The 'VKORC1' and 'CYP2C9' gene variants as pharmacogenetic factors in acenocoumarol therapy in Serbian patients: Consideration of hypersensitivity and resistance

Uvod/Cilj Terapija kumarinima predstavlja jedan od najboljih modela za primenu farmakogenetike. Doprinos faktora koji utiču na terapiju kumarinima može značajno da varira između etničkih grupa, što opravdava sprovođenje studija specifičnih za populaciju. Cilj ove studije je bio da se analizira uticaj najvažnijih genetičkih faktora (geni VKORC1 i CYP2C9) koji utiču na terapiju kumarinima kod bolesnika iz Srbije. Metode Sprovedena je retrospektivna studija koja je obuhvatila 207 bolesnika na terapiji acenokumarolom. Genetičke analize su vršene direktnim sekvenciranjem. Analiziran je uticaj na dozu acenokumarola varijanti (VKORC1*2, CYP2C9*2, CYP2C9*3) koje izazivaju preosetljivost i varijanti gena VKORC1 koje izazivaju rezistenciju na kumarine. Višestruka regresiona analiza je korišćena u cilju dizajniranja matematičkog modela za predviđanje individualne doze leka na osnovu kliničko-demografskih i genetičkih podataka. Rezultati Studija je potvrdila značajan uticaj analiziranih genetičkih faktora na održavanje doze acenokumarola. Dizajniran je matematički model za predviđanje individualne doze acenokumarola i njegov nekorigovani R2 je bio 61,8. Prilikom testiranja, naš model je dao R2 vrednost od 42,6 i pokazao bolje predviđanje u poređenju sa modelom koji su dali drugi autori. Kod analiziranih bolesnika pronađeno je devet različitih varijanti u kodirajućem regionu gena VKORC1. Među nosiocima ovih varijanti 78% je bilo potpuno rezistentno, te nije bilo moguće postići terapeutski efekat čak ni sa visokim dozama acenokumarola. Zaključci Populacioni model za predviđanje individualne doze acenokumarola može pokazati prednosti u odnosu na modele koji se koriste na generalizovan način. Takođe, VKORC1 varijante koje izazivaju rezistenciju na kumarin treba uzeti u obzir prilikom planiranja terapije.Introduction/Objective Coumarin therapy represents one of the best models for applying pharmacogenetics. The contribution of factors influencing coumarin therapy can vary significantly between ethnic groups, which justifies conducting population-specific studies. The aim of this study was to analyze the influence of the most important genetic factors (VKORC1 and CYP2C9 genes) that affect coumarin therapy in patients from Serbia. Methods A retrospective study involving 207 patients on acenocoumarol therapy was conducted. Genetic analyses were performed by direct sequencing. Influence on acenocoumarol dose of variants (VKORC1, CYP2C9*2, CYP2C9*3) causing hypersensitivity and VKORC1 variants causing resistance to acenocoumarol were analyzed. Multiple regression analysis was used to design a mathematical model for predicting individual drug dosage based on clinical-demographic and genetic data. Results The study confirmed significant influence of the analyzed genetic factors on acenocoumarol maintenance dose. We designed mathematical model for predicting individual acenocoumarol dose and its unadjusted R2 was 61.8. In the testing cohort, our model gave R2 value of 42.6 and showed better prediction in comparison with model given by other authors. In the analyzed patients, nine different variants in the VKORC1 coding region were found. Among carriers of these variants 78% were completely resistant, and it was not possible to achieve therapeutic effect even with high doses of acenocoumarol. Conclusions Population-specific model for prediction individual dose of acenocoumarol, may show advantages over protocols that are used in a generalized manner. Also, VKORC1 variants which cause coumarin resistance should be considered when planning therapy

Marketing planning in a construction company

Diplomová práce řeší problematiku marketingového plánování ve stavební firmě. Práce je rozdělena na teoretickou a praktickou část. Teoretická část práce definuje problematiku marketingového plánování v oblasti stavebnictví. Popisuje stavební trh, marketingový mix, analýzu mikro a makro prostředí, mezinárodní marketing a aplikování marketingového plánu ve stavební firmě. Praktická část diplomové práce se skládá ze dvou částí. První část obsahuje analýzu zákonů o zadávání veřejných zakázek v Bosně a Hercegovině a v České republice. V druhé části je popsána konkrétní stavební firma. Na základě jednotlivě provedených analýz je zpracován marketingový plán pro firmu XYZ, která plánuje rozšířit okruh působnosti na trh Bosny a Hercegoviny. Cílem práce bylo analyzovat a popsat, jak marketingové plánování ovlivňuje účast stavební firmy ve veřejných zakázkách. Výstupem diplomové práce je definování návrhu marketingového plánu pro konkrétní stavební firmu na základě zjištěného stavu této firmy a prostředí.The thesis deals with the issue of marketing planning in a construction company. The thesis is divided into theoretical and practical parts. The theoretical part of the thesis defines the problem of marketing planning in the construction industry. It describes the construction market, marketing mix, analysis of micro and macro environment, international marketing and application of marketing plan in a construction company. The practical part of the thesis consists of two parts. The first part contains an analysis of the public procurement laws in Bosnia and Herzegovina and the Czech Republic. The second part describes a specific construction company. On the basis of the individual analyses, a marketing plan is prepared for the company XYZ, which plans to expand its scope of activity to the market of Bosnia and Herzegovina. The aim of the thesis was to analyze and describe how marketing planning affects the participation of a construction firm in public procurement. The output of the thesis is the definition of a marketing plan proposal for a specific construction firm based on the identified state of this firm and the environment.

Bone turnover markers in medicamentous and physiological hyperprolactinemia in female rats

Background/Aim. There is a lack of data on the effects of prolactin on calcium metabolism and bone turnover in hyperprolactinemia of various origins. The aim of this study was to compare the influence of medicamentous and physiological hyperprolactinemia on bone turnover in female rats. Methods. Experimental animals (18 weeks old, Wistar female rats) were divided as follows: the group P - 9 rats, 3 weeks pregnant; the group M3-10 rats that were intramuscularly administrated sulpirid (10 mg/kg) twice daily for 3 weeks, the group M6 - 10 rats that were intramuscularly administrated with sulpirid (10 mg/kg) twice daily for 6 weeks, and age matched nulliparous rats as the control group: 10 rats, 18-week-old (C1) and 7 rats, 24 weeks old (C2). Laboratory investigations included serum ionized calcium and phosphorus, urinary calcium and phosphorous excretion, osteocalcin and serum procollagen type 1 N-terminal propeptide (P1NP). Results. Experimental animals in the group P compared to the control group, displayed lower mean serum ionized calcium (0.5 ± 0.2 vs 1.12 ± 0.04 mmol/L; p < 0.001); higher mean serum phosphorus (2.42 ± 0.46 vs 2.05 ± 0.2 mmol/L; p < 0.05); increased urinary calcium (3.90 ± 0.46 vs 3.05 ± 0.58; p < 0.01) and significantly increased P1NP (489,22 ± 46,77 vs 361.9 ± 53,01 pg/mL; p < 0.001). Experimental animals in the group M3 had significantly decreased P1NP, compared to the control group. Prolongated medicamentous hyperprolactinemia (the group M6) induced increased serum ionized calcium (1.21 ± 0.03 vs 1.15 ± 0.02 mmol/L; p < 0.001); decreased serum phosphorus (1.70 ± 0.13 vs 1.89 ± 0.32 mmol/L; p < 0.001); decreased osteocalcin and P1NP. Conclusions. Physiological hyperprolactinemia does not have such harmful effect on bone metabolism as medicamentous hyperprolactinemia. Chronic medicamentous hyperprolactinemia produces lower serum levels of bone formation markers. Assessment of bone turnover markers in prolongated medicamentous hyperprolactinemia provides an opportunity for earlier diagnosis of bone metabolism disturbances and should be considered as mandatory

Reversal of FLT3 Mutational Status and Sustained Expression of NPM1 Mutation in Paired Presentation, and Relapse Samples in a Patient with Acute Myeloid Leukemia

We report a case of de novo acute myeloid leukemia (AML) with unstable FLT3 gene mutations and stable NPM1 mutation. FLT3/D835 and NPM1 (Type A) mutations were detected upon diagnosis. During the relapse, the FLT3/D835 mutation changed to an FLT3/ITD mutation while the NPM1 (Type A) mutation was retained. Cytogenetic analyses showed the normal karyotype at diagnosis and relapse. Our findings raise interesting questions about the significance of these mutations in the leukemogenic process, about their stability during the evolution of the disease, and regarding the selection of appropriate molecular markers for the monitoring of minimal residual disease

Мијелодиспластични синдром са приближном тетраплоидијом удруженом са мутацијом гена TP53 – редак случај

Introduction. Chromosomal numerical aberrations are very common in hematological malignancies, but near-tetraploidy (80-104 chromosomes) is rare in myeloid lineage malignancies, with only a few cases reported in myelodysplastic syndrome (MDS). Due to a small number of cases with this rare cytogenetic abnormality, clinicopathological significance of near-tetraploidy in MDS is still unknown. In this case report we present a case of de novo MDS patient with near-tetraploidy in association with TP53 mutation, and we aimed to elucidate the prognostic significance of this rare genetic feature. Case Outline. In August of 2018 a 71-year-old male presented with severe anemia, thrombocytopenia, and leucopenia and enlarged spleen. Laboratory data were as follows: hemoglobin (Hb) 93 g/L, white blood cells (WBC) 2.8×109/L and platelets 23x109/L. The bone marrow aspirate was hypercellular, megakaryocytes were not found, granulocytic cells were 15% with signs of dysplasia, with 16% of blast cells without Auer rods. The finding was in correlation with diagnosis of MDS, type RAEB2 which was also confirmed by immunophenotyping. Cytogenetic finding was near-tetraploidy (48,XY+mar[10]/92,XXYY[10]), and TP53 mutational analysis showed the presence of mutation in exon 8 (p.D281A; c.842 A>C). The patient received from time-to-time packed red blood cells and platelets, and died four months after initial diagnosis. Conclusion. Near-tetraploidy associated with TP53 mutation has been described only in few MDS cases. Results of these reports including ours suggest that the association of TP53 mutation and near-tetra polyploidy is a poor prognostic factor.Нумеричке аберације хромозома су веома честе код хематолошких малигнитета, али су приближне тетраплои- дије (80–104 хромозома) ретке у малигнитетима мијелоидне лозе, са само неколико случајева пријављених у мијелоди- спластичком синдрому (МДС). Због малог броја случајева са овом ретком цитогенетском абнормалношћу, клиничко- -патолошки значај приближне тетраплоидије у МДС-у је још увек непознат. Овим приказом de novo болесника са МДС-ом, са приближном тетраплоидијом и мутацијом у гену TP53, циљ нам је био да расветлимо прогностички значај ове рет- ке генетске карактеристикe. Приказ болесника Приказан је 71-годишњи болесник који је у августу 2018. године развио симптоме тешке анемије, тромбоцитопеније, леукопеније и увећане слезине. Лабо- раторијске анализе су показале следеће: хемоглобин 93 g/L, леукоцити 2,8 × 109 /L и тромбоцити 23 × 109 /L. Аспират коштане сржи је био хиперћелијски, мегакариоцити нису на- ђени, 15% гранулоцитa је било са знацима дисплазије, 16% бластa без Ауерових штапића. Налаз је одговарао дијагнози МДС-а, типа рефракторне анемије са вишком бласта 2, што је потврђено и имунолошком фенотипизацијом. Цитогенет- ском анализом утврђено је присуство приближне тетрапло- идије (48,XY+mar10/92,XXYY[10]), а анализа мутација у гену TP53 показала је присуство мутације у егзону 8 (p.D281A; c.842 A > C). Болесник је по потреби примао трансфузију еритроцита и тромбоците, а умро је четири месеца након почетне дијагнозе. Закључак Присуство приближне тетраплоидије удружене са мутацијама у гену TP53 описано је само у неколико слу- чајева МДС-а. Резултати ових случајева, као и наши резул- тати, указују на то да приближна тетраплоидија повезана са присуством мутација у гену TP53 представља фактор лоше прогнозе

The assessment of genetic potential in performance tested gilts by means of selection indexes method

The objective of this research paper was to make an assessment of breeding value of performance tested gilts of Swedish Landrace and F-1 crossbreds of Swedish Landrace and Great Yorkshire by the method of selection index. The traits on whose basis the breeding value was estimated were: daily liveweight gain, average backfat thickness measured at two sites and carcass meat percentage. These traits were corrected for body mass of 100kg by the method of base indexes and the following average values were determined: corrected daily liveweight gain (KZDP) 408.93g/day, corrected average backfat thickness measured at two sites (KSL) 9.77mm and corrected carcass meat percentage (KPM) 61.08%. Studying the effect of genotype, year and birth season of gilts a statistically significant variation (P>0.05) of these traits provoked by the mentioned factors was not determined while the gilts sire statistically highly significantly (P lt 0.001) influenced all studied traits. Heritability coefficients were: h(2)= 0.255 for KZDP, h(2)= 0.356 for KSL and h(2)= 0.349 for KPM. The four selection index equations were constructed among which as the most optimal was chosen the one which includes all three traits (KZDP, KSL and KPM) and whose coefficient of the correlation of selection index and aggregate genotype was r(IAG) = 0.594

Hydroxyurea Induces Bone Marrow Mesenchymal Stromal Cells Senescence and Modifies Cell Functionality In Vitro

Hydroxyurea (HU) is an antineoplastic agent that functions as an antimetabolite compound by inhibiting the ribonucleotide reductase. HU acts mainly as a cytostatic drug that through DNA replication stress may trigger a premature senescence-like cell phenotype, though its influence on bone marrow-derived mesenchymal stem/stromal cell (BMMSC) functions has not elucidated yet. Our results indicate that HU inhibits the growth of human BMMSC alongside senescence-like changes in both morphology and replicative potential, provokes cell cycle arrest at the S phase without affecting cellular viability and induces the expression of senescence-associated β-galactosidase and p16INK4. Moreover, HU-induced senescent BMMSC, although they did not change MSC markers expression, exhibited reduced capacity osteogenic and adipogenic differentiation. Conversely, HU treatment increased immunoregulatory functions of BMMSC compared with untreated cells and determined by T-cell proliferation. Interestingly, HU did not influence the capacity of BMMSC to induce monocytic myeloid-derived suppressor cells. Thus, these results suggest that HU improves the BMMSC functions on the T-cell inhibition and preserves their interaction with myeloid cell compartment. Mechanistically, BMMSC under HU treatment displayed a downregulation of mTOR and p38 MAPK signaling that may explain the reduced cell differentiation and increased immunomodulation activities. Together, the results obtained in this investigation suggest that HU by inducing senescence-like phenotype of BMMSC influences their cellular differentiation and immunoregulatory functions

Carcass Quality and Fatty Acids Profile of the Fatteners of Swallow-Belly Mangalitsa Breed Reared in Outdoor System

The objective of this study was to determine carcass quality of fatteners of Swallow-belly Mangalitsa breed reared in outdoor system. In m. longissimus dorsi (MLD) samples chemical composition, cholesterol content and fatty acids profile was determined. The study included 22 castrated males reared in the oak forest from the early spring to late autumn, with minimal corn harvest (approx. 0.5 kg per day). During the winter, the fatteners were kept in wooden stables and fed with cereals (corn and barley mixture in a ratio of 70:30, approx. 3 kg per day). The data was carried out by statistical package SAS 9.1.3 (SAS Inst. Inc., 2002-2003) and basic statistical parameters are showed (Mean±SD). At the end of fattening, the pigs were about one year old, and slaughtered at 92.0±16.7 kg live body weight. The average lean content in the carcasses was 35.9±2.8 %, while the content of intermuscular fat in MLD was 6.2±1.2 %. In the MLD samples average content of proteins, water, fat and ash were 20.7±0.5 %, 70.4±0.5 %, 7.7±0.5 % and 1.0±0.1 %, respectively. Average cholesterol content was 41.4±3.8 mg/100g. Shares of saturated fatty acids (ΣSFA) in MLD fat was 35.6±1.2 %, monounsaturated fatty acids (ΣMUFA) was 56.6±1.8 % and polyunsaturated fatty acids (ΣPUFA) was 6.9±1.2%. Within ΣSFA the most common was palmitic acid (C16:0, 24.7±0.6 %), wile within ΣMUFA it was oleic acid (C18:1c-9, 46.9±1.1 %) and within ΣPUFA it was linoleic acid (C18:2n-6, 5.6±1.0 %). ΣPUFA and ΣSFA ratio was 0.2±0.0, while omega-6 and omega-3 ratio was 25.1±5.6. Showed ratios are not desirable from the nutritional aspect of human diet