Bao-Tyan Wang

Antenatal treatment of chylothorax and cystic hygroma with OK-432 in nonimmune hydrops fetalis.

Objectives: To present our experience of using OK-432 in treating fetal cystic hygroma and chylothorax complicated with nonimmune hydrops fetalis. Methods: OK-432 (Picibanil®) was injected into the fetal pleural cavity or fetal cystic hygroma. Results:Patient 1: A 23-year-old, gravida 2, para 1, was found to have a recurrent fetal chylothorax at GA 29 weeks. Serial amnioreduction and thoracocentesis was performed at GA 31, 32, 33, and 34 weeks. Intrapleural OK-432 injection was performed twice at GA 33 and 34 weeks. Cyanosis and respiratory distress were noted immediately after birth (GA 34 weeks). The baby expired despite of aggressive neonatal resuscitation. Patient 2: A 26-year-old, gravida 2, para 1, was found to have a cystic hygroma of her fetus at GA 17 weeks. Karyotype of the cystic fluid and the amniocytes were 46, XY. Fetal ascites developed at GA 22 weeks. OK-432 injection into the tumour was performed at GA 23 weeks. Stabilization of the cystic hygroma was noted throughout the pregnancy (about 3.5 cm in diameter). Serial fetal paracentesis and/or amnioreduction were performed. Karyotype of the ascites was again 46, XY. Maternal dietary modification with medium chain triglyceride was also prescribed. Chylothorax developed and the baby was born by cesareans at GA 32 weeks. Resolution of pleural effusion, ascites, and regression of cystic hygroma were noted since the 2nd day after birth. The baby had survived beyond 4 months of age at submission. Conclusion: Combination of antenatal OK-432 injection, maternal dietary modification, serial thoracocentesis plus paracentesis, together with amnioreduction and tocolysis, appeared to contribute to the success of antenatal treatment. Fetal pulmonary expansion may determine the immediate neonatal survival.

Fetal OK‐432 pleurodesis: complete or incomplete?

D. Paladini*†, G. Sglavo†, M. Quarantelli‡, M. R. D’armiento§, P. Martinelli† and M. Salvatore‡ †Fetal Cardiology Unit, Department of Gynecology and Obstetrics, University Federico II of Naples, ‡Biostructure and Bioimaging Institute, National Research Council, Naples and §Department of Pathology, University Federico II of Naples, Italy *Correspondence. (e-mail: paladinid@unina.it) DOI: 10.1002/uog.2621

Two novel null mutations in a Taiwanese cystic fibrosis patient and a survey of East Asian CFTR mutations

Cystic fibrosis (CF; OMIM number 219700) is one of the most common and life-shortening autosomal recessive disorders in Caucasians, with an incidence of 1 in 2,500 newborns [Welsh et al., 2001]. Its incidence is significantly reduced in Asian populations (1 in 90,000) [Wright and Morton, 1968]. Little is known about the spectrum of CFTR mutations in Asians. Data from the small number of reports indicated that a potentially large pool of Asian CF mutations are novel. The commercially available mutation panels are not suitable for screening Asian CF mutations. A more comprehensive mutation detectionmethod is necessary to search for the unknown mutations. Using recently developed temporal temperature gradient gel electrophoresis (TTGE) method [Wong et al., 2001; Alper et al., in press (to be published elsewhere)], a CF patient from Taiwan was found to be compound heterozygote for E7X and 989992insA novel null mutations. The patient was a 11⁄2-year-old boy who was born at 41 weeks of gestation with normal weight and unremarkablemedical history until 7months of agewhen he was admitted to hospital in Taiwan with severe vomiting and abdominal distension. He was diagnosed with acute gastroenteritis. At 1 year of age, he had severe cough, poor appetite, and sputum colonization with Pseudomonas aeruginosa. The diagnosis of CF was made when the patient was 2 years old, based on the recurrent respiratory tract infection with pneumonia, sputum colonization with Pseudomonas and Staphylococcus epidermis, mucosal pluggings, bronchiectasis, echogenic bowl, developmental delay, steatorrhea, and elevated sweat chloride (89mmol/L).Molecular analysis by Genzyme Genetics pan-ethnic 87 mutation panel did not reveal anyCFTRmutations. Since autopsy revealed evidence of CF in his deceased older sister, a comprehensive mutational analysis of the CFTR gene was felt necessary. CFTR exons and their flanking intron regions were amplified using 27 primer pairs followed by TTGE analysis [Wong et al., 2001]. Exons 1 and 6b that revealed abnormal TTGE banding patterns were sequenced (data not shown). Twomutationswere identified;G151T in exon 1 changes a glutamate at amino acid residue 7 to a stop codon, and 989-992 insertion A causes frameshift and a truncated CFTR protein of 306 amino acids. The intron 8 polythymidine and GT tract variants were found to be homozygous 7T and heterozygous 11/12 GT, both were unremarkable. Although the parents were unavailable for carrier testing, the finding of two deleterious mutations and the positive family history substantiates the diagnosis of CF in this patient. Unfortunately, the patient died before the mutations were identified. He was pancreatic insufficient. Since CF cases were rarely seen in Taiwan, the patient was not properly treated with enzyme supplement. The two mutations found in our patient have never been reported. The patient’s father is from Taiwan and the mother from Vietnam. A literature search found more than 80 documented cases of CF patients in East Asia [Macek et al., 1992; Wang et al., 1993; Crawford et al., 1995; Zielenski et al., 1995a; Hojo et al., 1997; Macek et al., 1997; Suwanjutha et al., 1998; Seki et al., 1999; Wagner et al., 1999; Yoshimura et al., 1999; Morokawa et al., 2000; Wu et al., 2000]. Approximately 50 caseswere reported in Japan and about 10 cases have been reported in patients from Taiwan and Mainland China [Seki et al., 1999]. Most of the patients have not been analyzed at themolecular level. Only 15 (including this study) had identified CFTR mutations (Table I). It is remarkable that the East Asian CF patients do not share any mutations with patients of other ethnic backgrounds. Even within the East Asians, the CFTR mutation spectrum of Chinese patients is distinct from that of Japanese patients (Table I) [Hojo et al., 1997; Macek et al., 1997; Seki et al., 1999; Yoshimura et al., 1999; Morokawa et al., 2000]. Most of the mutations are frameshift, splice sitemutations, or nonsensemutations that predict a CFTR protein with null activity. All patients presented classical CFphenotype (Table I)with elevated sweat chloride and pancreatic insufficiency. *Correspondence to: Dr. Lee-Jun C. Wong, Ph.D., Associate Professor, Institute for Molecular and Human Genetics, P.O. Box 571477, Georgetown University Medical Center, 3800 Reservoir Rd, NW M4000, Washington, DC 20057. E-mail: wonglj@georgetown.edu

Microchip, reverse transcription-polymerase chain reaction and culture methods to detect enterovirus infection in pediatric patients

Background: Enterovirus infection usually presents with mild and self‐limited illness in children. However, Enterovirus type 71 can be characterized by neurotropism and may cause severe illness or even sudden death. Early detection of the virus will allow a physician to provide intensive or aggressive intervention. The purpose of the present study was to compare sensitivity of two innovative laboratory methods, that is, the DR.EV microchip method (DR. Chip Biotechnology, Shin‐Tsu, Taiwan) and the reverse transcription‐polymerase chain reaction (RT‐PCR) method following conventional virus culture in detecting enterovirus infection in pediatric patients with herpangina or hand–foot–mouth disease.

Detection of Novel CFTR Mutations in Taiwanese Cystic Fibrosis Patients

BACKGROUND AND PURPOSE Cystic fibrosis (CF) in Asian populations is very rare. We performed molecular genetic analysis in 2 Taiwanese CF patients for detection of cystic fibrosis transmembrane conductance regulator (CFTR) mutations. METHODS Temporal temperature gradient gel electrophoresis (TTGE) was used for mutation detection, and direct sequencing was used for identification of mutations. RESULTS In one patient, 2 novel mutations, E7X and 989-992insA, were identified and the carrier status of his parents was confirmed. In the other patient, 3 mutations, S895N, 2215insG, and 1898+5G>T, were found. The 2215insG and S895N were found cis in the same chromosome. These splice site, frameshift, and nonsense mutations produce severely truncated CFTR polypeptides which lack a transmembrane domain, nucleotide binding folds, and the regulatory region, and are predicted to be null in CFTR function. CONCLUSIONS These cases underscore the importance of comprehensive mutation analysis of Taiwanese CF patients. Definitive molecular findings can confirm the clinical diagnosis and facilitate patient management, carrier testing, and genetic counseling. Furthermore, there is an urgent need to understand the mutation spectrum and the clinical features of the CFTR gene in Asian patients in order that a mutation panel can be established for effective screening of CF chromosomes.

Subtelomeric rearrangements and 22q11.2 deletion syndrome in anomalous growth-restricted fetuses with normal or balanced G-banded karyotype.

To determine the frequencies of cryptic subtelomeric rearrangements and 22q11.2 deletion in anomalous growth‐restricted fetuses with normal or balanced G‐banded karyotypes.

Persistent fifth aortic arch associated with 22q11.2 deletion syndrome.

BACKGROUND Chromosome 22q11.2 deletion is frequently associated with conotruncal malformations and aortic arch anomalies. This study investigated the association of chromosome 22q11.2 deletion with clinical manifestations in four pediatric patients with persistent fifth aortic arch. METHODS Four patients with persistent fifth aortic arch treated between July 1997 and June 2004 were included in this retrospective study. There were two girls and two boys, aged 2 days to 11.3 years, with persistent fifth aortic arch and cardiac conotruncal malformations. Chart recordings, plain chest films, two-dimensional and Doppler echocardiograms, cardiac catheterization with angiograms, surgical findings, and cytogenetic study were analyzed. RESULTS Clinically, all four patients had the cardinal phenotypic features of 22q11.2 deletion syndrome, including cardiovascular malformations (conotruncal malformations and aortic arch anomalies), abnormal facies, thymic hypoplasia, canopy anomaly of the palate (high-arched palate, rather than cleft palate), and hypocalcemia (or hypoparathyroidism). All four patients were confirmed to have chromosome 22q11.2 deletion. CONCLUSION Congenital conotruncal malformations, including tetralogy of Fallot with pulmonary atresia or stenosis, and aortic arch anomalies including a persistent fifth aortic arch or a right aortic arch, should lead to suspicion of chromosome 22q11.2 deletion when manifested together with any one of the other four cardinal phenotypic features.

Redundant skin over the nape in a girl with monosomy 1p36 caused by a de-novo satellited derivative chromosome: a possible new feature?

We present a case of monosomy 1p36 who, in addition to delayed growth and development, and mild facial dysmorphism also had redundant skin over the nape. Chromosome analysis showed that the monosomy 1p36 was caused by a de-novo satellited derivative chromosome. We propose that the redundant skin over the nape should be considered as a feature of this condition.

Maternally inherited unbalanced translocation of chromosome 22 in a 5-day-old neonate with persistent fifth aortic arch and tetralogy of Fallot

a , b c c d * Meng-Luen Lee , Lon-Yen Tsao , Bao-Tyan Wang , Mei-Huei Lee , Ing-Sh Chiu Department of Pediatrics, Division of Pediatric Cardiology and Director of Pediatric Intensive Care Unit, Changhua Christian Hospital, No. 135, Nanhsiao St., Changhua 50050, Taiwan Division of Neonatology and Director of Neonatal Intensive Care Unit, Changhua 50050, Taiwan Laboratory of Medical Genetics, Changhua Christian Hospital, Changhua 50050, Taiwan Department of Surgery, Division of Cardiovascular Surgery, College of Medicine, National Taiwan University Hospital, Taipei 10002, Taiwan

Interstitial Deletion 13q31 Associated with Normal Phenotype: Cytogenetic Study of a Family with Concomitant Segregation of Reciprocal Translocation and Interstitial Deletion

Gain or loss of a fragment in human chromosomes has been associated with abnormal phenotypes in numerous genetic disorders. However, it is also possible that lack or excess of a particular chromosomal segment is a neutral polymorphism among populations and thus does not cause obvious abnormal phenotype. In this study, conventional GTG-banded karyotyping and molecular cytogenetic analyses (including fluorescence in situ hybridization, spectral karyotyping and comparative genomic hybridization) were applied to study the genotype-phenotype correlation in a Taiwanese family, in which a concomitant segregation of del(13)(q31q31) interstitial deletion and t(13;18)(q32;p11.2) reciprocal translocation in a 2-year-old girl (the proband) was noticed. Two family members (the father and grandmother of the proband) who carried the del(13)(q31q31) but not the translocation t(13;18) both revealed a normal phenotype at adulthood. The finding, which appears novel, that interstitial deletion 13q31 could be associated with a normal phenotype, is therefore valuable in genetic counseling.