Genotype-phenotype corelation in Serbian patients with hyperphenylalaninemia

Cilj rada: Ispitivana je korelacija genotipa i fenotipa bolesnika sa hiperfenilalninemijom. Utvrðena je incidencija hiperfenilalninemije u našoj populaciji. Prikazana je učestalost najčešćih mutacija u PAH genu i njihov uticaj na fenotip. Posebna pažnja je poklonjena ispitivanju metaboličkog i kliničkog fenotipa najčešće mutacije u našoj populaciji. Izneti su zaključci praćenja metaboličke kontrole kod naših bolesnika. Materijal i metode: Za ispitivanje incidencije hiperfenilalaninemije korišćeni su rezultati Republičkog zavoda za statistiku. Korišćena je medicinska dokumentacija Instituta za zdravstvenu zaštitu majke i deteta i rezultati Instituta za molekularnu genetiku i genetičko inženjerstvo. Biohemijske analize raðene su Guthrievim testom, Enzimskom metodom i Aminoanalizatorom. Za analizu dobijenih rezultata korišće su deskriptivne i analitičke statističke metode. Rezultati: Incidencija hiperfenilalninemije na našim prostorima iznosi 1: 15 130. Najveći broj bolesnika (62%) ima klasičnu fenilketonuriju. Najčešće mutacije PAH gena u našoj populaciji su: p.L48S, p.R4OW, p.P281L, p.E3906 i p.1306V, i javljaju se u 2/3 svih bolesnika. Interesantno je da je naša najčešća mutacija L48S prisutna kod 31% obolelih, što do sada nije zabaleženo ni u jednoj populaciji. Homozigoti i funkcionalni hemizigoti za L48S mutaciju se mogu javiti kod svih kategorija hiperfenillaninemije (blaga hiperfenilalaninemija, srednje teška fenilketonurija i klasična fenilketonurija). Pokazali smo i da tip mutacije nije značajan za konačan IQ bolesnika sa hiperfenilalaninemijom, već kontinuirano održavanje koncentracije fenilalnina u krvi u dozvoljenim granicama. Zaključak: Naša zemlja pripada grupi sa srednjom incidencijom hiperfenilalaninemija u svetu. S obzirom da naša najčešća mutacija pripada grupi mutacija sa rezidualnom enzimskom aktivnosti, koja izgleda zavisi od brojnih faktora, tolerancija fenilalanina je najbolji način za kategorizaciju ovih bolesnika. Odluka oko primene terapije i dalje treba da se zasniva na koncentraciji fenilalnina u krvi, a ne na osnovu mutacije u PAH genu. Na osnovu analize genotipa, preko 50% naših bolesnika su kandidati za BH4 terapiju.Goal: Correlation between genotype and phenotype was tested in patients with hyperphenylalaninemia. Incidence of hyperphenylalaninemia in our population was calculated. Frequency of most common mutations in PAH gene was presented, along with it's impact on phenotype. Special focus was put directed on exploration of metabolic and clinical phenotype of the most prevalent mutation in our population. Conclusions derived from metabolic follow-up of our patients were presented as well. Matherials and methods: To determine incidence of hyperphenylalaninemia, data from Statistical Office of the Republic of Serbia. Medical documentation of Mother and Child Health Care Institute for used along with genetic findings of Institute for Molecular Genetics and Genetical Engineering. Biochemical analysis were performed using Guthrie test, Enzyme assay and Aminoacid analyser. Statistical analysis included both descriptive and analytical methods. Results: Incidence of hyperphenylalaninemia in our population was estimated at 1: 15 130. Majority of patients (62%) have classic phenylketonuria. Most common mutations in PAH gene in Serbia are: p.L48S, p.R4OW, p.P281L, p.E3906 i p.1306V, found in 2/3 of all patients. Interesting finding refers to L48S being the most prevalent mutation (found in 31%), which is unparalleled finding worldwide. Homozygous and functionaly hemizigous L48S carriers are found among all types of hyperphenylalaninemia (mild hyperphenylalaninemia, moderate hyperphenylalaninemia and classic phenylketonuria). It was also shown in these results that mutation type is not a valuable predictor for final IQ status of patients with hyperphenylalaninemia; on the other hand, metabolic control of disease was found as significant factor of final outcome. Conclusion: Our country can be considered as having an average incidence of hyperphenylaninemia. Considering the fact that most common mutation in Serbia relates to residual enzyme activity (determined by multitude of factors), tolerance of phenylalanine is the best way to classify severity of the disease. Thus, decision over treatment should be based on phenylalanine blood concentrations, and not on PAH gene mutation. According to genotype profiling, more than 50% of our patients could be candidates for tetrahydrobiopterine (BH4) treatment

Association of mitochondrial DNA variants and cognitive impairment of phenylketonuria patients

Uvod: Fenilketonurija (PKU) je metaboličko oboljenje uzrokovano mutacijama u genu za fenilalanin hidroksilazu (PAH). Ukoliko se ne leče, pacijenti sa fenilketonurijom razvijaju tešku mentalnu retardaciju, koja može biti i posledica neurodegeneracije. Ovo je prva studija koja istražuje prisustvo mitohondrijalnih DNK varijanti kod pacijenata sa fenilketonurijom, m. 10398A, za koju je pokazana povezanost sa neurodegenerativnim oboljenjima, kao i m. 10410T. Metode: U ovoj studiji je analizirano 64 pacijenta sa fenilketonurijom i 50 zdravih kontrola iz srpske populacije. Pacijenti su podeljeni u grupe prema uzrastu u kome je postavljena dijagnoza i odnosu prema dijeti sa niskim sadržajem fenilalanina. IQ je određen pomoću odgovarajućih uzrasnih skala. Rezultati: Mitohondrijalne DNK varijante, m. 10398A i m. 10410T, su detektovane sekvenciranjem. Učestalost m. 10398A je bila jednaka kod pacijenata i zdravih kontrola (82,81% i 82,00%), što ukazuje na isto etničko poreklo ovih ispitanika. U slučaju m. 10410, nisu detektovane različite varijante. U grupi pacijenata kojima je kasno postavljena dijagnoza i koji su neadekvatno lečeni, nije pronađena statistička značajnost u srednjoj vrednosti IQ između pacijenata sa m. 10398A i m. 10398G. Isto je pokazano i za pacijente sa višim IQ koji su otkriveni prilikom neonatalnog skrininga i koji su pravilno lečeni. Međutim, kada su iz ove grupe isključeni pacijenti koji nose p. L48S mutaciju u PAH genu, koja ima nekonzistentan uticaj na fenotip, prisustvo m. 10398A alela je povezano sa nižim IQ. Zaključak: Ova studija je potvrdila važnost neonatalnog skrininga i pravilnog sprovođenja dijete kod pacijenata sa fenilketonurijom. Statističke analize nisu jasno utvrdile uticaj mitohondrijalne DNK varijante, m. 10398A, na IQ ovih pacijenata, osim kad je i PAH genotip uključen u analizu. Studije u većim grupama ce rasvetliti povezanost između mutacija u PAH genu, mitohondrijalnih DNK varijanti i složenog kognitivnog fenotipa kod pacijenata sa PKU.Background: Phenylketonuria (PKU) is a metabolic disorder caused by phenylalanine hydroxylase gene (PAH) mutations. If left untreated, PKU patients develop severe mental retardation potentially due to neurodegeneration. This is the first study that investigates presence of mitochondrial DNA variants in PKU patients, m.10398A, reportedly associated with neurodegenerative diseases and m.10410T. Methods: We analyzed 64 PKU patients and 50 healthy controls from Serbian population. PKU patients were categorized into groups according to time of diagnosis and compliance to low-phenylalanine diet. The IQ was determined according to age-appropriate scales. Results: We detected m.10398A and m.10410T variants by direct sequencing. Frequency of m.10398A was similar in patients and healthy controls (82.81% and 82.00% respectively) suggesting their identical ethnic background. No variation was detected for m.10410. In group with late diagnosis and poorly controlled diet, no statistically significant difference in average IQ was found between patients with m.10398A and m.10398G. The same was shown for PKU patients with higher IQ, diagnosed at neonatal screening and treated with low-phenylalanine diet. However, when patients carrying p.L48S, a PAH mutation with inconsistent effect, were excluded from the study, presence of m.10398A variant was associated with lower IQ. Conclusions: This study emphasizes the importance of neonatal screening and good control of low-phenylalanine diet in PKU patients. Statistical analysis did not indicate clear impact of mitochondrial DNA variant m.10398A on IQ of PKU patients, except when PAH genotype was also considered. Studies in larger cohorts will elucidate the association between PAH gene mutations, mitochondrial DNA variants and complex PKU cognitive phenotype

Chemical Analysis of Mortars of Archeological Samples form Mediana

The aim of this study was mineralogical and chemical analysis of mortar from the floor, ceiling and wall of Stibadium B, from the archaeological site of Mediana. ICP-OES and FTIR-spectroscopy were used to determine chemical composition and some major mineralogical species. The obtained results show that lime mortar is probably used. Large contribution of silicon- and aluminum-oxides, indicate the presence of quartz and clay minerals derived from the aggregate, river sand and crushed bricks. The obtained results also show large amount of iron, manganese and copper. The determinated metals in samples from floor and wall of Stibadium B, are mostly present in oxide fraction, while in sample from ceiling, they are mostly found in silicate fraction

Sintactic indicators of genderlect in later language development

Men and women use language means in various ways, the means of the same language, when they speak and write. This is the essence of genderlect theory which tries to explain what these differences are, and such theory incited numerous researches in current years. The aim of this work is to investigate syntactic features of youngest school age students' texts and differences in that production between boys and girls. 181 pupils participated in this research from the first, up to the fourth grade (96 boys and 85 girls). Each student wrote three essays, so there were 543 texts in this corpus. Our study included moth quantitative (the text length presented by a number of words, communicative sentences and clauses; the length of the sentence presented by a number of words and clauses) and qualitative (various sentences types) indicators. Differences between genders were analysed in comparison to researched variables. The results proved that there exist statistically significant differences in text length and sentence length in comparison to all measured parameters, i.e. girls produce longer texts and sentences than boys. When text structure is in question, it is important that the greatest number of complex sentences (compound, complex and complex-compound), are also constructed by girls. However, differences are not significant in relation, though they occur more often in the girls' texts. Our results show that there are certain quantitative differences between girls and boys at younger elementary school age with respect of some syntactic features, while in terms of syntactic maturity, i.e. the complexity of the produced texts there are no significant differences.Kada govore i pišu, muškarci i žene na različite načine koriste jezička sredstva. Na tome se zasniva teorija o polnolektima koja pokušava da objasni u čemu se te razlike ogledaju, što je poslednjih godina podstaklo mnoga istraživanja na ovu temu. Cilj rada je da se ispitaju sintaksička svojstva tekstova učenika mlađeg osnovnoškolskog uzrasta i razlike u njihovoj produkciji između dečaka i devojčica. U istraživanju je učestvovao 181 učenik (96 dečaka i 85 devojčica) od I do IV razreda. Svi su pisali po tri sastava, tako da su u korpus za analizu ušla ukupno 543 teksta. Analiza je obuhvatila kvantitativne (dužina teksta izražena brojem reči, brojem komunikativnih rečenica i brojem klauza) i kvalitativne (različiti tipovi rečenica) sintaksičke pokazatelje. Analizirane su razlike među polovima u odnosu na ispitivane varijable. Na osnovu dobijenih rezultata može se zaključiti da se razlike između tekstova koje su pisali dečaci i devojčice na mlađem osnovnoškolskom uzrastu odnose na kvantitativne pokazatelje, dok u stepenu sintaksičke složenosti, tj. zrelosti produkovanih tekstova nema značajnih razlika

Primena Veberovih fokalno-direktrisnih ravanskih krivih u aproksimaciji konturne krive osnove arhitektonskih objekata

One of the major aims when researching some problems in architectural design of buildings is to fully understand and adequately apply the underlying scientific foundations that architects use in their projects. In this paper we analyze the application possibilities of the Weber's focal-directorial curves in the approximation of ground-base contour line of architectural objects i.e. buildings. Thus, a Weber's curve with m foci and n directrices was defined. Furthermore, particular qualifiers were introduced in order to estimate the level of adequacy of the conducted approximation. The importance of the research can be sought in the fact that the exact procedure has been created with its applicability in architectural-urban design of contemporary forms as well as in the domain of the historical heritage and conservation in the sense of the creating proper geometrical models for further computer aided use.Jedan od glavnih ciljeva istraživanja nekih problema u arhitektonskom dizajniranju zgrada je potpuno razumevanje i adekvatno primenjivanje naučnih načela koje arhitekte koriste u svojim projektima. U ovom radu analiziramo mogu nosti primene eberovih fokalno-direktrisnih krivih u aproksimaciji konture osnove arhitektonskih objekata, tj. zgrada. U vezi sa tim, definisana je Veberova kriva sa m fokusa i n direktrisa. Osim toga, uvedeni su posebni kvalifikatori kako bi se procenio nivo preciznosti izvršene aproksimacije. ažnost istraživanja se posebno iskazuje u činjenici da je postupak kreiran sa mogu noš u primene u arhitektonsko-urbanističkom dizajniranju savremenih oblika, kao i u domenu zaštite i revitalizacije istorijskog nasle a u smislu stvaranja odgovaraju ih geometrijskih modela za dalju upotrebu pomo u računara

Industrial by-products as a sources high-value polisaccharide compounds

Agricultural wastes and industrial by-products represent a major origin of carbohydrates. Obtaining hemicelluloses such as arabinoxylan and pectin from these sources presents attractive utilization of waste material. In additions, these polysaccharide products gained much attention because of their excellent biocompatibility, biodegradation and non-toxicity as well as extensive spectrum of their biological and functional properties. Polysaccharides with a molecular weight of about 20 kDa were isolated from the wheat chaff and sugar beet shreds. Obtained pectin produced gel coacervats with gelatin while arabinoxylan from wheat chaff formed a gel by mixing with β-glucan in the presence of laccase. Both of prepared gels under simulated gastric and intestinal conditions indicated a strong stability. Furthermore, biological investigation of extracted polysaccharides revealed good antioxidative activities

Early-onset ischaemic stroke in a patient with the novel F2 c.1824C>T gene variant and PAI-1 4G/4G, MTHFR 677TT genotype

Introduction. Ischemic stroke (IS) is a heterogeneous dis-order caused by several genetic and environmental risk factors. It was suggested that coagulation disorders cause 1-4% of cases with IS, especially in patients with early onset of IS. Case report. We describe a case of a young adult male who developed an unprovoked IS. Biochemical, immunological, and thrombophilia screening, as well as DNA sequencing, were performed in order to reveal molecular pathology underlying the stroke of the patient. Thrombophilia testing showed that patient was a homozygous carrier for PAI-1 4G/5G and MTHFR C677T mutations. Additional genetic analysis revealed the presence of the recently reported F2 c.1824C>T gene variant, located in the last exon of the pro-thrombin gene and has previously been shown to cause hy-perprothrombinemia, hypofibrinolysis, and altered fibrin clot phenotype. Conclusion. Our results suggest that the newly reported F2 c.1824C>T gene variant might have a synergistic effect with PAI 4G/4G and MTHFR 677TT genotype in the formation of altered fibrin clot phenotype characterized by thin, densely packed fibrin fibers, which makes clot less susceptible to fibrinolysis and greatly in-creases the risk for early ischemic stroke onset.Ishemijski moždani udar (IMU) je heterogeni poremećaj koji može biti uzrokovan genetskim faktorima rizika i faktorima sredine. Poremećaji koagulacije mogu biti uzročnici u 1-4% slučajeva IMU, naročito kod bolesnika kod kojih se IMU dogodi u mlađem životnom dobu. Prikaz bolesnika. Prikazan je slučaj bolesnika koji je u mlađem životnom dobu razvio IMU nepoznatog uzroka. Urađeni su biohemijski, imunološki i testovi za trombofiliju kao i sekvenciranje DNK sa ciljem da se utvrdi molekularna patologija koja je mogla biti u osnovi moždanog udara kod tog bolesnika. Testovima za trombofiliju utvrđeno je da je bolesnik homozigotni nosilac mutacija PAI-1 4G/5G i MTHFR C677T. Dodatnom genetičkom analizom otkriveno je prisustvo nedavno opisane F2 c.1824C>T genske varijante, koja se nalazi u poslednjem egzonu gena za protrombin i za koju je prethodno pokazano da izaziva hiperprotrombinemiju, hipofibrinolizu i izmenjeni fenotip fibrinskog ugruška. Zaključak. Naši rezultati ukazuju na to da bi nova F2 c.1824C>T genska varijanta mogla imati sinergistički efekat sa PAI 4G/4G i MTHFR 677TT genotipom u nastanku fibrinskog ugruška sa izmenjenim fenotipom, koji se odlikuje tankim, gusto upakovanim fibrinskim vlaknima, što čini ugrušak manje podložnim fibrinolizi i povećava rizik od nastanka IMU u ranijem životnom dobu

Neonatal outcome following exposure to organophosphorous pesticides

The aim of our study was to determine the neonatal outcome in mothers and children exposed to organophosphorous pesticides (OP). We found that 22.4% pregnant women were exposed to organophosphorous pesticides. OP pesticide concentration was higher in breast milk, newborn sera than maternal sera. Newborn parameters such as birth weight, birth length, head circumference, Apgar score and presence of meconium, as well as gestational age of delivery, showed no significant difference between the two groups. However, postpartum weight loss, hospitalization duration, levels of newborn bilirubin and glycaemia differed significantly between the two groups. Morbidity and presence of CNS disorders were six times and more than twelve times higher, respectively, in the OP-exposed than in the OP pesticide non-exposed group

Crevna limfangiektazija kod pasa, teško do dijagnoze - 4 slučaja

Intestinal lymphangiectasia is an uncommon disease which can cause severe, chronic protein-losing enteropathy in dogs. Four dogs were presented at the Belgrade Clinic for Small Animals with clinical signs of chronic diarrhea, lethargy, anorexia, vomiting and weight loss. Abnormal physical examination findings included dehydration, signs of pain on abdominal palpation, and ascites. The most important clinicopathological findings were lymphopenia and hypoproteinemia with hypoalbuminemia. Abdominal ultrasound revealed intestinal abnormalities in all dogs. To establish an undoubted diagnosis of intestinal lymphangiectasia, endoscopy and histopathology were conducted.Crevna limfangiektazija predstavlja retko oboljenje kod pasa koje može izazvati ozbiljne, hronične enteropatije sa gubitkom proteina. Na Klinici za male životinje u Beogradu su primljena četiri psa sa simptomima hronične dijareje, letargije, anoreksije, povraćanja i gubitka na težini. Opštim pregledom su ustanovljene promene u smislu dehidratacije, bolnosti na palpaciju abdomena i ascitesa. Najznačajnije kliničko-patološke promene su bile limfopenija i hipoproteinemija sa hipoalbuminemijom. Ultrazvučnim pregledom abdomena su ustanovljene promene na crevima kod svih pasa. Da bismo postavili nesumljivu dijagnozu crevne limfangiektazije urađena je endoskopija i histopatologija

Sex dimorphism of postural parameters of the human acetabulum

The aim of this investigation was to examine normal acetabular morphometry, its sex dimorphism and the acetabular dysplasia rate in Serbian adults. For each hip, the centre-edge angle of Wiberg, the acetabular angle of Sharp, acetabular depth and acetabular roof obliquity were measured. The center-edge angle of Wiberg correlated negatively with the acetabular angle of Sharp and acetabular roof obliquity, but positively correlated with acetabular depth. Our results suggest that the prevalence of acetabular dysplasia in the Serbian population is lower than in Western countries. We confirmed the existence of significant gender differences in acetabular morphology among the subjects of our study. These sex-related differences in acetabular morphology were the cause for more dysplastic female acetabula compared with male acetabula