Serum response factor (SRF) is a transcription factor known to mediate phenotypic plasticity in smooth muscle cells (SMCs). Despite the critical role of this protein in mediating intestinal injury response, little is known about the mechanism through which SRF alters SMC behavior. Here, we provide compelling evidence for the involvement of SRF-dependent microRNAs (miRNAs) in the regulation of SMC apoptosis. We generated SMC-restricted Srf inducible knockout (KO) mice and observed both severe degeneration of SMCs and a significant decrease in the expression of apoptosis-associated miRNAs. The absence of these miRNAs was associated with overexpression of apoptotic proteins, and we observed a high level of SMC death and myopathy in the intestinal muscle layers. These data provide a compelling new model that implicates SMC degeneration via anti-apoptotic miRNA deficiency caused by lack of SRF in gastrointestinal motility disorders.

The aim of this study was to analyze the antenatal characteristics of HLHS, its association with increased nuchal translucency, extracardiac anomalies and other obstetric outcomes. The medical records of antenatally diagnosed HLHS cases managed at Seoul National University Hospital between January 2007 and April 2012 were reviewed. The main policy of fetuses with HLHS in our institution is to maintain pregnancy and intention to treat with staged surgical palliation. Twenty two fetuses (n = 22) were diagnosed antenatally during this period. Two cases had increased NT (9.09%). Only minor extracardiac anomalies were found in 2 cases (megacisterna magna, unilateral multicystic dysplastic kidney). Most fetuses (90.9%) had an isolated HLHS. Eleven cases performed karyotyping antenatally and all fetuses had normal karyotype. The 4 cases resulted in termination of pregnancy at other hospitals as parents wanted. In 18 cases who maintained pregnancy, there was no intrauterine fetal death and 2 cases (11.1%) developed cardiomegaly at near term. Seventeen cases attempted vaginal delivery, in 3 cases (17.6%), emergent cesarean section occurred due to intrapartum fetal distress. In 14 (82%) cases, labor progressed without any fetal distress. All cases had normal cord ABGA (Cord pH > 7.1). Postnatally three cases had diagnosed as Cornelia de lange syndrome, 2q duplication, unilateral MCDK. Most fetuses with HLHS in our institution in Korea were isolated HLHS. And the incidence of increased NT, extracardiac anomalies and abnormal karyotype in HLHS were much less than previous reports in western country. There could be racial difference in characteristics of HLHS fetuses in Asian people.

Although screening ultrasound for detection of major fetal anomaly is usually done in mid-trimester, fetal anomalous structure can be also found in the third trimester during subsequent routine ultrasound. To objective of this study is to evaluate the frequency and spectrum of fetal anomalies diagnosed in the third trimester. Cases with fetal anomalies in singleton pregnancy which were confirmed by postnatal diagnosis was evaluated between Jan 2008 and Dec 2009 in Seoul National University Hospital. The gestational age of detection of fetal anomaly was classified as 1st/2nd trimester and 3rd trimester. We excluded cases in which routine screening ultrasound for detection of major fetal anomaly in mid-trimester was not done. There were a total of 162 cases of fetal major anomalies during the study period. Twenty-nine cases (18%) of fetal anomalies were diagnosed subsequently in the third trimester, and 14 cases (48%) had CNS (central nervous system) or heart anomalies. In 6 cases with CNS anomalies, 4 cases were those with intracranial hemorrhage or destructive lesion and 2 cases were Dandy-Walker variant. Among 11 cases with heart anomalies, 4 cases were those with arrhythmia, 2 cases with coarctation of aorta/hypoplastic left heart syndrome, and 1 case with pulmonary stenosis. Three cases has both CNS and heart anomalies. Even though the result of routine ultrasound in mid-trimester is normal, fetal anomalies can be additionally detected in about 20% of cases during the third trimester. About half of these anomalies diagnosed in third trimester are CNS or heart anomalies, and spectrum of anomalies might be different from that diagnosed in the first/second trimester. P30.19: Table

To determine if there is a difference in pregnancy outcomes among triplet pregnancies which were reduced to twin pregnancies, ongoing triplet pregnancies, and primary twin pregnancies. This retrospective cohort study was conducted in consecutive multi-fetal pregnancies who were delivered after 20 weeks of gestation in Seoul National University Hospital between January 2007 and June 2011. Study population was divided into three groups: 1) ongoing triplet pregnancies (primary triplet pregnancies), 2) triplet pregnancies which were reduced to twin pregnancies after selective fetal reduction during the first trimester (reduced twin pregnancies), and 3) primary twin pregnancies. Pregnancy outcomes included gestational age at delivery and the rate of preterm delivery (< 34 weeks). Data analysis was performed using SPSS version 19.0. During the study period, we identified 65 primary triplet pregnancies, 44 reduced to twin pregnancies, and 795 primary twin pregnancies. Gestational age at delivery (GAD) in reduced twin pregnancies (median [range]: 37.4 weeks [24.4–39.7]) was significantly higher than in primary triplet pregnancies (median [SD]: 34.4 weeks [24.1–39.0) (P < 0.001). However, there was no difference in GAD between reduced twin pregnancies and primary twin pregnancies (median [range]: 37.3 weeks [20.4–40.6]). The rate of preterm delivery (< 34 weeks) in reduced twin pregnancies was not different from that in primary triplet pregnancies (22.7% vs. 32.3%, P > 0.2), but tended to be higher than in primary twin pregnancies (22.7% vs. 13.6%, P = 0.09). The rate of preterm delivery (< 34 weeks of gestation) of ongoing triplet pregnancies was comparable to that of triplet pregnancies reduced to twin pregnancies after selective fetal reduction. Women with triplet pregnancies should be informed of complete pregnancy outcomes when they make a decision about selective fetal reduction.

Maternal hypertension is an important medical complication associated with fetal growth restriction. The purpose of this study is to show whether there is a difference of catch-up growth potential in growth restricted fetuses according to the presence of absence of maternal hypertension. Singleton fetuses on suspicion of growth restriction by antenatal ultrasound and whose birth weight was less than 10th percentile for their gestational age (more than 34 weeks) were included. Postnatal body weights were monitored at around 1, 3, 6 and 12 months. Newborns with aneuploidy or multiple and/or severe structural anomalies were excluded. Maternal hypertensive disease was defined as chronic or gestational hypertension, preeclampsia or superimposed preeclampsia. The degree of growth restriction was measured by a z-score for each age and gender of the infant using the Korean growth standards. The gap between z-score at specific month and z-score at birth was interpreted as a degree of catch-up growth. The women with hypertensive disease were delivered at lower gestational age than those without hypertension (median [range]: 36.1 [34.3–40.9] vs. 38.5 [34.7–41.9], P < 0.05), but there were no differences of maternal height, follow-up duration or gender ratio of newborns between the two groups. The gap between z-score at 3, 6, and 12 months and z-score at birth was significantly higher in newborn born to mothers with hypertensive disease than those without hypertension for each. The difference of the gap of z-score at 12 months between the two groups was still significant after adjusting for gestational age at birth (median [range]: 4.41 [2.69–6.87] vs. 2.08 [1.79–6.26], P < 0.05). Secondary growth restriction caused by maternal hypertension may be transient in utero and can be overcome postnatally. However, constitutionally small babies lag behind normal growth.

Previous studies reported that oligohydramnios is associated with adverse perinatal outcome in preterm premature rupture of membranes (PPROM). However, patients with initially normal amniotic fluid volume (AFV) can maintain adequate AFV or progress to have oligohydramnios, and the clinical outcome of patients who maintain normal AFV has not been well examined. This study was conducted to address this issue. The AF index (AFI) was serially measured in patients with PPROM (< 34 weeks) with an interval of several days. Patients who delivered within 2 days of admission (n = 66) or in whom AFI was not measured serially were excluded from analysis. Oligohydramnios was defined as AFI < 5 and cases were divided into 3 groups according to the change of AFI during 1st and 2nd ultrasonography (USG): Group 1, maintaining normal AFI; Group 2, normal AFI at 1st USG but oligohydramnios at 2nd USG (developing oligohydramnios); Group 3, oligohydramnios at 1st USG. The AF index (AFI) was serially measured in 188 patients with PPROM with a median interval of 3 days. The rate of oligohydramnios at 1st USG was 35% (66/188). Among 122 patients who had normal AF at 1st USG, 84 of patients (69%) maintained normal AFV and 38 patients (31%) developed oligohydramnios in 2nd USG. Cases in groups 2 and 3 had shorter mean interval-to-delivery than those in group 1 (group 1, 4.8 ± 5.3 weeks; group 2, 1.9 ± 2.1 weeks; group 3, 1.6 ± 1.6 weeks; P < 0.001) and these differences remained significant after adjustment for gestational age. In 141 patients, amniocentesis was performed within 1 week of 1st USG measure and AF matrix metalloproteinase-8 was measured after delivery. Cases in groups 2 and 3 had higher rate of intra-amniotic infection and/or inflammation than those in group 1 (group 1, 28% [21/75]; group 2, 55% [16/29]; group 3, 70% [26/37]; P < 0.001). Cases with persistent normal AFI had better outcome than those with developing oligohydramnios. Serial measurement of AFI is needed in cases with PPROM.

To evaluate whether fetal small bowel perforation in utero affects perinatal outcomes in fetus with jejunal or ileal atresia. All neonates with surgically confirmed jejunal or ileal atresia between January, 2000 and February, 2011 were enrolled in this study. The neonates were divided into two groups according to the presence of fetal bowel perforation, suspected by prenatal sonography and finally confirmed by surgery: P(+), neonates with perforation (n = 6) and P(−), neonates without perforation groups (n = 20). Medical records of two groups were retrospectively analyzed for comparison of neonatal birth characteristics and perinatal outcomes. There were a total of 26 jejunal or ileal atresia. Compared to P(−) group, maternal age was younger and male was more frequent in P(+) group. These differences, however, were not statistically significant, maybe due to small sample size. Other neonatal birth characteristics were similar in both groups. On prenatal sonographic examination, ascites (66.7%), pseudocyst (86%) and intra-abdominal calcification (50%) were frequently found in P(+) group. There is no neonatal death. Median hospital stay was not different between two groups. Bowel perforation in utero might not affect perinatal outcomes in fetus with jejunal or ileal atresia. Therefore, early intervention may be unnecessary only reason of bowel perforation in utero. P08.10: Table 1. Clinical characteristics and perinatal outcomes

대부분의 항암화학요법은 임신부에게 사용되었을 때 유의한 기형 발생 가능성이 있는 것으로 알려져 있다. 비록 일부 항암약제의 경우 기관 발생시기를 지나고 투여하였을 때 태아에게 안전하다는 연구결과가 보고되고 있으나, 여전히 임신중의 암환자에게 항암화학요법을 시행하는 것에는 논란이 있으며, 동물실험이 아닌 사람에게 항암화학요법의 영향을 알아볼 수 있는 연구를 하는 것은 한계가 있다. 저자들은 폐암을 진단받아 오랜 기간 지속적으로 다양한 항암화학요법으로 치료해오던 중 만삭으로 추정되는 시기에 임신 사실을 알게 된 환자1예를 경험하였기에 문헌 고찰과 함께 보고하는 바이다. 환자는 25세의 출산력이 없는 여성으로 6년 전에 비소세포폐암을 진단받았으며, 임신 사실을 모른 채 임신 1, 2 삼분기에 해당하는 시기에도 지속적으로 근치적 항암화학요법을 받았으나, 마지막 월경일로부터 약 41주가 되었을 때 임신사실을 확인하였으며, 지연임신으로 입원하여 산부인과 및 종양내과 협진하에 유도분만이 결정되었다. 자연분만으로 2,450 g의 여아를 특별한 합병증 없이 출산하였으며, 아기에게서 육안적 기형은 발견되지 않았다.

We compared the role of the cervical length (CL) at 19–21 vs. 22–24 gestational weeks in triplet pregnancy to predict preterm birth. The maternal records of triplet pregnancies managed at Seoul National University Hospital from November 2005 to February 2011 were reviewed. Women undertaken cervical cerclage operation were excluded. The relationship between CL ⩽ 20 mm and preterm delivery before 34 weeks' gestation was assessed. All patients didn't use any prophylactic progesterone. Thirty five women were included. CL was measured at 19–21 weeks (n = 25) and at 22–24 weeks (n = 26). CL of 16 women were measured at both periods. At 19–21 weeks, among 24 women with CL > 20 mm, 9 (37.5%) women delivered before 34 weeks and 15 (62.5%) women delivered after 34 week. Negative predictive value was 62.5%. At 22–24 weeks (n = 26), all 6 women with CL ⩽ 20 mm delivered before 34 weeks. And among 20 women with CL > 20 mm, 2 (10%) women delivered before 34 weeks, 18 (90%) women delivered after 34 weeks. Negative predictive value was 90%. Women with CL at both periods (n = 16), all 16 were CL > 20 mm at 19–21 weeks. Five (31.3%) showed CL change from > 20 mm at 19–21 weeks to CL ⩽ 20 mm at 22–24 weeks. Eleven (68.8%) did not show cervical shortening between this period. All women with CL shortening group (n = 5) delivered before 34 weeks. In CL maintaining group (n = 11), 10 (90.9%) delivered after 34 weeks and only one (9.09%) delivered before 34 weeks (P = 0.001). On the basis of CL > 20 mm, optimal timing of screening cervical length to predict preterm delivery in the triplet pregnancy is 22–24 weeks' gestation. Earlier cervical length measurement shows low negative predictive value.

Uterus didelphys with obstructed hemivagina and ipsilateral renal agenesis is rare, sometimes referred to as Herlyn-Werner-Wunderlich syndrome (HWW). It usually diagnosed after menarche with pelvic pain or abdominal mass. We report 2 cases of Herlyn-Werner-Wunderlich syndrome in fetus. One of that the dilated vagina and uterus mimicking dilated ureter was identified by prenatal sonography, prior to symptoms development. And the other was diagnosed by postnatal MRI due to highly suspicion of mullerian anomaly with renal agenesis in female fetus. It would be advisable to look for mullerian anomaly, whenever unilateral renal agenesis or dysplastic kidney in a fetus for early and accurate diagnosis.

To investigate the prevalence of intrauterine death (IUD) after 20 weeks in monochorionic twins and to evaluate the perinatal outcome of the surviving co-twins compared to dichorionic twins. We retrospectively reviewed medical records for 869 twin pairs born from 2000 to June 2009 in the Seoul National University Hospital. Nineteen twin pregnancies (11 monochorionic diamniotic and 8 dichorionic) were complicated by IUD after 20 weeks of gestation. Cranial ultrasound scans were performed most of surviving co-twins complicated IUD. A 195 monochorionic diamniotic (MCDA) and 674 dichorionic diamniotic (DCDA) twin pairs were included in the study. Single fetal demise occurs in 5.6% (11/195) of MCDA twins, and 1.2% (8/674) of DCDA twins. 92.3% (180/195) MCDA twin pairs resulted in 2 survivors, 2.1% (4/195) in no survivor, while 98.7% (665/674) DCDA twin pairs resulted in 2 survivors, 0.1% (1/674) in no survivor. Gestational age (GA) at delivery and GA at IUD was not statistically different between MCDA and DCDA twins (35.0 (range, 30.1–40.7) vs. 32.1 (range, 30.1–41.3) weeks) (P = 0.55) and (29.0 (range, 20.3–38.6) vs. 29.9 (range, 22.4–33.1) weeks) (P = 0.60), respectively. The incidence of cerebral injury and cardiac complication was 18.2% (2/11) (intraventricular hemorrhage, encephlaomalacia), 9.1% (1/11) (cardiomyopathy) in MCDA twin pregnancies compared to DCDA ones 0%. Our study confirmed the higher risk of perinatal morbidity for the co-twins in case of single fetal demise after 20 weeks in MCDA twin pregnancies. MCDA twin pregnancies have a much higher risk ischemic lesion in vital organs such as brain and heart compared to DCDA ones, limited by small number of cases.

To evaluate the outcomes in isolated agenesis of corpus callosum (ACC). We retrospectively reviewed data for 34 cases of ACC born between 2000 and 2010 in the Seoul National University Hospital. The following variables were assessed: maternal age, gestational age at delivery, birth weight, gender, type of ACC (complete or partial), associated abnormalities and clinical outcomes including developmental delay, seizure, visual problem, hearing impairment and death. Developmental delay was analyzed for patients with a minimum follow-up period of 1 year. There were no significant differences in the maternal age, rate of small for gestational age, male and complete ACC and clinical outcomes. However, preterm birth was significantly higher in nonisolated ACC. Concerning the patients with isolated ACC, 2 out of 7 (28.6%) had developmental delay. 2 out of 13 patients (15.4%) had seizure and 1 patient (7.7%) had visual problem. The outcome of isolated ACC is not favorable. The developmental delay, seizure and visual problem were not uncommon in isolated ACC. The prognosis is not always good even in fetuses with isolated ACC. P02.02: Table 1. Characteristics and outcome data of the isolated and nonisolated ACC

To describe the discrepancy in Doppler waveforms between two umbilical arteries (UAs) that may make a clinically significant impact on perinatal outcome and recommend that Doppler waveforms of UA should be obtained from both UAs, not just one. This was a prospective observational study involving 50 high-risk pregnant women who were admitted to Seoul National University Hospital for fetal surveillance between July 2010 and March 2011. High-risk pregnancy included intrauterine growth restriction with or without preeclampsia, twin-to-twin transfusion syndrome, or oligohydramnios. UA Doppler waveforms were obtained at intra-abdominal cord insertion site in order to distinguish one from the other. Doppler findings of one UA were expressed as present end-diastolic flow (EDF), absent end-diastolic flow (AEDF), or reversed end-diastolic flow (REDF). The findings were recorded at both UAs and into four patterns: EDF/AEDF, AEDF/AEDF, AEDF/REDF, and REDF/REDF. There was no EDF/REDF case. A total of 155 observations were undertaken. Twenty-seven of 50 patients (54%) showed AEDF or REDF in at least one umbilical artery during surveillance period and were included for analysis. Of these 27, 11 patients (40.7%) revealed discrepancy in Doppler waveforms between two umbilical arteries. The proportion of patients according to UA Doppler patterns was 30% in EDF/AEDF (8/27), 37% in AEDF/AEDF (10/27), 11% in AEDF/REDF (3/27), 22% in REDF/REDF (6/27). (Table 1). Doppler waveforms have discrepancy between two umbilical arteries in more than one third of high-risk pregnancies beginning to show AEDF or REDF in at least one UA. When Doppler waveforms of an UA were revealed as AEDF or REDF, clinicians must be cautioned to examine the contralateral UA.

Fetus-in-fetu (FIF), a rare congenital anomaly, is a fetus incorporating the well-differentiated tissue of its twin. The authors report a FIF case prenatally mimicking meconium peritonitis (MP). At the gestational age of 34 weeks 4 days, a complex of intra-abdominal heteroechogenic cystic mass with irregular margin and calcification, ascites, and polyhydarmnios, was found on sonographic examination, which was suggestive of MP in utero. On sonography and computered tomography after birth, however, a tumorous condition was suspected rather than meconium peritonitis. An irregular fetiform mass partially covered with skin was found in the retroperitoneum during abdominal exploration. Pathologic exam revealed two fingers, small intestine, large intestine, liver, pancreas and cartilage in the mass, indicated FIF. FIF often is overlooked in the differential diagnosis of a prenatal abdominal cystic mass. Prenatal sonographic finding similar to that of this case might be confused with complicated MP or teratoma with malignant potential. Because this diagnosis is finally made after pathological analysis, all parts of the mass should be removed considering malignant potential.

L. E. Romine1, M. Hwang5, N. Trivedi2, J. Wan4, Y. LaCoursiere2, P. Do4, K. Benirschke3, D. Schrimmer2, D. H. Pretorius1 1Radiology, University of California, San Diego, CA, USA; 2Reproductive Medicine, University of California, San Diego, CA, USA; 3Pathology, University of California, San Diego, CA, USA; 4School of Medicine, University of California, San Diego, CA, USA; 5School of Medicine, Vanderbilt University, Nashville, TN, USA

Duodenal obstruction and atrioventicular septal defect are classic markers of Down syndrome. According to previous reports, trisomy 21 occurs in approximately 30% of neonates with duodenal obstruction and 40% of those with atrioventricular septal defect, respectively. However, on the basis of our clinical experience, it may be different in Korean population, especially who had duodenal obstruction. Therefore, we carried on a retrospective chart review to clarify our opinion. Forty-one cases of duodenal obstruction and 140 cases of atrioventricular septal defects were found at the Seoul National University Hospital from September 1999 to February 2010. Diagnosis was confirmed by surgery, postnatal echocardiography, or autopsy. Medical record of each case was reviewed retrospectively. In 41 neonates with duodenal obstruction, two (4.9%) had trisomy 21 (one from duodenal atresia, and the other from duodenal web). Thirty-nine neonates were live-born, and they all underwent operations. According to operative finding, 26 had duodenal atresia, 9 had duodenal web, and 4 had annular pancreas. In 140 neonates with atrioventricular septal defect, forty-three (30.7%) had trisomy 21. Among them, 139 were live-born and 115 underwent operations. The occurrence of Down syndrome in cases with atrioventricular septal defect is similar to preexisting data, but that in those with duodenal obstruction is only one-sixth. Duodenal obstruction in Down syndrome is far less common in Korean (Asian) population than in western population. The significance of some antenatal sonographic markers for Down syndrome may be different among races or ethnic groups.

Previous study suggested different cutoff value of pelvis antero-posterior diameter (APD) of kidney for pyelectasia for each trimester; 4 mm for 2nd and 7 mm for 3rd trimester, and 10 mm as pathologic. The objective of this study was to examine the cutoff value of pelvis APD for prediction of surgical management in renal units with antenatally diagnosed pyelectasia. Pelvis APD was serially measured in 91 renal units of 74 patients with pyelectasia from 2nd trimester. Pyelectasia was defined as pelvis APD of more than 4 mm in 2nd trimester and that of more than 7 mm in 3rd trimester. ROC curves were constructed to select the cutoff value for prediction of surgical management, and 10 mm was selected as the cutoff value for both 2nd and 3rd trimester. Renal units were divided into 4 groups according to change of pelvis APD during 2nd and 3rd trimester: group 1, normalized APD (4 < APD < 10 mm in 2nd trimester and APD⩽7 mm in 3rd trimester, n = 33); group 2, persistent but without progressive change (4 < APD < 10 mm in 2nd trimester and 7 < APD < 10 mm in 3rd trimester, n = 21); group 3, progressive change (4 < APD < 10 mm in 2nd trimester and pelvis APD ≥ 10 mm in 3rd trimester, n = 21); group 4, APD ≥ 10 mm in 2nd trimester (n = 16). Renal units were divided into 2 groups according to the postnatal outcome; surgical and conservative management. In total, 11% (10/91) of renal units underwent surgical management: 0% of renal units in group 1 (0/33); 0% of renal units in group 2 (0/21); 5% of renal units in group 3 (1/21); and 56% of renal units in group 4 (9/16) (P < 0.001). Renal units in group 4 had higher rate of surgical management than those in group 3 (P < 0.005). Renal units with pelvis APD ≥ 10 mm in 2nd trimester had worse outcome than those with progressive change. Ten or more (mm) of pelvis APD at 2nd trimester may be a powerful predictor for surgical treatment.

Nuchal translucency (NT) is the most powerful screening tool for Down syndrome and cardiac anomaly. Because of its clinical impact, strict guidelines were established to get accepted NT measurement. However, to stick to the guidelines in most pregnant women is time-consuming and superfluous. We undertook this study whether the simplified protocol enables to select low risk group and skip the recommended NT measurement in them. A prospective study was conducted. The measurements of NT and crown–rump length (CRL) were performed by one author in 315 fetuses between 11 + 0 and 13 + 6 weeks of gestation. First, CRL was measured in the ordinary view that was mid-sagittal section of fetus in neutral position, and NT was measured at the same frozen screen (1st measured value, 1MV). Then, NT was measured again according to the Fetal Medicine Foundation (FMF) guidelines (2nd measured value, 2MV). Data were analyzed using ROC curve (95% CI, P < 0.05). There was good correlation between 1MV and 2MV in each case (r = 0.89). ROC curve was conducted to examine the relationship between 1MV and over the 95th percentile (p) 2MV (AUC: 0.98, P < 0.001). NT value of 2 mm in 1MV could be used as a cut-off to obtain over the 95p 2MV (sensitivity of 100%, specificity of 80.5%, PPV 22.7%, NPV 100%). All the 95p or more 2MV could be detected with this simplified protocol. NT ≥ 2 mm in 1MV was 22.2% of all cases, in other words, we had only to measure 2MV in 22.2% patients. If NT is less than 2 mm at ordinary CRL view, we may skip the recommended NT measurement. Supporting information can be found in the online version of this abstract. ROC curve for identification of over the 95p 2MV by 1MV. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.