Влијанието на Ц677Т полиморфизмот на генот за МТХФРврз инциденцијата на токÑичните ефекти од виÑоките дози МТХ кај деца Ñо акутна лимфоблаÑтна леукемија

In current protocols for treatment of acute lymphoblastic leukemia in childhood methotrexate (MTX) is one of the crucial cytostatics. The occurrence of MTX toxicity is still  Ð° great problem because of the interpatient differences in drug metabolism. These differences may  be due  to polymorphisms of genes involved in the folate metabolism. The present study was carried out to determine the prevalence of MTHFR  C677T polymorphism in children with acute lymphoblastic leukemia (ALL). Also the effect of the genotype on the toxic  effects during therapy with high doses of МТХ in 45 patients with ALL treated in accordance with the protocol ALL BFM 95 and ALL BFM 2000 was evaluated. All 45 patients with ALL were genotyped for MTHFR C677T polymorphism. Correlation with the presence of a certain polymorphism and toxic effects of the chemotherapy with high doses of МТХ was  made in the Department  for hematology and oncology at the University Clinic for children's diseases – Skopje. The control group included 32 healthy patients. In the study group 24 (53.3%) children had a wild type of polymorphism (CC), 15 (33.33%) children were heterozygous (CT) for MTHFR C677T polymorphism, and 6 (13.33%) children were homozygous for the variant type  of the polymorphism (ТТ). The  correlation of the genotype with МТХ toxicity indicated a statistical significance only for oral mucositis, while for the other toxic effects there was no statistically significant correlation. In our study the results indicated that oral mucositis was statistically significantly more frequently identified in the case of the variant carriers for this polymorphism. For the other toxic effects caused by the therapy with high doses of МТХ no statistically  significant correlation with MTHFR C677T polymorphism was  identified. This  occurrence maybe due  to the small number of patients analyzed in this study and the possible protective influence of other genetic polymorphisms included in the folate metabolism, which were not subject to consideration of this study.Во актуелните протоколи за третман на акутна лимфоблаÑтна леукемија во  Ð´ÐµÑ‚Ñката возраÑÑ‚ метотрекÑатот (МТХ) е еден од круцијалните цитоÑтатици. Предвидувањето на појавата на токÑични ефекти во тек на терапијата Ñо МТХ претÑтавува Ñè уште голем проблем заради индивидуалните разлики во метаболизмот на лекови кај пациентите. Овие разлики можеби Ñе должат на приÑуÑтво на полиморфизми на гените вклучени  Ð²Ð¾  Ñ„олатниот метаболизам. Целта на  Ñтудијата беше  Ð´Ð°  Ñе одреди инциденцијата на МТХФР Ц677Т полиморфизмот кај децата Ñо акутна лимфоблаÑтна леукемија (ÐЛЛ) и да Ñе анализира влијанието на генотипот врз  Ð¼Ð°Ð½Ð¸Ñ„еÑтацијата на токÑичните ефекти во тек на терапија Ñо виÑоки дози МТХ кај пациенти Ñо ÐЛЛ третирани по  Ð¿Ñ€Ð¾Ñ‚околот ÐЛЛ БФМ  95. Беше вклучена и контролна група  Ð¾Ð´ 32 здрави иÑпитаници. Беше  Ð½Ð°Ð¿Ñ€Ð°Ð²ÐµÐ½Ð° генотипизација за полиморфизмот Ц677Т кај 45 пациенти Ñо ÐЛЛ и негова корелација Ñо токÑичните ефекти од хемотерапијата Ñо виÑоки дози  ÐœÐ¢Ð¥ анализирани од болничките иÑтории на пациенти Ñо ÐЛЛ лекувани на Одделот за хематологија и онкологија при  ÐˆÐ—У УниверзитетÑка клиника за детÑки болеÑти - Скопје.  Ð’о ÑтудиÑката група 15 (33,33%) пациенти беа хетерозиготи (ЦТ) за полиморфизмот Ц677Т на генот MTHFR, а 6 (13,33%) пациенти хомозиготи (ТТ). Во контролната група  10 иÑпитаници  (31,25%) беа  хетерозиготи за полиморфизмот (ЦТ), а 6 иÑпитаници (18,75%) хомозиготи (ТТ). Корелацијата на генотипот Ñо токÑичните ефекти од виÑоките дози  Ð½Ð° МТХ покажа ÑтатиÑтичка ÑигнификантноÑÑ‚ Ñамо за орален мукозит, додека беше ÑтатиÑтички неÑигнификантна за оÑтанатите токÑични ефекти. Оваа појава можеби Ñе должи на малата група пациенти којашто беше анализирана во оваа Ñтудија и можното протективно влијание на  Ð´Ñ€ÑƒÐ³Ð¸  Ð³ÐµÐ½Ñки полиморфизми вклучени во фолатниот метаболизам, а кои не беа предмет на оваа Ñтудија

ТокÑични ефекти предизвикани од хемотерапија Ñо виÑоки дози метотрекÑат кај деца Ñо акутна лимфоблаÑтна леукемија

Intensification of systemic chemotherapy with incinclusionof high doses methotrexate (MTX) has contributed to the improvement of event-free survival in children with acute lymphoblastic leukemia (ALL). Despite this benefit, this agent might cause serious toxicity, even life-treating events during treatment. Therefore, prediction, early detection and management of toxic effects during therapy with high doses of MTX is still a great challenge for every pediatric oncologist.The aim of our study was to evaluate the incidence of toxic effects of chemotherapy with high doses of MTX (5 g/m^2) and to compare them with toxicity during application of lower doses of MTX (2 g/m^2).Retrospective record review of 77 children with medium risk ALL was done. Patients were treated in the Department of hematology and oncology at the University children's hospital in Skopje. Forty-five of them were treated with 5 g/m^2 and 32 of them were treated with 2 g/m^2 (historic group). Toxicity was registered according to the protocol for acute toxicity, part of the ALL BFM 95 protocol.Toxic effects were predominant in the group treated with higher doses of MTX. The  most significant toxic  effects were hepatotoxicity, oral mucositis and myelosuppression.  More  severe grade of hepatotoxicity and oral mucositis were present in the study group. In our study toxic  effects were more common in the study group due to application of higher doses of MTX.Variations in toxicity between the patients of the study group are probably due  to the genetic differences in the drug absorption, their excretion and cellular transport. Current studies are dedicated on discovering genetic markers which will  be able  to predict the risk  of appearance of MTX toxicity.Intensification of systemic chemotherapy with in- clusion of high doses methotrexate (MTX) has contributed to the improvement of event-free survival in children with acute lymphoblastic leukemia (ALL). Despite this benefit, this agent might cause serious toxicity, even life-treating events during treatment. Therefore, prediction, early detection and management of toxic effects during therapy with high doses of MTX is still a great challenge for every pediatric oncologist.The aim of our study was to evaluate the incidence of toxic effects of chemotherapy with high doses of MTX (5 g/m2) and to compare them with toxicity during application of lower doses of MTX (2 g/m2).Retrospective record review of 77 children with medium risk ALL was done. Patients were treated in the Department of hematology and oncology at the University children's hospital in Skopje. Forty-five of them were treated with 5 g/m2 and 32 of them were treated with 2 g/m2 (historic group). Toxicity was registered according to the protocol for acute toxicity, part of the ALL BFM 95 protocol. Toxic effects were predominant in the group treated with higher doses of MTX. The  most significant toxic  effects were hepatotoxicity, oral mucositis and myelosuppression.  More  severe grade of hepatotoxicity and oral mucositis were present in the study group. In our study toxic  effects were more common in the study group due to application of higher doses of MTX. Variations in toxicity between the patients of the study group are probably due  to the genetic differences in the drug absorption, their excretion and cellular transport. Current studies are dedicated on discovering genetic markers which will  be able  to predict the risk  of appearance of MTX toxicity

KIR Gene Frequencies in Women with Infertility Problems

Introduction: Natural killer (NK) cells are the predominant lymphocyte population in the decidua. Being the most abundant leucocytes, the activity of NK cells is important in different immuno-pathological conditions, such as recurrent spontaneous abortions, infertility and problems in implantation. The NK cells recognize HLA class I molecules on trophoblasts trough killer immunoglobulin-like receptors (KIRs) found on their surface. The KIRs are classified as either activating or inhibitory, regarding the effect they produce on NK cells upon interaction with corresponding ligand. Since KIR genes exhibit extensive polymorphism and individuals differ in both the number and kind (activating vs. inhibitory) of KIR genes, it is hypothesized that the KIR gene content might influence the pregnancy outcome. Aim: The aim of this pilot study is to analyze the frequency of different KIR genes in women with infertility problems, and compare them to healthy women. Material and Methods: Total of 122 healthy women (Control) and 25 women with reproductive problems (MISSC) participated in this study. After signing of written consent DNA was isolated from peripheral blood using phenol/chloroform method. The genotyping of 16 KIR genes was performed using commercially available kit from Dynal Biotech, (Pel-Freez Clinical Systems, Brown Deer, WI, USA), based on SSP method. Results: We found that inhibitory KIR are present in similar observed frequency in both control and patients with MISSC, except KIR2DL5 which was found in lower frequency in patients with MISSC. There are no significant differences of all noninhibitory KIR between control and patients with MISSC. The number of inhibitory KIR genes in patients with MISSC was lover, except for seven inhibitory KIR genes which was almost doubled. The number of noninhibiotry (stimulatory) KIR genes was lower in patients with MISSC, except for those with three KIR genes which were almost four times more frequent. We found significantly bigger percentage of 0,34 – 0,60 activating/inhibitory KIR gene number ratio in the patients with MISSC. Conclusion: In conclusion, there are differences in the KIR gene distribution, gene number, and activating/inhibitory KIR gene number ratio between control and Macedonian patients with MISSC. Further analysis of frequencies of corresponding KIR genotypes or in the ratio of activating/inhibiting genes content in two groups are needed

Anti Citrullinated Protein / Peptide Antibody Assay, Rheumathoid Factor or Both as Shifted Test in Diagnostic and Prognostic Evaluation in Patients with Rheumathoid Arthritis

The aim of this study was to compare the diagnostic values of laboratory variables, to present quantitative evaluations of the anti citrullinated protein / peptide antibody (ACPA), or anti CCP ( anti-cyclic citrullinated peptide, anti-CCP 2) antibodies in second generation antibody assay diagnostic test with reference to sensitivity and specificity, the predictive value of the positive and negative test and precision of the test for ACPA antibodies, rheumatoid factor, C-reactive protein and DAS 28 index, in the early diagnosis of untreated rheumatoid arthritis

Referentni intervali na Apo A-1, Apo B i soodnosot Apo B/Apo A-1 vo primerok na makedonska populacija

Narusuvanjata vo serumskite koncentracii na apolipoproteinite A-1 i B (Apo A-1 i Apo B), kako i soodnosot apolipoprotein B/apolipoprotein A-1 (Apo B/Apo A-1), se smeta deka pretstavuvaat nezavisni rizik faktori za koronarna srceva bolest. Se preporacuva sekoja laboratorija da opredeli sopstveni referentni intervali za serumskite koncentracii na Apo A-1 i na Apo B, kako i za soodnosot Apo B/Apo A-1, bidejki tie zavisat od geografskata lokacija. Za taa cel gi opredelivme serumskite koncentracii na Apo A-1 i na Apo B i go presmetavme Apo B/Apo A-1 soodnosot kaj 122 zdravi individui od makedonska populacija (70 `eni i 52 ma`i) na vozrast od 18-60 godini. Pri opredeluvaweto koristevme komercijalen, standardiziran manuelen imunoturbidimetriski test (Randox). Izmerenite koncentracii (x ± SD) iznesuvaa: Apo A-1 = 1,39 ± 0,28 g/l vo zenskata populacija i 1,42 ± 0,27 g/l vo maskata populacija, a Apo B = 0,95 ± 0,24 g/l vo zenskata i 1,00 ± 0,23 g/l vo maskata populacija. Presmetaniot Apo B/Apo A-1 soodnos iznesuvase 0,70 ± 0,17 kaj zenskata populacija i 0,72 ± 0,18 kaj maskata populacija. Studentoviot t-test pokaza deka ne postoi signifikantna razlika (r>0,05) vo srednite vrednosti za Apo A-1 i za Apo B, kako i za Apo B/Apo A-1 soodnosot, me|u zenskata i maskata podgrupa, odnosno deka podelbata spored polot ne e potrebna od dijagnosticka gledna tocka. Koncentraciite na Apo A-1 i na Apo B presmetani za mesanata grupa (N=122) iznesuvaa 1,40 ± 0,28 g/L i 0,97 ± 0,23 g/L. Soodnosot Apo B/Apo A-1 vo celata grupa iznesuva{e 0,71 ± 0,17. Tipot na distribucija na vrednostite za serumskite Apo A-1 i Apo B, kako i soodnosot Apo B/Apo A-1 be{e testiran preku opredeluvanje na vrednostite na koeficientot na asimetrija (Skewness), na koeficientot na splosnatost (Kurtosis) i na standardnite devijacii na ovie dva koeficienti. Vrednostite na ovie koeficienti za site tri parametri pokazaa deka distribucijata na vrednostite e spored Gausovata (Gauss) kriva. Vo soglasnost so normalnata distribucija, referentnite granici za Apo A-1, za Apo B, kako i za Apo B/Apo A-1 soodnosot bea opredeleni parametarski (sredna aritmeticka golemina ± 2 standardni devijacii). Referentnite intervali za Apo A-1 iznesuvaa 0,84 - 1,96 g/l, 0,51 - 1,43 g/l za Apo B i 0,37 - 1,05 za soodnosot Apo B/Apo A-

Cholesteryl ester transfer protein, low density lipoprotein particle size and intima media thickness in patients with coronary heart disease

Cholesteryl ester transfer protein (CETP) plays a key role in reverse cholesterol transport and high density lipoprotein (HDL) metabolism. Predominance of small, dense LDL particles is associated with an increased risk of atherosclerosis and coronary heart disease (CHD). The aim of the study was to determine the potential relationship between the CETP concentration and low density lipoprotein (LDL) particle size and their association with intima media thickness (IMT) in patients with CHD. Lipid parameters, CETP concentration and LDL particle size were determined in 100 healthy subjects (control group) and in 100 patients with CHD, aged 43 to 77 years. Plasma CETP concentrations were measured by an enzyme-linked immuno-sorbent assay with two different monoclonal antibodies. LDL subclasses were separated by nondenaturing polyacrilamide 3-31% gradient gel electrophoresis. CETP concentration was higher in patients compared to controls (2.02 ± 0.75 mg/ml vs. 1.74 ± 0.63 mg/ml, p<0.01). Mean LDL particle size (nm) was significantly smaller in patients than in controls (24.5 ± 1.1 vs. 26.1 ± 0.9; p<0.001). There was no relation between LDL particle size and CETP concentration (r=-0.1807, p=0.072). Age, diastolic blood pressure, CETP concentration and LDL particle size were independent factors for determing IMT by multiple linear regression analysis. They accounted for 35.2 % of the observed variability in IMT. CETP is not an independent contributor of LDL particle size. CETP might play a role in determining lipoprotein distributions, but did not seem to be the sole factor in the formation of small LDL particles

THE EFFECT OF PLASMA PREPARATION RICH IN GROWTH FACTORS ON PATELLAR STABILITY AFTER MEDIAL PATELLOFEMORAL LIGAMENT REEFING

Introduction: Although more than 100 operative procedures have been described for the treatment of patellar instability, there is no single universally successful procedure. For the most patients with lateral patellar instability medial patellofemoral ligament (MPFL) reefing is recommended. When we perform MPFL reefing we are not aware of the quality and strength of the MPFL tissue. In the presence of recurrent patellar instability,the quality and strength of MPFL tissue is often compromised and it disturbs patellar stability after MPFL reefing. Biomedicine development,recognizing the ligament healing process show us that autologous blood products, particularly PRP can enhance healing in soft tissue injuries. Purpose: The purpose of this study was to determine the potential effect of Plasma preparation rich in growth factorson patellar stability after MPFL reefing. Material and methods: Plasma preparationrich in growth factors was produced from a unit of autologous whole blood using Arthrex ACP double syringe system.Platelet gel was prepared by adding bovine thrombin and 10% solution of calcium chloride.The platelet gel was applied locally into the place where suturing of the MPFL was performed. In this prospective, randomized and double blindstudy12 patients were included:6 patients in the PG group who received platelet gel and 6 patients in the control group who were not treated with platelet gel. Patellar stability was evaluated before surgery and 3 months after surgery with Axial stress radiographs. Results: The calculated 3 month improvement was 12.67 ± 2.51 in the control group and 17.33 ± 1.52 in the PG group, (p=0.064).Although there was greater improvement in patellar stability in PG group comparing to the control group, the difference was not statistically significant (p>0.05).The main reason for this was probably the small number of patients included in the study. Conclusion: Results showed that growth factors from the plasma preparation rich in growth factorshave positive effect on patellar stability after MPFL reefing.We believe that they stimulate and accelerate physiological healing and reparative tissue processes in ligament healing. More studies should be made, including more patients, if we want to get more relevant results

Symmetric dimethyl arginine and N-acetyl-β-Dglucosaminidase lysozimuria of proximal renal tubules as a target for nephrotoxicity in patients with rheumatoid arthritis treated with disease modifying antirheumatic drugs

The aim of this study was to determine the effect of initial therapy with some disease modifying antirheumatic drugs (DMARDs) (Methotrexate and Ketoprofen) on glomerular and tubular integrity in patients with Rheumatoid arthritis (RA). To determine whether there is a change in clinical and laboratory indicators of renal function in course of the follow up of treatment and whether that change correlates with the dynamics of the quantity of enzymes excreted in urine and reactants of the acute phase. Using colorimetric method for determination of NAG, samples of 70 participants were examined (35 RA patients treated with Ketoprofen only, 35 RA patients treated with combined use of Methotrexate and Ketoprofen). The follow up was 5 time-intervals in the course of 24 weeks. There was moderate correlation between NAG and microalbuminuria (r=0,34) in the group of patients treated with Ketoprofen only, while statistically significant correlation (r=0,21) was seen in group of patients with combined use of Methotrexate and Ketoprofen. NAG enzymuria in size, number of patients registered, and time of appearance were greater and appears earlier in the group with the combined use of Methotrexate and Ketoprofen compared with the mono-therapy with Ketoprofen. Mean urinary NAG induction was increasing with the concomitant use of Methotrexate and Ketoprofen. Methotrexate is more potent NAG inductor than Ketoprofen and provokes greater tubular enzymuria than Ketoprofen

Determination of the Diagnostic Values of Asymmetric Dimethylarginine as an Indicator for Evaluation of the Endothelial Dysfunction in Patients with Rheumatoid Arthritis

Introduction. To compare the diagnostic values of laboratory variables, to present evaluations of the diagnostic test for asymmetric dimethyl arginine (ADMA), rheumatoid factor (RF), C-reactive protein (CRP), and DAS28 index, and to define the effect of untreated rheumatoid arthritis on endothelial function. In order to determine whether ADMA changes depending on the disease evolution, ADMA was used as an indicator for endothelial dysfunction. Methods. Using an ELISA technology of DLD-Diagnostika-GMBH for the detection of ADMA, the samples of serum and urine have been examined in 70 participants (35 RA who were not treated, 35 healthy controls). RF was defined with the test for agglutination (Latex RF test) in the same participants. Results. Out of 35 examined patients with RA, RF appeared in 17 patients (sensitivity of the test, 51.42%). In 20 of the 35 examined patients with RA, we found the presence of ADMA (sensitivity of the test, 57.14%). Anti-CCP antibody was present in 24 examined patients with RA (sensitivity of the test, 68.57%). Conclusion. ADMA has equal or very similar sensitivity and specificity to RF in untreated RA (sensitivity of 57.14% versus 48.57%, specificity of 88.57% versus 91.42%) in the detection of asymptomatic endothelial dysfunction in untreated RA

Measles outbreak in Macedonia: epidemiological, clinical and laboratory findings and identification of susceptible cohorts.

OBJECTIVES: Despite a 92-99% national vaccination coverage since 2000, the former Yugoslav Republic of Macedonia experienced a large measles outbreak between 2010 and 2011. Here we investigate the characteristics of patients hospitalized during this outbreak at the Clinic of Infectious Diseases in Skopje. METHODS: Epidemiological, clinical and laboratory data of 284 measles patients, including 251 from Skopje (43.80% of the 573 reported cases) and 33 from elsewhere in Macedonia were collected. RESULTS: The most affected age groups were children up to 4 years of age and adolescents/adults of 15 years and older. Most patients were unvaccinated (n=263, 92.61%) and many had non-Macedonian nationalities (n=156, 54.93%) or belonged to the Roma ethnicity (n=73, 25.70%). Bronchopneumonia and diarrhea were the most common complications. Eighty-two out of 86 tested patients (95.35%) had measles-specific IgM antibodies. The outbreak was caused by the measles variant D4-Hamburg. CONCLUSIONS: The epidemic identified pockets of susceptibles in Skopje and indicated that additional vaccination opportunities in particular for people with non-Macedonian nationality and traveler communities are warranted to ensure efficient measles control in Macedonia. The high attack rate among children of less than 1 year suggests that vaccination before 12 months of age should be considered in high risk settings