Direct purification and immobilization of his-tagged enzymes using unmodified nickel ferrite NiFe2O4 magnetic nanoparticles.

Purification of valuable engineered proteins and enzymes can be laborious, costly, and generating large amount of chemical waste. Whilst enzyme immobilization can enhance recycling and reuse of enzymes, conventional methods for immobilizing engineered enzymes from purified samples are also inefficient with multiple-step protocols, regarding both the carrier preparation and enzyme binding. Nickel ferrite magnetic nanoparticles (NiFe2O4 MNPs) offer distinct advantages in both purification and immobilization of enzymes. In this work, we demonstrate the preparation of NiFe2O4 MNPs via a one-step solvothermal synthesis and their use in direct enzyme binding from cell lysates. These NiFe2O4 MNPs have showed an average diameter of 8.9 ± 1.7 nm from TEM analysis and a magnetization at saturation (Ms) value of 53.0 emu g-1 from SQUID measurement. The nickel binding sites of the MNP surface allow direct binding of three his-tagged enzymes, D-phenylglycine aminotransferase (D-PhgAT), Halomonas elongata ω-transaminase (HeωT), and glucose dehydrogenase from Bacillus subtilis (BsGDH). It was found that the enzymatic activities of all immobilized samples directly prepared from cell lysates were comparable to those prepared from the conventional immobilization method using purified enzymes. Remarkably, D-PhgAT supported on NiFe2O4 MNPs also showed similar activity to the purified free enzyme. By comparing on both carrier preparation and enzyme immobilization protocols, use of NiFe2O4 MNPs for direct enzyme immobilization from cell lysate can significantly reduce the number of steps, time, and use of chemicals. Therefore, NiFe2O4 MNPs can offer considerable advantages for use in both enzyme immobilization and protein purification in pharmaceutical and other chemical industries.

Predictors of Postpartum Depression Among Women with Low Incomes in the United States.

PURPOSE: To evaluate the predictors of postpartum depression and to examine the effects of maternal racial identity on postpartum depression among women with low incomes in the United States. STUDY DESIGN AND METHODS: We conducted a secondary data analysis using baseline data from the Baby's First Years study, including postpartum women living below the federal poverty line who were recruited from four diverse communities in the United States. Postpartum depression symptoms were assessed using the Center for Epidemiologic Studies Depression Scale (CES-D). Logistic regressions were performed to identify associations between pre-identified postpartum depression risk factors and postpartum depression among participants, followed by predictive margins analyses to elaborate on probability of postpartum depression across different racial identity groups in the sample. RESULTS: The sample included 1,051 postpartum women. Almost one-half of participants identified as Black (45.9%), followed by White (20.7%), American Indian (1.8%), and Asian (1.3%). Prevalence of postpartum depression in the sample was 24%. Financial insecurity and alcohol use were positively associated with postpartum depression, whereas level of education and reported physical health were negatively associated with postpartum depression. Mothers who identified as Black had an 8.3% higher probability of postpartum depression than that of White mothers in the sample. CLINICAL IMPLICATIONS: Nurses working with populations with low income should assess social determinants of health to provide holistic mental health care. Women with low incomes should be referred to resources which account for their financial burden. Racial disparities exist in perinatal care, and birth and postpartum mental health outcomes. It is crucial to address the systemic racism faced by Black mothers experiencing postpartum depression.

Knowledge of Postbirth Warning Signs and Postpartum Education Among Women in Ghana.

OBJECTIVE: To assess the relationship between postpartum education and knowledge of postbirth warning signs among women in Ghana. DESIGN: Cross-sectional survey. SETTING: Tamale West Hospital in Tamale Metropolitan Area, Ghana. PARTICIPANTS: Women (N = 151) who gave birth to healthy newborns and were admitted to the postnatal ward. METHODS: We collected data from surveys distributed in the hospital. The survey included items for sociodemographic characteristics, obstetric history, postpartum education provided, and knowledge of nine common postbirth warning signs. We used descriptive statistics and multivariate logistic regression models to analyze the data. RESULTS: Participants reported knowing an average of 5.2 of 9 (SD = 2.84) postbirth warning signs. Severe bleeding (94.70%, n = 143), fever (82.12%, n = 124), and severe headache (72.19%, n = 109) were the postbirth warning signs most frequently identified by participants. Swelling in the leg (37.09%, n = 56) and thoughts of hurting oneself (33.11%, n = 50) were the postbirth warning signs least frequently identified by participants. Knowledge about postbirth warning signs was positively associated with reports of receiving educational handouts on the postnatal ward (adjusted OR = 4.64, 95% confidence interval [1.27, 17.04]) and reports that four or more postpartum complications were taught before hospital discharge (adjusted OR = 27.97, 95% confidence interval [7.55, 103.57]) compared to zero to three postpartum complications. CONCLUSION: All women need comprehensive discharge education on the warning signs of complications after birth. Promoting knowledge of postbirth warning signs can decrease delays in seeking care and contribute to the reduction of maternal mortality in Ghana.

Gold-iron oxide (Au/Fe3O4) magnetic nanoparticles as the nanoplatform for binding of bioactive molecules through self-assembly.

Nanomedicine plays a crucial role in the development of next-generation therapies. The use of nanoparticles as drug delivery platforms has become a major area of research in nanotechnology. To be effective, these nanoparticles must interact with desired drug molecules and release them at targeted sites. The design of these "nanoplatforms" typically includes a functional core, an organic coating with functional groups for drug binding, and the drugs or bioactive molecules themselves. However, by exploiting the coordination chemistry between organic molecules and transition metal centers, the self-assembly of drugs onto the nanoplatform surfaces can bypass the need for an organic coating, simplifying the materials synthesis process. In this perspective, we use gold-iron oxide nanoplatforms as examples and outline the prospects and challenges of using self-assembly to prepare drug-nanoparticle constructs. Through a case study on the binding of insulin on Au-dotted Fe3O4 nanoparticles, we demonstrate how a self-assembly system can be developed. This method can also be adapted to other combinations of transition metals, with the potential for scaling up. Furthermore, the self-assembly method can also be considered as a greener alternative to traditional methods, reducing the use of chemicals and solvents. In light of the current climate of environmental awareness, this shift towards sustainability in the pharmaceutical industry would be welcomed.

Discharge Readiness and Associated Factors Among Postpartum Women in Tamale, Ghana.

Ghana experiences a relatively high maternal mortality ratio, with the majority of maternal deaths occurring in the postpartum period. Discharge readiness is a reliable indicator of maternal health outcomes and involves a postpartum woman's perception of preparedness to leave the hospital following delivery. We measured the discharge readiness of postpartum women in Ghana through an institutional-based cross-sectional study involving 151 participants. Participants completed an interviewer-administered survey, and data analyses included linear regression models. The study sample demonstrated relatively high discharge readiness, with a mean score of 177.57 on a scale from 0.00 to 220.00. Higher gravidity was positively associated with discharge readiness score, while longer length of hospital stay and receiving educational handouts were negatively associated with discharge readiness score. Clinical interventions addressing the factors found to be associated with discharge readiness have significant potential to improve postpartum care and maternal outcomes in Ghana.

Cross-Cultural Adaptation and Validation of the Dagbani Readiness for Hospital Discharge Scale-New Mother.

The Readiness for Hospital Discharge Scale-New Mother (RHDS-NM), originally developed for use in English-speaking populations, is an instrument designed to measure women's perceptions of readiness for hospital discharge after birth. We translated and cross-culturally adapted the RHDS-NM into Dagbani and conducted reliability and validity assessments of the new Dagbani RHDS-NM in the Tamale Metropolitan Area, Ghana. The average scale-level content validity index was excellent at 1.00. The Dagbani RHDS-NM had a Cronbach's α reliability coefficient of .94. Exploratory factor analysis indicated four factors with Cronbach's α reliability coefficients of .958, .915, .899, and .667 that represented the Personal Status, Knowledge, Expected Support, and Coping Ability subscales, respectively. Our findings provide initial evidence to support the reliability and validity of the Dagbani RHDS-NM.

Antibacterial Activities of Ga(III) against E. coli Are Substantially Impacted by Fe(III) Uptake Systems and Multidrug Resistance in Combination with Oxygen Levels.

The continued emergence and spread of antimicrobial resistance (AMR), particularly multidrug resistant (MDR) bacteria, are increasing threats driving the search for additional and alternative antimicrobial agents. The World Health Organization (WHO) has categorized bacterial risk levels and includes Escherichia coli among the highest priority, making this both a convenient model bacterium and a clinically highly relevant species on which to base investigations of antimicrobials. Among many compounds examined for use as antimicrobials, Ga(III) complexes have shown promise. Nonetheless, the spectrum of activities, susceptibility of bacterial species, mechanisms of antimicrobial action, and bacterial characteristics influencing antibacterial actions are far from being completely understood; these are important considerations for any implementation of an effective antibacterial agent. In this investigation, we show that an alteration in growth conditions to physiologically relevant lowered oxygen (anaerobic) conditions substantially increases the minimum inhibitory concentrations (MICs) of Ga(III) required to inhibit growth for 46 wild-type E. coli strains. Several studies have implicated a Trojan horse hypothesis wherein bacterial Fe uptake systems have been linked to the promotion of Ga(III) uptake and result in enhanced antibacterial activity. Our studies show that, conversely, the carriage of accessory Fe uptake systems (Fe_acc) significantly increased the concentrations of Ga(III) required for antibacterial action. Similarly, it is shown that MDR strains are more resistant to Ga(III). The increased tolerance of Fe_acc/MDR strains was apparent under anaerobic conditions. This phenomenon of heightened tolerance has not previously been shown although the mechanisms remain to be defined. Nonetheless, this further highlights the significant contributions of bacterial metabolism, fitness, and AMR characteristics and their implications in evaluating novel antimicrobials.

Localization of Coated Iron Oxide (Fe3O4) Nanoparticles on Tomato Seeds and Their Effects on Growth.

Food demand due to the growing global population has been stretching the agriculture sector to the limit. This demands the cultivation of plants in shrinking land areas which makes the search for highly effective systems for plant nutrition and pest control important. In this context, the application of nanoparticles (NPs) in agriculture can have a transformative effect on food production techniques as it can enable the delivery of bioactive agents (including growth factors, pesticides, and fungicides) directly to plants. Herein, we report the application of unfunctionalized as well as amine-functionalized and polycaprolactone-coated Fe3O4 NPs to seed treatment in tomato (Solanum lycopersicum). The study reveals that the treatment has no side effects on plant germination and development. Furthermore, the translocation of NPs in seeds and seedlings posttreatment depends on the surface functionalization of the NPs. X-ray fluorescence spectroscopy analysis of seedlings suggested that around 66% of unfunctionalized Fe3O4 NPs were translocated in the cotyledons, while only 50% of functionalized NPs (both amine and polycaprolactone) were translocated. Our results demonstrate that all particles were taken up by the seeds, thus suggesting that the functionalized NPs can act as a versatile platform for delivering of active compounds, such as fungicides and growth factor agents.

A fetus coexisting with a complete hydatidiform mole with trisomy 9 of maternal origin.

A complete hydatidiform mole (CHM) coexisting with a viable fetus is a rare finding in pregnancies. Accurate diagnosis often relies on ultrasonographic, histopathological and molecular techniques in the definite diagnosis. To the best of our knowledge, a liveborn fetus coexisting with CHM with trisomy 9 has not been described. The use of molecular genotyping and immunohistochemical laboratory investigations enabled the CHM to be fully characterized. Postzygotic diploidization of a triploid conception arising from dispermy is the proposed mechanism of its formation.

Combining child social skills training with a parent early intervention program for inhibited preschool children.

BACKGROUND: Previous studies have demonstrated the efficacy of early intervention for anxiety in preschoolers through parent-education. The current study evaluated a six-session early intervention program for preschoolers at high risk of anxiety disorders in which a standard educational program for parents was supplemented by direct training of social skills to the children. METHODS: Seventy-two children aged 3-5 years were selected based on high behavioural inhibition levels and concurrently having a parent with high emotional distress. Families were randomly assigned to either the intervention group, which consisted of six parent-education group sessions and six child social skills training sessions, or waitlist. After six months, families on waitlist were offered treatment consisting of parent-education only. RESULTS: Relative to waitlist, children in the combined condition showed significantly fewer clinician-rated anxiety disorders and diagnostic severity and maternal (but not paternal) reported anxiety symptoms and life interference at six months. Mothers also reported less overprotection. These gains were maintained at 12-month follow-up. Parent only education following waitlist produced similar improvements among children. Quasi-experimental comparison between combined and parent-only interventions indicated greater reductions from combined intervention according to clinician reports, but no significant differences on maternal reports. CONCLUSIONS: Results suggest that this brief early intervention program for preschoolers with both parent and child components significantly reduces risk and disorder in vulnerable children. The inclusion of a child component might have the potential to increase effects over parent-only intervention. However, future support for this conclusion through long-term, randomised controlled trials is needed.

Effects of multielectrode renal denervation on elevated sympathetic nerve activity and insulin resistance in metabolic syndrome.

OBJECTIVE: This study aimed to investigate the effects of renal denervation (RDN) on sympathetic nerve activity and insulin resistance in patients with metabolic syndrome. METHODS: Seventeen patients fulfilled at least four of five criteria for metabolic syndrome and under stable use of at least two antihypertensive drugs were randomized in 3 : 1 ratio to RDN (n = 13, 12 men, age: 58 ±â€Š7 years) and control groups (n = 4, three men, age: 60 ±â€Š5 years) and followed up for 3 months. Muscle sympathetic nerve activity (MSNA) at rest and during standard 75 g oral glucose tolerance test (OGTT) was assessed. RESULTS: In the RDN group, office and average 24-h blood pressures reduced by 16 ±â€Š21/10 ±â€Š11 mmHg (P = 0.01/0.007) and 14 ±â€Š16/5 ±â€Š8 mmHg (P = 0.008/0.03) respectively; waist circumference reduced by 3.1 ±â€Š3.6 cm (P = 0.008); and resting MSNA reduced from 55 ±â€Š9 bursts per minute to 46 ±â€Š8 bursts per minute (P = 0.0008) at month 3 post-RDN. During OGTT, although blunted MSNA responses were noted at baseline throughout the 120-min test, improved MSNA responses with burst frequency increased to 52 ±â€Š8 bursts per minute (P < 0.001 vs. the resting MSNA, n = 13) at 30 min and to 54 ±â€Š8 bursts per minute (P = 0.004 vs. the resting MSNA, n = 10) at 120 min and were observed at month 3 post-RDN. No such improvements were observed in the controls. No significant change was observed in the HOMA-IR in both groups at month 3. CONCLUSION: In this pilot study of patients with metabolic syndrome and associated hypertension, RDN reduced elevated sympathetic nerve activity and restored the normal neural response to oral glucose loading.

Renal Nerve Stimulation-Induced Blood Pressure Changes Predict Ambulatory Blood Pressure Response After Renal Denervation.

Blood pressure (BP) response to renal denervation (RDN) is highly variable and its effectiveness debated. A procedural end point for RDN may improve consistency of response. The objective of the current analysis was to look for the association between renal nerve stimulation (RNS)-induced BP increase before and after RDN and changes in ambulatory BP monitoring (ABPM) after RDN. Fourteen patients with drug-resistant hypertension referred for RDN were included. RNS was performed under general anesthesia at 4 sites in the right and left renal arteries, both before and immediately after RDN. RNS-induced BP changes were monitored and correlated to changes in ambulatory BP at a follow-up of 3 to 6 months after RDN. RNS resulted in a systolic BP increase of 50±27 mm Hg before RDN and systolic BP increase of 13±16 mm Hg after RDN (P<0.001). Average systolic ABPM was 153±11 mm Hg before RDN and decreased to 137±10 mm Hg at 3- to 6-month follow-up (P=0.003). Changes in RNS-induced BP increase before versus immediately after RDN and changes in ABPM before versus 3 to 6 months after RDN were correlated, both for systolic BP (R=0.77, P=0.001) and diastolic BP (R=0.79, P=0.001). RNS-induced maximum BP increase before RDN had a correlation of R=0.61 (P=0.020) for systolic and R=0.71 (P=0.004) for diastolic ABPM changes. RNS-induced BP changes before versus after RDN were correlated with changes in 24-hour ABPM 3 to 6 months after RDN. RNS should be tested as an acute end point to assess the efficacy of RDN and predict BP response to RDN.

Diagnostic yield of array CGH in patients with autism spectrum disorder in Hong Kong.

BACKGROUND: Chromosomal microarray offers superior sensitivity for identification of submicroscopic copy number variants (CNV) and it is advocated to be the first tier genetic testing for patients with autism spectrum disorder (ASD). In this regard, diagnostic yield of array comparative genomic hybridization (CGH) for ASD patients is determined in a cohort of Chinese patients in Hong Kong. METHODS: A combined adult and paediatric cohort of 68 Chinese ASD patients (41 patients in adult group and 27 patients in paediatric group). The genomic DNA extracted from blood samples were analysed by array CGH using NimbleGen CGX-135K oligonucleotide array. RESULTS: We identified 15 CNV and eight of them were clinically significant. The overall diagnostic yield was 11.8 %. Five clinically significant CNV were detected in the adult group and three were in the paediatric group, providing diagnostic yields of 12.2 and 11.1 % respectively. The most frequently detected CNV was 16p13.11 duplications which were present in 4 patients (5.9 % of the cohort). CONCLUSIONS: In this study, a satisfactory diagnostic yield of array CGH was demonstrated in a Chinese ASD patient cohort which supported the clinical usefulness of array CGH as the first line testing of ASD in Hong Kong.

Persistent Increase in Blood Pressure After Renal Nerve Stimulation in Accessory Renal Arteries After Sympathetic Renal Denervation.

Blood pressure response to renal denervation is highly variable, and the proportion of responders is disappointing. This may be partly because of accessory renal arteries too small for denervation, causing incomplete ablation. Renal nerve stimulation before and after renal denervation is a promising approach to assess completeness of renal denervation and may predict blood pressure response to renal denervation. The objective of the current study was to assess renal nerve stimulation-induced blood pressure increase before and after renal sympathetic denervation in main and accessory renal arteries of anaesthetized patients with drug-resistant hypertension. The study included 21 patients. Nine patients had at least 1 accessory renal artery in which renal denervation was not feasible. Renal nerve stimulation was performed in the main arteries of all patients and in accessory renal arteries of 6 of 9 patients with accessory arteries, both before and after renal sympathetic denervation. Renal nerve stimulation before renal denervation elicited a substantial increase in systolic blood pressure, both in main (25.6±2.9 mm Hg; P<0.001) and accessory (24.3±7.4 mm Hg; P=0.047) renal arteries. After renal denervation, renal nerve stimulation-induced systolic blood pressure increase was blunted in the main renal arteries (Δ systolic blood pressure, 8.6±3.7 mm Hg; P=0.020), but not in the nondenervated renal accessory renal arteries (Δ systolic blood pressure, 27.1±7.6 mm Hg; P=0.917). This residual source of renal sympathetic tone may result in persistent hypertension after ablation and partly account for the large response variability.

A Fetus with Hb Bart's Disease Due to Maternal Uniparental Disomy for Chromosome 16.

We here report an unusual case of Hb Bart's (γ4) disease. Thalassemia screening of a couple showed that the wife was an α(0)-thalassemia (α(0)-thal) carrier and her husband's mean corpuscular volume (MCV) was normal. Chorionic villus sampling (CVS) was performed at 13 weeks' gestation for positive Down syndrome screening and chromosomal study of the cultured CVS showed a normal karyotype. Ultrasound examination at 22 weeks' gestation showed fetal cardiomegaly and raised middle cerebral artery peak systolic velocity. Cordocentesis confirmed fetal anemia and showed Hb Bart's disease. Multiplex gap-polymerase chain reaction (gap-PCR) for α-thal deletions on DNA extracted from the CVS showed the presence of a homozygous α(0)-thal - -(SEA) (Southeast Asian) deletion. The husband was found to be a carrier of the α(+)-thal -α(3.7) (rightward) deletion. Non paternity was excluded by fluorescent PCR using short tandem repeat (STR) markers on chromosomes 13, 18 and 21. A de novo terminal deletion of chromosome 16 was excluded by array comparative genomic hybridization (aCGH). Detection of uniparental disomy (UPD), using STR markers on chromosome 16 showed maternal uniparental isodisomy from 16pter to 16p13.2, and uniparental heterodisomy from 16p13.13 to 16qter.

Effects of multielectrode renal denervation on cardiac and neurohumoral adaptations in resistant hypertension with cardiac hypertrophy: an EnligHTN I substudy.

OBJECTIVE: This EnligHTN I nonrandomized substudy investigated the effect of multielectrode renal denervation (RDN) on cardiac and neurohumoral adaptations. METHODS: Eighteen patients with true drug-resistant hypertension [age: 56 ±â€Š10 years, 12 men, BMI: 33.6 ±â€Š5.4 kg/m, office blood pressure (BP) by automatic device (Omron): 182 ±â€Š19/97 ±â€Š18 mmHg and ambulatory BP (Spacelabs): 153 ±â€Š16/87 ±â€Š15 mmHg receiving 4.5 antihypertensive drugs/day] and left ventricular hypertrophy underwent multielectrode RDN (EnligHTN system; St. Jude Medical), whereas 10 patients served as controls. Both groups were followed-up for 6 months. RESULTS: Demographic data were homogenous between both patient groups. In addition to reduction of office (-42/-17 mmHg, P < 0.001) and ambulatory (-19/-9 mmHg, P < 0.001) BP, RDN contributed to attenuation of left ventricular mass index from 140.0 ±â€Š17.0 g/m (57.9 ±â€Š7.9 g/m) to 126.7 ±â€Š19.2 g/m (52.6 ±â€Š8.4 g/m) (P < 0.01 for both) and left atrial diameter from 42.4 ±â€Š4.3 to 40.6 ±â€Š3.6 mm (P = 0.004) at 6 months. Up to 56% of the RDN-group patients achieved a target of less than 140/90 mmHg in the office BP; proportion of RDN-group patients with concentric left ventricular hypertrophy had decreased by 39%; mitral lateral E/E' ratio decreased from 14.8 ±â€Š6.1 to 12.0 ±â€Š3.2 (P = 0.016); isovolumic relaxation time shortened from 109.8 ±â€Š16.2 to 100.8 ±â€Š17.1 ms (P = 0.003); and N-terminal pro B-type natriuretic peptide levels reduced from 84.9 ±â€Š35.9 to 57.2 ±â€Š38.8 pg/ml (P < 0.001) significantly at 6 months post-RDN. Control patients exhibited no significant changes in all the above parameters (P > 0.05) at 6 months. CONCLUSION: Multielectrode RDN contributes to improvement of diastolic dysfunction, reduction of left ventricular mass and attenuation of NT-proBNP, suggesting additional cardiovascular benefits in drug-resistant hypertension associated with left ventricular hypertrophy.

Thoracic Spinal Cord Stimulation for Heart Failure as a Restorative Treatment (SCS HEART study): first-in-man experience.

BACKGROUND: Preclinical studies suggest that neuromodulation with thoracic spinal cord stimulation (SCS) improves left ventricular (LV) function and remodeling in systolic heart failure (HF). OBJECTIVE: The purpose of this study was to evaluate the safety and efficacy of a SCS system for the treatment of systolic HF. METHODS: We performed a prospective, multicenter pilot trial in patients with New York Heart Association (NYHA) class III HF, left ventricular ejection fraction (LVEF) 20%-35%, and implanted defibrillator device who were prescribed stable optimal medical therapy. Dual thoracic SCS leads were used at the T1-T3 level. The device was programmed to provide SCS for 24 hours per day (50 Hz at pulse width 200 µs). RESULTS: We enrolled 22 patients from 5 centers:17 patients underwent implantation of a SCS device and 4 patients who did not fulfill the study criteria served as nontreated controls. No deaths or device-device interactions were noted during the 6-month period in the 17 SCS-treated patients. Fifteen of 17 completed the efficacy endpoint assessments: composite score improved by 4.2 ± 1.3, and 11 patients (73%) showed improvement in ≥4 of 6 efficacy parameters. There was significant improvement in NYHA class (3.0 vs 2.1, P = .002; 13/17 improved); Minnesota Living with Heart Failure Questionnaire (42 ± 26 vs 27 ± 22, P = .026; 12/17 improved); peak maximum oxygen consumption (14.6 ± 3.3 vs 16.5 ± 3.9 mL/kg/min, P = .013; 10/15 improved); LVEF (25% ± 6% vs 37% ± 8%, P<.001; 14/16 improved); and LV end-systolic volume (174 ± 57 vs 137 ± 37 mL, P = .002; 11/16 improved) but not in N-terminal prohormone brain natriuretic peptide. No such improvements were observed in the 4 nontreated patients. CONCLUSION: The results of this first-in-human trial suggest that high thoracic SCS is safe and feasible and potentially can improve symptoms, functional status, and LV function and remodeling in patients with severe, symptomatic systolic HF.

The clinical impact of chromosomal microarray on paediatric care in Hong Kong.

OBJECTIVE: To evaluate the clinical impact of chromosomal microarray (CMA) on the management of paediatric patients in Hong Kong. METHODS: We performed NimbleGen 135k oligonucleotide array on 327 children with intellectual disability (ID)/developmental delay (DD), autism spectrum disorders (ASD), and/or multiple congenital anomalies (MCAs) in a university-affiliated paediatric unit from January 2011 to May 2013. The medical records of patients were reviewed in September 2013, focusing on the pathogenic/likely pathogenic CMA findings and their "clinical actionability" based on established criteria. RESULTS: Thirty-seven patients were reported to have pathogenic/likely pathogenic results, while 40 had findings of unknown significance. This gives a detection rate of 11% for clinically significant (pathogenic/likely pathogenic) findings. The significant findings have prompted clinical actions in 28 out of 37 patients (75.7%), while the findings with unknown significance have led to further management recommendation in only 1 patient (p < 0.001). Nineteen out of the 28 management recommendations are "evidence-based" on either practice guidelines endorsed by a professional society (n = 9, Level 1) or peer-reviewed publications making medical management recommendation (n = 10, Level 2). CMA results impact medical management by precipitating referral to a specialist (n = 24); diagnostic testing (n = 25), surveillance of complications (n = 19), interventional procedure (n = 7), medication (n = 15) or lifestyle modification (n = 12). CONCLUSION: The application of CMA in children with ID/DD, ASD, and/or MCAs in Hong Kong results in a diagnostic yield of ∼ 11% for pathogenic/likely pathogenic results. Importantly the yield for clinically actionable results is 8.6%. We advocate using diagnostic yield of clinically actionable results to evaluate CMA as it provides information of both clinical validity and clinical utility. Furthermore, it incorporates evidence-based medicine into the practice of genomic medicine. The same framework can be applied to other genomic testing strategies enabled by next-generation sequencing.

Spread of X inactivation on chromosome 15 is associated with a more severe phenotype in a girl with an unbalanced t(X; 15) translocation.

We report on a baby girl with multiple congenital abnormalities, including cleft palate, intrauterine growth restriction, and double outlet right ventricle (DORV) with ventricular septal defect. She had an unbalanced chromosome translocation t (X;15) resulting in monosomy 15pter → p10 and trisomy Xq13.1 → q28. All three copies of Xq encompass the XIST gene. It is known that X chromosome inactivation could spread to the autosome part of an unbalanced translocation involving chromosome X and an autosome. To confirm the spread of X chromosome inactivation on chromosome 15, we evaluate the methylation change by the HumanMethylation450 BeadChip, a whole genome DNA methylation micorarray that includes 15,259 probes spanning 717 genes on chromosome 15. Results showed there was gain in DNA methylation of more than 20% in 586 CpG sites spanning the long arm of chromosome 15. We further examined the hypermethylated CpG sites located in CpG-island promoter, because genes subjected to X chromosome inactivation will have an increase in DNA methylation level in this region. A total of 75 sites representing 24 genes were hypermethylated. Nearly all of these probes are located in region proximal to the breakpoint, from 15q11.2 to 15q21.3 (35Mb) suggesting that X inactivation was spread to the proximal region of 15q. Gain of DNA methylation, especially in the CpG-island promoter, can result in functional inactivation of genes, and therefore could potentially worsen the phenotype of our patient.