The importance of determining carrier status in Duchenne's and Beker's muscular dysrophy in the population of Serbia

Distrofinopatije su bolesti koje nastaju kao posledica mutacija u genu zadistrofin. Dišenova mišićna distrofija (DMD) predstavlja najteži oblik iz ove grupebolesti. Karakteriše je rani početak bolesti, progresivna mišićna slabost koja dovodi dogubitka pokretljivosti bolesnika i kardio-pulmonalne slabosti zbog zahvatanja srčanog irespiratornih mišića. Bekerova mišićna distrofija (BMD) se javlja kasnije, ima blaži tokbolesti, ali sa velikom varijabilnošću u kliničkoj slici. Nasleđujuju se X-vezanorecesivno, oboljevaju muškarci dok su žene uglavnom zdravi prenosioci bolesti.Procenjeno je da su 2/3 majki nosioci, 5-10% ima gonadni mozaicizam, dok 25-30%nema mutaciju. Gen za distrofin (DMD gen) je najveći opisani gen u humanom genomui često je podložan promenama. Najčešće su prisutne intragenske delecije (65%-70%) iduplikacije (5-15%) jednog ili više egzona, dok su tačkaste mutacije prisutne u 20%slučajeva. 1/3 bolesnika ima de novo mutaciju. Za sada nema uspešne terapijedistrofinopatija, pa je utvrđivanje statusa nosioca kod ženskih članova u familiji odznačaja za davanje genetskog saveta i prenatalnu dijagnozu.Cilj rada je bio da se utvrde i analiziraju delecije i duplikacije u genu za distrofin kodprobanada; da se u slučajevima potvrđenih delecija i duplikacija kod probanada utvdistatus prenosioca kod njihovih ženskih srodnka; u slučajevima bez dokazanih delecija iduplikacija kod probanada, da se ispita mogućnost indirektne genetičke analize zaodređivanje statusa ženskih prenosioca; da se u indikovanim slučajevima izvršiprenatalna molekularno genetička analiza DMD gena, primenom adekvatne metode zadatu porodicu.Material i metode: Uzorak su činila 72 DMD/BMD probanda, 69 ženskih članova iz44 porodice probanada i 11 trudnica (15 trudnoća). Genomska DNK za analizu jeizolovana iz limfocita periferne krvi ispitanika metodom isoljavanja prema standardnojproceduri, a za prenatalnu dijagnozu, DNK je izolovana iz uzorka horionskih resica,plodove vode ili krvi pupčanika ploda primenom komercijalnog kita. Za detetekcijudelecija i duplikacija u DMD genu kod probanda primenjene su metoda lančane reakcijepolimeraze (PCR) i metoda istovremenog umnožavanja vezanih proba (MLPA); zadetekciju ženskih nosioca primenjene su MLPA metoda i analiza vezanosti; zaprenatalnu dijagnozu primenjene su PCR metoda, analiza vezanosti i MLPA metoda...Dystrophinopathies are diseases that result from mutations in thedystrophin gene. Duchenne muscular dystrophy (DMD) is the most severe form of thisgroup of diseases. It is characterized by an early onset of the disease, a progressivemuscle weakness that leads to loss of mobility of the patient, spreading to the heart andrespiratory muscles, causing cardio-pulmonary weakness. Becker muscular dystrophy(BMD) occurs in late childhood or adolescence, has a milder course of the disease, butwith widely variable clinical presentations. It has an X-linked recessive inheritedpattern, whereby males are affected, while females are mostly healthy carriers of thedisease. It is estimated that 2/3 of the mothers are carriers, 5-10% have gonadalmosaicism and 25-30% have no mutation. The gene for dystrophin (DMD gene) is thelargest known gene in the human genome and is often subject to change. The mostcommon changes are intragenal deletions (65% -70%) and the duplication (5-15%) ofone or more exons, as well as point mutations in 20% of cases. 1/3 of patients have denovo mutation. There is no successful therapy for dystrophinopathy, therefore thedetection of female carriers in a family is important for genetic counseling and prenataldaignosis.The aim of work was to determine and analyze the deletions and duplications of thedystrophin gene in probands; in the cases of confirmed deletions and duplications in thetrial, to determine the status of the carrier in their female relatives; in the cases with noproven deletions and duplications in the trial, the possibility of indirect genetic analysisfor determining the status of female carriers; in the indicated cases to perform prenatalmolecular genetic analysis of the DMD gene, using an appropriate method for aparticular family.Material and methods: The sample consisted of 72 DMD/BMD probands, 69 femalemembers from 44 proband families and 11 pregnant women (15 pregnancies). Thegenomic DNA for analysis was isolated from the peripheral blood lymphocytes of thesubjects, according to the standard procedure, and for prenatal diagnosis, the DNA wasisolated from the sample of chorionic villi, amniotic fluid or blood of the umbilical cordusing a commercial kit. For the detection of deletions and duplications in the DMDgene, polymerase chain reaction (PCR) method and multiplex ligation-dependent probeamplification (MLPA) method were applied; for the detection of female carriers, theMLPA method and linkage analysis were used; for prenatal diagnosis, the PCR method,linkage analysis and MLPA methods were applied..

Covid-19 and vaccination

Nakon prvog slučaja infekcije SARS-CoV-2 virusom u Kini krajem 2019. godine, COVID-19 brzo prerasta u epidemiju, a zatim i pandemiju. Prema najnovijim po- dacima COVID-19 pandemija beleži oko 250 miliona zaraženih ljudi u svetu i preko 5 miliona smrtnih slučajeva kao posledica COVID-19 infekcije. U radu su, na osnovu pregleda relevantne literature, prikazani podaci o vak- cinama dostupnim u našoj zemlji, njihovim karakteristikama, vakcinaciji trudnica i dece, kao i preporuke Svetske zdravstvene organizacije i Centra za kontrolu i prevenciju bolesti, za primenu vakcina. I pored dostupnosti vakcina, stepen vakcinacije u našoj zemlji još uvek nije doveo do stvaranja kolektivnog imuniteta. Razlog leži u nepoverenju prema vakcinama zbog njihovog brzog puštanja u upotrebu. To nameće potrebu za jasnim i pouzdanim preporukama zdravstvenih radnika kako bi se ublažila zabrinu- tost u vezi sa bezbednošću vakcina.After the first case of SARS-CoV-2 virus infection in China at the end of 2019, COVID-19 quickly grew into an epidemic, and then a pandemic. According to the latest data from the COVID-19 pandemic, there are about 250 million infected and over 5 million deaths in the world as a result of the COVID-19 infection. Based on a review of relevant literature, the paper presents data on vaccines available in our country, their characteristics, vaccination of pregnant women and children, as well as the recommendations of the World Health Organization and the Center for Disease Control and Prevention for vaccines. Despite the availability of vaccines, the level of vaccination in our country has not yet led to the collective immunity. The main reason lies in the distrust of vaccines due to their rapid release. This imposes the need for clear and reliable recommendations from healthcare professionals to alleviate concerns about vaccine safety

Application of modern methods of molecular genetics In practice

Introduction: The development of new methods of molecular genetics, especially in the last two decades, has led to enormous progress in the field of medicine and enabled more comprehensive prenatal and postnatal diagnosis of hereditary diseases and other disorders, as well as preimplantation testing. Aim: The aim of the work is to present modern methods of molecular genetics and their application in the diagnosis of hereditary diseases and developmental disorders. Methods: Having reviewed the relevant literature, insight was gained into the current development of molecular genetic methods and their practical application. Results: Chromosome microarray is an analysis of all chromosomes in the genome, which, thanks to microchip technology, enables the detection of chromosomal microdeletions and microduplications, as well as changes in the number of copies, in just one reaction. This method has found application in all areas of clinical genetics, and especially in the detection of genomic changes in patients with intellectual disabilities, developmental delays, autism spectrum disorders and congenital anomalies. The gene microarray is also based on microchip technology, and depending on the type of DNA chip used, it is used to detect mutations and DNA variations at the level of the entire genome or to study gene expression. The rapid development of biotechnology and bioinformatics has enabled the simultaneous analysis of a large number of genes through parallel (deep) sequencing, known as a new generation of DNA sequencing methods. This technology detects already known gene variants, new ones, as well as the presence of a predisposition. Today, in clinical practice, differently designed gene panels are used for postnatal diagnosis of monogenic and multifactorial diseases. Particularly noteworthy are the “clinical exome” panels with over 6,000 genes, the whole exome (about 22,000 genes) or the whole genome.Conclusion: Modern times have marked the development and improvement of molecular genetics methods for quick and more accurate diagnosis of hereditary diseases and developmental disorders, which gives a new perspective on the possibilities of prenatal, postnatal and preimplantation genetic testing

Application of modern methods of molecular genetics In practice

Introduction: The development of new methods of molecular genetics, especially in the last two decades, has led to enormous progress in the field of medicine and enabled more comprehensive prenatal and postnatal diagnosis of hereditary diseases and other disorders, as well as preimplantation testing. Aim: The aim of the work is to present modern methods of molecular genetics and their application in the diagnosis of hereditary diseases and developmental disorders. Methods: Having reviewed the relevant literature, insight was gained into the current development of molecular genetic methods and their practical application. Results: Chromosome microarray is an analysis of all chromosomes in the genome, which, thanks to microchip technology, enables the detection of chromosomal microdeletions and microduplications, as well as changes in the number of copies, in just one reaction. This method has found application in all areas of clinical genetics, and especially in the detection of genomic changes in patients with intellectual disabilities, developmental delays, autism spectrum disorders and congenital anomalies. The gene microarray is also based on microchip technology, and depending on the type of DNA chip used, it is used to detect mutations and DNA variations at the level of the entire genome or to study gene expression. The rapid development of biotechnology and bioinformatics has enabled the simultaneous analysis of a large number of genes through parallel (deep) sequencing, known as a new generation of DNA sequencing methods. This technology detects already known gene variants, new ones, as well as the presence of a predisposition. Today, in clinical practice, differently designed gene panels are used for postnatal diagnosis of monogenic and multifactorial diseases. Particularly noteworthy are the “clinical exome” panels with over 6,000 genes, the whole exome (about 22,000 genes) or the whole genome.Conclusion: Modern times have marked the development and improvement of molecular genetics methods for quick and more accurate diagnosis of hereditary diseases and developmental disorders, which gives a new perspective on the possibilities of prenatal, postnatal and preimplantation genetic testing

Genetic aspect of hearing impairment

Razvoj slušnog aparata je veoma složen proces koga uslovljava me�����usobno delovanje genomskog programa tj. genetičkih instrukcija za anatomo-funkcionalno strukturiranje aparata čula sluha i faktora sredine, koji ga omogućavaju i usmeravaju u toku embrionalnog, fetalnog i nakon toga tokom postnatalnog razvoja. Greške ili deficijencije, i štetno faktorsko delovanje jednog i/ili drugog, iskazuje se u kompleksu oštećenja sluha. Najčešći uzročni faktori potiču iz genoma, genetičke strukture individue. Procenjeno je da više od 50% svih vrsta gubitaka sluha ima substancijalno genetičku komponentu. Oštećenje sluha, kao najčešći senzorni poremećaj, genetički je vrlo heterogen. Mapiranja i identifikacija gena sa osobenim ulogama u razvoju struktura i funkcija ovog čula, podvrgnuta DNK probama, metodama direktne ili indirektne detekcije mutacija – signiraju vezu greške i oštećenja. Više stotina gena je otkriveno u mehanizmu razvoja slušnog aparata i njegove funkcije. Jedni su odgovorni svojom aktivnošću za jasno prepoznate komponente strukture i funkcije čula, drugi – preko enkodiranih proteina sadejstvuju, od modifikacija, sinteze regulatora transkripcije drugih gena, faktora rasta i dr. Identifikovani su i neki geni koji uzrokuju dominantne i recesivne oblike sindromske i/ili nesindromske gluvoće, što je u kliničkoj genetici već u domenu moguće detekcije nosioca genetičkog opterećenja, prenatalnog nalaza i predmet genetičkog savetovališta. Skrining novoro�����enčadi na slušna oštećenja dao je nedvosmisleno veliki učinak, od rane detekcije i tretmana, uz brzi napredak genetike slušnih oštećenja – svrsishodnijim pristupom, prekoncepcijskoj i prenatalnoj dijagnostici, promenom kliničkog pristupa u obradi i lečenju tih oštećenja.Development of hearing apparatus is very complex process dependent on genome program, i.e. genetic instructions for anatomical and functional structuring of hearing sense and environmental factors, both of which enable and direct it through embriological, fetal, and postnatal period. Defects or deficiencies, and harmful consequences of one and/or both, are evident in hearing impairment. The most comon causes are genome and genetic structure of an individual. It is estimated that over 50% of all hearing losses are substantially genetical. Hearing impairment, being the most common sensorial disorder, is genetically very heterogenous. Mapping and identification of genes with specific roles in the development of structures and functions of this sense, when tested with DNA analysis, with direct or indirect mutation detection – point out the link between defect and impairment. Many hundreds of genes were discovered in the mechanism of development of hearing apparatus and its function. Some are responsible for clearly recognized components of structure and function of the sense, others – through encoded proteins act together in modifications, synthesis of regulators of other gene transcriptions, growth factors, etc. Furthermore, some genes were identified which cause dominant and recessive forms of syndrome and/or nonsyndrome deafness, which is in the domain of possible detection of carrier of genetic predisposition, prenatal positive finding and the subject of genetic counseling. Screening of newborns for hearing impairment has brought a great deal, from early detection and treatment, along with fast improvement of genetics in hearing impairment – with more appropriate approach, preconceptual and prenatal diagnostics, and changes in clinical approach in treatment of those impairments

Implementation of the next generation sequencing (ngs) methods in early diagnosis of heritable diseases

U cilju rane dijagnostike i prevencije naslednih poremećaja, proteklih decenija su na raspolaganju bile različite metode. Analize genetičkog materijala su se kretale od klasične citogenetičke obrade kariotipa radi uočavanja numeričkih i strukturnih aberacija hromozoma, do najfinijih ispitivanja za detektuju genskih mutacija na molekularnom nivou. Poslednjih godina razvijaju se potpuno nove metode za brzu, efikasnu i dostupnu analizu naslednog materijala, koje su poznate kao “next generation sequencing” (NGS) ili nova generacija metoda za sekvenciranje DNK. Ove metode omogućavaju ispitivanje ne samo pojedinačnih gena ili delova gena nego i većeg broja segmenata, sve do kompletne nasledne osnove tj. čitavog genoma čoveka. Primena ovakvog pristupa dovodi do prave tihe revolucije u medicinskoj genetici i disciplinama sa kojima ona sarađuje, nagoveštavajući promenu u konceptu dijagnostike naslednih poremećaja. U prenatalnoj dijagnostici NGS je već našla primenu u potpuno neinvazivnoj detekciji najčešćih hromozomskih aberacija (Daunov, Edvardsov, Patau sindrom, aberacije polnih hromozoma) analizom fetalnih ćelija prisutnih u krvi majke. Test NIFTY već je dostupan i trudnicama u našoj sredini. U postnatalnom periodu NGS se koristi za ispitivanje odabranih panela gena, ili, po potrebi, čitavog genoma/ egzoma, sve sa ciljem što efikasnije dijagnostike pre svega monogenskih, ali i oligogenskih i poligenskih bolesti. Predlaže se čak da analiza kompletnog genoma postane deo neonatalnog skriniga, ali za sada to nije prihvaćeno. Nesumnjivo je da rezultati NGS donose veliki napredak medicinsko − genetičkoj praksi, ali i rađaju nove etičke dileme u oblasti rane detekcije naslednih poremećaja i intervencije kod ovih stanja.In order to early diagnostics and prevention of hereditary disorders, past decades have been brought different methods. Analysis of genetic material ranged from classical cytogentic karyotype analysis for processing numerical and structural chromosomalaberratios, by most sophisticated examination of manor gene mutation on molecular level. In recent years develop completely new methods for rapid, efficient and publicly available analysis of inheritance material, that are known as the “next” button generation sequencing” (NGS). These methods allow for examination not only individual genes or parts of genes but also a larger number of segments, all up to complete inherited basis i.e. the entire human genome research. Implementation of this approach leads to genuine silent revolution in medical genetics and disciplines with which it cooperate, suggesting a change in the concept of heritage disorders diagnosing. In prenatal diagnostics NGS has already found application in completely noninvsive detection of chromosomal aberrations (Down, Edwards, Patau syndrome, sex chromosomes aberration) by analysis of fetal cells present in mother’s blood. Such tests (i.e. NIFTY) are already available and for pregnant women in our country. In postnatal period NGS is used for the examination of selected genes by gene panels, or, if necessary, the entire genome/exome research, all with the aim as efficient diagnostics primarily monogenic, but oligogenic and polygenic diseases also. It is proposed that the analysis of even complete genome become part of neonatal screening, but for now it is not accepted yet. It is undeniable that the results of NGS make a great progress in medical − genetic practice, but gained new ethical dilemmas in the field of early detection of hereditary disorders and intervention in these situation

Studying elipse with GeoGebra

U radu je prikazano proučavanje elipse pomoću računala i rav{c}unalnog programa GeoGebra. Detaljno je opisana mogućnost izvođenja jednadžbe elipse pomoću GeoGebre. Prikazan je način rješavanja zadaća uz uporabu računala i GeoGebre. Ponuđene su i računalne aplikacije za dalje proučavanje elipse.The paper presents a way to study the ellipse using a computer and GeoGebra software. The possibility of performing the equation of an ellipse using GeoGebra is described in detail. A method of solving examples with the use of computers and GeoGebra is shown. Applets for further study of the ellipse are offered

Genetic aspect of hearing impairment

Razvoj slušnog aparata je veoma složen proces koga uslovljava me�����usobno delovanje genomskog programa tj. genetičkih instrukcija za anatomo-funkcionalno strukturiranje aparata čula sluha i faktora sredine, koji ga omogućavaju i usmeravaju u toku embrionalnog, fetalnog i nakon toga tokom postnatalnog razvoja. Greške ili deficijencije, i štetno faktorsko delovanje jednog i/ili drugog, iskazuje se u kompleksu oštećenja sluha. Najčešći uzročni faktori potiču iz genoma, genetičke strukture individue. Procenjeno je da više od 50% svih vrsta gubitaka sluha ima substancijalno genetičku komponentu. Oštećenje sluha, kao najčešći senzorni poremećaj, genetički je vrlo heterogen. Mapiranja i identifikacija gena sa osobenim ulogama u razvoju struktura i funkcija ovog čula, podvrgnuta DNK probama, metodama direktne ili indirektne detekcije mutacija – signiraju vezu greške i oštećenja. Više stotina gena je otkriveno u mehanizmu razvoja slušnog aparata i njegove funkcije. Jedni su odgovorni svojom aktivnošću za jasno prepoznate komponente strukture i funkcije čula, drugi – preko enkodiranih proteina sadejstvuju, od modifikacija, sinteze regulatora transkripcije drugih gena, faktora rasta i dr. Identifikovani su i neki geni koji uzrokuju dominantne i recesivne oblike sindromske i/ili nesindromske gluvoće, što je u kliničkoj genetici već u domenu moguće detekcije nosioca genetičkog opterećenja, prenatalnog nalaza i predmet genetičkog savetovališta. Skrining novoro�����enčadi na slušna oštećenja dao je nedvosmisleno veliki učinak, od rane detekcije i tretmana, uz brzi napredak genetike slušnih oštećenja – svrsishodnijim pristupom, prekoncepcijskoj i prenatalnoj dijagnostici, promenom kliničkog pristupa u obradi i lečenju tih oštećenja.Development of hearing apparatus is very complex process dependent on genome program, i.e. genetic instructions for anatomical and functional structuring of hearing sense and environmental factors, both of which enable and direct it through embriological, fetal, and postnatal period. Defects or deficiencies, and harmful consequences of one and/or both, are evident in hearing impairment. The most comon causes are genome and genetic structure of an individual. It is estimated that over 50% of all hearing losses are substantially genetical. Hearing impairment, being the most common sensorial disorder, is genetically very heterogenous. Mapping and identification of genes with specific roles in the development of structures and functions of this sense, when tested with DNA analysis, with direct or indirect mutation detection – point out the link between defect and impairment. Many hundreds of genes were discovered in the mechanism of development of hearing apparatus and its function. Some are responsible for clearly recognized components of structure and function of the sense, others – through encoded proteins act together in modifications, synthesis of regulators of other gene transcriptions, growth factors, etc. Furthermore, some genes were identified which cause dominant and recessive forms of syndrome and/or nonsyndrome deafness, which is in the domain of possible detection of carrier of genetic predisposition, prenatal positive finding and the subject of genetic counseling. Screening of newborns for hearing impairment has brought a great deal, from early detection and treatment, along with fast improvement of genetics in hearing impairment – with more appropriate approach, preconceptual and prenatal diagnostics, and changes in clinical approach in treatment of those impairments

Implementation of the next generation sequencing (ngs) methods in early diagnosis of heritable diseases

U cilju rane dijagnostike i prevencije naslednih poremećaja, proteklih decenija su na raspolaganju bile različite metode. Analize genetičkog materijala su se kretale od klasične citogenetičke obrade kariotipa radi uočavanja numeričkih i strukturnih aberacija hromozoma, do najfinijih ispitivanja za detektuju genskih mutacija na molekularnom nivou. Poslednjih godina razvijaju se potpuno nove metode za brzu, efikasnu i dostupnu analizu naslednog materijala, koje su poznate kao “next generation sequencing” (NGS) ili nova generacija metoda za sekvenciranje DNK. Ove metode omogućavaju ispitivanje ne samo pojedinačnih gena ili delova gena nego i većeg broja segmenata, sve do kompletne nasledne osnove tj. čitavog genoma čoveka. Primena ovakvog pristupa dovodi do prave tihe revolucije u medicinskoj genetici i disciplinama sa kojima ona sarađuje, nagoveštavajući promenu u konceptu dijagnostike naslednih poremećaja. U prenatalnoj dijagnostici NGS je već našla primenu u potpuno neinvazivnoj detekciji najčešćih hromozomskih aberacija (Daunov, Edvardsov, Patau sindrom, aberacije polnih hromozoma) analizom fetalnih ćelija prisutnih u krvi majke. Test NIFTY već je dostupan i trudnicama u našoj sredini. U postnatalnom periodu NGS se koristi za ispitivanje odabranih panela gena, ili, po potrebi, čitavog genoma/ egzoma, sve sa ciljem što efikasnije dijagnostike pre svega monogenskih, ali i oligogenskih i poligenskih bolesti. Predlaže se čak da analiza kompletnog genoma postane deo neonatalnog skriniga, ali za sada to nije prihvaćeno. Nesumnjivo je da rezultati NGS donose veliki napredak medicinsko − genetičkoj praksi, ali i rađaju nove etičke dileme u oblasti rane detekcije naslednih poremećaja i intervencije kod ovih stanja.In order to early diagnostics and prevention of hereditary disorders, past decades have been brought different methods. Analysis of genetic material ranged from classical cytogentic karyotype analysis for processing numerical and structural chromosomalaberratios, by most sophisticated examination of manor gene mutation on molecular level. In recent years develop completely new methods for rapid, efficient and publicly available analysis of inheritance material, that are known as the “next” button generation sequencing” (NGS). These methods allow for examination not only individual genes or parts of genes but also a larger number of segments, all up to complete inherited basis i.e. the entire human genome research. Implementation of this approach leads to genuine silent revolution in medical genetics and disciplines with which it cooperate, suggesting a change in the concept of heritage disorders diagnosing. In prenatal diagnostics NGS has already found application in completely noninvsive detection of chromosomal aberrations (Down, Edwards, Patau syndrome, sex chromosomes aberration) by analysis of fetal cells present in mother’s blood. Such tests (i.e. NIFTY) are already available and for pregnant women in our country. In postnatal period NGS is used for the examination of selected genes by gene panels, or, if necessary, the entire genome/exome research, all with the aim as efficient diagnostics primarily monogenic, but oligogenic and polygenic diseases also. It is proposed that the analysis of even complete genome become part of neonatal screening, but for now it is not accepted yet. It is undeniable that the results of NGS make a great progress in medical − genetic practice, but gained new ethical dilemmas in the field of early detection of hereditary disorders and intervention in these situation

Studying elipse with GeoGebra

U radu je prikazano proučavanje elipse pomoću računala i rav{c}unalnog programa GeoGebra. Detaljno je opisana mogućnost izvođenja jednadžbe elipse pomoću GeoGebre. Prikazan je način rješavanja zadaća uz uporabu računala i GeoGebre. Ponuđene su i računalne aplikacije za dalje proučavanje elipse.The paper presents a way to study the ellipse using a computer and GeoGebra software. The possibility of performing the equation of an ellipse using GeoGebra is described in detail. A method of solving examples with the use of computers and GeoGebra is shown. Applets for further study of the ellipse are offered