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The purpose of this study was to determine the prevalence of allele and genotype variants of the follicle-stimulating hormone receptor (FSHR) gene polymorphic region at position Asn680Ser in the Albanian male population and associate them with the clinical parameters of infertility. The study included 114 infertile men (mean age 35.04±5.85 years) stratified according to the level of spermatogenetic impairment (oligoasthenozoospermia, asthenozoospermia and normospermia) and 112 fertile men (mean age 36.44±7.05 years) with normal semen parameters. Genotyping of the FSHR gene at position 680 was performed by TaqMan genotyping assay. All the participants underwent semen analysis, and serum reproductive hormones (FSH, luteinizing hormone, prolactin and testosterone) were also measured. The FSHR Asn680Ser genotype frequencies were as follows: Asn/Ser 42%, Ser/Ser 33.9% and Asn/Asn 24.1% in the control group, and Asn/Ser 56.1%, Ser/Ser 22.8% and Asn/Asn 21.1% in the whole group of infertile men (χ2-test: P=0.08). There was no statistically significant correlation between serum hormone levels and semen characteristics or between fertility status and FSHR Asn680Ser gene variants in the control group and the group of infertile men. However, adjusted logistic regression analysis (age, body mass index, smoking and alcohol as covariates) revealed increased odds ratio for male infertility among heterozygous Asn/Ser genotype carriers associated with lower values of semen parameters (normal morphology, concentration, total sperm count and motility). In conclusion, our case-control study further confirmed previous reports on no significant association between the FSHR Asn680Ser polymorphisms and male infertility. Nevertheless, the data presented herein indicate that the Asn/Ser genotype may increase the risk of male infertility in Albanian population., Cilj ovoga istraživanja bio je odrediti pojavnost alela i varijante genotipa receptora folikularno stimulirajućeg hormona (FSHR) na poziciji Asn680Ser kod muškaraca albanske populacije u odnosu na kliničke parametre neplodnosti. Istraživanje je obuhvatilo 114 neplodnih muškaraca (srednja dob 35,04±5,85 godina) svrstanih prema razini oštećenja spermiograma (oligoastenozoospermija, astenozoospermija i normospermija) te 112 plodnih muškaraca (srednja dob 36,44±7,05 godina) s urednim nalazom spermiograma. Genotipizacija gena FSHR na poziciji 680 učinjena je primjenom TaqMan probe. Kod svih sudionika istraživanja učinjena je analiza sjemena i reprodukcijskih hormona uključujući FSH, luteinizirajući hormon, prolaktin i testosteron. U kontrolnoj skupini ispitanika kod FSHR Asn680Ser genotipa utvrđena pojavnost Asn/Ser bila je 42%, Ser/Ser 33,9% i Asn/Asn 24,1%, dok se u skupini neplodnih ispitanika incidencija kretala od 56,1% za Asn/Ser, 22,8% za Ser/Ser i 21,1% za Asn/Asn (χ2-test; p=0,08). Nije ustanovljena značajna statistička povezanost između razine hormona, karakteristika sjemena, stanja plodnosti u varijanti gena FSHR Asn680Ser u kontrolnoj skupini u odnosu na ispitanike u skupini neplodnih muškaraca. Ipak, primjenom prilagođene, logističke i regresijske analize (dob, indeks tjelesne mase, pušenje i alkohol kao kovarijable) utvrđeno je da postoji veća vjerojatnost javljanja muške neplodnosti kod nositelja heterozigota Asn/Ser koji su povezani sa sniženim vrijednostima parametara sjemena (morfologija, koncentracija, ukupan broj i pokretljivost). Zaključno možemo utvrditi da ovo istraživanje potvrđuje ranija izvješća kako ne postoji značajna povezanost između polimorfizma FSHR Asn680Ser i muške neplodnosti. Ipak, navedeni podaci upućuju na to da Asn/Ser genotip može povisiti rizik muške neplodnosti u albanskoj populaciji.

The aim of the study was to assess sleep architecture and breathing in sleep in children with Down syndrome. The study was conducted by using overnight video-polysomnography (V-PSG) in children with Down syndrome and age-matched children from the general population. Analysis of polysomnographic parameters revealed that compared to the norms of healthy age- and maturitymatched children from the general population, children with Down syndrome had significantly shorter sleep latency (p=0.007), shorter total sleep time (p=0.004), lower sleep efficiency (p=0.010), less NREM1 sleep phase (p=0.0002), less NREM3 sleep phase (p=0.034), less REM sleep (p=0.034) in favour of more NREM2 phase but not significantly (p=0.069), and spent more time awake after sleep onset (p=0.0002). Children with Down syndrome had significantly more obstructive sleep apnoeas and hypopnoeas per hour (higher obstructive sleep apnoeas and hypopnoeas index) (p=0.008), but less central sleep apnoea per hour (lower central apnoeas index) (p=0.041), which led to the nonsignificantly lower total apnoea-hypopnoea index (p=0.762) in children with Down syndrome. The mean and longest apnoea duration did not differ significantly between these two groups. Children with Down syndrome had a significantly lower mean and nadir oxygen saturation (p=0.008 and p=0.001, respectively). In conclusion, the majority of respiratory complications in children with Down syndrome can be prevented by raising awareness of sleep disturbances in children with Down syndrome among their parents and health care providers and by introducing early routine V-PSG in the follow up of these children., Cilj: Istražiti arhitektoniku spavanja i disanja tijekom sna u djece s Downovim sindromom. Metode: Cjelonoćna videopolisomnografija (V-PSG) u skupini djece s Downovim sindromom i u onoj iz opće populacije odgovarajuće dobi i zrelosti. Rezultat: Analiza polisomnografskih parametara pokazala je da djeca s Downovim sindromom, u usporedbi s normativima u zdrave djece u općoj populaciji, odgovarajuće dobi i zrelosti, imaju značajno kraću latenciju spavanja (SL) p=0,007, kraće ukupno vrijeme spavanja (TST) p=0,004, nižu efikasnost spavanja (SE) p=0,010, manje NREM1 faze spavanja p=0,0002, manje NREM3 faze spavanja p=0,034, manje REM spavanja p= 0,034 (na račun više površnog NREM2 spavanja, ali ne statistički značajno, p=0,069) i da provode više vremena budni nakon započimanja spavanja (p=0,0002). Djeca s Downovim sindromom imaju značajno više opstruktivnih apneja i hipopneja po satu spavanja (viši OAHI) p=0,008, ali imaju manje centralnih apneja po satu spavanja (niži CAI) p=0,041, što pridonosi statistički neznačajno nižem ukupnom AHI-u (p=0,762) u djece s Downovim sindromom. Prosječno i najdulje trajanje apneja nije se značajno razlikovalo između dviju skupina. Djeca s Downovim sindromom su imala značajno niže i prosječne i nadir saturacije kisika (p=0,008 i p=0,001). Zaključak: Možemo prevenirati većinu respiratornih komplikacija u djece s Downovim sindromom podizanjem svjesnosti njihovih roditelja o poremećajima spavanja u te djece, ali i zdravstvenih djelatnika, te uključivanjem rutinske cjelonoćne videopolisomnografije u njihovo praćenje.

Prenatalna dijagnostika kromosomskih poremećaja podrazumijeva citogenetičku analizu stanica ploda dobivenih jednom od rutinskih invazivnih tehnika - biopsijom korionskih resica, placentocentezom, amniocentezom ili kordocentezeom. Izbor tipa invazivne dijagnostike ovisi ponajprije o individualnoj procjeni čimbenika rizika za rađanje kromosomski bolesnog djeteta, a zatim i o riziku lošeg ishoda, odnosno gubitka trudnoće kao najteže komplikacije invazivnog zahvata. Problemi koji se mogu javiti u prenatalnoj dijagnostici kromosomskih poremećaja su pojava kromosomskog mozaicizma te razlikovanje pravog od pseudomozaicizma, rizik neuočavanja suptilnih kromosomskih razmještanja zbog ograničenja razlučivanja svjetlosnog mikroskopa, nemogućnost detekcije kriptičkih mozaičnih oblika te zagađenje majčinim stanicama. U tim se slučajevima uz klasičnu citogenetičku analizu provodi i neka od metoda molekularne dijagnostike. Poseban izazov u prenatalnoj dijagnostici su blizanačke trudnoće. Uvođenjem metoda potpomognute oplodnje i novih protokola stimulacije, značajno se povećao broj višeplodnih trudnoća. Razlikovanje monozigotnih od dizigotnih blizanaca kod dikorijalnih diamnijskih blizanaca istog spola može se precizno odrediti usporednom analizom mikrosatelitnih lokusa. Mehanizmima kao što je postzigotno nerazdvajanje može doći do nastajanja različitih kromosomskih statusa, čak i kod monozigotnih blizanaca. Zato je amniocenteza tehnika izbora, a uzorci se trebaju uzeti iz svake amnijske vreće, posebice ako jedan ili oba blizanca imaju ultrazvučno dijagnosticirane abnormalnosti., Prenatal diagnosis of chromosomal abnormalities comprises cytogenetic analysis of fetal cells obtained after one of the routine invasive techniques, i.e. chorionic villus sampling, placentocentesis, amniocentesis, or cordocentesis. The choice of the type of invasive procedure is primarily based on individual combined calculation of all risk factors of having a chromosomally abnormal child. An important factor for the patient to decide to undergo an invasive procedure is the chance of pregnancy loss as the worst scenario. The problems that can arise in prenatal diagnosis of chromosomal disorders are diff erent types of mosaic forms and diffi culty to distinguish true fetal mosaicism from pseudomosaicism, the risk of missing a subtle anomaly due to the limitation of resolution of light microscopy, and maternal cell contamination. In these cases, some of molecular diagnostic procedures are performed. There is an increasing number of twin pregnancies after stimulation protocols and use of some of the techniques of in vitro fertilization procedures in the infertility treatment. Diff erentiation of monoyzgotic and dizygotic pregnancies is possible by applying microsatellite analysis. Chromosomal discordance in twin pregnancies in most cases is caused by postzygotic nondisjunction, and is recorded both in monozygotic and dizygotic twins. Amniocentesis is the technique of choice, and karyotyping from both amniotic sacs is recommended, especially in cases where one fetus has major structural malformations.

Although, deletion of short arm of chromosome 18 is one of the most frequent autosomal terminal deletions, mosaic form of 18p deletion is infrequently observed. Furthermore, prenatally detected cases of 18p deletion and isochromosome 18q mosaicism are extremely rare. Herein, we present a case of del(18p)/i(18q) mosaicism, prenatally detected after chorionic villus sampling. A 37-year-old woman was referred for prenatal diagnosis because of fetal septated cystic hygroma measuring 4.3 mm. Cytogenetic analysis showed a mosaic 46,XX,del(18)(p11.2)/46,XX,i(18)(q10) karyotype in both, short- and long-term culture. Parents elected to terminate the pregnancy. Fetal mosaic karyotype was confirmed by chromosomal analysis of cultured skin fibroblasts. Molecular characterization of chromosome 18 structural aberrations was performed by fluorescence in situ hybridization (FISH). Considering variable ultrasound findings among cases with del(18p)/i(18q) mosaicism, we emphasized that first and second trimester ultrasound screening examinations for fetal malformations, followed by cytogenetic and molecular evaluations, are very important in the management of prenatally detected cases.

Clinical features in patients with segmental aneuploidy often vary depending on the size of the chromosomal segment involved. Monosomy 2p is usually observed as a part of more complex syndromes among probands of balanced reciprocal translocation carriers. Patients with dup4q syndrome have variable clinical features, which are both related to the size of duplicated segment of the 4q and specific associated monosomy. Clinical findings of our patient were compatible with those previously reported in dup4q and del2p patients. Herein are presented the clinical and cytogenetic findings in a 4-year-old female with an unbalanced karyotype 46,XX,der(2)t(2;4)(p25.1;q31.3)pat. Clinical phenotypes of 2p;4q translocation cases are variable, because the involved breakpoints vary case-by-case. We also compare similarity of the clinical features of our proband and other patients carrying either duplication of the distal part of 4q and patients carrying a deletion of distal part of 2p as described in the literature. To our knowledge, this is the first case of partial trisomy 4q accompanied with partial monosomy 2p.

The extent of clinical expression in cases of segmental aneuploidy often varies depending on the size of the chromosomal region involved. Here we present clinical and cytogenetic findings in a 5-month old boy with a duplication of a chromosomal segment 4p16.1→4pter and a deletion of a chromosomal segment 8p23.1→8pter. His karyotype was determined by applying classical GTG banding and FISH method (WHCR region, centromere 4, centromere 8, telomere 8p) as 46,XY,der(8)t(4;8)(p16.1;p23.1).ish der(8)t(4;8)(D8S504-,WHCR+,D8Z2+)dn. Parents are not related and have normal karyotypes, indicating de novo origin. We have compared similarity of the clinical features in our proband to other patients carrying only a duplication of the distal part of 4p or a deletion of distal part of 8p or similar combination described in the literature.

The aim of the study is to investigate the efficiency of the second-trimester biochemical screening, with maternal serum alpha-fetoprotein (MS-AFP) and free b-subunit of human chorionic gonadotropin (free b-hCG), during the ten-year period. The study included 11,292 of pregnant women between the 15th and 18th gestational week, who underwent screening from November 1996 to December 2006. The risk for trisomy 21 and trisomy 18 were calculated by computer software, based on a model which generated the final risk for fetal aneuploidies from the pregnant woman’s a priori age risk and the likelihood ratio of the distribution of the biochemical markers, according to the second-trimester gestation. With the cut-off value of the final risk 1:250, the detection rate for trisomy 21 was 75% (21/28). In women less than or equal to 35, the detection was 57.1% (8/14) and 92.9% (13/14) in those over 35 years, respectively. The detection rate of trisomy 18 was 50% (2/4). The results confirmed that the implementation of double-test, as non-invasive screening for fetal aneuploidies, should be accepted as a complementary method of antenatal care.

Hypophosphatasia is a metabolic bone disease characterized by bone and teeth hypomineralization due to defective function of tissue-nonspecific alkaline phosphatase (TNSALP). The disorder is caused by various mutations in the TNSALP gene localized on short arm of chromosome 1. Infantile hypophosphatasia is a severe form of the disease inherited as an autosomal recessive trait which presents before age of six months and often has fatal outcome. We report a patient with typical clinical course for infantile hypophosphatasia who was homozygous for the c.1402G>A mutation. The same mutation has been previously associated with a more severe perinatal form also in a Croatian family what indicates a possible common ancestral origin and phenotypic variability potential of c.1402G>A mutation of TNSALP gene.

Autosomal trisomies account for more than 80 % of significant chromosomal disorders and are routinely detected by the cytogenetic analysis of cultivated amniotic fluid cells. However, this approach is time-consuming and requires a significant level of training and expertise. The main aim of our work was to introduce QF-PCR to our lab, a quicker, simpler and cheaper method. We also aimed to evaluate the usefulness of the chosen marker set in the Croatian population and the reliability and accuracy of the obtained results. STR loci from chromosomes 13, 18 and 21 were co-amplified, separated by capillary electrophoresis and analysed. Characteristic triplets and/or 2:1 patterns were detected for trisomic samples while normal samples were either homozygous or heterozygous. The tested set of loci showed high heterozygosity and therefore a good potential for analyzing the Croatian population. The results of QF-PCR were in full compliance with the cytogenetic analysis which was also performed for cultivated amniotic fluid cell samples.

The aim of this study was to determine if insertion-deletion polymorphism of angiotensin-converting enzyme is a risk factor for the development of preeclampsia. Sixty women with preeclampsia and 50 normotensive pregnant women were included in this study. Preeclampsia was defined as blood pressure > 140/90 mmHg in a previously normotensive women with proteinuria >300 mg/L in a 24-hours. Twelve women also had preeclampsia in previous pregnancy. The genotyping of polymorphism in the intron 16 of the angiotensin-converting enzyme was performed by the polymerase chain reaction followed by the agarose electrophoresis. The patients were divided into three groups according to the presence (I) or absence (D) of insertional polymorphism (II, ID, and DD). Genotype distribution and allele frequencies were compared by Mantel-Haenszel c 2 testing. The frequency of DD genotype was not significantly higher in women with preeclampsia (26/60)than in the control group (14/50, p=0.096). The D allele frequency was significantly higher in 17 women with preeclampsias who required delivery before 34 weeks of pregnancy (0.735), than in 43 women in whom obstetric complications took place after 34 weeks of pregnancy (0.56, p=0.036). The D allele frequency was 0.83 in women having recurrent preeclampsia, i.e. significantly higher compared with women, who were for the first time, experienced preeclampsia (0.57, p=0.013). This study showed a significantly positive association between D allele frequency and risk of recurrent preeclampsia and preterm delivery before 34 weeks of pregnancy. The deletion genotype could be an important contributing factor for an early onset and recurrent preeclampsia.

SAŽETAK Posljednjih godina došlo je do naglog razvoja dijagnostičkih metoda kojima se može otkriti genetski uzrok neplodnosti, a time procijeniti i rizik prijenosa genetskog poremećaja. Razvojem tehnika potpomognute oplodnje moguće je liječiti veoma teške oblike neplodnosti koji nose značajno veći rizik prijenosa određenih molekularnih promjena na potomstvo. Kod takvih slučajeva veoma je važno otkriti mutacijsku promjenu, jer se time pruža mogućnost davanja precizne genetske informacije o rizicima nasljeđivanja i načina prenatalnog određivanja genotipa ploda. S obzirom na to da ne postoje jedinstvene smjernice u pristupu dijagnostike i liječenja neplodnosti, nastoje se izdvojiti skupine visokorizičnih neplodnih bolesnika prema njihovim kliničkim osobitostima koje će biti podvrgnute rutinskim dijagnostičkim postupcima u svrhu otkrivanja uzroka neplodnosti. Za određene genske poremećaje, kao što su cistična fibroza i sindrom fragilnog X, poznata je uloga u etiopatogenezi neplodnosti. Najnovija molekularna istraživanja genskih mutacija, posebice u β-podjedinici FSH i LH gena, pokazuju svoju dijagnostičku i kliničku vrijednost kod otkrivanja genetskih uzroka neplodnosti., During the last few years, there is a rapidly expansion of new diagnostic techniques improving the detection of genetic causes of infertility. It is followed by a precise risk assessment of genetic change transmission and possibility to chose the most adequate methode of medically assisted reproduction. The treatment of severe forms of both male and female infertility became possible by the development of very sofisticated in vitro fertilising techniques. Such patients carry the significantly higher risk of having mutational change in one of the disease genes involved in regulation of human reproduction. The detection of mutation in these patients improve the treatment and prognostic assessment in potential future pregnancies. The unique guidelines for the diagnosis and the treatment of infertile couples still doesn’t exist. There are several categories of patients selected by clinical features and associated with some forms of genetic abnormalities. For particular genetic disorders such as cystic fibrosis and fragile X syndrome, their role in pathogenesis of infertility is very well known. Every day, new genetic mutations associated with infertility are discovered. The recent studies involving investigations of significance of molecular mutations, particulary in the gene for the β-subunit of FSH and LH , showed thier diagnostic and clinical value in evaluation both male and female infertility.

Objective. To estimate the degree of risk of the echogenic intracardiac foci (IEF) for fetal chromosomopathies and to determine its association with structural anomalies of the fetus. Material and methods. During the period of two years 190 pregnant patients had been send for fetal echocardiography. Examination had been performed by transvaginal¬ (12–17 weeks of gestation) or transabdominal approach (18 weeks or more of gestation). Results. IEF was observed in 17 fetuses, multifocal appearance was found in 2 out of 17 fetuses. In 3 cases IEF had resolved during the 8 weeks period of time. Additional structural anomalies were detected in 11 fetuses. In 2 fetuses trisomy 21 had been confirmed. Conclusion. A single soft marker as IEF is commonly encountered during the second trimester among the fetuses with chromosomal aberation. As do many sonographic markers IEF can be resolved during the pregnancy and often can be found in normal fetuses., Cilj rada je bio na vlastitom uzorku utvrditi u kojoj mjeri ultrazvučni nalaz hiperehogenih intrakardijalnih žarišta (IEF) pridonosi dijagnostici kromosomopatija i strukturalnih anomalija. Uzorak i metode. Tijekom dvije godine 190 trudnica između 12. i 39. tjedna trudnoće je primljeno radi fetalne ehokardiografije. Pregled je obavljen vaginalnom sondom od 5 MHz pri trudnoćama 12.–17. tjedna ili zavinutom abdominalnom sondom od 3,5 MHz nakon 17. tjedna trudnoće. Rezultati. IEF su nađeni u 17 fetusa, multifokalni u 2 od njih. U 3 fetusa su IEF u roku od osam tjedana nestali. U 11 fetusa su nađene dodatne strukturalne anomalije. Trisomija 21 je potvrđena u 2 fetusa. Zaključak. IEF su »meki« ultrazvučni biljezi fetalne aneuploidije, često su prolazni, a nalaze se i u eukariotičnih fetusa.