Application of risk score analysis to low-coverage whole genome sequencing data for the noninvasive detection of trisomy 21, trisomy 18, and trisomy 13.

OBJECTIVES: Clinical performance of a low coverage, low cost, massively parallel sequencing (MPS)-based assay to stratify risk of trisomy 21, 18, and 13 pregnancies was determined. METHODS: The study included 1100 samples with birth outcome or karyotype results, comprising low-risk patients (84.2%) negative for risk indications from maternal age, serum screening, ultrasound, or family history, and high-risk patients (15.8%) with at least one of the aforementioned indications. Cell free DNA (cfDNA) was extracted from maternal plasma. Library preparation incorporated 96 index barcodes to enable sequencing on a HiSeq 2000 or 2500. Risk scores were calculated using chromosomal representation, fetal fraction, and maternal age at the estimated date of delivery. A risk score greater than or equal to 1 in 100 was used to stratify samples as high risk for trisomy 21, trisomy 18, or trisomy 13. RESULTS: Sensitivity and specificity were calculated based on risk group stratification. Trisomy 21, trisomy 18, and trisomy 13 were detected with greater than 99% sensitivity and 99.9% specificity. Fetal sex classification accuracy was 99.3%. CONCLUSIONS: We conclude that simplified MPS can be used to stratify the risk of pregnancies for trisomy 21, trisomy 18, and trisomy 13 and accurately determine fetal sex. © 2015 John Wiley & Sons, Ltd.

The utilization of circulating cell-free fetal DNA testing and decrease in invasive diagnostic procedures: an institutional experience.

OBJECTIVE: To characterize the patient population utilizing circulating cell-free fetal DNA (ccffDNA) testing at a large academic center and evaluate trends in the performance of invasive diagnostic procedures. STUDY DESIGN: A retrospective cohort study of all patients who underwent cell-free DNA testing from May to December 2012 was performed. RESULT: During the study period, 206 patients had cell-free DNA testing. Of those, 75% (155/206) were of ages ⩾ 35 years. Of those undergoing ccffDNA testing, 41% had positive aneuploidy screening and 38% had abnormal ultrasound findings. Only 7% of the patients with negative ccffDNA testing opted for an invasive diagnostic procedure compared with 60% with positive testing (P<0.01). The rate of invasive procedures decreased from 5.9% of all visits to the center during a similar 8-month period in 2010 to 4.1% of all visits during the study period (P<0.01). CONCLUSION: Our data suggest that ccffDNA testing leads to reduced uptake of invasive procedures.

The controllability beliefs scale used with carers of people with intellectual disabilities: psychometric properties.

BACKGROUND: Beliefs about the controllability of behaviour have been consistently shown to be important in understanding the responses of carers to the challenging behaviour of people with intellectual disabilities (IDs). This paper reports the reliability and validity of the controllability beliefs scale (CBS), a 15-item measure of beliefs regarding the controllability of challenging behaviour when used with carers of people with IDs. METHODS: Two hundred and sixty-four carers of people with IDs completed the CBS, 74 people also completed the modified attributional style questionnaire and the self-injury behavioural understanding questionnaire scale to determine concurrent and convergent validity and 34 people completed the scale twice within a 2- to 4-week period to determine test-retest reliability. RESULTS: The scale has a two-factor structure and has adequate internal reliable. The scale is significantly correlated with the controllability, internality and stability items from the Modified Attributional Style Questionnaire, showed expected associations with behavioural and internal emotional understanding items from the self-injury behavioural understanding questionnaire. The scale has good test-retest reliability. CONCLUSIONS: The data support use of the CBS in clinical practice and research to assess carers' beliefs regarding challenging behaviour of people with IDs.

The PRaCTICaL study of nurse led, intensive care follow-up programmes for improving long term outcomes from critical illness: a pragmatic randomised controlled trial.

OBJECTIVES: To test the hypothesis that nurse led follow-up programmes are effective and cost effective in improving quality of life after discharge from intensive care. DESIGN: A pragmatic, non-blinded, multicentre, randomised controlled trial. SETTING: Three UK hospitals (two teaching hospitals and one district general hospital). PARTICIPANTS: 286 patients aged >or=18 years were recruited after discharge from intensive care between September 2006 and October 2007. INTERVENTION: Nurse led intensive care follow-up programmes versus standard care. Main outcome measure(s) Health related quality of life (measured with the SF-36 questionnaire) at 12 months after randomisation. A cost effectiveness analysis was also performed. RESULTS: 286 patients were recruited and 192 completed one year follow-up. At 12 months, there was no evidence of a difference in the SF-36 physical component score (mean 42.0 (SD 10.6) v 40.8 (SD 11.9), effect size 1.1 (95% CI -1.9 to 4.2), P=0.46) or the SF-36 mental component score (effect size 0.4 (-3.0 to 3.7), P=0.83). There were no statistically significant differences in secondary outcomes or subgroup analyses. Follow-up programmes were significantly more costly than standard care and are unlikely to be considered cost effective. CONCLUSIONS: A nurse led intensive care follow-up programme showed no evidence of being effective or cost effective in improving patients' quality of life in the year after discharge from intensive care. Further work should focus on the roles of early physical rehabilitation, delirium, cognitive dysfunction, and relatives in recovery from critical illness. Intensive care units should review their follow-up programmes in light of these results. TRIAL REGISTRATION: ISRCTN 24294750.

Reducing the cost of detection of congenital heart disease in fetuses of women with pregestational diabetes mellitus.

OBJECTIVE: To provide a cost minimization analysis to support a paradigm shift in the use of comprehensive ultrasound and echocardiography in the prenatal diagnosis of congenital heart disease (CHD) in fetuses of women with diabetes mellitus (DM). METHODS: In this retrospective cohort study, the diabetic clinic service database of the University of California San Diego was searched from January 2001 to June 2004 for pregnant women with Type I or II DM and HbA1c >6.3%. Subjects underwent comprehensive ultrasound examination (with four-chamber views and outflow tracts) and fetal echocardiography according to a standard protocol. Newborns were examined for cardiac defects and underwent postnatal echocardiography as indicated. The cost of screening was evaluated. RESULTS: Of 115 neonates and two terminations of pregnancy there were 20 (17%) cases of CHD. Six of these CHD were major and all six were detected prenatally by both ultrasound and echocardiography. Three additional clinically insignificant cases of CHD were identified by fetal echocardiography. Eleven cases of CHD were identified by postnatal echocardiography only, all of which were clinically insignificant lesions. The prenatal detection rate of major CHD was 100% (6/6) for both ultrasound and echocardiography. The sensitivites of ultrasound (30% (95% CI, 13-54%)) and echocardiography (45% (95% CI, 24-68%)) were similar (P = 0.32). A cost minimization analysis was done using the published Medicaid (California) system's relative value unit and conversion factors. Accordingly, our current protocol costs $6503.43 per case of major CHD detected. If echocardiography had been performed only as indicated and postnatal echocardiography had been performed on all neonates, the cost would have been $7056.83 per case of major CHD detected. Alternatively, combined targeted ultrasound with indicated prenatal and postnatal echocardiography would have been associated with a cost of $4996.05 per case of major CHD detected. CONCLUSION: Detection of major CHD was excellent with both comprehensive prenatal ultrasound and echocardiography. Echocardiography added little to the prenatal diagnosis of CHD if the comprehensive ultrasound examination was normal. The highest cost-benefit ratio and most efficacious protocol for screening based on our data would be comprehensive ultrasound with prenatal and postnatal echocardiography only as indicated. Further prospective studies are warranted.

Can prenatal ultrasound detect the effects of in-utero alcohol exposure? A pilot study.

OBJECTIVES: The aim of this pilot study was to explore possible ultrasound parameters for the early detection of alcohol-mediated fetal somatic and central nervous system (CNS) maldevelopment. Maternal alcohol ingestion during pregnancy may lead to fetal alcohol spectrum disorders (FASD), which encompass a broad range of structural abnormalities including growth impairment, specific craniofacial features and CNS abnormalities. Early detection of fetuses at risk of FASD would support earlier interventions. METHODS: We performed a longitudinal prospective pilot study from 2004 to 2006 at two sites in Ukraine. A sample of pregnant women who reported consuming moderate-to-heavy amounts of alcohol participated in a comprehensive maternal interview, and received ultrasound evaluation of fetal growth and specific fetal brain measurements during the second and third trimesters. These measurements were compared with those collected from a group of pregnant women who consumed little-to-no alcohol during pregnancy, and who were recruited and followed in the same manner. RESULTS: From 6745 screened women, 84 moderate-to-heavy alcohol users and 82 comparison women were identified and ultrasound examinations performed. After controlling for maternal smoking, alcohol-exposed fetuses had shorter mean femur length, caval-calvarial distance and frontothalamic measurements in the second trimester (P < 0.05), and alcohol-exposed fetuses also had shorter frontothalamic distance measurements in the third trimester relative to comparison fetuses (P < 0.05). In addition, after controlling for maternal smoking, both mean orbital diameter and biparietal diameter measurements were significantly smaller on average in the alcohol-exposed group in the third trimester relative to comparison fetuses (P < 0.05). CONCLUSIONS: Significant differences in selected somatic and brain measurements were noted between alcohol-exposed and comparison fetuses, suggesting these markers may be further explored for clinical utility in prenatal identification of affected children. Further study correlating these findings with alcohol-related physical features of the newborn and subsequent comparisons of neuro-developmental outcomes will help define potential uses of prenatal ultrasound for intervention and prevention of FASD.

Prenatal three-dimensional ultrasound: perception of sonographers, sonologists and undergraduate students.

OBJECTIVE: To assess the perception of non-pregnant sonographers, sonologists and undergraduate students on the use of three-dimensional (3D) ultrasound technology in fetal medicine. METHODS: This was a study of two groups of non-pregnant subjects. Group I included 520 (305 female, 215 male) medical professionals who completed a questionnaire after attending a lecture on 3D imaging. Factors such as gender, career and having children were analyzed with respect to the attendee's responses about use of 3D ultrasound for medical purposes and for reassurance. Group II included 137 (75 female, 60 male, two unknown) undergraduate students from bioengineering, psychology and physiology classes who completed a questionnaire after attending a brief presentation on two-dimensional and 3D fetal imaging. Factors such as gender and area of educational interest were analyzed with respect to the students' responses about the use of 3D ultrasound for medical purposes and for parental-fetal attachment. RESULTS: In Group I, 63% said that they would like to have a 3D ultrasound examination in the future, while 14% said that they would not. Common reasons given for wanting a 3D ultrasound exam in the future were for medical purposes (39%) or reassurance (18%). The main differences perceived between two-dimensional (2D) and 3D ultrasound were medical advantages (65%) and parental reassurance (28%). 62.4% of Group I thought 3D technology should be in wide use in obstetric ultrasound and 73.6% thought that 3D ultrasound would reassure parents carrying normal fetuses. Gender, age and career did not have a significant influence on perception of 3D ultrasound. In Group II, the majority (91%) said they could see a remarkable difference between 2D and 3D ultrasound. 83% responded that they would like to have a 3D ultrasound examination of their own baby in the future for the following reasons: 34% for the detailed picture, 31% for increased abnormality detection, 13% for reassurance or curiosity; 8% thought it would be unnecessary or a negative experience. Concerning parental-fetal attachment, 72% thought 3D ultrasound would have a positive effect. The majority of Group II (93%) thought 3D ultrasound would be valuable and 56% thought 3D ultrasound would assist in diagnosing fetal abnormalities. There was no significant relationship between gender, age or area of interest and the perception of 3D ultrasound. CONCLUSIONS: Responses by sonographers and physicians suggest that 3D ultrasound will have a role in the future for medical indications and in reassuring patients carrying normal fetuses. Our results also suggest that undergraduate students believe that 3D ultrasound will be a valuable technique in obstetrics and that it will have a positive effect on parental-fetal attachment.

RIZ1 repression is associated with insulin-like growth factor-1 signaling activation in chronic myeloid leukemia cell lines.

RIZ1 is a histone methyltransferase whose expression and activity are reduced in many cancers. In chronic myelogenous leukemia (CML), blastic transformation is associated with loss of heterozygosity in the region where RIZ1 is located and with decreased RIZ1 expression. Forced RIZ1 expression in model CML blast crisis (BC) cell lines decreases proliferation, increases apoptosis and enhances differentiation. We characterized molecular mechanisms that may contribute to potential CML tumor suppressor properties of RIZ1. Several RIZ1-regulated genes involved in insulin-like growth factor-1 (IGF-1) signaling were identified using cDNA microarrays. RIZ1 was shown to associate with promoter regions of IGF-1 and to increase histone H3 lysine 9 methylation using chromatin immunoprecipitation assays. IGF-1-blocking antibody was used to demonstrate the importance of autocrine IGF-1 signaling in CML-BC cell line viability. Forced RIZ1 expression in CML-BC cell lines decreases IGF-1 receptor activation and activation of downstream signaling components extracellular signal-regulated kinase 1/2 and AKT. These results highlight the therapeutic potential of inhibiting IGF-1 pathway in the acute phase of CML.

Prenatal detection of associated anomalies in fetuses diagnosed with cleft lip with or without cleft palate in utero.

OBJECTIVE: The aim of this study was to determine the prenatal detection rate of associated anomalies in fetuses with a suspected cleft lip with or without cleft palate. METHODS: Fetuses with a suspected cleft lip with or without cleft palate, determined by prenatal ultrasound, were prospectively enrolled. Additional anomalies suspected by ultrasound or genetic testing were recorded. Postnatal outcome was obtained. RESULTS: Forty-five fetuses with a cleft lip with or without cleft palate, diagnosed prenatally with either two-dimensional and/or three-dimensional ultrasound, were studied. Postnatal follow-up revealed that 16 (35.6%) of these 45 fetuses had an additional structural or syndromic abnormality. Of the 37 fetuses with prenatally determined 'isolated' cleft lip with or without cleft palate, eight (21.6%) had an additional malformation identified after delivery. CONCLUSION: In pregnancies complicated by a cleft lip with or without cleft palate, patients should be informed of the risks of associated anomalies, some of which may be undetected prenatally.

The importance of pancreatic embryonic epithelium for mesenchyme-to-epithelial transition during islet development.

Stem or progenitor cells are a promising potential alternative source of pancreatic islets for transplantation in the treatment of juvenile-onset diabetes. However, to derive islets from such cells, it is important to elucidate the mechanisms of normal pancreatic development. Previous work in our laboratory has shown that, contrary to previous thinking, pancreatic mesenchyme when combined with pancreatic epithelium can contribute cells to islets. However, the signals and role of individual tissues involved in this mesenchyme-to-epithelial transition (MET) have yet to be elucidated. The aim of this study was to investigate whether MET can occur in the absence of pancreatic epithelium. Chick and quail eggs were incubated for 4 days and the dorsal pancreatic buds and stomach rudiments were microdissected. Mesenchyme and epithelium of the organ rudiments were separated after collagenase treatment. Separated pancreatic mesenchyme were cultured alone and in combination with stomach (nonpancreatic). After 7 days of culture, the specimens were analysed using immunohistochemistry for quail-specific nucleolar antigen (QCPN), insulin, and islet precursor cell marker (ISL-1). Pancreatic mesenchyme when cultured in the absence of epithelium did not differentiate into islets, but differentiated into fibroblast-like cells. When pancreatic mesenchyme were cultured in combination with stomach epithelium, there was no evidence of mesenchymally derived islets. We have demonstrated that pancreatic mesenchyme require pancreatic epithelium to differentiate into islet cells. These findings further increase our understanding of normal pancreatic islet development and may help to elucidate the molecular mechanisms of MET in islet development.

Effects of ultrasound on maternal-fetal bonding: a comparison of two- and three-dimensional imaging.

OBJECTIVES: The purpose of this study was to evaluate the effect of two-dimensional (2DUS) compared to three-dimensional ultrasound (3DUS) imaging on the maternal-fetal bonding process. METHODS: Fifty mothers who had 2DUS and 50 who had 2DUS and 3DUS were included in the study. A postpartum survey by telephone interview was carried out to assess maternal-fetal bonding. Bonding was evaluated by analysis of extent of prenatal image sharing, maternal ability to form a mental picture of the baby and mother's comments about their ultrasound images. Data were analyzed using the independent t-test, Chi-square and Mann-Whitney U-tests. RESULTS: Mothers who received 3DUS showed their ultrasound images to more people (median, 27.5; interquartile range, 14.5-40.0) than mothers receiving 2DUS alone (median, 11.0; interquartile range, 5.0-25.5) (P < 0.001, Z = -3.539). Eighty-two percent of the subjects screened with 3DUS had a greater tendency to form a mental picture of the baby postexamination compared to 39% of the 2DUS subjects (P < 0.001, Z = -3.614). Mothers receiving a 3DUS study were more likely to receive comments on the similarities/differences of the neonate compared to those having 2DUS studies. Furthermore, 70% of the mothers receiving 3DUS felt they 'knew' the baby immediately after birth vs. 56% of the mothers receiving 2DUS (P = 0.009, Z = -2.613). Both 2DUS and 3DUS experiences were positive, however, the comments made by the mothers undergoing 3DUS (n = 18) were more exclamatory (amazed, wonderful, fabulous) than those undergoing 2DUS (n = 4). Patients having a 3DUS examination consistently scored higher than those having a 2DUS examination alone for all categories of maternal-fetal bonding. CONCLUSION: 3DUS appears to more positively influence the perceptions of mothers to their babies postbirth compared to 2DUS. Specifically, mothers who had 3DUS showed their ultrasound images to a greater number of people compared to mothers who had 2DUS alone and this may represent mother's social support system. 3DUS may have a greater impact on the maternal-fetal bonding process.

Psychological consequences of maxillofacial trauma: a preliminary study.

AIMS: To identify the prevalence of post-traumatic psychological symptoms after maxillofacial trauma and prognostic factors related to poor outcome. METHODS: Thirty-nine patients were assessed within 10 days of injury and 24 again 4-6 weeks later using five standardised self-report measures on each occasion and a short structured interview at the time of initial contact. RESULTS: Specific post-traumatic psychological symptoms were present at initial assessment in 21 patients (54%), with 9 (41%) meeting diagnostic criteria for post-traumatic stress disorder at review 4-6 weeks later. Other psychiatric problems, such as anxiety and depression, were identified by the General Health Questionnaire and the Hospital Anxiety and Depression Scale. Characteristics associated with poorer outcome included: a previous history of psychological distress; fear of the unknown, and female sex. CONCLUSION: These findings highlight the adverse psychological effect of maxillofacial trauma both immediately after the event and 4-6 weeks after injury. Proper assessment of injured patients must include psychological aspects and further research is needed to identify the most appropriate response.

The innervation of FGF-induced additional limbs in the chick embryo.

Motoneurones that supply the vertebrate limb innervate their muscle targets in a highly reproducible manner. As development proceeds, these limb-specific motoneurones send out axons, which grow towards the developing limb and then congregate at its base to form the plexus. In the plexus, in response to unknown positional cues, these axons rearrange, often changing their original spatial relationships, before sorting out to emerge in the defined nerve trunks that innervate the limb. Several proposals have been put forward to explain how this reproducible innervation pattern is achieved. These include (1) that early differences in the motoneurone identity dictate their future axonal trajectories, (2) that axons actively respond to attractive or repulsive positional cues provided by the limb bud itself, or (3) that motor axons are passively deployed, following pathways of least mechanical resistance. We have addressed the question of the relative roles of motoneurone identity and the signals that the axons encounter on their journey towards the limb bud. Using the developing chick embryo as our experimental model we tested the effect of providing an additional limb target for motor axons leaving the flank level of the spinal cord. To do this we placed FGF-soaked beads in the presumptive flank of 2-day-old chick embryos. This treatment induces an additional limb containing muscles. We investigated whether such additional limbs are innervated and by which neurones. We show that rather than the additional limbs being solely supplied by axons diverted from the two existing limb plexuses, motoneurones that normally supply the flank alter their trajectories to enter the induced limb. Once in the limb, axons respond to positional cues within the bud to generate the stereotypical innervation pattern. Our results show that the tendency of 'flank' motoneurones to innervate flank can be overcome by the presence of an additional limb.

A waste of time: non-attendance at out-patient clinics in a Scottish NHS Trust.

UNLABELLED: The Patient's Charter states that patients have a responsibility to attend out-patient appointments or to notify the hospital if they are unable to do so. Non-attendance without notification has substantial financial costs for the NHS and may have clinical implications to the non-attender and other patients on the waiting list. OBJECTIVE: To identify reasons for non-attendance of patients for their first appointment after referral. DESIGN: A survey by questionnaire of a random sample of non-attenders of an NHS trust. SETTING: Aberdeen Royal Hospitals NHS Trust. SUBJECTS: Ten per cent of all non-attenders to the Trust out-patient clinics over a twelve month period. RESULTS: One hundred and fifty five (32%) patients contributed to the survey. Cancellations accounted for 22% (34) of missed appointments with factors relating to illness or treatment, being the most common reason (14; 44%). Patients failing to attend without prior notification stated that hospital administrative problems (75; 57%) and personal administrative problems (31; 23%) were the primary reasons. Clinical speciality, day of the week, the month, availability of a telephone or car, and socioeconomic group were not significantly associated with non-attendance. CONCLUSION: The majority of patients show a responsible attitude to attendance at outpatients when appointments were received. Non-attendance was found to be due to a combination of institutional factors (commonly administrative) and patient factors such as forgetting about the appointment.

Erythrocyte and erythropoietin responses to hemorrhage in the immature and near term ovine fetus.

OBJECTIVE: Previous reports suggested that immature ovine fetuses have a greater erythropoietin response to hemorrhage than those near term. This study tested the hypothesis that immature ovine fetuses would expand their red cell mass more rapidly than near term fetuses after hemorrhage. STUDY DESIGN: Chronically catheterized immature ovine fetuses at 109.5 +/- 0.3 (mean +/- SE) days' gestation (term = 150 days) were studied over a 10-day period. They either underwent hemorrhage of 40% of their measured blood volume on day 3 or were in a time control group monitored without hemorrhage. Red cell mass, hematocrit, blood volume, plasma volume, and plasma erythropoietin concentrations were measured at 24- and 48-hour intervals. Responses in the immature fetuses were compared with responses in near term fetuses. RESULTS: In the control group red cell mass, hematocrit, blood, and plasma volumes increased significantly, whereas plasma erythropoietin concentration decreased significantly with advancing gestational age. In immature fetuses that underwent hemorrhage, the relative changes in red cell mass, hematocrit, and plasma erythropoietin concentration were not significantly different from those seen in the near term fetuses. The only significant posthemorrhage difference was that the increases in blood and plasma volumes were greater in the immature compared with the near term fetuses. CONCLUSION: Immature and mature fetuses have similar erythrocyte and erythropoietin responses to moderately severe hemorrhage. The larger blood and plasma volume responses in the immature fetuses are consistent with the concept that they have a greater extracellular fluid volume.

A monkey model for enterohemorrhagic Escherichia coli infection.

Adult Macaca radiata (n=22) were infected intragastrically with 10(12) Escherichia coli O157:H7 strain 84-01, which produces Shiga toxins 1 and 2. Clinical symptoms and bacterial excretion were documented in each monkey for a specified time period before they were killed. At necropsy, samples were obtained for culture and histologic and ultrastructural examination. Seventeen monkeys had diarrhea: E. coli O157 was isolated from postinfection stool samples from all monkeys and from autopsy cultures for 14 of 22 monkeys. Histologic examination showed attaching-effacing lesions, which appeared at 12 h and persisted for 7 days, in 12 monkeys. Widening of the intercellular spaces, degeneration and vacuolization of the epithelial cells, epithelial tufting, extrusion of epithelial cells, and neutrophilic infiltration were characteristic features seen in 20 of the 22 infected monkeys but not in 4 control monkeys. This monkey model closely parallels the early stages of the disease produced by E. coli O157:H7 and would be useful in the further study of pathogenic mechanisms and prevention methods in enterohemorrhagic E. coli infections.

Detection of Jarcho-Levin syndrome at 12 weeks' gestation by nuchal translucency screening and three-dimensional ultrasound.

Jarcho-Levin syndrome (JLS) or spondylothoracic dysotosis causes early newborn death from respiratory insufficiency. Prenatal diagnosis is possible using fetal ultrasound, but requires a high level of suspicion and definitive diagnosis may be delayed well into the second trimester. We present a case in which a combination of three-dimensional ultrasound and measurement of nuchal translucency thickness allowed successful detection of recurrent JLS at 12 weeks' gestation.