Sturgeon osteocalcin shares structural features with matrix gla protein evolutionary relationship and functional implications

Osteocalcin (OC) and matrix Gla protein (MGP) are considered evolutionarily related because they share key structural features, although they have been described to exert different functions. In this work, we report the identification and characterization of both OC and MGP from the Adriatic sturgeon, a ray-finned fish characterized by a slow evolution and the retention of many ancestral features. Sturgeon MGP shows a primary structure, post-translation modifications, and patterns of mRNA/protein distribution and accumulation typical of known MGPs, and it contains seven possible Gla residues that would make the sturgeon protein the most gamma-carboxylated among known MGPs. In contrast, sturgeon OC was found to present a hybrid structure. Indeed, although exhibiting protein domains typical of known OCs, it also contains structural features usually found in MGPs (e. g. a putative phosphorylated propeptide). Moreover, patterns of OC gene expression and protein accumulation overlap with those reported for MGP; OC was detected in bone cells and mineralized structures but also in soft and cartilaginous tissues. We propose that, in a context of a reduced rate of evolution, sturgeon OC has retained structural features of the ancestral protein that emerged millions of years ago from the duplication of an ancient MGP gene and may exhibit intermediate functional features.Portuguese Science and Technology Foundation [POCTI/MAR/57921/2004, SFRH/BD/9077/2002]; Fundo Europeu De Desenvolvimento Regional (FEDER) (Portugal); National Funding; Center of Marine Sciences (CCMAR

Safety and toxicological evaluation of a synthetic vitamin K2, menaquinone-7

Menaquinone-7 (MK-7) is part of a family of vitamin K that are essential co-factors for the enzyme γ-glutamyl carboxylase, which is involved in the activation of γ-carboxy glutamate (Gla) proteins in the body. Gla proteins are important for normal blood coagulation and normality of bones and arteries. The objective of this study was to examine the potential toxicity of synthetic MK-7 in BomTac:NMRI mice and in Sprague-Dawley rats. In an acute oral toxicity test, mice were administered a single oral dose of 2000 mg/kg body weight (limit dose) and no toxicity was observed during the 14-day observation period. In the subchronic oral toxicity test in rats, animals were administered MK-7 for 90 days by gavage at the following doses: 0 (vehicle control, corn oil), 2.5, 5, and 10 mg/kg body weight/day. All generated data, including clinical observations, ophthalmology, clinical pathology, gross necropsy, and histopathology, revealed no compound-related toxicity in rats. Any statistically significant findings in clinical pathology parameters and/or organ weights noted were considered to be within normal biological variability. Therefore, under the conditions of this experiment, the median lethal dose (LD50) of MK-7 after a single oral administration in mice was determined to be greater than the limit dose level of 2000 mg/kg body weight. The no observed adverse effect level (NOAEL) of MK-7, when administered orally to rats for 90 days, was considered to be equal to 10 mg/kg body weight/day, the highest dose tested, based on lack of toxicity during the 90-day study period

Исследование влияния частоты и длительности импульсов на свойства покрытий, осаждаемых на титановые сплавы методом микродугового оксидирования

Объектом исследования являлись кальций-фосфатные покрытия, сформированные методом микродугового оксидирования, при различных специально подобранных режимах работы. Целью данной работы являлось исследование влияния частоты следования и длительности импульсов на физико-химические и механические свойства покрытий, сформированных методом микродугового оксидирования. В результате исследования были выявлены утверждения, которые могут быть полезны при подборе режимов для формирования специальных покрытий для разного рода применений.The object of the study is calcium-phosphate cotaing formed by microarc oxidation method on different parameters. The aim of the work is to study the effect of the repetition frequency and pulse duration on the physical, chemical and mechanical properties of coatings formed by microarc oxidation. As a result of the research, statements were identified that can be useful in selecting modes for forming special coatings for various types of applications

Novel mutations in the VKORC1 gene of wild rats and mice – a response to 50 years of selection pressure by warfarin?

<p>Abstract</p> <p>Background</p> <p>Coumarin derivatives have been in world-wide use for rodent pest control for more than 50 years. Due to their retarded action as inhibitors of blood coagulation by repression of the vitamin K reductase (VKOR) activity, they are the rodenticides of choice against several species. Resistance to these compounds has been reported for rodent populations from many countries around the world and poses a considerable problem for efficacy of pest control.</p> <p>Results</p> <p>In the present study, we have sequenced the <it>VKORC1 </it>genes of more than 250 rats and mice trapped in anticoagulant-exposed areas from four continents, and identified 18 novel and five published missense mutations, as well as eight neutral sequence variants, in a total of 178 animals. Mutagenesis in <it>VKORC1 </it>cDNA constructs and their recombinant expression revealed that these mutations reduced VKOR activities as compared to the wild-type protein. However, the <it>in vitro </it>enzyme assay used was not suited to convincingly demonstrate the warfarin resistance of all mutant proteins</p> <p>Conclusion</p> <p>Our results corroborate the <it>VKORC1 </it>gene as the main target for spontaneous mutations conferring warfarin resistance. The mechanism(s) of how mutations in the <it>VKORC1 </it>gene mediate insensitivity to coumarins <it>in vivo </it>has still to be elucidated.</p

2D-electrophoresis and multiplex immunoassay proteomic analysis of different body fluids and cellular components reveal known and novel markers for extended fasting

Proteomic technologies applied for profiling human biofluids and blood cells are considered to reveal new biomarkers of exposure or provide insights into novel mechanisms of adaptation

Dietary reference values for vitamin K

Following a request from the European Commission, the EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) derives dietary reference values (DRVs) for vitamin K. In this Opinion, the Panel considers vitamin K to comprise both phylloquinone and menaquinones. The Panel considers that none of the biomarkers of vitamin K intake or status is suitable by itself to derive DRVs for vitamin K. Several health outcomes possibly associated with vitamin K intake were also considered but data could not be used to establish DRVs. The Panel considers that average requirements and population reference intakes for vitamin K cannot be derived for adults, infants and children, and therefore sets adequate intakes (AIs). The Panel considers that available evidence on occurrence, absorption, function and content in the body or organs of menaquinones is insufficient, and, therefore, sets AIs for phylloquinone only. Having assessed additional evidence available since 1993 in particular related to biomarkers, intake data and the factorial approach, which all are associated with considerable uncertainties, the Panel maintains the reference value proposed by the Scientific Committee for Food (SCF) in 1993. An AI of 1 mu g phylloquinone/kg body weight per day is set for all age and sex population groups. Considering the respective reference body weights, AIs for phylloquinone are set at 70 mu g/day for all adults including pregnant and lactating women, at 10 mu g/day for infants aged 7-11 months, and between 12 mu g/day for children aged 1-3 years and 65 mu g/day for children aged 15-17 years. (C) 2017 European Food Safety Authority. EFSA Journal published by John Wiley and Sons Ltd on behalf of European Food Safety Authority

New perspectives on rare connective tissue calcifying diseases

Connective tissue calcifying diseases (CTCs) are characterized by abnormal calcium deposition in connective tissues. CTCs are caused by multiple factors including chronic diseases (Type II diabetes mellitus, chronic kidney disease), the use of pharmaceuticals (e.g. warfarin, glucocorticoids) and inherited rare genetic diseases such as pseudoxanthoma elasticum (PXE), generalized arterial calcification in infancy (GACI) and Keutel syndrome (KTLS). This review explores our current knowledge of these rare inherited CTCs, and highlights the most promising avenues for pharmaceutical intervention. Advancing our understanding of rare inherited forms of CTC is not only essential for the development of therapeutic strategies for patients suffering from these diseases, but also fundamental to delineating the mechanisms underpinning acquired chronic forms of CTC

Circulating Matrix Gla-protein:a biomarker for vascular disease

Outline of this thesis This thesis focuses on measurements of circulating MGP species and their potential diagnostic and clinical value. Chapter 1 gives an introduction of MGP and vitamin K. In Chapter 2, the mono-antibody MGP ELISA measuring total uncarboxylated MGP (t-ucMGP) levels is presented and characterised, with application of t-ucMGP measurements in patient populations with cardiovascular disease (angioplasty and aortic valve stenosis) and vascular calcification (hemodialysis and calciphylaxis). Further investigation of t-ucMGP as a biomarker in dialysis patients is described in Chapters 3 and 4. Chapter 3 describes the correlation between t-ucMGP levels and the extent of coronary artery calcification in hemodialysis patients. In Chapter 4, the association between t-ucMGP levels and (cardiovascular) mortality in a large Dutch cohort of hemodialysis and peritoneal dialysis patients is investigated. In Chapter 5, we compared the four available MGP assays, including two novel dual-antibody MGP ELISA. All assays were applied in patients with cardiovascular disease, rheumatic diseases, and end-stage renal disease. To determine which assay would be suited to assess vascular vitamin K status, the effect of changes in vitamin K status on circulating MGP species was also investigated. For this, the assays were applied in two groups of healthy subjects with opposite vitamin K status: subjects receiving vitamin K supplementation and those receiving vitamin K antagonists. In Chapter 6, we investigated the functions of MGP in humans by phenotypic characterization of a newly-identified Dutch patient with Keutel syndrome. In order to further elucidate the difference in phenotype between MGP deficient mice and men, we characterized the circulating MGP species of this patient and his first-degree relatives. Additionally, MGP species were determined in tissues from the first described Keutel syndrome patient in the literature. Chapter 7 describes the investigation of the vitamin K intake and the hepatic and extra-hepatic vitamin K status in a Dutch cohort of hemodialysis patients