Simona Bucerzan

Alterations in Lipid and Carbohydrate Metabolism in Patients with Classic Congenital Adrenal Hyperplasia due to 21-Hydroxylase Deficiency

Background: Classic 21-hydroxylase deficiency (21HD) presents some traits of the metabolic syndrome. Aim: To characterize discrete alterations of lipid and carbohydrate metabolism in children and young adults with classic 21HD, which could predict early atherogenesis. Patients and Methods: Twenty-seven Caucasian patients with classic 21HD (4–31 years); 27 sex-, age- and BMI-matched controls. Clinical parameters, hormonal status and genotype were assessed in all patients. Lipid parameters, including relative (%) and absolute (mg/dl) small-dense low-density lipoproteins subfractions (sd-LDL) were measured in patients and controls. Oral glucose tolerance tests were performed in both groups. Results: sd-LDL (%) was significantly higher in patients than controls (39.7 ± 5.9 vs. 35.5 ± 5.7%; p = 0.008). The same applies for absolute sd-LDL (mg/dl) (42.6 ± 11.9 vs. 36.4 ± 7.5; p = 0.029). HDL-cholesterol was lower in patients (p = 0.032). Fasting glucose and insulin were significantly higher in patients. Similar differences were noticed for HOMA-IR (p = 0.001), IRI (p = 0.001) and HOMA-B (p = 0.002). IRI correlated directly and significantly with the total hydrocortisone dose and the duration of treatment. Fasting glucose correlated with absolute sd-LDL. No obvious differences were seen between clinical forms or genotype groups. Conclusions: Substitution therapy should be adapted particularly at young ages to prevent early atherogenesis and cardiovascular risk in later life.

Baseline characteristics and outcome in Romanian patients with Gaucher disease type 1

BACKGROUND/AIM To present clinical and genetic characteristics of all Romanian patients with Gaucher disease type 1, in whom specific diagnosis has been confirmed by enzymatic and molecular methods and to analyze their outcome with and without enzymatic replacement therapy (ERT). PATIENTS, METHODS There are fifty patients (F/M - 1.63/1) with Gaucher disease type 1. Clinical status, haemoglobin, thrombocytes, hepatic/splenic volume, bone mineral density and severity score were assessed at baseline and every six months thereafter. Thirty-nine patients (78%) received imiglucerase (44.4+/-13.6 U/kg/2 weeks) for 3.1+/-1.4 years. RESULTS Based on general prevalence data, our group represents 22.7% of the expected total number of patients with Gaucher disease type 1 in Romania. Mean age was 15.5 years at clinical onset and 28.9 years at confirmation of diagnosis. The genotype N370S/L444P was frequent in our group (35.9% of alleles). Anaemia, thrombocytopenia, splenomegaly and bone disease were present at 38%, 70%, 100% and 84%, respectively. Mean values for haemoglobin, thrombocytes, hepatic volume and chitotriosidase normalized after 0.5, 1.5, 2.5 and 3 years of ERT, respectively. Splenomegaly regressed from 14.4 x N (normal) to 3.06 x N over four years of treatment. Bone disease was ameliorated under ERT, yet bone mineral density worsened in patients treated with 30 U/kg/2 weeks. CONCLUSIONS The genotype N370S/L444P is frequent in our patients, in line with the severe phenotypes. ERT improved haematological parameters and visceromegaly, without a clear benefit for bone mineral density. To attain therapeutic goals, an early treatment start with optimal dosage is mandatory.

Clinical and genetic characteristics in a group of 45 patients with Turner syndrome (monocentric study)

Introduction Recent years have seen a shift in perspective on Turner syndrome, as it is no longer considered a significant disability due to therapeutic advances. The delay of diagnosis and the underdiagnosis are common in Turner syndrome, especially because of the great phenotypic variability and lack of firm diagnostic criteria. Aim Our first aim was to assess the clinical and the cytogenetic characteristics and growth rate in growth hormone (GH)-treated patients as compared to those with spontaneous growth. The second aim was to analyze the Y chromosomal sequences. Materials and methods We analyzed 45 patients diagnosed with Turner syndrome in Genetic Pathology Centre of Cluj Emergency Children’s Hospital. We carried out a study of the clinical features, the correlations between the karyotype and the phenotype, and we also made a research of Y chromosome sequences. Results The average age at diagnosis was 8.9±5.4 years. A significant association was observed between the number of external phenotypical abnormalities and internal malformations (r=0.45), particularly the cardiovascular ones (r=0.44). Patients treated with GH showed improvement in growth rate, with final stature significantly better than in untreated patients; benefits following treatment were greater if diagnosis was made before the age of 5 years. Thirteen percent of patients experienced spontaneous and complete puberty, whereas 30% experienced incomplete puberty. Patients with the 45,X genotype had a greater stature deficit and a higher incidence of cardiac malformations, compared with patients with 45,X/46,XX mosaic karyotype. Y chromosome sequences were found in only one patient, who subsequently underwent gonadectomy. Conclusion The importance of this study resides, to the best of our knowledge, in the fact that the largest group of patients in Romania was analyzed and assessed. To draw firm conclusions on the most valuable clinical indicators for Turner syndrome diagnosis in clinical practice, studies on large groups of patients should be conducted.

The c.301_302delAG PROP1 gene mutation in Romanian patients with multiple pituitary hormone deficiency.

Abstract Objective: To establish the frequency of the c.301_302 delAG mutation of the PROP1 gene in Romanian patients with multiple pituitary hormone deficiency (MPHD). Subjects and methods: Somatic assessment, hormonal test, bone age, magnetic resonance imaging of the pituitary gland, and molecular diagnosis were performed in 26 patients with MPHD (7 patients with familial form of MPHD and 19 patients with sporadic form of MPHD). Results: The c.301_302delAG mutation was detected in the homozygous state in 10 patients belonging to 5 unrelated families (7 patients with familial history of MPHD and 3 patients with sporadic form of MPHD). Those 10 patients presented variable pituitary hormone deficiency and pituitary morphology. Conclusions: The c.301_302delAG homozygous genotype had a high frequency of 38% (10/26), reaching 100% (7/7) in group with familial cases of MPHD and 16% (3/19) in group with sporadic forms of MPHD.

Evaluation of c677t and a1298c polymorphism of the methylenetetrahydrofolate reductase gene as a maternal risk factor for trisomy 21 (a monocentric study)

Abstract Aim: To assess the risk for trisomy 21 in children, depending on the polymorphisms C677T and A1298C of the methylenetetrahydrofolate reductase (MTHFR) gene in mothers. Methods: For 93 mothers who have children with trisomy 21 and 202 mothers of healthy children (control group), genotyping of MTHFR polymorphisms C677T and A1298C was performed. Results: For each polymorphism, three genotypes were identified (normal homozygous, heterozygous and mutant homozygous). For the polymorphism C677T, the frequencies of the three genotypes (CC, CT and TT) were 50.5%, 40.8% and 8.6% in mothers of children with trisomy 21, versus 42.6%, 46% and 11.4% in mothers of healthy children, with no statistically significant differences. The frequency of the polymorphism A1298C was not statistically significant between the two groups for the genotype (AA) (48.4% vs 56.4%) or the genotype (AC) (39.8% vs 38.6%), but the genotype TT was more frequent in mothers of children with trisomy 21 (11.8% vs 4.9%; p = 0.033; OR = 2.57). Conclusion: Women with genotype CC for the polymorphism A1298C of the MTHFR gene have a 2.57 times higher risk of offspring with trisomy 21.

OC-88 Clinical and genetic characteristics, diagnosis, treatment and outcome in patients with mucopolysaccharidosis tipe i and tipe ii in our country

Introduction Mucopolysaccharides (MPS) are a group of rare diseases with chronic progression caused by excessive intralysosomal accumulation of glycosaminoglycans (GAG), due to certain acid hydrolases deficiencies. The unchanged mucopolysaccharides accumulate in the lysosomes of reticuloendothelial system cells, resulting in severe functional and structural alterations in various tissues and organs. Aim of the study was to assess the clinical, genetic characteristics and outcome under enzyme replacement treatment (ERT) of Romanian MPS type I and type II patients. patients and methods. The study group included five MPS type I and nineteen MPS type II patients (4 girls and 20 boys), aged between 1–14 years. The study methods consisted in: clinical and standard auxological assessment; goniometry; neurological and psychological evaluation; otorhinolaryngology examination with audiogram, ophthalmological examination, cardiology evaluation (ECG, Doppler echocardiography), spirometry, ultrasonography, specific enzyme activity assay and molecular analysis (PCR and sequencing). Results All patients presented coarse facial features, organomegaly, arthropathy, cardiac involve-ment and respiratory difficulties, but variable neurological impairment. The clinical onset was at 1.53±0.74 years; the unspecific diagnosis was established at 4.86±8.6 years and specific diagnosis at 5.84±6.2 years. Molecular analysis revealed 11 unreported mutations. ERT was started at 7.7±5.28 years (limits between 1.5 years and 20.25 years). ERT reduces organomegaly and ameliorates rate of growth, joint mobility, reduces the number of episodes of respiratory infections, but does not prevent deterioration of cognition and other neurologic functions. Conclusions An important gap between the clinical onset and specific diagnosis resulted in this group. The ERT should be started at an early age prior to irreversible injuries installation.

OC-87 Gaucher disease in romania – baseline characteristics, specific diagnosis. treatment and outcome

Gaucher disease is a autosomal recessive inherited monogenic disease caused by beta-glucocerebrosidase deficiency. Clinically, there are three types: type 1 (non-neuronopathic), type 2 (acute neuronopathic) and type 3 (chronic neuronopathic), in 92%, 1% and respectively 7% of patients. Specific diagnosis has been possible in Romania since 1997 and enzyme replacement therapy since 2002. The aim of the study is to present the epidemiologic, clinical and molecular data of the Romanian patients with Gaucher disease ant their evolution. Patients and methods Seventy-nine patients (76 patients with Gaucher disease type 1 and 3 patients with Gaucher disease type 3; F/M=1.37/1) were evaluated clinically and by measurement of haemoglobin, thrombocytes, hepatic and splenic volume, chitotriosidase, bone density (Z score) and severity index at diagnosis and every each 6–12 months. Sixty-nine patients were treated with imiglucerase for a period of 1–13 years. Results Gaucher disease prevalence in our country, evaluated according to the number of patients diagnosed, is 1/250,000 vs. the potential prevalence of 1/100,000. Mean age at clinical onset was 15.4 years and 28.8 years at specific diagnosis. We found a high incidence of genotype N370S/L444P (38%) and of genotype N370S/? (32%), predictive for a severe phenotype vs 15.3% and 9.6% respectively in patients reported in International Gaucher Registry. Splenectomy was effectuated in 30% patients before the enzyme replacement therapy. Anaemia, thrombocytopenia, splenomegaly and bone disease was present in 52%, 37.7%, 100% and 91% of patients. The status of untreated patients was progressively worsening and four of the patients died. Enzyme replacement therapy led to normalisation of haemoglobin, thrombocytes, hepatic volume and chitotriosidase after 0.5, 1.5, 2 and 3 years respectively. After 4 years of treatment, splenomegaly has been reduced from 14.4 of normal to 3.06 of normal. Clinical evolution of bone disease was favourable. Conclusion Gaucher disease is still undiagnosed in our country. Severe forms of disease are more prevalent. The patients have access to specific, enzyme and molecular diagnosis. Evolution under enzyme replacement therapy is very good.

Ultrasonographic evaluation of the median nerve at the level of the carpal tunnel outlet and mid forearm in patients with type II Mucopolysaccharidosis

AIMS The ultrasonographic (US) evaluation of the median nerve at the level of the carpal tunnel outlet (CTO) and mid forearm in pediatric patients with mucopolysaccharidosis type II (MPS II) and comparison with healthy subjects. MATERIAL AND METHOD Fifteen children with MPS II and 44 healthy children were included in the study and they were divided into three age groups. The cross-sectional area, the appearance of the nerve, and the ratio of the cross-sectional areas were evaluated by US. RESULTS At the level of the CTO the mean area of the nerve was increased in all MPS II groups compared with the correspondent healthy age groups and the differences were statistically significant (p<0.01). At the level of the mid forearm the differences were statistically significant only for the first age group. Other US findings at the level of the CTO in the MPS II groups were represented by hypoechogenicity (86.67 % on the right and 93.33% on the left), thickened fascicles (80% bilaterally), irregular contour (53.33% bilaterally) and the presence of the Doppler signal including the nerve (26.67 % on the right and 33.33 % on the left). The CTO/mid forearm cross-sectional area ratio was higher in all MPS II age groups and the differences were statistically significant (p<0.001). CONCLUSION In patients with MPS II there are significant US changes in the size and aspect of the median nerve.

Clinical and Genetic Characteristics of Romanian Patients with Mucopolysaccharidosis Type II

BACKGROUND Mucopolysaccharidosis type II (MPS II, Hunter syndrome) is a rare X-linked disorder caused by deficiency of iduronate-2-sulfatase (I2S) enzyme, which leads to the accumulation of partially digested glycosaminoglycans (GAGs) in the lysosomes and induces multisystemic alteration (coarse facial features; skeletal dysplasia; hepatosplenomegaly; joint stiffness and contractures; heart, lung, vision, and hearing disability; profound neurological decline).The purpose of this study is to present the clinical and genetic characteristics of Romanian patients with Hunter syndrome and the genotype-phenotype correlation. MATERIAL AND METHODS 15 unrelated patients, with MPS II ranging from mild (4 subjects) to severe phenotype (11 subjects) aged 2 to 20 years, were evaluated clinically, cognitive development, enzyme assay and molecular analysis. RESULTS The molecular analysis of the 15 unrelated Romanian MPS II patients has identified 15 different mutations (2 major genetic defects (13%) and 13 minor genetic defects (87%)): microdeletions and point mutations (missense, nonsense), seven of them described for the first time-deletion encompassing 3 to exon 7; c823G>T, pD275Y; c.1600A>C (pN534H); c.102_10delAG (p.D5Cfs*11); c.448_471del (p.P150_P157del); c.421delA (p.I141Yfs*72); and c.419-1G>C. The major genetic defects were correlated with a severe course of disease. CONCLUSION This is the first study on the clinical and molecular characterization of the MPS II Romanian patients. This study supports the evidence of the mutational heterogeneity of the I2S gene as well as the difficulty to correlate genotype and phenotype in the patients with MPS II.

A Novel Nonsense Mutation of the AGL Gene in a Romanian Patient with Glycogen Storage Disease Type IIIa.

Background. Glycogen storage disease type III (GSDIII) is a rare metabolic disorder with autosomal recessive inheritance, caused by deficiency of the glycogen debranching enzyme. There is a high phenotypic variability due to different mutations in the AGL gene. Methods and Results. We describe a 2.3-year-old boy from a nonconsanguineous Romanian family, who presented with severe hepatomegaly with fibrosis, mild muscle weakness, cardiomyopathy, ketotic fasting hypoglycemia, increased transaminases, creatine phosphokinase, and combined hyperlipoproteinemia. GSD type IIIa was suspected. Accordingly, genomic DNA of the index patient was analyzed by next generation sequencing of the AGL gene. For confirmation of the two mutations found, genetic analysis of the parents and grandparents was also performed. The patient was compound heterozygous for the novel mutation c.3235C>T, p.Gln1079⁎ (exon 24) and the known mutation c.1589C>G, p.Ser530⁎ (exon 12). c.3235 >T, p.Gln1079⁎ was inherited from the father, who inherited it from his mother. c.1589C>G, p.Ser530⁎ was inherited from the mother, who inherited it from her father. Conclusion. We report the first genetically confirmed case of a Romanian patient with GSDIIIa. We detected a compound heterozygous genotype with a novel mutation, in the context of a severe hepatopathy and an early onset of cardiomyopathy.