Ambiguous Genitalia and Lissencephaly in A 46,XY Neonate with a Novel Variant of Aristaless Gene.

Introduction: Disorders of sexual development can present isolated or as a part of complex genetic syndromes. Case presentation: A newborn with ambiguous genitalia and prenatally diagnosed brain malformations was referred to our hospital. Prenatal ultrasound examination and MRI showed lissencephaly and absence of the corpus callosum. At admission, physical examination revealed microphallus, hypospadia and complete fusion of labioscrotal folds with nonpalpable gonads, normal blood pressure and serum biochemistry. Cortisol level was normal (201 nmol/L), testosterone elevated (14.4 nmol/L), FSH 0.1 IU/L, LH 0.7 IU/L, estradiol 241 pmol/L. Seizures were noted on the 2nd day and the child was started on anticonvulsives. When 17-OHP level results came back elevated (200 nmol/L), ACTH test was performed and the child was started on hydrocortisone and fludrocortisone treatment. Congenital adrenal hyperplasia became unlikely when karyotype result showed normal male karyotype (46, XY, SRY+) with no Mullerian structures seen on ultrasonographic exam. As association of ambiguous genitalia and lissencephaly strongly suggested a mutual genetic background, diagnosis of X-linked lissencephaly with ambiguous genitalia (X-LAG) became apparent. Conclusions: The presented case highlights the importance of looking at the whole clinical picture instead of separate isolated findings with emphasis on patient-centered approach guided by clinical findings and patient history.

Alignment of dynamic networks.

MOTIVATION: Network alignment (NA) aims to find a node mapping that conserves similar regions between compared networks. NA is applicable to many fields, including computational biology, where NA can guide the transfer of biological knowledge from well- to poorly-studied species across aligned network regions. Existing NA methods can only align static networks. However, most complex real-world systems evolve over time and should thus be modeled as dynamic networks. We hypothesize that aligning dynamic network representations of evolving systems will produce superior alignments compared to aligning the systems' static network representations, as is currently done. RESULTS: For this purpose, we introduce the first ever dynamic NA method, DynaMAGNA ++. This proof-of-concept dynamic NA method is an extension of a state-of-the-art static NA method, MAGNA++. Even though both MAGNA++ and DynaMAGNA++ optimize edge as well as node conservation across the aligned networks, MAGNA++ conserves static edges and similarity between static node neighborhoods, while DynaMAGNA++ conserves dynamic edges (events) and similarity between evolving node neighborhoods. For this purpose, we introduce the first ever measure of dynamic edge conservation and rely on our recent measure of dynamic node conservation. Importantly, the two dynamic conservation measures can be optimized with any state-of-the-art NA method and not just MAGNA++. We confirm our hypothesis that dynamic NA is superior to static NA, on synthetic and real-world networks, in computational biology and social domains. DynaMAGNA++ is parallelized and has a user-friendly graphical interface. AVAILABILITY AND IMPLEMENTATION: http://nd.edu/∼cone/DynaMAGNA++/ . CONTACT: tmilenko@nd.edu. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Molecular genetic study of congenital adrenal hyperplasia in Serbia: novel p.Leu129Pro and p.Ser165Pro CYP21A2 gene mutations.

PURPOSE: Congenital adrenal hyperplasia (CAH) is an autosomal recessive disease characterized by impaired adrenal steroidogenesis and most often caused by CYP21A2 gene mutations. For the first time, we reported complete spectrum and frequency of CYP21A2 gene mutations in 61 unrelated patients with classical and non-classical CAH from Serbia. METHODS: Direct DNA sequencing of whole CYP21A2 gene and polymerase chain reaction with sequence-specific primers for detection of CYP21A1P/CYP21A2 chimeras were combined. RESULTS: We identified 18 different pathogenic alleles-two of them novel. Mutation detection rate was highest in patients with salt-wasting form of CAH (94.7%). The most prevalent mutation was intron 2 splice site mutation, c.290-13A/C>G (18.5%). Other mutation frequencies were: CYP21A1P/CYP21A2 chimeras (13%), p.P30L (13%), p.R356W (11.1%), p.G110fs (7.4%), p.Q318X (4.6%), p.V281L (4.6%), p.I172N (2.8%), p.L307fs (2.8%), p.P453S (1.9%), etc. Mainly, frequencies were similar to those in Slavic populations and bordering countries. However, we found 6.5% of alleles with multiple mutations, frequently including p.P453S. Effects of novel mutations, c.386T>C (p.Leu129Pro) and c.493T>C (p.Ser165Pro), were characterized in silico as deleterious. The effect of well-known mutations on Serbian patients' phenotype was as expected. CONCLUSIONS: The first comprehensive molecular genetic study of Serbian CAH patients revealed two novel CYP21A2 mutations. This study will enable genetic counseling in our population and contribute to better understanding of molecular landscape of CAH in Europe.

Exploring the structure and function of temporal networks with dynamic graphlets.

MOTIVATION: With increasing availability of temporal real-world networks, how to efficiently study these data? One can model a temporal network as a single aggregate static network, or as a series of time-specific snapshots, each being an aggregate static network over the corresponding time window. Then, one can use established methods for static analysis on the resulting aggregate network(s), but losing in the process valuable temporal information either completely, or at the interface between different snapshots, respectively. Here, we develop a novel approach for studying a temporal network more explicitly, by capturing inter-snapshot relationships. RESULTS: We base our methodology on well-established graphlets (subgraphs), which have been proven in numerous contexts in static network research. We develop new theory to allow for graphlet-based analyses of temporal networks. Our new notion of dynamic graphlets is different from existing dynamic network approaches that are based on temporal motifs (statistically significant subgraphs). The latter have limitations: their results depend on the choice of a null network model that is required to evaluate the significance of a subgraph, and choosing a good null model is non-trivial. Our dynamic graphlets overcome the limitations of the temporal motifs. Also, when we aim to characterize the structure and function of an entire temporal network or of individual nodes, our dynamic graphlets outperform the static graphlets. Clearly, accounting for temporal information helps. We apply dynamic graphlets to temporal age-specific molecular network data to deepen our limited knowledge about human aging. AVAILABILITY AND IMPLEMENTATION: http://www.nd.edu/∼cone/DG.

MAGNA++: Maximizing Accuracy in Global Network Alignment via both node and edge conservation.

MOTIVATION: Network alignment aims to find conserved regions between different networks. Existing methods aim to maximize total similarity over all aligned nodes (i.e. node conservation). Then, they evaluate alignment quality by measuring the amount of conserved edges, but only after the alignment is constructed. Thus, we recently introduced MAGNA (Maximizing Accuracy in Global Network Alignment) to directly maximize edge conservation while producing alignments and showed its superiority over the existing methods. Here, we extend the original MAGNA with several important algorithmic advances into a new MAGNA++ framework. RESULTS: MAGNA++ introduces several novelties: (i) it simultaneously maximizes any one of three different measures of edge conservation (including our recent superior [Formula: see text] measure) and any desired node conservation measure, which further improves alignment quality compared with maximizing only node conservation or only edge conservation; (ii) it speeds up the original MAGNA algorithm by parallelizing it to automatically use all available resources, as well as by reimplementing the edge conservation measures more efficiently; (iii) it provides a friendly graphical user interface for easy use by domain (e.g. biological) scientists; and (iv) at the same time, MAGNA++ offers source code for easy extensibility by computational scientists. AVAILABILITY AND IMPLEMENTATION: http://www.nd.edu/∼cone/MAGNA++/

Risk assessment of cardiovascular mortality in Macedonia type 2 diabetes patients based on decode model.

AIM: To estimate the absolute risk (%) of 5- and 10-years cardiovascular mortality in Macedonian type 2 diabetes patients based on DECODE model, and the gender difference of the estimated risk. METHODS AND MATERIALS: Observational, cross-sectional study including a cohort of 1,404 type 2 diabetes patients; inclusion criteria: aged 25 to 65 years, absence of confirmed arterial disease, history of ischaemic heart disease, cerebrovascular disease or peripheral arterial disease; and absence of life-threatening conditions, such as cancer; at the time of risk assessment. Absolute risk was assessed based on the following risk factors: gender, age, known diabetes, smoking status, systolic blood pressure and total cholesterol. RESULTS: From the study cohort, 884 were identified as eligible for analysis, 503 (56.9%) of these were women. The estimated absolute risk (%) of 5- and 10-year cardiovascular mortality, based on DECODE model, was 1.1±1.3% and 5.5±6.1%, respectively; significantly higher absolute risk was estimated in men (1.7±1.6 vs 0.6±0.8, p<0.001 and 8.9±7.6 vs 2.9±2.5, p<0.001, for 5- and 10-years absolute risk, respectively). DISCUSSION AND CONCLUSION: This study is a first assessment of cardiovascular mortality in the Macedonian type 2 diabetic population based on DECODE model. It would be of both clinical and scientific interest to assess the risk prediction accuracy of the model, and to compare it with other diabetes-specific and diabetes non-specific models.

Graphlet-based edge clustering reveals pathogen-interacting proteins.

MOTIVATION: Prediction of protein function from protein interaction networks has received attention in the post-genomic era. A popular strategy has been to cluster the network into functionally coherent groups of proteins and assign the entire cluster with a function based on functions of its annotated members. Traditionally, network research has focused on clustering of nodes. However, clustering of edges may be preferred: nodes belong to multiple functional groups, but clustering of nodes typically cannot capture the group overlap, while clustering of edges can. Clustering of adjacent edges that share many neighbors was proposed recently, outperforming different node clustering methods. However, since some biological processes can have characteristic 'signatures' throughout the network, not just locally, it may be of interest to consider edges that are not necessarily adjacent. RESULTS: We design a sensitive measure of the 'topological similarity' of edges that can deal with edges that are not necessarily adjacent. We cluster edges that are similar according to our measure in different baker's yeast protein interaction networks, outperforming existing node and edge clustering approaches. We apply our approach to the human network to predict new pathogen-interacting proteins. This is important, since these proteins represent drug target candidates. AVAILABILITY: Software executables are freely available upon request. CONTACT: tmilenko@nd.edu SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Type 2 diabetes mellitus and impaired glucose regulation in overweight and obese children and adolescents living in Serbia.

An increase in the prevalence of pediatric type 2 diabetes mellitus (T2DM) has been reported by numerous studies in the United States during the past two decades. Available data from Europe are scarce, but also suggest the rising prevalence of this disease in overweight children. The aim of this study was to determine the prevalence of previously undiagnosed T2DM, impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) in a clinic cohort of otherwise healthy overweight and obese Caucasian children and adolescents living in Serbia. The study group consisted of 301 subjects (176 girls, 125 boys) aged 5.2-18.9 years, with body mass index >90th percentile. Oral glucose tolerance test was performed in all subjects. Previously undiagnosed T2DM was discovered in 0.3% (n=1) and impaired glucose regulation in 15.9% (n=48) of the subjects. Isolated IFG was detected in 4.3% (n=13), isolated IGT in 8.3% (n=25) and combined IFG and IGT in 3.3% (n=10) of the subjects. Disturbances of glucose metabolism were present in a substantial number of the subjects, which emphasizes the need for prevention and treatment of childhood obesity.

Molecular analysis of ring y chromosome in a 10-year-old boy with mixed gonadal dysgenesis and growth hormone deficiency.

Ring Y chromosome is a very rare chromosomal aberration. The published mixed gonadal dysgenesis (MGD) patients with a ring Y chromosome are short in stature, but are not growth hormone (GH) deficient. We present the molecular cytogenetic and molecular characterization of ring Y chromosome mosaicism in a 10-year-old boy with MGD whose short stature could be explained by the high percentage of cells monosomic for the X-chromosome, but also by the presence of severe GH deficiency. The ring Y chromosome in our patient is a de novo structural aberration. The father's karyotype was normal.

Insulin pump therapy with continuous glucose monitoring improves metabolic control in brittle type 1 diabetes.

AIM: to evaluate the combination of insulin pump therapy and continuous glucose monitoring in outcome on metabolic control in patients with brittle type 1 diabetes. MATERIALS AND METHODS: Insulin pump therapy was initiated in eleven brittle type 1 diabetics with poor metabolic control (mean Hba1c = 9.6%). Metabolic control was evaluated with CGMS and HbA1c in the following 6 months. RESULTS: Glycated haemoglobin showed a reduction in 1.4% in the 6 months following initialisation of pump therapy. Physical activity, various foods and insulin were tested with CGMS. There were no severe hypoglycaemia and occasional postprandial hyperglycaemia, where patients and their family learned the practical issues of carbohydrate counting. During the next 6 months on pump therapy, the patients successfully managed their diabetes. CONCLUSIONS: Insulin pump therapy can be initiated and used effectively in brittle type 1 diabetics to improve metabolic control and quality-of-life. When diabetes and pump management are appropriately individualized, this kind of therapy can help type 1 diabetics to achieve and to sustain metabolic control. Lifestyle flexibility, quality-of-life improvement, and independence can be maintained throughout young adulthood.

[Hypertension and diabetes mellitus]. / Hipertensija i dijabetes melitus.

AIM: To evaluate hypertension in patients with Diabetes Mellitus (DM) and its correlation with age, duration of DM, Body Mass Index (BMI) and HbA1C). MATERIALS AND METHODS: A retrospective study was made on 1211 patients with DM (male 554 and female 657), hospitalized at Clinic of Endocrinology between January 2001 and December 2002. Patients were divided in two groups: Control group (CG)-subdivided into 3 groups patients with DM type 1 (CG-1), DM type 2 on oral anti-hyper-glycemic agents (CG-2)and DM type 2 on insulin therapy (CG-3) and Examined Group (EG), the same groups for diabetes, including hypertension. RESULTS: We found hypertension in 12.6% patients with DM type 1, 30.5% in DM type 2 on oral anti-hyper-glycemic agents and 33.4% in DM type 2 on insulin therapy. CONCLUSION: Hypertension is mostly presented in DM type 2 patients (33,4%), instead of 12.6% in DM type 1. There is statistical significance (p<0.05) between duration of DM in patients with and without hypertension.

[Causes of ambiguous external genitalia in neonates]. / Uzroci dvopolnog izgleda spoljasnjih genitalija kod novoredjencadi.

INTRODUCTION: The classification of disorders such as ambiguous genitalia in newborns is difficult because similar or identical phenotypes could have several different aetiologies. In most cases it was impossible to correlate the aetiology of the disorder and the appearance of the external genitalia [1-3]. A newborn with ambiguous genitalia needs prompt evaluation that will permit gender assignment and detection of life-threatening conditions (salt-losing crisis due to congenital adrenal hyperplasia or Wilms' tumour). We studied the causes and characteristics of ambiguous genitalia in newborn infants over the period from 1990 to 1999. PATIENTS AND METHODS: The following genital phenotypes are considered as ambiguous: 1. Hypospadias with no palpable gonads; 2. Hypospadias with micropenis and no palpable gonads or one palpable gonad; 3. Newborn with female external genitalia and a gonadal mass in labia or labial fusion and/or clitoral enlargement [1, 4]. The diagnostic evaluation of newborns with ambiguous genitalia consisted of history and physical examination, determination of serum electrolytes, plasma 17-hydroxyprogesterone (17-OHP), chromosome analysis on cultured lymphocytes, sonogram of the abdomen in connection with a genitogram; and whenever it was necessary, basal plasma concentrations of testosterone and, after the stimulation with human chorionic gonadotropin (hCG), laparotomy for definitive determination of gonadal histology. All disorders with ambiguous genitalia have been classified in four groups: [6]: 1. Female pseudohermaphroditism (FPH); 2. Male pseudoherma phroditism (MPH); 3. True hermaphroditism (TH); 4. Asymmetrical gonadal dysgenesis (ASGD). RESULTS: The causes of sexual differentiation disorders in a group of 38 newborns with ambiguous genitalia are presented in Table 1. Main criteria for the diagnosis of FPH were normal female karyotype 46, XX, masculinization of external genitalia and no palpable gonads. Genitography revealed urogenital sinus and vagina, and ultrasound examination the uterus. During initial examination seven of 15 newborns with congenital adrenal hyperplasia (CAH) (Table 2) due to 21-hydroxylase (P450c21) deficiency (21-OHD) had clinical or laboratory signs of adrenal crisis. Two children had a simple virilizing form of 21-OHD. The female gender was chosen for these children. In other three patients with FPH isolated clitoral hyperplasia or labial fusion was the main reason for the studies. The common characteristics of newborns with MPH were as follows: normal male karyotype 46,XY with normally developed or dysgenetic testes, and/or good response to hCG stimulation. The complete androgen insensitivity (testicular feminization) was detected in two children (Table 3) with female external genitalia and palpable gonads in the labial folds, and female gender was chosen. The Denys-Drash syndrome was detected in one newborn with ambiguous genitalia, no palpable gonads, and normal response to hCG, and ultrasound findings of multiple bilateral renal tumours were identified as Wilms' tumour. In other newborns with MPH incomplete masculinization consisted of hypospadias, mostly of perineoscrotal type and of micropenis (penile size less than 2 cm) and/or bilateral or unilateral cryptorchidism (Table 3). In all children male sex was chosen. Asymmetrical gonadal dysgenesis was detected in two newborn infants. Both children had 46,XY/46,XX karyotype, testes on one side of the abdomen, and streak gonad on the other, developed vagina, uterus and unilateral Fallopian tube, and were raised as females. True hermaphroditism was established in one newborn with 46,XX karyotype, with a testis on one side of the abdomen and an ovotestis on the other side. The parents decided for male gender. The aetiology of ambiguous genitalia was not established in five children; in two children with 46,XY and one with 46,XX karyotype (with palpable gonads) the diagnostic study was not completed. CONCLUSIONS: The most common cause of ambiguous genitalia in our newborn patients was CAH due to 21-OH deficiency [2, 4, 6, 7]; 87 percent of patients had salt wasting form of the disease. In the majority of patients the appearance of the external genitalia made possible the detection of the disease immediately after the birth. So, the relative high incidence of adrenal crisis in our patients with CAH (38%) seems unreasonable. The decision for gender assignment was possible after the appropriate study of the nature of the disorder. The causes of MPH are numerous and heterogeneous [1, 3, 8]. With the exception of two patients with complete form of androgen insensitivity, in all newborns with MPH the male gender predominated. The appearance of external genitalia with severe perineoscrotal hypospadia and/or micropenis suggested the possibility of incomplete androgen resistance. If a male assignment is being considered, the response of the phallic size to treatment with testosterone was recommended. If penile size did not reach the 2.5 cm range or above, a male sex assignment was not advisable [1]. It is important for the paediatric surgeon to be involved in the diagnostic evaluation of these infants to plan the timing and techniques of the surgical reconstruction [6]. The decision to raise a patient with sex chromosome mosaicism, true hermaphroditism, or mixed gonadal dysgenesis as either a male or a female was based on the appearance of the external genitalia and possible fertility [1, 9]. The parental decision of male sex in our patients with true hermaphroditism could not be considered as optimal.

Transient neonatal diabetes mellitus in a child with paternal uniparental disomy of chromosome 6.

We report a male infant with transient neonatal diabetes mellitus (TNDM; MIM 601410), macroglossia, hypertelorism, umbilical hernia, inguinoscrotal hernia and onychomycosis. Diabetes mellitus was diagnosed 10 days after birth and resolved after 6.5 months of treatment. Genetic investigation indicated the presence of paternal uniparental disomy of chromosome 6 (UPD 6). The finding of paternal UPD 6 allows prediction of a transient, rather than permanent NDM, and no increased recurrence risk of TNDM in subsequent pregnancies. Therefore, finding of NDM should be a strong indicator for genetic testing.

[Ectopy of the thyroid gland as cause of neck or tongue base tumour]. / Ektopija tireoidne zlezde kao uzrok tumora na vratu ili na bazi jezika.

One of the most important causes of congenital hypothyroidism found in 35-42% of cases, is ectopy of the thyroid gland. Thyroid gland can be displaced at base or under the tongue, or under the hyoid bone. Ectopic gland is also hypoplastic, secreting not enough amounts of thyroid hormones. By negative feedback mechanism that cause elevation of TSH. Under the permanent TSH stimulation ectopic gland can enlarge appearing as a neck or tongue base "tumour". In this way, by measuring TSH level in a newborn, all children with thyroid gland ectopy can be detected. Ectopy of the thyroid gland as cause of congenital hypothyroidism was present in three patients as a tumour of neck or tongue base. After surgical removal of the "tumour", histopathologic analysis revealed that it was the thyroid tissue. No patient passed the thyroid function test nor identification of the thyroid tissue (ultrasound or scintigraphy) before surgery. All were born in the regions of Serbia where screening for congenital hypothyroidism was not carried out at all or only temporary. Screening of newborns for congenital hypothyroidism is based on measuring TSH level. By this method all patients with thyroid gland ectopy can be detected. Scintigraphic examination after surgery detected no thyroid tissue and replacement therapy with Na L-thyroxine started.

[Relationship between height velocity and growth hormone secretion in normal and short prepubertal children]. / Brzina rastenja i sekrecija hormona rasta kod dece niskog i normalnog rasta u prepubertetskom uzrasty.

In 60 prepubertal children of both sexes, aged between 6.4 and 15.5 years, with normal or short stature and growing with heigh velocity standard aviation scores (SDS) between 2.69 and -5.75, we has performed 12h noctunral growh hormone (GH) profiles. We found a statistically significant relationship between growth velocity and growth hormone secreation in the group of 60 children with different degrees of GH deficiency, short normal children and children with normal stature. The relationship was expressed with a logarithmic type of curve. In the gruop of 13 children with constituonal growth delay or in the group of 9 children with the familial short stature, we did not find a significant relationship between growth velocity and GH secretion. In the group of short children with complete GH deficiency there were marked individual differences in growth velocity after the beginning of thyroid replacement therapy; children with secondary hypothyroidsm were growing significantly faster than children with normal thyreotrpic function. In our opinion the height velocity is controlled with thyroid hormones, genetic influences, and possibly, with other unknown factors which operate independntly of GH secreation during prepubertal years.