may be used in other animal models, particularly rodents, as well as in pediatric samples where sample amount is always a problem. Thus, the proposed new method offers mild enzymatic processing that greatly facilitates oxysterol determinations to delineate their role in physiopathology.

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“…Fasting blood glucose was determined monthly. Levels of serum triglycerides(TG), total cholesterol (TC), high density lipoprotein (HDL-c), low-density lipoprotein cholesterol (LDL-c), were determined by enzymatic method [40][41][42][43][44][45][46]. Glycated Hemoglobin (HbA1c) and blood glucose were determined using high-performance liquid chromatography and glucose oxidase method respectively [47,48].…”

Section: Laboratory Assaymentioning

confidence: 99%

Effect of a nutrition education programme on the metabolic syndrome in type 2 diabetes mellitus at a Level 5 Hospital in Kenya: “a randomized controlled trial”

THUITA-Wambugu

Kiage

Onyango

et al. 2020

Preprint

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Background Type 2 diabetes mellitus (T2D), is a life-threatening condition of global public health concern. It worsens in the presence of the metabolic syndrome (MetS), a complex disorder characterized by co-occurrence of at least three of such factors as hypertension, obesity, dyslipidemia and insulin resistance. However, lifestyle interventions reduce the risk of both MetS and T2D, and nutrition education can empower individuals on the appropriate lifestyle changes. The aim of the current study was to evaluate the effect of a nutrition education programme, with and without inclusion of peer to peer support, on MetS in T2D patients. Methods This was a randomized controlled trial with two intervention groups and one control. One of the intervention groups involved a nutrition education programme with peer-to-peer support (NEP); the other involved only the education program, while the control received standard care. Each group had 51 participants. The nutrition education programme was conducted for 2h per week for 8 weeks. In addition, the NEP had weekly peer-to-peer interactions for 8 weeks. All groups had follow-up sessions for 6 months. Data on MetS risk factors as well as food intake patterns and physical activity levels were taken at baseline and at different time points during the study. Analysis of Co-variance and regression were used in the analysis. Results: The MetS prevalence improved in the NEP (90% to 52%) and NE (86% to 69%),while it worsened in C (88% to 91%). There was improvement in the mean values of the anthropometric parameters in the NEP and NE which worsened in the control group. There was a general improvement in mean values of blood lipids, fasting blood glucose and HbA1c in all the groups, with NEP showing the greatest improvements, followed by NE, except for triglycerides and HDL where the control group had better improvement than the NE. Changes in the anthropometric and metabolic indicators mirrored the changes in food intake patterns and physical activity, where the greatest improvements occurred in the NEP. Conclusions: Nutrition education with inclusion of peer to peer support was of clinical benefit in improving metabolic outcomes and reducing MetS in T2D patients.

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A simple method for the determination of the cholesterol esterase activity.

Cited by 12 publications

References 15 publications

The Fluorophore 4′,6‐Diamidino‐2‐phenylindole (DAPI) Induces DNA Folding in Long Double‐Stranded DNA

The Fluorophore 4′,6‐Diamidino‐2‐phenylindole (DAPI) Induces DNA Folding in Long Double‐Stranded DNA

Determination of total plasma oxysterols by enzymatic hydrolysis, solid phase extraction and liquid chromatography coupled to mass-spectrometry

Effect of a nutrition education programme on the metabolic syndrome in type 2 diabetes mellitus at a Level 5 Hospital in Kenya: “a randomized controlled trial”

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