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

Security and defence research in the European Union: a landscape review

This landscape report describes the state of play of the European Union’s policies and activities in security and defence and the EU-funded research aimed at supporting them, with an exclusive focus on intentional harm. It is organised around several thematic building blocks under the umbrella of the three core priorities defined in the European agenda on security. The report reviews the current main risks and threats but also those that may emerge within the next 5 years, the policy and operational means developed to combat them, the main active stakeholders and the EU legislation in force. In this context, a short history of EU research on security and defence is presented, followed by an inventory of relevant research and development projects funded under the Horizon 2020 framework programme during the period 2014-2018. The specific contributions of the Joint Research Centre to security research are also highlighted. Finally, future avenues for security and defence research and development are discussed. Please note that the executive summary of this landscape report has been published simultaneously as a companion document.JRC.E.7-Knowledge for Security and Migratio

ИМУНОЛОШКИ ПРАКТИКУМ: Привремен учебник за практична настава од имунологија

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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

Имунолошки практикум: - лабораториска имунодијагностика -

The subject of immunology was introduced for the first time in the six-year study system of the Faculty of Medicine in Skopje in the academic year 1993/94. The subject was taught in the winter semester of the third year (5th semester) with a total fund of 30 hours (15 hours of theoretical teaching and 15 hours of practical teaching). In the beginning, the entire theoretical and practical teaching was carried out by teachers and assistants in physiology from the Institute of Physiology and teachers and assistants in transfusiology from the Republic Institute of Transfusiology. During 1994, elections were announced and the first teacher and the first assistants for the subject of immunology were chosen. On 02.03. In 1995, the first Department of Immunology was established at the Faculty of Medicine in Skopje. Since 2001, the subject Immunology has been taught in the fourth semester (second year) with a fund of 15 plus 15 hours and with an additional 15 hours of seminars. From the academic year 2005/06, the European Credit Transfer System (ECTS) of the Faculty of Medicine in Skopje started, according to which the subject Immunology is taught with a pool of hours 24 and 21 (45) in the third semester (second year). In June 1995, the first textbook on immunology was published, that is, the book Basic immunological methods intended for the practical teaching of this subject. In 2005, the book Immunologic practicum for the practical teaching of the immunology subject was published by the Institute of Immunobiology and Human Genetics. The employees of the Institute were busy with the task of writing an Immunology practicum - immuno-diagnostics according to the ECTS program and making it available to the students. In this practicum, seven blocks of exercises from immunodiagnostics to immunology have been introduced. Each block is composed of three exercises, the first two of which are laboratory methods for immunodiagnostics, and the third exercise is the interpretation of the results of the corresponding area of immunology. Along with these seven blocks, i.e. 21 exercises, additional materials are offered in the form of video clips (on compact disc) for easier mastering of the material. Case analyzes are added to each exercise on the interpretation of immunological results in order for students to connect the immunological results with future clinical disciplines in which they will apply the acquired knowledge of immunology. Students should master the provided materials before attending exercises in order to win the highest number of points and thus a higher grade. I hope that the practicum will be useful to the students and that it will be a good basis for the adoption of the new curriculum of this subject.   Prof. Dr Mirko Spiroski Skopje, November 2006Предметот имунологиjа за прв пат е воведен во шестогодишниот систем на студии на Медицинскиот факулет во Скопjе во учебната 1993/94 година. Предметот се слушаше во зимскиот семестар од трета година (5. семестар) со вкупен фонд од 30 часови (15 часа теоретска настава и 15 часа практична настава). Во почетокот целокупната теоретска и практична наставата jа изведуваа наставници и асистенти по физиологиjа од Институтот за физиологиjа и наставници и асистенти по трансфузиологиjа од Републичкиот завод за трансфузиологиjа. Во текот на 1994 година беа распишани избори и беа избрани првиот наставник и првите асистенти по предметот имунологиjа. На 02.03. 1995 година е конституирана првата Катедра за имунологиjа на Медицинскиот факултет во Скопjе. Од 2001 предметот Имунологиjа се слушаше во четвртиот семестар (втора година) со фонд на часови 15 и 15 и со дополнителни 15 часови семинари. Од учебната 2005/06 започна Европскиот кредитниот трансфер систем (ЕКТС) на Медицинскиот факултет во Скопjе, според коj предметот Имунологиjа се слуша со фонд на часови 24 и 21 (45) во третиот семестар (втора година). Во jуни месец 1995 година излезе од печат првиот учебник по имунологиjа, односно книгата Основни имунолошки методи наменета за практичната настава од овоj предмет. Во 2005 излезе од печат книгата Имунолошки практикум за практичната настава од предметот имунологиjа во издание на Институтот за имунобиологиjа и хумана генетика. Вработените од Институтот се зафатиjа со задача да напишат Имунолошки практикум - имуно-диjагностика според програмата за ЕКТС и да им го стават на располагање на студентите. Во овоj практикум се внесени седум блокови од вежби од имунодиjагностиката во имунологиjата. Секоj блок е сотавен од три вежби, од кои првите две се лабораториски методи за имунодиjагностика, а третата вежба е толкување на резултатите од соодветаната област на имунологиjата. Кон овие седум блокови, односно 21 вежба се понудени дополнителни материjали во облик на видео исечоци (на компакт диск) за полесно совладување на материjалот. Кон секоjа вежба за толкување на имунолошките резултати додадени се анализи на случаи со цел студентите да ги поврзат имунолошките резултати со идните клинички дисциплини во кои ќе го применат стекнатото знаење од имунологиjата. Студентите треба да ги совладаат предвидените материjали пред да присуствуваат на вежби за да освоjат наjголем броj поени и со тоа повисока оценка. Се надевам дека практикумот ќе им биде од корист на студентите и дека ќе претставува добра основа за донесување на новата наставна програма од овоj предмет.   Проф. д-р Мирко Спироски Скопjе, ноември 2006 годин

АНАЛИЗА НА СЛУЧАИ ОД ИМУНОЛОГИЈАТА

One of the most important goals of the modernization of Medical Faculties in the European Union is the unification of basic knowledge from pre-clinical courses with clinical judgment and that as early as possible (even in the first semester). In that sense, various novelties are introduced in the contents of the subjects and their connection in logical units. The main weight that immunology has as a subject for medical students is how to master the many types of cells and their specific ratios, the even greater number of immunological mechanisms, and especially the fitting of that knowledge into immunological deviations. The most important goal of the immunology course is to enable medical students to think immunologically so that they can successfully apply immunological knowledge when they encounter a specific clinical condition. This is achieved by analyzing immunology cases in order to explain basic immunological knowledge to students and specifically relate it to clinical practice. In clinical courses, immunological case presentations are used where students are introduced to the clinical approach in solving specific immunological diseases, starting from anamnesis, family history, current condition, clinical examinations, laboratory and other diagnostic examinations, differential diagnosis, treatment, rehabilitation and forecast. In this book, already published analyses of immunology cases are selected from the highest quality books used for that purpose in the world, grouped into seven blocks of two cases each, as provided in the Immunology Practicum. The names of the cases are invented so that students can remember them more easily. Although I expect reactions to this approach to the education of medical students by clinically oriented teachers, I believe that this book will be well received by students and that it will encourage the writing of similar and better textbooks with this approach for other subjects at the Faculty of Medicine in Skopje.   Prof. Dr Mirko Spiroski October, 2007Една од најзначајните цели од модернизацијата на Медицинските факултети во Европската Унија е обединувањето на базичните знаења од предклиничките предмети со клиничкото просудување и тоа што е можно порано (уште во првиот семестар). Во таа смисла се воведуваат различни новини во содржините на предметите и нивното поврзување во логични целини. Главната тежина што ја има имунологијата како предмет за студентите по медицина е како да се совладаат многуте видови клетки и нивните специфични соодноси, уште поголемиот број имунолошки механизми, а особено вклопувањето на тие знаења во имунолошките отстапувања. Најзначајна цел на предметот имунологија е да ги оспособи студентите по медицина имунолошки да размислуваат за да можат успешно да ги применат имунолошките сознанија кога ќе се сретнат со конкретна клиничка состојба. Тоа се постигнува со анализа на случаи од имунологијата со цел на студентите да им се објаснат базичните имунолошки знаења и конкретно да ги поврзат со клиничката пракса. Во клиничките предмети се користат прикази на имунолошки случаи каде што студентите се запознаваат со клиничкиот приод во разрешување на конкретни имунолошки заболувања, почнувајќи од анамнеза, фамилна анамнеза, сегашна состојба, клинички испитувања, лабораториски и други дијагностички испитувања, диференцијална дијагноза, лекување, рехабилитација и прогноза. Во оваа книга се одбрани веќе објавени анализи на случаи од имунологијата од најквалитетните книги кои се користат за таа намена во светот, групирани во седум блока од по два случаи, како што се предвидени во Имунолошкиот практикум. Имињата на случаите се измислени за да можат студентите полесно да ги запаметат. Иако очекувам реакции кон ваквиот приод за образование на студентите по медицина од страна на клинички ориентираните наставници, верувам дека книгава ќе биде добро прифатена од студентите и дека таа ќе поттикне пишување на слични и подобри учебници со ваков приод и за другите предмети на Медицинскиот факултет во Скопје.   Проф. д-р Мирко Спироски Октомври, 2007 годин

Association of Cytokine Gene Polymorphisms with Chronic Obstructive

The aim of this study was to examine the association of 22 cytokine gene polymorphism in Macedonians with chronic obstructive pulmonary disease (COPD). The sample of the population comprised of 301 normal respondents and 62 patients with COPD. Cytokine genotyping was performed by polymerase chain reaction with sequence-specific priming (PCR-SSP). Positive (susceptible) association was found between patient with COPD and IL-1α -889/C allele; where as negative (protective) association among was found for the following alleles IL-1β +3962/C; IL-12B -1188/A; IFNγ +874/T; IL-2 -330/G; IL-4 -1098/G and IL-4-33/C. We found positive (susceptible) association between patients with COPD and following genotypes: IL4 -33/T:T; IFNγ +874/A:A; IL-4 -1098/T:T ; IL-1α -889/C:C; IL-1β +3962/C:T; IL-12B -1188/C:C; IL-4Rα +1902/G:G; IL-10 -1082/G:G; IL-2 -330/T:T; IL-4 -590/C:C; and IL-1α -889/C:T. Negative (protective) association between patients with COPD and following genotypes was found: IFNγ +874/A:T; IL-4 -33/C:T; IL-4 -1098/G:T; IL-2 -330/G:T; IL-1β +3962/C:T; IL-4 -590/C:T; IL-10 -1082/A:G; and IL-4 -33/C:C. Positive (susceptible) association between patients with COPD and following haplotypes was found: IL-4/TCT; IL-10/ATC; and IL-2/TG, and negative (protective) association was found between the patients with COPD and haplotypes for: IL-4/TTC; and IL-4/GCC. It could be concluded that several cytokine polymorphisms are positively (susceptible), or negatively (protective) associated with COPD in Macedonians

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

Methylenetetrahydrofolate reductase (MTHFR-677 and MTHFR-1298) genotypes and haplotypes and plasma homocysteine levels in patients with occlusive artery disease and deep venous thrombosis

The aim was to investigate different genotypes and haplotypes of methylenetetrahydrofolate reductase (MTHFR-677, -1298) and plasma concentration of total homocysteine (tHcy) in Macedonian patients with occlusive artery disease (OAD) and deep venous thrombosis (DVT). Investigated groups consists of 80 healthy, 74 patients with OAD, and 63 patients with DVT. Plasma tHcy was measured with Microplate Enzyme Immunoassay. Identification of MTHFR genotypes and haplotypes was done with CVD StripAssay. The probability level (P-value) was evaluated by the Student's t-test. Plasma concentration of tHcy in CC and CT genotypes of MTHFR C677T was significantly increased in patients with OAD and in patients with DVT. Plasma concentration of tHcy in AC genotype of MTHFR A1298C was increased in patients with OAD and in patients with DVT. Plasma concentration of tHcy was significantly increased in AA genotype of patients with OAD, but not in patients with DVT. We found a significant increase of plasma tHcy in patients with OAD in comparison with healthy respondents for normal:heterozygote (CC:AC), heterozygote:normal (CT:AA), and heterozygote:heterozygote (CT:AC) haplotypes. Plasma concentration of tHcy in patients with DVT in comparison with healthy respondents was significantly increased for normal:normal (CC:AA), normal heterozygote (CC:AC), and heterozygote:heterozygote (CT:AC) haplotypes. We conclude that MTHFR C677T and MTHFR A1289C genotypes and haplotypes are connected with tHcy plasma levels in Macedonian patients with OAD and DVT