Frequency of the most common CYP3A5 polymorphisms in the healthy population of the Republic of Macedonia

The genetic polymorphism affecting the CYP3A5 enzyme is responsible for inter-individual and interethnic variability in the metabolism of CYP3A5 substrates. The aim of this study was to analyze the distribution of the most common CYP3A5*3 allelic variants in the healthy population of R. Macedonia and to investigate if the allelic frequency falls within the assumed range for European Caucasians. The total of 174 healthy volunteers from the general population were included. The genotyping of the CYP3A5*3 variant alleles, *3A (rs15524) and *3E (rs28365095), was performed with Real-Time PCR based on the allelic discrimination method using a TaqMan SNP genotyping assay according to the manufacturer’s instructions. The CYP3A5*3 allele is abundantly present displaying an allelic frequency of 0.922. We estimate that 0.82 of the Macedonian population are homozygotes for the variant and do not have a CYP3A5 enzymatic activity. Our study demonstrated a high prevalence of CYP3A5*3 allele in the Macedonian population. The distribution of CYP3A5 alleles was similar to that found in other European Caucasians. As the goals of personalized medicine are beginning to be realized, this provides basic information on the CYP3A5 allele frequency for the future pharmacogenetic research in R. Macedonia

Effects of single nucleotide polymorphisms and haplotypes of the SLCO1B1 gene on the pharmacokinetic profile of atorvastatin in healthy Macedonian volunteers

OATP1B1 is an influx transporter known to mediate the uptake of various endogenous compounds and xenobiotics. Several sequence variations have been discovered in the SLCO1B1 gene encoding OATP1B1. The aim of this study was to investigate the effects of SLCO1B1 polymorphisms on the pharmacokinet-ics of atorvastatin in healthy volunteers of Macedonian origin. Twenty three participants, genotyped for SLCO1B1 c.388A>G, c.521T>C, c.571T>C, c.597C>T, c.1086C>T, c.1463G>C and c.*439T>G polymorphisms using TaqMan allelic discrimination assay, ingested a single 80mg dose of atorvastatin. The plasma concentrations of atorvastatin were measured for 48h using Tandem Liquid Chromatography-Mass Spectrometry, LC-MS-MS, and the peak plasma concentration (Cmax), time to peak plasma concentration (Tmax), elimination half-life (t1/2), constant rate of elimination (kel), mean residence time (MRT, expo), volume of distribution (Vd/kg), clearance (CL/kg), area under curve AUC0-48h and AUC0-∞ were determined.Our data confirmed that the SLCO1B1 gene is highly polymorphic, with a frequency of the c.521T>C single-nucleotide polymorphism (SNP) being the lowest (app. 15%) and of all other SNPs alleles above 40%. Exceptions were c.1463G>C and c.1086C>T SNPs for which variant alleles were not identified. The strongest correlation was observed between the c.521T>C and c.571T>C SNPs pair. The haplotype analysis revealed 10 different haplotypes, with *1J/*1K/*1L being the dominant, with a frequency of app. 40%. The haplotype *15/*16/*17, containing both variant alleles of the functionally most distinguished SNPs, c.388A>G and c.521T>C, occurred with a frequency of 13%. However, *15/*16/*17 homozygotes were not identified in the study group. In this study, no significant differences in the kel,t1/2,Cmax,Tmax,AUC0-48h, AUC0-∞, MRT expo, Vd and CL between the carriers of different c.388A>G, c.597C>T andc.*439T>G genotypes were observed. Subject with a variant allele C in the c.521T>C SNP, c.521CC genotype, had markedly higher values for Cmax and AUC0-48h, 140% and 67%, respectively, in comparison with the carriers of the c.521TT genotype. Also, the carriers of the variant allele C at c.571T>C SNP, c.571 CC genotype, had 55% and 43% lower mean Cmax and AUC0-48h in comparison with the carrier of c.571TT.These differences lacked statistical significance due to the size of the sample. In addition, no significant differences in the pharmacokinetic parameters of atorvastatin between the *15/*16/*17 heterozygotes and *15/*16/*17 non-carriers were observed. In conclusion, this extensive analysis of the effect of SLCO1B1 polymorphisms on the pharmacokinetic profile of atorvastatin showed that c.521T>C and c.571T>C SNPs may affect the inter-individual response to atorvastatin. Additional studies, with a large sample size, are needed to confirm this finding

Efektite na prostaciklin vo tretmanot na dijabeticnata nefropatija kaj staorci

Mikrovaskularnite komplikacii, a pred se dijabeticnata nefropatija, se edni od najteskite komplikacii na dijabetot, od koi vo golema mera zavisi i prognozata na dijabetot kaj ovie pacienti. Etiopatogenezata na ovaa komplikacija e multifaktorijalna i za sega se uste ne kompletno rasvetlena, a vklucuva morfoloski, patolosko-anatomski i biohemiski metabolni narusuvanja. Se smeta deka narusuvanjeto na modularnata funkcija na endoteliumot moze da bide kriticen i inicijalen faktor vo razvojot na dijabeticnite vaskularni komplikacii. Vrz osnova na farmakodinamskite efekti koi sto gi poseduva prostaciklinot (PGI2) i negovite analozi, se smeta deka istite moze da bidat korisni vo tretmanot na dijabeti~nata nefropatija. Osnovna cel na ovaa studija be{e da se procenat efektite na prostaciklin (PGI2) vo tretmanot na dijabeticnata nefropatija, eksperimentalno predizvikana so streptozocin. Kaj normotenzivni staorci od sojot Wistar, eksperimentalno bese induciran najprvin dijabet so ednokratna i.p. administracija na streptozocin (STZ), a kako komplikacija na dijabetot i jasni znaci i simptomi na dijabeticna nefropatija (proteinurija, zgolemeno serumsko nivo na urea i kreatinin, poliurija, zgolemena aktivnost na NAG vo mockata). Tretman so prostaciklin (p.o.) vo doza od 0.1 mg/kg /t.t./den, vo tekot na 4 nedeli, dovede do signifikantno namaluvanje na simptomite i znacite na bubreznite ostetuvanja, vo odnos na grupata zivotni koi ne primaa prostaciklin. Vrz osnova na dobienite rezultati moze da se zakluci deka prostaciklinot moze da ima znacajna uloga vo tretmanot na dijabeticnata nefropatija, eksperimentalno inducirana so streptozocin

Ulogata na endoetelin-1 vo razvojot na dijabeticna nefropatija inducirana so streptozocin

Dijabeticnata nefropatija pretstavuva edna od hronicnite mikrovaskularni komplikacii na dijabetot, so multifaktorijalna i ne do kraj rasvetlena etiopatogeneza. So ogled na toa sto kaj pacientite so dijabet, osobeno kaj onie so dijabeticna nefropatija, se najdeni zgolemeni vrednosti na endotelin-1, se pretpostavuva deka istiot moze da ima znacajna uloga vo razvojot na dijabeticnata nefropatija. Osnovna cel na nasata studija bese da se detektiraat promenite vo plazmatskoto nivo na endotelin-1 po eksperimentalno induciran dijabet, i dijabeticna nefropatija kaj staorci so streptozocin. So ogled na dobro poznatite efekti na AKE-inhibitorite, vo ovaa studija go ispituvavme i vlijanieto na enalapril (AKE inhibitor) na plazmatskite koncentracii na endotelin-1, kako i negovite efekti vo tretmanot na dijabeti~na nefropatija. Ednokratnata i.p. administracija na streptozocin (STZ) predizvika signifikantno zgolemuvanje na plazmatskite koncentracii na endotelin-1, proprateni so jasno izrazeni simptomi i znaci na dijabeticna nefropatija (mikroalbuminurija, zgolemeni urinarni vrednosti na N-acetyl-fl-D-glucosamidase, zgolemeni serumski koncentracii na urea, poliurija). Cetiri nedelniot tretman so enalapril dovede do signifikantno namaluvanje na plazmatskite koncentracii na endotelin-1 i do podobruvanje na simtomite i znacite na dijabeticnata nefropatija. Dobienite rezultati potvrduvaat deka endotelin-1 moze da ima znacajna uloga vo razvojot i progresijata na dijabeticnata nefropatija, a AKE inhibitorite, odnosno enalapril, mozat da ja ublazat i usporat progresijata na dijabeticnata nefropatij

Pharmacokinetic profile of atorvastatin in relation to SLCO1B1 C.521T>C and C.388A> variants in healthy volunteers

OATP1B1 is an influx transporter known to be implicated as important determinant of the intestinal absorption and hepatobiliary clearance of hydrophilic statins, such as atorvastatin. Several sequence variations have been discovered in the SLCO1B1 gene encoding OATP1B1, with some of them, such as c.388A>G (p.Asn130Asp) and c.521T>C (Val174Ala) associated with increased and reduced OATP1B1 activity, respectively. The aim of the study was to investigate the effects of these two SLCO1B1 SNPs on the pharmacokinetics of atorvastatin. Twenty three healthy Macedonian volunteers were genotyped for these two SNPs using TaqMan allelic discrimination assay. After ingestion of a single dose of 80 mg, the plasma concentrations of atorvastatin were measured for 48 h using LC-MS-MS and the Cmax, Tmax, t1/2, kel, MRT, Vd, CL, AUC0-48h and AUC0-~ were determined. Allele frequencies of the variants were in Hardy-Weinberg Equilibrium, with 39 and 15% for c.388A>G and c.521T>C, respectively. Low correlation between this SNP pair (R2=0.137; D’=0.700) was observed. No significant differences in the kel, t1/2, Cmax, Tmax, AUC0-48h, AUC0-~, MRT, Vd and CL between the carriers of different c.388A>G genotypes were observed. Subject with a c.521CC genotype had markedly higher values for Cmax and AUC0-48h, 140 and 67% respectively, in comparison with the carriers of the c.521TT genotype. These differences lacked statistical significance due to the size of the sample. In addition, the effects of SLCO1B1 diplotypes on pharmacokinetic parameters were investigated comparing the effects of *15 non-carriers (n=17) and *15 heterozygotes (n=6), as *15 homozygotes were not identified in the study. The dominant effect of the c.521T>C SNP was confirmed. Marginal statistical differences were observed in Cmax, AUC0-48h, AUC0-~ and CL, with Cmax and AUC0-~ 45% (p=0.062) and 38% (n=0.09) higher, and CL 30% (p=0.07) lower in *15 heterozygotes/carriers of c.521C allele. Additional studies, with a large sample size are needed to confirm this finding

Epoetin alfa ja namaluva nefrotoksicnosta inducirana so cisplatin kaj staorci

Klinickata efikasnost na cisplatin kako antitumorski lek e nesomnena, no dozno-limitiracki faktor za negova upotreba pretstavuva izrazitata nefrotoksicnost. Najnovite istrazuvawa pokazuvaat deka epoetin alfa moze da ima znacajna uloga ne samo vo terapiski celi za korekcija na razni vidovi na anemii, tuku istiot moze da bide efikasen i kako nevroprotektiv, hepatoprotektiv, kardioprotektiv i osobeno znacajno kako nefroprotektiv kaj nefrotoksicnost inducirana od preparati na baza na platina. Glavna cel na ovaa studija bese da se utvrdi efektot na epoetin alfa vo prevencijata na nefrotoksicnost eksperimentalno inducirana so dolgotrajna administracija na cisplatin vo doza od 2 mg/kg/t.t./nedela vo tek na 8 nedeli, kaj Wistar staorci. Dobienite rezultati od ovaa studija pokazuvaat deka epoetin alfa signifikantno gi ublazuva funkcionalnite bubrezni poremetuvawa inducirani so dolgotrajna administracija na cisplatin, ja podobruva opstata sostojba i go namaluva mortalitetot kaj ispituvanite zivotni

Expression of matrix metalloproteinases 2, 7 and 9 in patients with colorectal cancer

Background/Aim. Matrix metalloproteinases (MMPs) are perceived to play a key role in tumor invasion and metastasis by their capacity to degrade basement membranes and extracellular matrix proteins. The aim of this study was to investigate the expressions of MMP-2, MMP-7 and MMP-9 in tumor tissue and their relation to clinicopathologic features in patients with colorectal cancer. Methods. Specimens of resected colorectal cancer and surrounding normal tissue of 82 patients were immunohistochemically stained for MMP-2, MMP-7 and MMP-9. The results of immunohistochemical expression of MMPs were correlated with some clinical and pathologic parameters. Results. Immunohistochemical expression of MMP-2 was more frequent in the patients with higher preoperative serum levels of carcinoembryonic antigen (CEA) (p = 0.047), MMP-2 (p = 0.018), MMP-9 (p = 0.036) and in those with lymph node metastasis (p = 0.018) and the advanced stage of the disease (p = 0.046). Expression of MMP-7 was more frequent in the patients with elevated preoperative serum levels of: CEA (p = 0.012), MMP-7 (p = 0.036), MMP-9 (p = 0.023) and with deeply invasive neoplasms (p = 0.027). MMP-9 cell expression was in a positive correlation with elevated preoperative serum levels of: CEA (p = 0.013), MMP-2 (p = 0.012), MMP-9 (p = 0.018) and depth of CRC invasion, i.e. Tparameter (p = 0.027). Conclusion. Immunohistochemical expression of MMPs is a useful indicator of the disease development and progression in patients with colorectal cancer

Frequencies of single-nucleotide polymorphisms and haplotypes of the SLCO1B1 gene in selected populations of the Western Balkans

As a membrane influx transporter, organic anion-transporting polypeptide 1B1 regulates the cellular uptake of a number of endogenous compounds and drugs. The aim of this study was to characterize the diversity of the SLCO1B1 gene encoding this transporter in two ethnic groups populating the Western Balkans. The distribution of SCLO1B1 alleles was determined at seven variant sites (c.388A>G, c.521T>C, c.571T>C, c.597C>T, c.1086C>T, c.1463G>C and c.*439T>G) in 266 Macedonians and 94 Albanians using the TaqMan allelic discrimination assay. No significant difference in the frequencies of the single nucleotide polymorphisms (SNPs) was observed between these populations. The frequency of the c.521T>C SNP was the lowest (C and c.1086C>T SNPs were not identified in either ethnic group. The haplotype analysis revealed 20 and 21 different haplotypes in the Macedonian and Albanian population, respectively. The most common haplotype in both ethnic groups, *1J/*1K/*1L, had a frequency of 39.0% and 26.6%, respectively. In both populations, the variant alleles of the functionally significant c.521T>C and c.388A>G SNPs existed in one major haplotype (*15/*16/*17), with a frequency of 8.6 and 2.4% in the Macedonian and Albanian subjects, respectively. In conclusion, sequence variations of the SLCO1B1 gene in the studied populations occur at high frequencies, which are similar to that of the Caucasian population. Further studies are needed to evaluate the clinical significance of these SNPs and/or the major SLCO1B1 haplotypes they form for a large number of substrates and for susceptibility to certain diseases

Serum Matrix Metalloproteinase-2, -7 and -9 (MMP-2, MMP-7, MMP-9) levels as Prognostic Markers in Patients with Colorectal Cancer

<p><strong>Introduction:</strong> Matrix metalloproteinases are produced by tumour cells, hence, they may be associated with tumour progression including invasion, migration, angiogenesis and metastasis. Finding prognostic markers to better identify patients with higher risk for poor survival would be valuable in order to customize pre- and postoperative treatment as well as to enable closer follow-up of these patients. Aim of our study was to examine<br />MMP-2, MMP-7 and MMP-9 serum levels and correlated them with pathological data such as stage of the colorectal cancer (CRC) and outcome.</p><p><strong>Methods: </strong>The investigation included 82 patients with operable CRC without distant metastases, who had underwent blood tests in order to determine the MMP-2, MMP-7 and MMP-9 serum levels in the following time periods: preoperatively, 3, 6, 9 and 12 months postoperatively.</p><p><br /><strong>Results</strong>: The values of the investigated MMPs decrease postoperatively and start to increase 6 month later in patients of all stages of the disease, reaching the highest value 12 month postoperatively with statistically important differences of MMP-2, MMP-7 and MMP-7 serum levels in terms of disease staging and defined points of time. Analysis of the results showed that the MMP-2 serum levels obtained 3 and 12 months postoperatively,<br />than MMP-7 serum levels 12 months postoperatively and the MMP-9 serum levels in all analyzed points in time were in significant association with the CRC patients’outcome.</p><p><br /><strong>Conclusion: </strong>The MMP-2, MMP-7 and especially MMP-9 serum values could be important indicators for diagnosis of the patients with CRC and for monitoring of disease progression.</p