genome-wide linkage scan in a moroccan family with autosomal-recessive exstrophy of the bladder...
TRANSCRIPT
S24 ESPU Programme 2009
DP70 camera. The Stereological analysis wasdone with the software Image J Pro usinga grid to determine volumetric densities(Vv). For the biochemical analysis, tissuesamples were fixed in acetone, delipidated,and dried. A hyroxyproline assay was thenperformed and total collagen content wasexpressed as mg hyroxyproline per mg drytissue. Means were statistically comparedusing the ANOVA and Unpaired T test(p< 0.05)
RESULTS
Gestational age ranged from 12 to 25 weekspost conception (WPC). Quantitative
analysis documented a increase in Vv smoothmuscle cells in male urachus (23.02%), whencompared to female (18.43%), there was nostatistically significance (p¼ 0.1478).Quantitative analysis documenteda increase in Vv of connective tissue infemale urachus (67.64%), when compared tomale (58.38%), there was a statisticallysignificance (p¼ 0.0439). Total collagenconcentration in male urachus(mean¼ 45656mg/mg), and in female(mean¼ 42308mg/mg) did not differsignificantly (p¼ 0.5912). As higher was thegestational age, smaller was the urachallumen area. In 13WPC fetuses the urachallumen was 16301mm2 and in 17WPC fetuses
the urachal lumen area was 1676mm2. Theurachal lumen was closed from the 18thWPCin male and female
CONCLUSIONS
In this work we determine that the urachallumen was closed from the 18WPC in allfetuses. The data obtained in the presentstudy can be used as base knowledge relatedto the development of the urachus.
# B02-4 (PP)
SERUM BASAL INHIBIN B AND ANTIMULLERIAN HORMONE LEVELS TO SHOW TESTICULAR TISSUE EXISTENCE: ANEXPERIMENTAL STUDY IN RATSMurat DAYANC1, Hasan Cem IRKILATA1, Yusuf KIBAR1, Ediz YESILKAYA2, Ugur MUSABAK3 and Seref BASAL1
1Gulhane Military Medical Academy, Urology, Ankara, TURKEY, 2Gulhane Military Medical Academy, Pediatric Endocrinology,Ankara, TURKEY, 3Gulhane Military Medical Academy, Immunology, Ankara, TURKEY
PURPOSE
Diagnostic laparoscopic is the mostappropriate method to approach childrenwith bilateral non-palpable testes asimaging studies are not considered to beeffective any more. However, some authorsadvocate endocrinological evaluationbeforehand since laparoscopy is an invasivemethod. The serum basal antimullerianhormone(AMH) and human chorionicgonadotropin stimulation test are the mostcommonly used two endocrinologicalmethods. We performed bilateralorchiectomy to simulate anorchia in newborn rats and compared inhibin B and AMHlevels of them with those of the bilateralcryptorchid and normal rats.
MATERIAL AND METHODS
Totally 108 Sprague-Dawley rats(3 groups of36 each) were included in this study. Bilateraltestis were placed and fixed to theintraabdominal region after birth in the firstgroup. Bilateral orchiectomy was performedafterbirth in thesecondgroup.Thirdgroupwasthecontrol group.All ratsweresacrified in1,3,7, 14, 21 and 35th day and serum samples werecollected with intracardiac aspiration.Seruminhibin B and AMH levels were measured.
RESULTS
Mean serum inhibin B and AMH levelswere 113,6 pg/ml and 3,38 ng/ml,
respectively in the first group and129,1 pg/ml and 3,4 ng/ml, respectivelyin the third group. On the 3rd day oforchiectomy, both inhibin B and AMHdecreased to unmeasurable levels (meaninhibin B 0,887 pg/ml and AMH 0,64 pg/ml) in the second group. There was nosignificant difference between those twohormone levels in terms of sensitivityand specifity (p< 0.005).
CONCLUSIONS
Testicular tissue existence could bedetermined with the measurement of serumbasal inhibin B and AMH levels.
# B02-5 (PP)
GENOME-WIDE LINKAGE SCAN IN A MOROCCAN FAMILY WITH AUTOSOMAL-RECESSIVE EXSTROPHY OF THE BLADDERIDENTIFIES A NOVEL SUSCEPTIBILITY LOCUS ON CHROMOSOME 3P25.3Michael LUDWIG1, Heiko REUTTER2, Franz RUSCHENDORF3, Markus DRAAKEN4, Regina BETZ4, Norbert HUBNER3,Kathrin SAAR3, Niklas SCHAFER4, Raimund STEIN5, Katja P. WOLFENBUTTEL6 and Markus M. NOTHEN4
1Dept. of Clinical Chemistry and Pharmacology, University of Bonn, Bonn, GERMANY, 2Institute of Human Genetics and Department ofNeonatology, Children’s Hospital, University of Bonn, Bonn, GERMANY, 3Max-Delbruck-Zentrum Berlin Buch, Berlin, GERMANY,4Institute of Human Genetics, University of Bonn, Bonn, GERMANY, 5Department of Urology, University of Mainz, Mainz, GERMANY,6Pediatric Urology, Sophia Children’s Hospital, ErasmusMC, Rotterdam, NETHERLANDS
PURPOSE
Exstrophy of the bladder (EB) is part of thebladder exstrophy-epispadias complex(BEEC) representing a spectrum of
urogenital anomalies in which part or all ofthe distal urinary tract fail to close and areexposed on the outer abdominal wall.Familial occurrence is rare and previousstudies are suggestive of an underlying
multifactorial mode of inheritance.However, no causally related genetic or non-genetic factor has been identified so far. Inthis study, we aim to identify potential risk/modifying loci that might contribute to EB.
ESPU Programme 2009 S25
MATERIAL AND METHODS
A genome-wide linkage scan wasperformed in a consanguineous kindred ofMoroccan origin where three affectedmales showed the same phenotype ofclassic EB.
RESULTS
Strongest evidence for linkage was obtainedfor chromosomal region 3p25.3 (parametricLOD score of 3.4). This region comprises atleast seven genes listed in the current NCBImap (Build 36.1) and sequence analysis of allthese genes has been initiated.
CONCLUSIONS
Our data provide a basis for the localizationand identification of a causally related geneimplicated in EB, most likely localized on3p25.3.
CHIP-BASED GENOME-WIDE SEARCH FO
# B02-6 (PP)R MICRO-ABERRATIONS IN PATIENTS WITHTHE EXSTROPHY-EPISPADIAS COMPLEX
Markus DRAAKEN1, Heiko REUTTER2, Thomas M. BOEMERS3, Simeon A. BOYADJIEV4,Per HOFFMANN5, Markus M. NOTHEN1 and Michael LUDWIG6
1Institute of Human Genetics, University of Bonn, Bonn, GERMANY, 2Institute of Human Genetics and Department of Neonatology,Children’s Hospital, University of Bonn, Bonn, GERMANY, 3Children’s Hospital, Cologne, Department of Pediatric Surgery and PediatricUrology, Koln, GERMANY, 4Section of Genetics, Department of Pediatrics, University of California Davis, Sacramento, USA, 5Life & BrainCenter, Department of Genomics, Bonn, GERMANY, 6Dept. of Clinical Chemistry and Pharmacology, University of Bonn, Bonn, GERMANY
PURPOSE
The current knowledge of genetic factorsunderlying the development of the exstrophy-epispadiascomplex (EEC) is very limited. Sincethis continous spectrum of malformations isassociated with reduced reproduction it isreasonable to assume that in a significantproportion of patients the resultantphenotype is caused be de novo mutations.
MATERIAL AND METHODS
We used a chip-based (Illumina HH610)genome-wide approach to systematicallysearch for micro-aberrations characterizedby loss or gain of genomic material. Thestudy sample comprises EEC patients ofEuropean origin with a negative family
history. Parallel investigation of thepatients’ parents will allow theidentification of de novo mutations ina highly efficient manner.
RESULTS
Initial data obtained from 10 patientsrevealed de novo micro-deletions in twocases. One patient carries a micro-deletionin intron 2 of the Leprecan-like protein 1gene, that is located in close vicinity ofp63. Interestingly, we recently observedreproducible dysregulation of various p63isoforms in eight of fourteen blood orbladder cDNA samples derived from NorthAmerican EEC patients. Another twopatients showed an intronic deletion in the
intestinal Maltase-glucoamylase gene at7q34-q36.1, a region that showed linkage(LOD 1.906) in a genome-wide scan ina German multiplex family.
CONCLUSIONS
These data indicate that a systematicgenomic approach has the potential ofidentifying causal genetic factors in EEC. It isexpected that the identification of thecausative genes will lead to a profoundunderstanding of the molecular mechanismsof normal and disturbed embryonicdevelopment of the human urogenital andrectal system.
BLADDER RECONSTRUCTION AND REGE
# B02-7 (V)NERATION OF A BLADDER EXSTROPHY IN A FETAL SHEEP MODEL
Barbara KORTMANN1, Paul GEUTJES2, Jane CREVELS1, Rene WIJNEN3, Alex EGGINK4, Toin VAN KUPPEVELT5,Katrien BROUWER5, Willeke DAAMEN5, Alex HANSSEN6, Luc ROELOFS1 and Wouter FEITZ1
1Radboud University Nijmegen Medical Centre, pediatric urology, Nijmegen, NETHERLANDS, 2Radboud University Nijmegen MedicalCentre, experimental pediatric urology, Nijmegen, NETHERLANDS, 3Radboud University Nijmegen Medical Centre, pediatric surgery,Nijmegen, NETHERLANDS, 4Radboud University Nijmegen Medical Centre, gynecology, Nijmegen, NETHERLANDS, 5Radboud UniversityNijmegen Medical Centre, biochemistry, Nijmegen, NETHERLANDS, 6Radboud University Nijmegen Medical Centre, central animallaboratory, Nijmegen, NETHERLANDS
PURPOSE
In this video we show the intra-uterinesurgery of a fetal lamb to create anexstrophy-like lesion and the postnatalbladder wall reconstruction using a collagen-based biomatrix.
MATERIAL AND METHODS
In 9 fetal sheep a bladder exstrophy wassurgically created at 79 days’ gestation. This
video shows the intra-uterine surgery ofa fetal lamb and the postnatalreconstruction of a new-born lamb. Theabdominal wall of the ewe was incised, theuterus exteriorized and opened. In the fetusa midline abdominal incision was made andthe bladder opened. The open bladder wallwas sutured to the abdominal wall and skinto create an exstrophy-like lesion. The lambwas replaced in the uterus and pregnancywas continued. After 140 days’ gestation thelamb was born by normal delivery.
Postpartum the lamb stayed with the motherfor 1 week. Then, bladder reconstructionwas performed: The bladder wall wasdissected from the abdominal wall, edges ofthe bladder wall were excised for pathologicevaluation, open bladder was covered witha round, 3 cm collagen-based biomatrixwhich was sutured to the bladder wall withMonocryl 6-0. A pigtail catheter was placedin the bladder and exteriorized through theabdominal wall. The construct was coveredwith omentum, loosely attached with