West Syndrome with Periventricular Leukomalacia: Ten-year Clinical Study

The aim of the study was to evaluate magnetic resonance imaging (MRI) findings in infants with periventricular leukomalacia (PVL) andWest syndrome (WS) and determine the neurodevelopmental outcome in children withWest syndrome and PVL. Ultrasound and brain MRI were performed in 37 infants with recognized PVL. PVL was categorized according to De Vries, whereas West syndrome was categorized according to International League Against Epilepsy 1989. West syndrome in our patients developed during the first 2 years of life. The most common interictal abnormality was hypsarrhythmia. All, except two patients had delayed development and various degrees of mental retardation.The most characteristic neuroimaging findings were major reduction in cerebral cortical gray matter volume, reduction in the volume of brain myelin, and delayed myelination. These findings may explain the anatomical association between the West syndrome onset and PVL and intellectual and cognitive deficit in premature infants with PVL

The association of allergy and otitis media with eff usion in children

Upala srednjeg uha s izljevom (OME) česta je bolest u djece s prevalencijom koja doseže do 20% i s utjecajem na provodni gubitak sluha, zakašnjeli razvoj govora i oštećenje sluznice srednjeg uha. Cilj ove studije je utvrditi povezanost alergija i perzistirajućih upala srednjeg uha s izljevom (OME) u djece. U istraživanje je uključeno sedamdeset šestero (76) djece između 2 i 14 godina s dijagnozom perzistirajuće upale srednjeg uha s izljevom, koja su podvrgnuta operativnom zahvatu postavljanja aerizacijskih cjevčica. Sedamdeset šestero (76) zdrave djece slične dobi koja su podvrgnuta različitim neotorinolaringološkim operativnim zahvatima uključena su u kontrolnu skupinu. Pozitivna anamneza alergijskog rinitisa, rezultati intradermalnih kožnih testova i vrijednosti ukupnog i specifi čnog IgE-a su dokumentirani.U skupini s dijagnozom upale srednjeg uha s izljevom 58-ero djece (76,3%) imalo je pozitivnu anamnezu alergijskog rinitisa, a njih 45-ero (59,2%) imalo je pozitivne nalaze intradermalnog kožnog testiranja. Vrijednosti ukupnog IgE-a bile su povišene u 46-ero djece (60.5%), a vrijednosti specifi čnog IgE-a bile su povišene u njih 39-ero (51,3%) u OME skupini. U kontrolnoj skupini 12-ero djece (15,8%) imalo je pozitivnu anamnezu alergijskog rinitisa, njih 10-ero (13,2%) imalo je pozitivne nalaze intradermalnog kožnog testiranja, vrijednosti ukupnog IgE-a bile su povišene u 17-ero djece (22,4%) iz kontrolne skupine, a njih 9-ero (11,8%) imalo je povišene vrijednosti specifi čnog IgE-a. Rad pokazuje značajno veću prevalenciju alergija u skupini djece s upalom srednjeg uha s izljevom u usporedbi s kontrolnom skupinom. Značajno veći broj pozitivnih nalaza alergoloških testiranja nalazimo u skupini djece s upalom srednjeg uha s izljevom u odnosu na kontrolnu skupinu.Otitis media with eff usion (OME) is a common disease in children with a prevalence of up to 20% and great impact on conductive hearing loss, delayed speech development and destruction of middle ear mucosa lining. The aim of the study was to evaluate the association of allergy and persistent OME in children. Seventy-six children aged 2 to 14 years diagnosed with OME were included in the study. Control group included 76 age-matched healthy children attending pediatric surgical unit for diff erent pediatric surgical procedures other than ENT surgery. Fifty-eight (76.3%) OME children had positive history of allergic rhinitis and 45 (59.2%) had positive results of intradermal skin tests. Total IgE levels were elevated in 46 (60.5%) children and specifi c IgE levels were elevated in 39 (51.3%) children in OME group. In control group, 12 (15.8) children had positive history of allergic rhinitis and only 10 (13.2%) had positive results of intradermal skin tests. Total IgE levels were elevated in 17 (22.4%) children and specifi c IgE levels were elevated in 9 (11.8%) control group children. This study showed the prevalence of allergic history to be higher in children with OME as compared with control group. The prevalence of positive results of diff erent allergy tests was also higher in the OME group as compared with control group

PHACES Syndrome with Intestinal Hemangiomatosis

We present a rare case of a neonate with PHACES syndrome (posterior fossa malformations, large facial hemangiomas, cerebral arterial anomalies, cardiovascular anomalies, eye anomalies and sternal clefting or supraumbilical raphe) and diffuse hemangiomatosis of the ileum, presenting with multiple intestinal perforations and peritonitis. The infant was successfully treated with propranolol and methylprednisolone as well as octreotide, tranexamic acid, and supportive therapy for massive intestinal bleeding

LYMPHEDEMA-DISTICHIASIS SYNDROME – A CASE REPORT

Limfedem-distihijazni sindrom rijedak je nasljedni oblik limfedema koji se obično počinje klinički manifestirati u kasnom pubertetu. Obilježen je asimetričnim limfedemom donjih ekstremiteta i dvostrukim redom trepavica koje se najčešće nalaze u lateralnom dijelu vjeđe. U radu prikazujemo četrdesetčetverogodišnjaka koji se prezentirao limfedemom nogu i hilotoraksom. S obzirom na opisanu kliničku prezentaciju, anamnestičkim podatkom operacije ventrikularnoga septalnog defekta i dvostrukim redom trepavica zadovoljeni su kriteriji limfedem-distihijaznog sindroma. Liječenje limfedema simptomatsko je, a uključuje elastične čarape i masažu čime se usporava njegova progresija.Lymphedema-distichiasis is a rare form of hereditary lymphedema which usually begins to manifest clinically in the late teens. It is marked with asymmetrical lymphedema of the lower limbs and a double row of eyelashes that is usually located at the lateral part of the eyelid. We are describing a 44-year-old patient that presented with lymphedema of the legs and chylothorax. With such a presentation, a history of ventricular septal defect surgery, and double row of eyelashes he fulfi lls the criteria for lymphedema-distichiasis syndrome. Treatment of lymphedema is symptomatic and includes elastic socks and massage, which slow down its progression

Analysis of the C609T Polymorphism of NQO1 Gene in South Croatian Patients with Hematological Malignancies

In this study we analyzed the effect of polymorphic variation of NAD(P)H: quinone oxidoreductase1 (NQO1) gene that encode enzyme which detoxifies harmful quinines and protect hematopoietic stem cells against oxidative stress. C609T polymorphism of NQO1 gene leads to loss of enzyme activity, which may be a risk factor in the etiology of specific types of hematopoietic malignancies.We analyzed C609T polymorphism in NQO1 gene in the group of 82 patients (56 adult and 26 children) with different type of hematopoietic malignancies and 99 healthy participants (61 adult and 38 children) using PCR and the RFLP method. We confirmed that the polymorphism C609T in NQO1 gene was more frequent in the adult patients’ group with myeloid disorders, (p=0.0267) compared with adult controls.We could not confirm the association C609T polymorphism with recurrent chromosome translocations (clonal karyotype changes) neither in the adult nor in pediatric group of patients

Dietary Factors Associated with Plasma Thyroid Peroxidase and Thyroglobulin Antibodies

The knowledge about dietary habits and their influence in the development of autoimmune thyroid disease is insufficient. The aim of this study was to analyse the association of dietary factors and plasma thyroid peroxidase antibodies (TPO-Ab) and/or thyroglobulin antibodies (Tg-Ab). The study enrolled 1887 participants originating from the South Croatia. Participants with elevated plasma TPO-Ab and/or Tg-Ab were defined as cases (n = 462) and those with TPO-Ab and/or Tg-Ab within referent values were defined as controls (n = 1425). Dietary intake was evaluated according to a food frequency questionnaire containing 58 food items. Principal component analysis was used to group food items into dietary groups. We used logistic regression analysis to examine dietary groups associated with positive plasma TPO-Ab and/or Tg-Ab. The results indicate that the dietary group with frequent consumption of animal fats and butter is associated with positive plasma TPO-Ab and/or Tg-Ab (p = 0.01). The dietary group with frequent consumption of vegetables as well as the dietary group with high consumption of dried fruit, nuts, and muesli are associated with negative findings of TPO-Ab and/or Tg-Ab (p = 0.048 and p = 0.02, respectively). We showed that the anti-inflammatory dietary groups are associated with the negative findings of plasma TPO-Ab and/or Tg-Ab

The HOXA1 FOXF1, OSR1, and MTRR Gene Polymorphisms as Risk Factors for Congenital Malformations

U ovom istraživanju proveli smo genetičku studiju povezanosti različitih tipova prirođenih anomalija s polimorfizmima gena uključenih u razvoj kao čimbenika rizika razvojnih poremećaja. Analizirali smo 140 uzoraka DNA izolirane iz tkiva parafinskih kocki od fetalno ili neonatalno umrle djece s prirođenim anomalijama a naspram kontrolne skupine zdrave djece kod četiri različita polimorfizma gena rs12329305 OSR1, rs10951154 HOXA1, rs9936833 u blizini FOXF1 i rs326119 MTRR. Spomenuti polimorfizmi analizirani su metodom real-time PCR. Analiza povezanosti napravljena je na razini alelske i genotipske distribucije između dvije ispitivane skupine. Značajnu alelsku (p=7×10−4) i genotipsku (p=0,0013) povezanost s prirođenim malformacijama našli smo u polimorfizmu rs12329305 gena OSR1. Dodatne analize povezanosti u istog polimorfizma pokazale su njegovu povezanost s podskupinom izoliranih anomalija (p=1.25×10−5) i to sa razvojnim anomalijama srca (p=5,12x10-8) i bubrega (p=4,18 x 10-5). Polimorfizam rs9936833 u blizini gena FOXF1 pokazao je genotipsku povezanost s prirođenim malformacijama (p=0,034), dok ostala dva polimorfizma nisu pokazala. Analizom distribucije genotipova ispitivanih polimorfizama uz uključen spol utvrdili smo da je OR za pojavnost prirođenih malformacija u skupini muškog spola za 1,8 puta veći nego u skupini ženskog spola (p=0,038). Rezultati ovog istraživanja po prvi put su pokazali da su polimorfizmi rs12329305 gena OSR1 i rs9936833 gena FOXF1, kao i muški spol, čimbenici rizika za nastanak različitih tipova prirođenih malformacija u mrtvorođene/neonatalno umrle djece.We tested the association of four development-related gene polymorphisms with congenital anomalies. We assumed that those polymorphic variants may be a risk factor for developmental disorders. We analyzed 140 DNA samples isolated from archived paraffin tissue of deceased patients in whom fetal/neonatal autopsy examination had shown congenital malformations versus a control group of healthy children for four different polymorphisms: OSR1 rs12329305, rs9936833 near FOXF1, HOXA1 rs10951154 and MTRR rs326119. These polymorphisms were genotyped using the TaqMan allelic discrimination assay. Association analysis was performed on the allelic and genotypic distribution between the two tested groups. Significant allelic and genotypic association with stillborn/neonatal death was observed for rs12329305 (p=7×10-4, p=0.0013, respectively). In addition, association analysis for the same polymorphism was shown in the subgroup with isolated anomalies (p=1.25×10−5), particularly in the subgroup of cases with kidney and heart anomalies (p=4.18 x 10-5, p=5.12×10-8, respectively). Polymorphism rs9936833 near the FOXF1 gene showed only genotypic association with congenital malformations (p=0.034), when we compared minor allele heterozygotes and homozygotes versus major allele homozygotes, while the other two polymorphisms were not revealed. We also found that the OR for congenital malformations occurrence was 1.8 times higher in males than in females (p = 0.038), when we analysed the genotypic distribution of four examined polymorphisms along with sex. This is the first study, as far as we know, that showed the OSR1 gene polymorphism rs12329305 and rs9936833 near FOXF1, as well as male sex, are risk factors for different types of congenital malformations in cases of stillborn/neonatal death

NQO1 and NBS1 gene polymorphisms as the risk factors in hematological malignancies

Zloćudne bolesti krvotvornog tkiva su klonalne bolesti koje nastaju zbog genetske promjene u jednoj krvotvornoj stanici koštane srži ili limfatičkog tkiva. Mehanizam nastanka klonalne promjene nije sasvim razjašnjen, ali se zna da je posljedica zajedničkog djelovanja genetskih i okolišnih čimbenika. Klonalna promjena nastaje kao krajnja posljedica nakupljanja naslijeđenih i stečenih somatskih mutacija u protoonkogenima i tumor-supresorskim genima. Treća skupina gena koja ima ključnu ulogu u patogenezi bolesti su geni za popravak oštećenja DNA, a njihova inaktivacija potiče klonalnu promjenu u smislu preživljenja i bujanja stanica. Produkt gena NQO1 sudjeluje u metaboličkom procesu detoksikacije kemijskih karcinogena, a produkt gena NSB1 sudjeluje u mehanizmu popravka oštećenja DNA, a time zajednički ostvaruju zaštitno djelovanje na stanice krvotvornog sustava. Istražili smo genske promjene (polimorfizam i delecijsku mutaciju) u dva konstitucijska gena NQO1 i NBS1. Polimorfizam C609T gena NQO1 i delecijsku mutaciju 657del5 gena NBS1 istraživali smo u 82 bolesnika koji boluju od različitih zloćudnih bolesti krvotvornog tkiva i 99 zdravih ispitanika standardnim postupcima. Za dokaz polimorfizma C609T proveli smo polimeraznu lančanu reakciju (PCR) i analizu duljine restrikcijskih ulomaka (RFLP) nakon cijepanja enzimom Hinf1 uz vizualizaciju produkata na agaroznom ili poliakrilamidnom gelu. Za dokaz delecijske mutacije 657del5 proveli smo postupak PCR i vizualizaciju PCR-produkata na visokorezolucijskom poliakrilamidnom gelu (kako bi razlikovali veličinu produkata od 5 parova baza). Dokazali smo da je polimorfizam C609T, genotipa T/T i C/T, gena NQO1 statistički značajno češće prisutan u skupini bolesnika (39%) nego u skupini zdravih kontrolnih ispitanika (21%) (p = 0,031, χ 2 ). Statistički značajna razlika uz povećani izgled (rizik) (omjer izgleda = 2,4) razvoja bolesti nađena je u bolesnika starije dobne skupine koji boluju od poremećaja mijeloidne loze (p = 0,026, χ 2 ). Analizirali smo i klonalne kariotipske promjene u bolesnika s obzirom na zastupljenost polimorfizma C609T gena NQO1. Nismo potvrdili značajnu razliku u zastupljenosti polimorfizma C609T, genotipa T/T ili C/T, gena NQO1 u bolesnika sa specifičnim citogenetskim klonalnim promjenama u kariotipu (41/77) od onih s normalnim kariotipom (36/77). Nadalje smo pokazali prisutnost Slavenske mutacije 657del5 gena NBS1 u 2 od 82 bolesnika. Ovu delecijsku mutaciju nismo našli u kontrolnih ispitanika, pa vjerujemo da bi ta mutacija mogla biti važna u leukemogenezi, tim više što se bolest očitovala već u ranoj dobi i to u dječaka (657del5/657del5), koji je obolio od MDS, i u djevojčice (657del5/N), koja je oboljela od ALL. Nismo mogli analizirati, niti ustvrditi važnost združenih (višestrukih) genotipskih promjena s nastankom zloćudne bolesti krvotvornog tkiva s obzirom na samo dva bolesnika sa Slavenskom mutacijom 657del5 gena NBS1, koji uz to nisu imali polimorfni genotip C609T (C/T ili T/T) gena NQO1 niti klonalne promjene u kariotipu. Zaključili smo da obje istraživane genotipske promjene imaju ulogu u patogenezi razvoja zloćudnih klonalnih bolesti krvotvornog tkiva. Buduća istraživanja bi mogla otkriti ulogu i drugih gena u održavanju stabilnosti genoma krvotvorne matične stanice, te međudjelovanje tih gena međusobno i s čimbenicima okoliša.Hematological malignancies are clonal diseases that develop due to the genetic aberration in a single hematopoietic cell in bone marrow or lymphoid tissue. The mechanism of this clonal aberration is not completely understood, but it is known to be caused by the combined effects of genetic factors and environmental influences. Clonal aberrations are result of the accumulation of inherited and acquired somatic mutations in proto-oncogenes and tumor suppressor genes. The third class of genes is the DNA reparation genes, which inactivation initiates clonal expansion due to the increased cell survival and proliferation. The NQO1 gene product participates in a metabolic process of detoxification of potential chemical carcinogens, whereas the NBS1 gene product participates in DNA reparation, thus both having the protective effects on hematopoietic cells. In our study we investigated genetic aberrations (polymorphism and deletional mutation) in two constitutional genes, NQO1 and NBS1. We investigated C609T polymorphism of the NQO1 gene and 657del5 mutation of the NBS1 gene in 82 patients with different hematopoietic malignancies and 99 healthy participants using standard methods. For detection of C609T polymorphism, we performed polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) after Hifn1 digestion. The products were visualized on agarose or polyacryamide gels. For detection of 657del5, we performed PCR with PCR-product visualization of high resolution polyacryamide gen (to distinguish between 5 base pair difference between PCR-products). We confirmed that the NQO1 gene C609T polymorphism, with T/T and T/C genotypes, significantly more frequent in the patients’ group (39%) compared with the healthy participants (21%) (p = 0.031, χ 2). The significant difference, with the increased likelihood (risk) (odd ratio = 2.4) of developing hematopoietic malignancy, was found in the group of adult (mostly elderly) patients with myeloid disorders (p=0.026, χ 2). Also, we analyzed clonal karyotype aberrations in patients in relation to the presence of NQO1 gene C609T polymorphism. Nevertheless, we could not confirm the significant difference regarding the frequency of C609T polymorphism, with T/T or C/T genotypes, between patients with the specific cytogenetic clonal aberrations in karyotype (41/77) and those with the normal karytype (36/77). Furthermore, we found the presence of the NBS1 gene 657del5 Slavic mutation in 2 out of 82 patients’ samples. This deletional mutation was not detected in the control group, therefore we believe that it could be important for leukemogenesis, particularly because disease developed in early age- in a boy (657del5/657del5) with MDS and a girl (657del5/N) with ALL. We could not analyze and evaluate the importance of the combined (multiple) genotype aberrations for the pathogenesis of hematopoieticl malignancies considering only two patients with the NBS1 gene 657del5 Slavic mutation, which was not accompanied neither with the NQO1 gene C609T polymorphism (C/T or T/T) nor with the karyotype clonal aberrations. Our study revealed that both of the analyzed genotype aberrations have a role in the pathogenesis of hematopoietic clonal malignancies. Further research should reveal the role of other genes in maintaining the stability of the genome of the hematopoietic stem cell, and the interaction of those genes with the environmental factors

NQO1 and NBS1 gene polymorphisms as the risk factors in hematological malignancies

Zloćudne bolesti krvotvornog tkiva su klonalne bolesti koje nastaju zbog genetske promjene u jednoj krvotvornoj stanici koštane srži ili limfatičkog tkiva. Mehanizam nastanka klonalne promjene nije sasvim razjašnjen, ali se zna da je posljedica zajedničkog djelovanja genetskih i okolišnih čimbenika. Klonalna promjena nastaje kao krajnja posljedica nakupljanja naslijeđenih i stečenih somatskih mutacija u protoonkogenima i tumor-supresorskim genima. Treća skupina gena koja ima ključnu ulogu u patogenezi bolesti su geni za popravak oštećenja DNA, a njihova inaktivacija potiče klonalnu promjenu u smislu preživljenja i bujanja stanica. Produkt gena NQO1 sudjeluje u metaboličkom procesu detoksikacije kemijskih karcinogena, a produkt gena NSB1 sudjeluje u mehanizmu popravka oštećenja DNA, a time zajednički ostvaruju zaštitno djelovanje na stanice krvotvornog sustava. Istražili smo genske promjene (polimorfizam i delecijsku mutaciju) u dva konstitucijska gena NQO1 i NBS1. Polimorfizam C609T gena NQO1 i delecijsku mutaciju 657del5 gena NBS1 istraživali smo u 82 bolesnika koji boluju od različitih zloćudnih bolesti krvotvornog tkiva i 99 zdravih ispitanika standardnim postupcima. Za dokaz polimorfizma C609T proveli smo polimeraznu lančanu reakciju (PCR) i analizu duljine restrikcijskih ulomaka (RFLP) nakon cijepanja enzimom Hinf1 uz vizualizaciju produkata na agaroznom ili poliakrilamidnom gelu. Za dokaz delecijske mutacije 657del5 proveli smo postupak PCR i vizualizaciju PCR-produkata na visokorezolucijskom poliakrilamidnom gelu (kako bi razlikovali veličinu produkata od 5 parova baza). Dokazali smo da je polimorfizam C609T, genotipa T/T i C/T, gena NQO1 statistički značajno češće prisutan u skupini bolesnika (39%) nego u skupini zdravih kontrolnih ispitanika (21%) (p = 0,031, χ 2 ). Statistički značajna razlika uz povećani izgled (rizik) (omjer izgleda = 2,4) razvoja bolesti nađena je u bolesnika starije dobne skupine koji boluju od poremećaja mijeloidne loze (p = 0,026, χ 2 ). Analizirali smo i klonalne kariotipske promjene u bolesnika s obzirom na zastupljenost polimorfizma C609T gena NQO1. Nismo potvrdili značajnu razliku u zastupljenosti polimorfizma C609T, genotipa T/T ili C/T, gena NQO1 u bolesnika sa specifičnim citogenetskim klonalnim promjenama u kariotipu (41/77) od onih s normalnim kariotipom (36/77). Nadalje smo pokazali prisutnost Slavenske mutacije 657del5 gena NBS1 u 2 od 82 bolesnika. Ovu delecijsku mutaciju nismo našli u kontrolnih ispitanika, pa vjerujemo da bi ta mutacija mogla biti važna u leukemogenezi, tim više što se bolest očitovala već u ranoj dobi i to u dječaka (657del5/657del5), koji je obolio od MDS, i u djevojčice (657del5/N), koja je oboljela od ALL. Nismo mogli analizirati, niti ustvrditi važnost združenih (višestrukih) genotipskih promjena s nastankom zloćudne bolesti krvotvornog tkiva s obzirom na samo dva bolesnika sa Slavenskom mutacijom 657del5 gena NBS1, koji uz to nisu imali polimorfni genotip C609T (C/T ili T/T) gena NQO1 niti klonalne promjene u kariotipu. Zaključili smo da obje istraživane genotipske promjene imaju ulogu u patogenezi razvoja zloćudnih klonalnih bolesti krvotvornog tkiva. Buduća istraživanja bi mogla otkriti ulogu i drugih gena u održavanju stabilnosti genoma krvotvorne matične stanice, te međudjelovanje tih gena međusobno i s čimbenicima okoliša.Hematological malignancies are clonal diseases that develop due to the genetic aberration in a single hematopoietic cell in bone marrow or lymphoid tissue. The mechanism of this clonal aberration is not completely understood, but it is known to be caused by the combined effects of genetic factors and environmental influences. Clonal aberrations are result of the accumulation of inherited and acquired somatic mutations in proto-oncogenes and tumor suppressor genes. The third class of genes is the DNA reparation genes, which inactivation initiates clonal expansion due to the increased cell survival and proliferation. The NQO1 gene product participates in a metabolic process of detoxification of potential chemical carcinogens, whereas the NBS1 gene product participates in DNA reparation, thus both having the protective effects on hematopoietic cells. In our study we investigated genetic aberrations (polymorphism and deletional mutation) in two constitutional genes, NQO1 and NBS1. We investigated C609T polymorphism of the NQO1 gene and 657del5 mutation of the NBS1 gene in 82 patients with different hematopoietic malignancies and 99 healthy participants using standard methods. For detection of C609T polymorphism, we performed polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) after Hifn1 digestion. The products were visualized on agarose or polyacryamide gels. For detection of 657del5, we performed PCR with PCR-product visualization of high resolution polyacryamide gen (to distinguish between 5 base pair difference between PCR-products). We confirmed that the NQO1 gene C609T polymorphism, with T/T and T/C genotypes, significantly more frequent in the patients’ group (39%) compared with the healthy participants (21%) (p = 0.031, χ 2). The significant difference, with the increased likelihood (risk) (odd ratio = 2.4) of developing hematopoietic malignancy, was found in the group of adult (mostly elderly) patients with myeloid disorders (p=0.026, χ 2). Also, we analyzed clonal karyotype aberrations in patients in relation to the presence of NQO1 gene C609T polymorphism. Nevertheless, we could not confirm the significant difference regarding the frequency of C609T polymorphism, with T/T or C/T genotypes, between patients with the specific cytogenetic clonal aberrations in karyotype (41/77) and those with the normal karytype (36/77). Furthermore, we found the presence of the NBS1 gene 657del5 Slavic mutation in 2 out of 82 patients’ samples. This deletional mutation was not detected in the control group, therefore we believe that it could be important for leukemogenesis, particularly because disease developed in early age- in a boy (657del5/657del5) with MDS and a girl (657del5/N) with ALL. We could not analyze and evaluate the importance of the combined (multiple) genotype aberrations for the pathogenesis of hematopoieticl malignancies considering only two patients with the NBS1 gene 657del5 Slavic mutation, which was not accompanied neither with the NQO1 gene C609T polymorphism (C/T or T/T) nor with the karyotype clonal aberrations. Our study revealed that both of the analyzed genotype aberrations have a role in the pathogenesis of hematopoietic clonal malignancies. Further research should reveal the role of other genes in maintaining the stability of the genome of the hematopoietic stem cell, and the interaction of those genes with the environmental factors

The HOXA1 FOXF1, OSR1, and MTRR Gene Polymorphisms as Risk Factors for Congenital Malformations

U ovom istraživanju proveli smo genetičku studiju povezanosti različitih tipova prirođenih anomalija s polimorfizmima gena uključenih u razvoj kao čimbenika rizika razvojnih poremećaja. Analizirali smo 140 uzoraka DNA izolirane iz tkiva parafinskih kocki od fetalno ili neonatalno umrle djece s prirođenim anomalijama a naspram kontrolne skupine zdrave djece kod četiri različita polimorfizma gena rs12329305 OSR1, rs10951154 HOXA1, rs9936833 u blizini FOXF1 i rs326119 MTRR. Spomenuti polimorfizmi analizirani su metodom real-time PCR. Analiza povezanosti napravljena je na razini alelske i genotipske distribucije između dvije ispitivane skupine. Značajnu alelsku (p=7×10−4) i genotipsku (p=0,0013) povezanost s prirođenim malformacijama našli smo u polimorfizmu rs12329305 gena OSR1. Dodatne analize povezanosti u istog polimorfizma pokazale su njegovu povezanost s podskupinom izoliranih anomalija (p=1.25×10−5) i to sa razvojnim anomalijama srca (p=5,12x10-8) i bubrega (p=4,18 x 10-5). Polimorfizam rs9936833 u blizini gena FOXF1 pokazao je genotipsku povezanost s prirođenim malformacijama (p=0,034), dok ostala dva polimorfizma nisu pokazala. Analizom distribucije genotipova ispitivanih polimorfizama uz uključen spol utvrdili smo da je OR za pojavnost prirođenih malformacija u skupini muškog spola za 1,8 puta veći nego u skupini ženskog spola (p=0,038). Rezultati ovog istraživanja po prvi put su pokazali da su polimorfizmi rs12329305 gena OSR1 i rs9936833 gena FOXF1, kao i muški spol, čimbenici rizika za nastanak različitih tipova prirođenih malformacija u mrtvorođene/neonatalno umrle djece.We tested the association of four development-related gene polymorphisms with congenital anomalies. We assumed that those polymorphic variants may be a risk factor for developmental disorders. We analyzed 140 DNA samples isolated from archived paraffin tissue of deceased patients in whom fetal/neonatal autopsy examination had shown congenital malformations versus a control group of healthy children for four different polymorphisms: OSR1 rs12329305, rs9936833 near FOXF1, HOXA1 rs10951154 and MTRR rs326119. These polymorphisms were genotyped using the TaqMan allelic discrimination assay. Association analysis was performed on the allelic and genotypic distribution between the two tested groups. Significant allelic and genotypic association with stillborn/neonatal death was observed for rs12329305 (p=7×10-4, p=0.0013, respectively). In addition, association analysis for the same polymorphism was shown in the subgroup with isolated anomalies (p=1.25×10−5), particularly in the subgroup of cases with kidney and heart anomalies (p=4.18 x 10-5, p=5.12×10-8, respectively). Polymorphism rs9936833 near the FOXF1 gene showed only genotypic association with congenital malformations (p=0.034), when we compared minor allele heterozygotes and homozygotes versus major allele homozygotes, while the other two polymorphisms were not revealed. We also found that the OR for congenital malformations occurrence was 1.8 times higher in males than in females (p = 0.038), when we analysed the genotypic distribution of four examined polymorphisms along with sex. This is the first study, as far as we know, that showed the OSR1 gene polymorphism rs12329305 and rs9936833 near FOXF1, as well as male sex, are risk factors for different types of congenital malformations in cases of stillborn/neonatal death