Elevated low-density lipoprotein (LDL) cholesterol levels lead to atherosclerosis and platelet hyperaggregability, both of which are known culprits of arterial thrombosis. Normalization of LDL cholesterol in familial hypercholesterolemia (FH) is not an easy task and frequently requires specific treatment, such as regularly performed lipid apheresis and/or novel drugs such as proprotein convertase subtilisin kexin 9 monoclonal antibodies (PCSK9Ab). Moreover, a high resistance rate to the first-line antiplatelet drug acetylsalicylic acid (ASA) stimulated research of novel antiplatelet drugs. 4-methylcatechol (4-MC), a known metabolite of several dietary flavonoids, may be a suitable candidate. The aim of this study was to analyse the antiplatelet effect of 4-MC in FH patients and to compare its impact on two FH treatment modalities via whole-blood impedance aggregometry. When compared to age-matched, generally healthy controls, the antiplatelet effect of 4-MC against collagen-induced aggregation was higher in FH patients. Apheresis itself improved the effect of 4-MC on platelet aggregation and blood from patients treated with this procedure and pretreated with 4-MC had lower platelet aggregability when compared to those solely treated with PCKS9Ab. Although this study had some inherent limitations, e.g., a low number of patients and possible impact of administered drugs, it confirmed the suitability of 4-MC as a promising antiplatelet agent and also demonstrated the effect of 4-MC in patients with a genetic metabolic disease for the first time.
- MeSH
- hyperlipoproteinemie typ II * farmakoterapie MeSH
- LDL-cholesterol MeSH
- lidé MeSH
- monoklonální protilátky farmakologie terapeutické užití MeSH
- proproteinkonvertasa subtilisin/kexin typu 9 MeSH
- proproteinkonvertasy terapeutické užití MeSH
- separace krevních složek * metody MeSH
- subtilisin MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Proprotein convertase subtilisin/kexin type 9 (PCSK9) is involved in the regulation of LDL receptors. Inhibition of PCSK9 increase uptake of LDL-particles and pathogen-associated molecular patterns (PAMPs). The aim of our study was to evaluate biological variation of serum PCSK9. METHODS: Within-subject (CVI) and between-subject (CVG) biological variations were assessed in 14 healthy volunteers in a 6-week protocol (7 samples, equidistant time intervals). Serum concentration of PCSK9 was measured by a Quantikine ELISA assay (R&D systems, Bio-Techne Ltd., UK) on a DS2 ELISA reader (Dynex Technologies GmbH, Germany). Precision (CVA) was assessed by duplicate measurements. Two methods with different levels of robustness were used for the estimation of CVI, SD-ANOVA and CV-ANOVA method. We calculated the index of individuality and reference change values. The experiment was fully compliant with EFLM database checklist. RESULTS: The within-subject values of PCSK9 in healthy persons, as calculated by two statistical methods, were 23.2% (SD-ANOVA with CVA of 5.6%) and 26.6% (CV-ANOVA with CVA of 4.8%). The CVG was 10.9% (SD-ANOVA), index of individuality and RCV were 2.13 and 66.3%, respectively. CONCLUSIONS: The high index of individuality indicates that common reference intervals can be used to interpret serum PSCK9 values.
- MeSH
- LDL-cholesterol MeSH
- LDL-receptory * MeSH
- lidé MeSH
- proproteinkonvertasa subtilisin/kexin typu 9 * MeSH
- subtilisiny MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Importance: Despite recent advances in treatment of severe aortic valve stenosis (AS), AS remains a life-threatening condition with no proven disease-modifying therapy. Low-density lipoprotein cholesterol (LDL-C) and lipoprotein(a) (Lp[a]) have been implicated in the pathobiology of AS. The proprotein convertase subtilisin/kexin type 9 inhibitor evolocumab reduces circulating LDL-C concentrations by 50% to 60% and Lp(a) by 20% to 30%. Objective: To determine whether evolocumab reduces the risk of AS events in patients with atherosclerotic cardiovascular disease. Interventions: Patients were randomized 1:1 to evolocumab or placebo. Design, Setting, and Participants: Exploratory analysis of the FOURIER trial, which enrolled 27 564 patients with stable atherosclerotic cardiovascular disease who were taking statin therapy at 1242 sites in 49 countries from February 2013 to November 2016. Patients were randomized to evolocumab or placebo and followed up for a median (interquartile range) of 2.2 (1.8-2.5) years. This post hoc analysis was performed from September 2019 to February 2020. Main Outcomes and Measures: Site-reported adverse events of new or worsening AS or aortic valve replacement (termed AS events). The adjusted risk of AS events was calculated with a multivariable model including concentrations of Lp(a) and LDL-C corrected for Lp(a) content, plus age, sex, diabetes, hypertension, current smoking, and estimated glomerular filtration rate. Evolocumab efficacy was tested using a Cox proportional hazards model. Results: Aortic stenosis events occurred in 63 patients (48 men [76%]; mean [SD] age, 69 [9] years) over a median of 2.2 years. Elevated Lp(a) concentration was associated with higher rates of AS events (adjusted hazard ratio [aHR], 1.55 [95% CI, 1.17-2.05] per SD; P = .002), including aortic valve replacement (aHR, 2.22 [95% CI, 1.38-3.58] per SD; P = .001), after multivariable adjustment. The corrected LDL-C concentration was not significantly associated with AS events (aHR, 1.23 [95% CI, 0.93-1.61] per SD; P = .14). The overall HR for AS events with evolocumab was 0.66 (95% CI, 0.40-1.09), with no apparent association in the first year (HR, 1.09 [95% CI, 0.48-2.47]) but an HR of 0.48 (95% CI, 0.25-0.93) after the first year of treatment. Conclusions and Relevance: In this exploratory analysis of the FOURIER trial, higher Lp(a) levels, but not Lp(a)-corrected LDL-C levels, were associated with a higher risk of subsequent AS events, including aortic valve replacement. Long-term therapy with evolocumab may reduce AS events, and this raises the possibility that specific pharmacologic lipid-lowering therapy could offer a means to prevent or slow the progression of AS. These exploratory findings merit further investigation with a dedicated randomized clinical trial. Trial Registration: ClinicalTrials.gov Identifier: NCT01764633.
- MeSH
- anticholesteremika terapeutické užití MeSH
- aortální stenóza krev farmakoterapie MeSH
- humanizované monoklonální protilátky terapeutické užití MeSH
- LDL-cholesterol krev MeSH
- lidé středního věku MeSH
- lidé MeSH
- PCSK9 inhibitory MeSH
- proproteinkonvertasa subtilisin/kexin typu 9 MeSH
- rizikové faktory MeSH
- subtilisin terapeutické užití MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- práce podpořená grantem MeSH
- randomizované kontrolované studie MeSH
- Klíčová slova
- studie FIELD, studie ACCORD, studie IMPROVE-IT,
- MeSH
- anticholesteremika * farmakokinetika farmakologie klasifikace metabolismus terapeutické užití MeSH
- antisense oligodeoxyribonukleotidy farmakokinetika farmakologie metabolismus terapeutické užití MeSH
- apolipoproteiny B MeSH
- azetidiny farmakokinetika metabolismus terapeutické užití MeSH
- cholestyraminová pryskyřice farmakokinetika metabolismus škodlivé účinky terapeutické užití MeSH
- deriváty kyseliny fibrové farmakokinetika metabolismus terapeutické užití MeSH
- dyslipidemie * farmakoterapie MeSH
- HDL-cholesterol účinky léků MeSH
- hypolipidemika * klasifikace terapeutické užití MeSH
- kardiovaskulární nemoci prevence a kontrola MeSH
- klinické zkoušky jako téma MeSH
- kombinovaná farmakoterapie MeSH
- LDL-cholesterol účinky léků MeSH
- lidé MeSH
- messenger RNA MeSH
- metaanalýza jako téma MeSH
- obezita patofyziologie MeSH
- proproteinkonvertasy ekonomika farmakokinetika farmakologie terapeutické užití MeSH
- statiny farmakokinetika klasifikace terapeutické užití MeSH
- subtilisin účinky léků MeSH
- transportní proteiny pro estery cholesterolu farmakokinetika farmakologie metabolismus terapeutické užití MeSH
- transportní proteiny farmakokinetika farmakologie metabolismus terapeutické užití MeSH
- triglyceridy farmakokinetika farmakologie metabolismus terapeutické užití MeSH
- Check Tag
- lidé MeSH
Whole whey hydrolyzed by Alcalase (WWH) was tested as a complex nitrogen source for the production of poly(3-hydroxybutyrate) (PHB) from waste frying oils by Cupriavidus necator H16. Addition of WWH (10 % (v/v) of cultivation media) supported the growth and PHB accumulation; PHB yields in Erlenmeyer flasks were more than 3.5-fold higher than in control cultivations. The positive influence of WWH on PHB production was confirmed in experiments performed in laboratory fermentor. C. necator cultivated with WWH produced 28.1 g PHB l(-1) resulting in a very high product yield coefficient of 0.94 g PHB per g oil. Since PHB yields were ~40 % higher than in the control cultivation, WWH can be considered as an excellent inexpensive nitrogen source for PHB production by C. necator.
- MeSH
- Cupriavidus necator růst a vývoj metabolismus MeSH
- dusík metabolismus MeSH
- hydrolýza MeSH
- hydroxybutyráty metabolismus MeSH
- kultivační média chemie MeSH
- mléčné bílkoviny metabolismus MeSH
- polyestery metabolismus MeSH
- subtilisiny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Several recombinant derivatives of serine protease inhibitor called silk protease inhibitor 2 (SPI2), which is a silk component in Galleria mellonella (Lepidoptera, Insecta), were prepared in the expression vector Pichia pastoris. Both the native and the recombinant protease inhibitors were highly active against subtilisin and proteinase K. The synthetic SPI2 gene with Ala codon in the P1 position was fused with mGFP-5 to facilitate detection of the transgene and its protein product. A construct of the fusion gene with plant regulatory elements (promoter 35S and terminator OCS) was inserted into the binary vector pRD400. The final construct was introduced into Agrobacterium tumefaciens that was then used for genetic transformation of the potato variety Velox. The transgene expression was monitored with the aid of ELISA employing polyclonal antibody against natural SPI2. In vitro tests showed increased resistance to the late blight Phytophthora infestans in several transformed lines. No effect was seen on the growth, mortality, life span or reproduction of Spodoptera littoralis (Lepidoptera, Insecta) caterpillars, while feeding on transformed potato plants expressing the fusion protein, indicating that the transformed potatoes may be harmless to non-target organisms.
- MeSH
- exprese genu MeSH
- extracelulární prostor enzymologie MeSH
- genetické inženýrství škodlivé účinky metody MeSH
- geneticky modifikované rostliny MeSH
- hmyzí proteiny genetika MeSH
- Lepidoptera genetika MeSH
- nemoci rostlin parazitologie MeSH
- odolnost vůči nemocem genetika MeSH
- Phytophthora infestans fyziologie MeSH
- Pichia genetika MeSH
- Solanum tuberosum cytologie genetika imunologie parazitologie MeSH
- Spodoptera MeSH
- subtilisin antagonisté a inhibitory metabolismus MeSH
- transformace genetická MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Comparison of cell-wall-bound extracellular proteinases (CEPs) from Lactobacillus paracasei (LBP) ssp. paracasei natural isolates BGHN14, BGAR75 and BGAR76 with Lactococcus lactis (LCL) ssp. cremoris Wg2, in their action on alpha(S1)-, beta- and kappa-casein was done. The CEPs of LBP strains were able to degrade alpha(S1)- and beta-caseins and their caseinolytic specificity depended on the type of buffer used. These CEPs, compared with LCL Wg2, differ in four amino acid residues in small segments predicted to be involved in substrate binding. The most striking features of this comparison are the presence of Ala instead of Ser(329) and the presence of Thr instead of Asn(256) and Ala(299), in the subtilisin-like region of the CEP in LBP natural isolates. Additional conservative amino acid substitution Leu to Ile(364) was found.
- MeSH
- bakteriální geny genetika MeSH
- bakteriální proteiny genetika metabolismus MeSH
- cysteinové endopeptidasy genetika metabolismus MeSH
- financování organizované MeSH
- kaseiny metabolismus MeSH
- katalytická doména genetika MeSH
- Lactobacillus enzymologie genetika MeSH
- Lactococcus lactis enzymologie genetika MeSH
- potravinářská mikrobiologie MeSH
- substituce aminokyselin MeSH
- substrátová specifita MeSH
- subtilisin genetika MeSH
- sýr mikrobiologie MeSH
- Publikační typ
- srovnávací studie MeSH
