Distribution of Killer Cell Immunoglobulin-Like Receptor Genes in Albanians from Republic of Macedonia

AIM: The aim of this study was to analyze Killer Ig-Like Receptor (KIR) gene polymorphisms in Albanians from Republic of Macedonia.MATERIAL NA METHODS: The studied sample consists of 104 healthy unrelated individuals, aged 20-45 years. All individuals are of Albanian nationality, residents of different geographical regions (Skopje, Gostivar, and Tetovo) in Republic of Macedonia. The population genetics analysis package, Arlequin, was used for analysis of the data.RESULTS: All 16 KIR genes known were observed in the Albanian individuals and framework genes (KIR3DL3, KIR3DP1, KIR2DL4, and KIR3DL2) were present in all individuals. The frequencies of other KIR genes were: KIR2DP1 (0.981), KIR2DL1 (1), KIR2DL2 (0.615), KIR2DL3 (0.865), KIR2DL5 (0.414), KIR3DL1 (0.933), KIR2DS1 (0.462), KIR2DS2 (0.606), KIR2DS3 (0.327), KIR2DS4 (0.875), KIR2DS5 (0.298), and KIR3DS1 (0.442). Tested linkage disequilibrium (LD) among KIR genes demonstrated that KIR genes present a wide range of linkage disequilibrium.CONCLUSION: This is the first study analyzing the polymorphism of KIR genes and genotype frequencies in Albanian individuals in the world. The results can be used for anthropological comparisons.

Food Allergy Based on Serum Specific IgE in Republic of Macedonia

ОСÐОВÐ:  Ðлергијата кон храна Ñе дефинира како неÑакан имун одговор кој Ñе јавува при повторувачки изложувања на било кој вид на храна и може да биде поÑредувана Ñо имуноглобулин Е (ИгЕ) противтела или не – ИгЕ поÑредувана. ÐлергиÑката реакција кон храна е доÑта чеÑта и Ñпоред поÑледните иÑтражувања Ñе јавува кај 6-8% од децата, а во поÑледните три декади забележан е пораÑÑ‚ во инциденцата на алергии кон храна. Доколку Ñе Ñомневаме за ИгЕ поÑредувана реакција на храната треба да Ñе направат лабораториÑки иÑпитувања и да Ñе одредат вкупните и Ñпецифичните ИгЕ противтела кон различните видови на храна.ЦЕЛ: Целта на овој труд е да Ñе преÑмета и прикаже процентот на алергии на храна во Република Македонија на оÑнова на позитивен наод на ИгЕ противтела во Ñерумот на пациенти упатени на ИнÑтитутот за имунобиологија и хумана генетика за алерголошки теÑтирања во деÑетгодишен период (2001-2011).ÐœÐТЕРИЈÐЛ И МЕТОДИ: Ðнализирани Ñе ретроÑпективно конÑекутивни пациенти во деÑет годишен период (од 01.01.2001 до 01.01.2011 година), упатени на ИнÑтитутот за имунобиологија и хумана генетика при МедицинÑкиот факултет во Скопје за иÑпитување на алергии. Ðнализите Ñе направени на 3312 иÑпитаника (пациенти) за кои имаме точни податоци за меÑтото од кое потекнуваат, за возраÑта и кај кои Ñе одредени вредноÑтите за вкупниот ИгЕ во Ñерумот, како и вредноÑтите за Ñпецифичните ИгЕ противтела наÑочени кон алергени од храната. Ðнализите за одредување на вкупен ИгЕ и Ñпецифичен ИгЕ во Ñерумот кај пациентите Ñе изработени на UniCAP100 System; Pharmacia, Uppsala, Sweden.РЕЗУЛТÐТИ: При анализата на диÑтрибуцијата на бројот на пациенти во Ñекоја група на концентрација на вкупниот ИгЕ покажуваат дека најголем дел од пациентите имале нормални вредноÑти за вкупниот ИгЕ во Ñерумот. Од вкупно 3312 пациенти Ñо Ñомневање за поÑтоење на ИгЕ поÑредувана алергија на храна кај 2367 Ñме добиле негативен наод за поÑтоење на зголемени нивоа на ИгЕ противтела, а Ñамо кај 945 иÑпитаници Ñме докажале поÑтоење на ИгЕ поÑредувана алергија на храна. ÐајчеÑтите позитивни алергени Ñе: белка од јајце, млеко, варено млеко, пченица, кикирики, протеини на млеко, лешник и јаболко, а од групно Ñпецифичните Ñе: Ñмешата од храна и Ñмешата од овошје.ЗÐКЛУЧОЦИ: Големиот број на негативни резултати ја покажува и потребата од зголемување на бројот на алерголошки лаборатории низ Република Македонија како и зголемување на едукацијата на матичните лекари преку Ñеминари, конференции и предавања. Ова е првата Ñтудија во Република Македонија за алергии кон храна во која Ñе прикажани резултати на оÑонова на добиените вредноÑти на Ñпецифични ИгЕ противтела кон различни алергени од храна во Ñерум.BACKGROUND: Food allergy is defined as an adverse immune response during repetitive exposure to any kind of food and may be mediated by immunoglobulin E (IgE) antibodies or not - IgE mediated. Food allergies are common and according to recent studies they occur in 6-8% of children, and in the last three decades there has been an increase in the incidence of food allergies. In IgE-mediated food reaction we should make laboratory tests to determine the total and specific IgE antibodies to various foods.AIM: The aim of this paper is to demonstrate the percentage of food allergies in the Republic of Macedonia based on positive IgE antibodies in the serum of patients referred to the Institute of Immunobiology and Human Genetics for allergies testing in ten year period (2001-2011).MATERIAL AND METHODS: Consecutive patients in ten year period (from 01.01.2001 to 01.01.2011), addressed to the Institute of Immunobiology and Human Genetics at the Medical Faculty tested for allergies were analyzed retrospectively. Three thousand and twelve (3312) patients with data for place of residence, age and serum values for  total IgE, specific IgE and group-specific IgE to food allergens were analyzed. The determination of total IgE and specific IgE for food allergens was performed with the UniCAP100 System (Pharmacia, Uppsala, Sweden).RESULTS: Analyzes of the distribution of the number of patients in each group of concentration of total IgE showed that most patients had normal serum total IgE levels. From a total of 3312 patients with suspected IgE-mediated food allergy, 2367 were found negative and only in 945 patients we determined the existence of IgE-mediated food allergy. The most common allergens in our study were: egg white, milk, boiled milk, wheat, peanuts, milk proteins, hazelnut and apple and from the group - specific allergens most common were food mixture and fruit mixture.CONCLUSIONS: The large number of negative results suggested the need for increasing the number of Allergologic Laboratories in Republic of Macedonia and increase of the need for better education of primary care physicians through seminars, conferences and lectures. This is the first study in the Republic of Macedonia for food allergies in which the presented results are based on the obtained specific IgE antibodies values in the sera of patients towards different food allergens

Total IgE Distribution in Food Allergy Suspected Patients in Republic of Macedonia (2001-2011)

BACKGROUND: IgE may be considered the hallmark of allergic disorders. It is easily detected in serum and can be measured as total IgE and as allergen-specific IgE. In fact, the serum IgE assay is used to diagnose an allergy.AIM: The aim of this study is to evaluate, investigate and present the distribution of total serum IgE levels, determined with UniCap system, in food-allergy suspected patients in a Republic of Macedonia.MATERIAL AND METHODS: In this study we analyzed retrospectively 8898 consecutive patients that were admitted for allergy testing at the Institute of Immunobiology and Human Genetics during the ten year period between 01.01.2001 and 01.01.2011. Total IgE levels in patient sera were detected with the in vitro system UniCAP100 (Pharmacia, Uppsala, Sweden).RESULTS: When we analyzed the number of patients according to the total IgE groups, we noted that most of the patients have normal levels of total IgE in serum. However, we also discovered a group of patients with elevated levels of total IgE that are greater than 200 kU/L. The average concentration of total serum IgE is higher in women in the age group 6 (6-7 years), followed by a steep decrease in the age group 9 (9-10 years), and after that the average concentrations of total IgE were mostly constant with the exception of a partial increase in the age group 21 (65-69 years). For men, the average serum concentrations of total IgE were highest in the age group of 6 (6-7 years), which was significantly higher than the average concentrations of total IgE in all other age groups.CONCLUSION: The large number of enrolled patients, a particular strength of this study, revealed that average concentrations of total IgE in men are higher than in women and that total IgE did not decrease with age. On the contrary, increased total IgE levels were found in patients aged 65 and 69 of both genders. We continue our work with analyses of the specific IgE antibodies values toward food and the correlation with total IgE values

Immunonephelometry and Reverse Hybrydization Genotyping in Diagnosis of Alpha-1-Antitrypsin Deficiency in Macedonians

BACKGROUND: With a frequency of 1:1600, the alpha-1-antitrypsin deficiency is one of the most frequent hereditary diseases and can be recessively inherited. AAT deficiency is most often caused by inheritance of the so-called PiZ allele. Inheritance of this allele increases the risk of developing chronic obstructive pulmonary diseases (COPD) and liver disease.AIM: The aim of this study was to present immunonephelometry and reverse hybridization genotyping in diagnosis of alpha-1-antitrypsin deficiency in Republic of Macedonia.MATERIAL AND METHODS: At the Institute of Immunobiology and Human Genetics, part of the Faculty of Medicine in Skopje, in the previous 7 years, total of 361 patients with suspected alpha-1-antitrypsin (AAT) deficiency were referred for analysis of AAT concentration using nephelometry (Dade Behring) and subsequent AAT genotyping of individuals with alpha-1-antytripsin deficiency at protein level, based on reverse hybridization technique.RESULTS: Measurement of AAT concentration (g/l) by nephelometry have shown normal level in the range of 1.37-1.41 g/l (88%), lower than normal AAT levels in the range of 0.70-0.83 g/l (8.03%), and concentration above the normal levels in the range of 2.28-2.4 g/l (3.88%).CONCLUSION: Diagnosis in the case of a suspicion of AAT deficiency is carried out by measuring the alpha-1-antitrypsin level in blood and by genotyping of alpha-1-antytripsin allele

Association of Methylenetetrahydrofolate Reductase (MTHFR-677 and MTHFR-1298) Genetic Polymorphisms with Occlusive Artery Disease and Deep Venous Thrombosis in Macedonians

Cilj Ispitati moguću povezanost genetičkog polimorfizma metilen-tetrahidrofolatne reduktaze (MTHFR-677, MTHFR-1298) s okluzivnom arterijskom bolešću i dubokom venskom trombozom u Makedonaca. Postupci Radili smo s 83 zdrave osobe, 76 bolesnika s okluzivnom arterijskom bolešću i 67 bolesnika s dubokom venskom trombozom. Od njih su prikupljeni su uzorci krvi i iz leukocita je izolirana DNA. Mutacije gena za MTHFR identificirane su testom CVD StripAssay (ViennaLab, Labordiagnostika GmbH, Beč, Austrija), a za analizu je uporabljen sustav za genetičku analizu PyPop. Potom su izračunani Pearsonove P vrijednosti, grubi omjer izgleda (odds ratio, OR) i Waldovi 95% intervali pouzdanosti (confidence intervals, CI). Rezultati Frekvencija alela C lokusa za MTHFR-677 bila je 0,575 u bolesnika s dubokom venskom trombozom, 0,612 u onih a okluzivnom arterijskom bolešću i 0,645 u zdravih osoba. Frekvencija alela T lokusa za MTHFR-677 bila je niža u zdravih osoba (0,355) nego u bolesnika s okluzivnom arterijskom bolešću (0,388) i dubokom venskom trombozom (0,425). Frekvencija alela A u lokusu MTHFR-1298 bila je 0,729 u zdravih osoba, 0,770 u bolesnika s okluzivnom arterijskom bolešću i 0,746 u bolesnika s dubokom venskom trombozom. Frekvencija alela C lokusa za MTHFR-1298 bila je 0,271 u zdravih osoba, 0,230 u bolesnika s okluzivnom arterijskom bolešću i 0,425 u bolesnika s dubokom venskom trombozom. Nije opažena povezanost polimorfizma MTHFR-677 i MTHFR-1289 s okluzivnom arterijskom bolešću ili dubokom venskom trombozom, nego se samo pokazao protektivni učinak diplotipa MTHFR/CA:CC za okluzivnu arterijsku bolest. Zaključak Osim protektivnoga učinka diplotipa MTHFR/CA:CC za okluzivnu arterijsku bolest, nismo našli značajnu povezanost polimorfizma lokusa MTHFR-677 i MTHFR-1289 s okluzivnom arterijskom bolešću i dubokom venskom trombozom.Aim To analyze the association of methylenetetrahydrofolate reductase polymorphisms (MTHFR-677 and MTHFR-1298) with occlusive artery disease and deep venous thrombosis in Macedonians. Methods We examined 83 healthy respondents, 76 patients with occlusive artery disease, and 67 patients with deep venous thrombosis. Blood samples were collected and DNA was isolated from peripheral blood leukocytes. Identification of MTHFR mutations was done with CVD StripAssay (ViennaLab, Labordiagnostika GmbH, Vienna, Austria) and the population genetics analysis package, PyPop, was used for the analysis. Pearson P values, crude odds ratio, and Wald’s 95% confidence intervals were calculated. Results The frequency of C alleles of MTHFR-677 was 0.575 in patients with deep venous thrombosis, 0.612 in patients with occlusive artery disease, and 0.645 in healthy participants. The frequency of T allele of MTHFR-677 was lower in healthy participants (0.355) than in patients with occlusive artery disease (0.388) and deep venous thrombosis (0.425). The frequency of A allele for MTHFR-1298 was 0.729 in healthy participants, 0.770 in patients with occlusive artery disease, and 0.746 in patients with deep venous thrombosis. The frequency of C allele of MTHFR-1298 was 0.271 in healthy participants, 0.230 in patients with occlusive artery disease, and 0.425 in patients with deep venous thrombosis. No association of MTHFR-677 and MTHFR- 1289 polymorphisms with occlusive artery disease and deep venous thrombosis was found, except for the protective effect of MTHFR/CA: CC diplotype for occlusive artery disease. Conclusion We could not confirm a significant association of MTHFR- 677 and MTHFR-1289 polymorphisms with occlusive artery disease or deep venous thrombosis in Macedonians, except for the protective effect of MTHFR/CA:CC diplotype against occlusive artery disease

ИМУНОЛОШКИ ПРАКТИКУМ: Привремен учебник за практична настава од имунологија

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ПРАКТИКУМ ЗА ХУМАНА ГЕНЕТИКА - 1: Учебник за практична настава од хумана гентика - второ издание -

The subject of human genetics was introduced for the first time with the European Credit Transfer System (ECTS) at the Faculty of Medicine in Skopje in the academic year 2005/2006. The subject is taken in the second semester with a total of 60 hours (30 hours of theory and 30 hours of exercises) and is valued at 4 credits. The content of the subject is divided into two functional units: Human Genetics-1, with a fund of 30 hours, in which the basics of human genetics are presented, and Human Genetics-2, with a fund of 30 hours, in which the genetic aspects of diseases are presented man, their diagnosis and treatment. In this practicum for Human Genetics-1, texts from 15 exercises have been entered, grouped into five blocks of three exercises each. At the beginning of the blocks are given the web objectives for each separate exercise and the points that bring the presence of the exercises and the knowledge gained from them. In each exercise, extensive material is given that is related to the performance of the corresponding exercise, mainly due to the lack of appropriate printed materials in the Macedonian language for their understanding. At the end of each exercise, several tasks are given that each student should perform, and in the corresponding spaces enter a part of the results that he received. In that way, students can recall the performance of the exercises during their studies. With the introduction of a course for Higher Vocational Schools at the Faculty of Medicine in Skopje, the subject Human Genetics was introduced for nurses with a fund of 15+0 hours, as well as for speech therapists with a fund of 30+30 hours. This practicum is intended for practical teaching for speech therapists, with a shortening of 5 exercises (one exercise from each block). The first edition of the Human Genetics Practicum-1 was printed in 2006. Comments from reviewers and students have been partially incorporated into this edition. We expect students to provide additional comments to improve the content and text of this practicum. I hope that the practicum will help students master the exercises from Human Genetics-1. Prof. Dr Mirko Spiroski Skopje, 2009Предметот хумана генетика за прв пат е воведен со Европскиот кредитен трансфер систем (ЕКТС) на Медицинскиот факултет во Скопје во учебата 2005/2006 година. Предметот се слуша во вториот семестар со вкупен фонд од 60 часови (30 часа теоретски и 30 часа вежби) и се вреднува со 4 кредити. Содржината на предметот е поделена во две функционални целини: Хумана генетика-1, со фонд од 30 часа, во која се изнесуваат основите на хумана генетика и Хумана генетика-2, со фонд од 30 часа, во која се изнесуваат генетски аспекти на болестите кај човекот, нивното дијагностицирање и лекување. Во овој практикум за Хумана генетика-1 внесени се текстови од 15 вежби, групирани во пет блока од по три вежби. На почетокот од блоковите се дадени уебните цели за секоја одделна вежба и поените кои ги носат присуството на вежбите и стекнатото знаење од нив. Во секоја вежба даден е обемен материјал кој е поврзан со изведувањето на соодветната вежба, главно заради недостаток од соодветни печатени материјали на македонски јазик за нивно разбирање. На крајот од секоја вежба дадени се неколку задачи кои треба секој студент да ги изведе, а во соодветните простори да внесе дел од резултатите кој ги добил. На тој начин студентите можат да се присетуваат на изведувањето на вежбите во текот на нивното учење. Со воведување насока за Високи стручни школи на Медицинскиот факултет во Скопје, воведен е предметот Хумана генетика за медицински сестри со фонд на часови 15+0, како и за логопеди со фонд на часови 30+30. Овој практикум е наменет за практичната настава за логопеди, со скратување на 5 вежби (по една вежба од секој блок).  Првото издание на Практикумот за хумана генетика-1 беше отпечатено во 2006 година. Забелешките од рецензентите и од студентите се делумно вградени во ова издание. Очекуваме студентите да дадат дополнителни забелешки за подобрување содржината и текстот на овој практикум. Се надевам дека практикумот ќе им помогне на студентите да ги совладаат вежбите од Хумана генетика-1.   Проф. д-р Мирко Спироски Скопје, 2009 годин

A case of a ten-year old girl with dominantly inherited Familial Mediterranean fever in Republic of Macedonia

The Familial Mediterranean fever (FMF, MIM249100) is an autoimflammatory genetic disease characterized with recurrent painful attacks in the abdomen, chest or joints, usually accompanied with high body temperature. It is classically inherited in an autosomal recessive manner. It is associated with mutations of the MEFV gene, coding for the protein pyrin. More than 140 mutations of the MEFV gene are defined worldwide. Despite the progress in establishing reliable tests practical for routine use, as much as 20% of the patients with FMF remain without a detectable mutation in the MEFV gene. This is the main reason why the diagnosis of FMF remains still a clinical one, according to Tel Hashomer criteria. A 10-year old girl admitted to the Clinic of Pediatrics at the Faculty of Medicine in Skopje for unexplained fever. After numerous laboratory analyses and specialist consultations were done, genetic testing for FMF was requested. The presence of an heterozygous mutation E148Q was confirmed at the Institute for Immunobiology and Human Genetics using a PCR based, reverse hybridization method. Administration of colchicine, the therapy of choice, in a dose of 1.5 mg/day, lead to complete resolution of the symptoms within some days following commencement. Although the disease is classically inherited in a recessive manner, some atypical cases of autosomal dominant inheritance are described. Our patient may be another example supporting the unusual dominant inheritance, since the heterozygous state for the E148Q mutation was the only positive finding in the genotyping of the 12 most frequent MEFV mutations

Immunoglobulin Classes and Subclasses in Macedonian Elderly People Maced

Abstract Aim: The aim of this study was to determine the association of HLA-A, -C and -B genes with ankylosing spondylitis in patients from the Republic of Macedonia. Material and Methods: This study included 307 subjects (250 healthy individuals and 57 patients with ankylosing spondylitis who were diagnosed at the University Clinic of Rheumatology in Skopje). The HLA typing of class 1 (HLA-A, HLA-C and HLA-B) genes was performed using the method of Reverse Line Strip, after isolation of DNK from the blood leucocytes with the standard phenol-chloroforme method. The HLA sub typing of HLA-B*27 was performed with high resolution single-strand polymorphism

Influence of the elevated ambient temperature on immunoglobulin G and immunoglobulin G subclasses in sera of Wistar rats

The aim of our research was to examine changes in the immune system of the rats influenced by the elevated ambient temperature. Male Wistar rats were divided, into 2 groups and housed at 20 ± 2°C (n=64, control group) and 35 ± 1°C (n=74, experimental group), during precise timing of 1, 4, 7, 14, 21, and 30 days. All the animals were given food and water ad libitum, and were lighted during 12 hours per day. We have measured IgG, IgG1, IgG2a, IgG2b and IgG2c. The obtained results showed significant elevation in the level of IgG after 4 and 7 days (+32%), IgG2a after 7th (+88%), 14th and 21nd day (+110%), IgG2b after 14 days (+60%) at 35 ± 1°C compared with the control group at 20 ± 2°C. IgG1 level was not affected and IgG2c showed significant decrease after 21st day at 35 ± 1°C. In conclusion, during the elevated ambient temperature the immune system is activated as one of the regulation mechanisms in homeostasis and survival of the population