FOXP1 Expression in Normal and Neoplastic Erythroid and Myeloid Cells

FOXP1 protein was firstly analyzed in normal tissues, and afterwards in different tumor tissues, mainly carcinoma and lymphoma. In B-cell malignancies, its role was well explored; its expression was shown to be connected with disease prognosis in certain B-non Hodgkin lymphomas. In this study, 16 bone marrow trephine samples from patients with no hematopoietic malignancies and 10 samples from peripheral blood of healthy individuals were immunostained with anti-FOXP1 antibody. Positive cells in bone marrows were not only lymphocytes, but also cells that are immunohistochemically positive for glycophorin C or myeloperoxidase. Peripheral blood samples showed no other positive cells, but small round lymphocytes. Additionally 60 samples from patients with myeloid lineage neoplasms were analyzed. 25 samples from patients with myelodisplastic syndrome (MDS) and 35 patients with myeloproliferative disease (MPD) were double immunostained with anti-FOXP1/anti-glycophorin C and anti-FOXP1/anti- myeloperoxidase antibodies. FOXP1 was found to be expressed in 22 cases of MDS and in none of MPD cases. Its expression in MDS was observed mostly in myeloperoxidase positive cells in contrast to gylcophorin C positive cells. Only two cases revealed both myeloperoxidase positive cells and gylcophorin C positive cells expressing FOXP1 transcription factor. Our results show that FOXP1 is present in normal cells of erythroid and myeloid linages and thus suggest its possible role in development of all hematopoetic cells as well as possible involvement in neoplasm development of myeloid disorders

Prognostic Markers and Gene Abnormalities in Subgroups of Diffuse Large B-cell Lymphoma: Single Center Experience

Aim To explore the association between FOXP1, BCL2, and BCL6 gene expression in diffuse large B-cell lymphoma tumor cells and their association with the presence of FOXP3 lymphocytes. Methods Samples of lymph nodes from 53 patients with newly diagnosed diffuse large B-cell lymphoma were taken at the time of the diagnosis and immunostained for CD10, MUM1, BCL6, BCL2, FOXP1, and FOXP3. Fluorescent in situ hybridization analysis was used for the detection of FOXP1, BCL2, and BCL6 gene abnormalities. The χ2 test was used for data analysis. Results FOXP1 protein was detected in 28 cases, genetic abnormalities involving the FOXP1 locus were found in 19 cases, and both were present in 13 cases (χ2 =7.157; P = 0.028). FOXP3 positive cells were detected in 37 cases. There was a significant relationship between BCL2 expression and FOXP1 genetic abnormalities (χ2 =5.858; P = 0.016) and between BCL2 expression and BCL2 genetic abnormalities (χ2=6.349; P = 0.012). There was also an association between BCL6 and FOXP1 genetic abnormalities (χ2 =8.497; P = 0.004). Conclusion Association was observed between additional FOXP1 gene copies and BCL2 protein expression as well as changes on both FOXP1 and BCL2 genes in samples of our DLBCL patients.. FOXP3 positive cells showed no association with presence of any of analyzed proteins considered as a prognostic markers in DLBCL neither with changes of their genes

Oncolytic Viruses

Maligni tumori danas su najčešći uzročnici smrtnosti u ljudi. Nakupljanje mutacija u stanicama uzrokuje nekontroliranu diobu stanica, kao i sprječavanje njihove programirane smrti, što dovodi do neoplastičnih promjena. Nakuplja se tumorska masa koja izbjegava imunosni odgovor domaćina. Moderan pristup terapiji malignih tumora, koji je još uvijek u istraživačkoj fazi, temelji se na primjeni bakterija i/ili virusa koji su genetički modificirani. Njima se potiče kemotaksijski učinak ili nastanak receptora koje će prepoznati imunosni sustav domaćina kako bi se potaknulo efektorski dio obrambenog sustava organizma na uništavanje tumorske mase. U ovom radu dan je sažet prikaz najnovijih metoda navedenog područja.Malignant tumors are the most common cause of mortality in humans. Accumulation of mutation in cells causes uncontrolled cell division as well as prevention of their programmed cell death, leading to neoplastic changes. Tumor mass which avoids the immune response of the host, grows and spreads. Modern approach to malignant tumor therapy, which is still in the research stage, is based on the use of genetically modified bacteria and/or viruses. They are used for promotion of the chemotactic effect or production of the receptors that will be recognized by the immune system of the host and in this way stimulate the effector part of the body’s defense system to destroy the tumor mass. This paper summarizes the most recent methods in the field. Keywords

Development of Immunohistochemical Staining Methods and Optimization of Immunohistochemical Protocol for BACH2 Transcription Factor

Imunohistokemijsko bojenje jest metoda detekcije prisutnosti i lokalizacije specifičnih antigena u stanici ili tkivu. Primjenjuje osnovni princip imunosne reakcije kod kojeg se specifično antitijelo veže na određeni antigen koji prepoznaje. Razvoj imunohistokemijskih metoda započeo je 1930-ih godina, od kada se, sve do danas, primjenjuju u različitim oblicima za mnoga znanstvena istraživanja i u rutinskim dijagnostičkim postupcima. U ovom radu opisan je razvoj metoda imunohistokemijskog bojenja te je prikazana optimizacija protokola za detekciju antigena BACH2 u ljudskom sekundarnom limfnom tkivu. Raspravljeni su pojedini koraci metode poput pripreme prereza tkiva koje se analizira, uvjeti demaskiranja antigena i inkubacije specifičnim antitijelom te je predložen optimalan protokol bojenja za naveden antigen primjenjiv u rutinskom hematopatološkom laboratoriju.Immunohistochemical staining is a method for detection of presence and localization of specific antigens in cells and/or tissues. It is based on the basic principle of immunoreactions: binding of specific antibody to a certain antigen. Development of immunohistochemical staining methods started during the 1930s and, in a number of variations, they are still used today both in research and routine diagnostic procedures. This paper offers presentation of immunohistochemical methods development and gives an example of protocol optimization for detection of BACH2 antigen in human secondary lymphoid tissue. Most important steps of the protocol are discussed (section preparation, antigen demasking and specific antibody incubation conditions). Optimal protocol for immunohistochemical staining for BACH2 antigen is given and can be used in any routine hematopathological laboratory

"Double hit" lymphoma or secondary MYC translocation lymphoma?

Chromosomal translocations that juxtapose different genes required for proliferation and differentiation are frequently associatedwith hematologic neoplasms. A term "double hit" lymphoma refers to a group of mature B-cell malignancies that harbor MYC rearrangement accompanied with another translocation commonly found in lymphomas (i.e. IGH/BCL2). A complex karyotype with multiple abnormalities and very aggressive course of disease are additional characteristics of this group. The response to currently available treatment regimens is unsatisfying, thus reported overall survival of these patients is usually very short. Today, the precise mechanisms of "double hit" lymphoma development are still unclear, although several possible pathways have been proposed in the literature. Similar oncogenic chain of events was also observed in another common hematological malignant neoplasm – multiple myeloma. MYC translocation as a secondary pathogenic phenomenon has been demonstrated in multiple myeloma, as well as complex cytogenetics and very aggressive course of the disease. Therefore, secondary translocation involving MYC gene might be a potential marker of aggressive neoplasms emerging from different cells of origin, and united under the umbrella of "MYC-plus" malignancies. That concept might, in the future, result in a novel therapy, targeting this distinct, but not unique cytogenetic aberration

MiR-7 in Cancer Development

MicroRNAs (miRNAs) are short non-coding RNA involved in the regulation of specific mRNA translation. They participate in cellular signaling circuits and can act as oncogenes in tumor development, so-called oncomirs, as well as tumor suppressors. miR-7 is an ancient miRNA involved in the fine-tuning of several signaling pathways, acting mainly as tumor suppressor. Through downregulation of PI3K and MAPK pathways, its dominant role is the suppression of proliferation and survival, stimulation of apoptosis and inhibition of migration. Besides these functions, it has numerous additional roles in the differentiation process of different cell types, protection from stress and chromatin remodulation. One of the most investigated tissues is the brain, where its downregulation is linked with glioblastoma cell proliferation. Its deregulation is found also in other tumor types, such as in liver, lung and pancreas. In some types of lung and oral carcinoma, it can act as oncomir. miR-7 roles in cell fate determination and maintenance of cell homeostasis are still to be discovered, as well as the possibilities of its use as a specific biotherapeutic

Genomische Bearbeitung und Auswahl auf der Grundlage von Genen, die mit sportlichen Leistungen in Verbindung stehen – einige bioethische Fragen

Projekt ljudskoga genoma završen je 2003. godine, a njegovi su rezultati otkrili dotada nepoznate detalje o našem genomu: skupinu informacija o tome kako ljudska bića izgledaju, kako djeluju, osjećaju, misle i razvijaju se. Uskoro su ostvarene i druge međunarodne suradnje poput projekta HapMap i Projekta 1000 genoma. Unatoč tomu što su primarno bili usmjereni na istraživanje varijabilnosti u ljudskoj populaciji, kao i moguće povezanosti različitih varijacija s različitim stanjima i bolestima, ti su projekti također znatno utjecali na razumijevanje utjecaja gena na sportski uspjeh. Usporedno su razvijene poboljšane metode genske analize i genskoga preinačavanja na temelju kojih je postalo moguće utvrditi kandidatske gene odgovorne za različite fenotipe uspješnosti u sportu te razviti protokole slične genskim terapijama za poboljšanje sportskih natjecateljskih performansi sportaša. Ovaj rad daje pregled razvoja u genetici, pregled kandidatskih gena povezanih sa sportskim natjecateljskim performansama te etičke dvojbe vezane uz preinačavanje genoma radi poboljšavanja sportskih natjecateljskih performansi.In 2003 the final results of the Human Genome Project revealed the details of our genome: a set of information about how human beings look, how we act, feel, think and develop. Soon after, other global collaborations such as the HapMap project and 1000 Genomes Project were conducted. Although the main focus was to investigate the variability in human populations and the possible connections of certain variations to different conditions and diseases, these projects also had a great impact on the understanding of the genetic influence on sports performance. In parallel, improved methods for gene analysis and gene editing were developed. Based on those methods, it became possible to detect candidate genes responsible for different performance phenotypes and develop protocols similar to gene therapies for performance enhancement in athletes. This review covers developments in genetics, the overview of candidate genes associated with athletic performance, and ethical dilemmas related to the modification of genome for sport performance enhancement.En 2003, les résultats finaux du Projet génome humain ont révélé les détails sur la structure de notre génome : un ensemble d’informations sur l’apparence physique de l’être humain, sur sa manière d’agir, de sentir, de penser, et de se développer. Peu de temps après, d’autres collaborations mondiales telles que le projet HapMap et le projet 1000 Genomes ont été menées à terme. Axés essentiellement sur la variabilité de la population humaine et sur les possibles liens de certaines variations avec différents états et maladies, ces projets ont toutefois eu un important impact sur notre manière de comprendre l’influence que les gènes exercent sur la performance sportive. En parallèle, des méthodes améliorées pour l’analyse du gène et la modification du gène ont été développées. Il est devenu possible sur la base de ces méthodes de détecter les gènes candidats responsables des différents phénotypes associés à la performance et de développer des protocoles similaires aux thérapies géniques afin d’améliorer les performances chez les athlètes. Cet article passe en revue les développements en génétique, donne un aperçu général des gènes candidats associés à la performance athlétique et des dilemmes éthiques liés à la modification du génome dans le but d’améliorer la performance sportive.Im Jahr 2003 enthüllten die endgültigen Ergebnisse des Humangenomprojekts die Details unseres Genoms: Eine Reihe von Informationen darüber, wie Menschen aussehen, wie wir handeln, fühlen, denken und uns entwickeln. Bald darauf wurden weitere weltweite Kooperationen wie das HapMap-Projekt und das 1000-Genome-Projekt in die Tat umgesetzt. Obgleich das Hauptaugenmerk auf der Variabilität der menschlichen Population und den möglichen Verquickungen bestimmter Variationen mit verschiedenen Zuständen und Krankheiten lag, übten diese Projekte auch eine große Wirkung auf das Verständnis des genetischen Einflusses auf die sportliche Leistung aus. Parallel dazu wurden verbesserte Methoden zur Genanalyse und Genbearbeitung entwickelt. Basierend auf diesen Methoden wurde es machbar, Kandidatengene zu detektieren, die für verschiedene Leistungsphänotypen verantwortlich sind, und Protokolle zu entwickeln, die Gentherapien zur Leistungssteigerung bei Sportlern ähneln. Dieser Aufsatz behandelt Entwicklungen in der Genetik, den Überblick über Kandidatengene im Zusammenhang mit sportlicher Leistung und ethische Dilemmata bezüglich der Modifizierung des Genoms zur Steigerung der sportlichen Leistung

Croatian vocabulary of molecular and cell biology

Nazivlje mnogih struka često se razvija neovisno o pravilima hrvatskoga jezika, posebice ako je riječ o mladim znanostima koje nastaju i razvijaju se na engleskome jeziku. Jedan je od takvih primjera leksik molekularne biologije. Iako je razvoj ovoga područja u Hrvatskoj započeo 1960-ih godina na Sveučilištu u Zagrebu i Institutu Ruđer Bošković, da bi 1984. godine bio pokrenut i studij molekularne biologije na Biološkome odsjeku Prirodoslovno-matematičkoga fakulteta, a 1989. osnovan Zavod za molekularnu biologiju pri istome odsjeku, sve donedavno nije postojao usustavljen način prilagodbe strukovnoga nazivlja. Budući da je molekularna biologija sve češće zastupljena u informativnim medijima zbog sve većega značenja rezultata istraživanja ovoga područja u kontekstu svakodnevnoga života, na Zavodu za molekularnu biologiju Prirodoslovno-matematičkoga fakulteta započeta su dva biološko-jezična projekta. Prvi je projekt prijevod rječnika molekularne i stanične biologije koji sadržava više od 12 000 unosa, čiji je cilj usustavljivanje hrvatskoga leksika područja kao rezultat suradnje jezikoslovaca i biologa. Drugi projekt – „Genetikon“ – financirala je Hrvatska zaklada za znanost putem projekta „Struna“, a on obuhvaća genetički leksik omogućujući da na jednome mjestu budu okupljeni najvažniji pojmovi i sustavno razrađeno hrvatsko nazivlje. U ovome radu dajemo kratak pregled razvoja jezika struke s posebnim naglaskom na šest osnovnih naputaka kojima nastojimo pridonijeti razvoju leksika genetike i stanične i molekularne biologije.The technical vocabulary of many areas is often developed independently of the rules of the Croatian language, especially in the case of young sciences that are developed in English. One such example is the vocabulary of molecular biology. The development of this research area in Croatia began in the 1960s at the University of Zagreb and the Institute Ruđer Bošković. In 1981 Molecular Biology study program started at the Department of Biology at the Faculty of Science, and in 1989 Divison of Molecular Biology at the same Department was founded. However, until recently there was no systematically elaborated development of professional vocabulary of this scientific area. Due to growing significance of the research results from molecular biology and genetics in the context of everyday life at the Department of Molecular Biology two projects were conducted. The first is a translation of the Dictionary of Molecular and Cell Biology that contains more than 12 000 entries, whose purpose is to make the professional vocabulary as result of cooperation between linguists and biologists. The second project –“Genetikon” – has been funded by the Croatian Science Foundation through the “Struna” project, and it covers genetic lexicon, enabling the most important concepts and systematically developed Croatian terminology to be gathered at one place. In this paper we give a short overview of the molecular biology vocabulary development with special emphasis on the six basic guidelines that we hope will contribute to the development of the technical vocabulary of genetics and cellular and molecular biology

Understanding of life cycles and fertilization during learning biology in primary school

Cilj ovog istraživanja je odrediti usvojenost i konceptualno razumijevanje makrokoncepta Razmnožavanje i razvoj organizma među učenicima 7. i 8. razreda osnovnih škola, na temelju učeničkih odgovora na pitanja uz makrokoncept Razmnožavanje i razvoj organizma na Županijskom natjecanju iz biologije 2015. i 2017. godine. Analiza učeničkih odgovora uključivala je specifično kodiranje odgovora i tumačenje biološkog značenja odgovora učenika. Izdvojeni su i objašnjeni problemi koje učenici pokazuju u svojim odgovorima kao i njihove miskoncepcije vezane uz koncept razmnožavanje. Utvrđeno je i opisano postojanje niza problema kod usvojenosti koncepta razmnožavanje te su kao najizraženije uočene miskoncepcije uz ključni koncept životni ciklus stanice i organizma te kod razumijevanja koncepta oplodnje. Utvrđene miskoncepcije ukazuju na nužnu promjenu organizacije nastavnih sadržaja biologije, kao i na potrebu uvođenja novih nastavnih strategija s naglaskom na aktivnost i samostalni rad učenika.The aim of this study was to determine the conceptual understanding of the macro concept of Reproduction and organism development among students of the 7th and 8th grade of primary schools, based on students\u27 answers to questions in relation to the macro concept of Reproduction and organism development at the regional competition in Biology in 2015 and 2017. Analysis of the students\u27 answers included specific coding of the answers and interpreting the biological importance and meaning of each answer. Problems and misconceptions in relation to the concept of reproduction were singled out and explained. The analysis showed there is a problem with understanding the concept of reproduction and the misconceptions based around the key concept of cell life cycle, as well as misunderstandings of the concept of fertilization were observed as most prominent. Misconceptions found in this study show the necessity of changing the organization of contents taught in primary school biology and the need for using new teaching strategies, based on the students’ activity and individual work

Prognostic Significance of B-cell Differentiation Genes Encoding Proteins in Diffuse Large B-cell Lymphoma and Follicular Lymphoma Grade 3

Aim To define prognostic significance of B-cell differentiation genes encoding proteins and BCL2 and BCL6 gene abnormalities in diffuse large B-cell lymphoma and follicular lymphoma grade 3 with >75% follicular growth pattern. Methods In 53 patients with diffuse large B-cell lymphoma and 20 patients with follicular lymphoma grade 3 with >75% follicular growth pattern the following was performed: 1) determination of protein expression of BCL6, CD10, MUM1/IRF4, CD138, and BCL2 by immunohistochemistry; 2) subclassification into germinal center B-cell-like (GCB) and activated B-cell-like (ABC) groups according to the results of protein expression; 3) detection of t(14;18)(q32;q21)/IgH-BCL2 and BCL6 abnormalities by fluorescent in situ hybridization in diffuse large B-cell lymphoma and follicular lymphoma grade 3 with >75% follicular growth pattern as well as in GCB and ABC groups; and 4) assessment of the influence of the analyzed characteristics and clinical prognostic factors on overall survival. Results Isolated BCL6 expression was more frequently found in follicular lymphoma grade 3 with >75% follicular growth pattern than in diffuse large Bcell lymphoma (P = 0.030). There were no differences in BCL2 and BCL6 gene abnormalities between diffuse large B-cell lymphoma and follicular lymphoma grade 3 with >75% follicular growth pattern. Diffuse large B-cell lymphoma and follicular lymphoma grade 3 with >75% follicular growth pattern patients were equally distributed in GCB and ABC groups. t(14;18)(q32;q21) was more frequently recorded in GCB group, and t(14;18)(q32;q21) with BCL2 additional signals or only BCL2 and IgH additional signals in ABC group (P = 0.004). The GCB and ABC groups showed no difference in BCL6 gene abnormalities. There was no overall survival difference between the patients with diffuse large B-cell lymphoma or follicular lymphoma grade 3 with >75% follicular growth pattern; however, GCB group had longer overall survival than ABC group (P = 0.047). Multivariate analysis showed that BCL6, CD10, and BCL2 expression, BCL2 and BCL6 abnormalities, and International Prognostic Index were not significantly related to overall survival. Conclusion Diffuse large B-cell lymphoma and follicular lymphoma grade 3 with >75% follicular growth pattern patients have very similar characteristics and their prognosis is more influenced by protein expression of B-cell differentiation stage genes than by tumor cells growth pattern, BCL2 and BCL6 abnormalities, and International Prognostic Index