Genomic array as compared to karyotyping in myelodysplastic syndromes in a prospective clinical trial

Karyotyping is considered as the gold standard in the genetic subclassification of myelodysplastic syndrome (MDS). Oligo/SNP‐based genomic array profiling is a high‐resolution tool that also enables genome wide analysis. We compared karyotyping with oligo/SNP‐based array profiling in 104 MDS patients from the HOVON‐89 study. Oligo/SNP‐array identified all cytogenetically defined genomic lesions, except for subclones in two cases and balanced translocations in three cases. Conversely, oligo/SNP‐based genomic array profiling had a higher success rate, showing 55 abnormal cases, while an abnormal karyotype was found in only 35 patients. In nine patients whose karyotyping was unsuccessful because of insufficient metaphases or failure, oligo/SNP‐based array analysis was successful. Based on cytogenetic visible abnormalities as identified by oligo/SNP‐based genomic array prognostic scores based on IPSS/‐R were assigned. These prognostic scores were identical to the IPSS/‐R scores as obtained with karyotyping in 95%‐96% of the patients. In addition to the detection of cytogenetically defined lesions, oligo/SNP‐based genomic profiling identified focal copy number abnormalities or regions of copy neutral loss of heterozygosity that were out of the scope of karyotyping and fluorescence in situ hybridization. Of interest, in 26 patients we demonstrated such cytogenetic invisible abnormalities. These abnormalities often involved regions that are recurrently affected in hematological malignancies, and may therefore be of clinical relevance. Our findings indicate that oligo/SNP‐based genomic array can be used to identify the vast majority of recurrent cytogenetic abnormalities in MDS. Furthermore, oligo/SNP‐based array profiling yields additional genetic abnormalities that may be of clinical importance.     

Adequate use of allele frequencies in Hispanics—a problem elucidated in nephrotic syndrome

Previous studies in children with focal segmental glomerulosclerosis (FSGS) and nephrotic syndrome (NS) in the USA have revealed inter-ethnic differences in their clinical presentation and outcome. However, ethnicity was based on self-identification rather than on molecular genetic data. Here, we show that genetic heterogeneity exists in self-identified Hispanic (Spanish-American) patients with steroid-resistant nephrotic syndrome (SRNS), as patients may be either of Caucasian or Mesoamerican (Native-American) genetic background. Twenty-one self-identified Hispanic patients with SRNS from 18 families were initially evaluated for mutations in the NPHS2 and WT1 genes. All patients resided and were cared for in the USA. We performed a total genome search for linkage in all Hispanic patients using 250K single nucleotide polymorphism microarrays, comparing Caucasian with Mesoamerican allele frequencies to determine regions of homozygosity by descent and to establish the correct allele frequency for each family. We found that only ten families (56%) of the 18 self-identified Hispanic families are genetically of Mesoamerican descent, whereas the other eight families (44%) are of Caucasian descent. Due to the small number of families examined, we were unable to draw any conclusion on the prevalence of NPHS2 and WT1 in this ethnic group, but the data do suggest that self-identification of ethnicity in Hispanic-American patients is not an adequate basis for genetic studies, as this cohort may represent not only patients of Mesoamerican origin but also patients of Caucasian origin. Thus, one needs to critically review previous studies of FSGS/SRNS patients that involved Hispanic patients as a group. Future larger studies may employ a total genome search for linkage to test self-identified Hispanic ethnicity for true Mesoamerican versus Caucasian ethnicity in order to generate valid genetic data.     

Right Ventricular Diastolic Function in Children With Pulmonary Regurgitation After Repair of Tetralogy of Fallot: Volumetric Evaluation by Magnetic Resonance Velocity Mapping

Objectives. We sought to assess right ventricular diastolic function in young patients with corrected tetralogy of Fallot and pulmonary regurgitation.

Background. Pulmonary regurgitation is an important problem in repair of tetralogy of Fallot. Its effects on right ventricular diastolic function in children are unknown.

Methods. Nineteen children with repair of tetralogy of Fallot (mean age [±SD] 12 ± 3 years, mean age at operation 1.5 ± 1) and 12 healthy children were studied. Summation of magnetic resonance velocity mapping pulmonary and tricuspid volume flow curves provided right ventricular time–volume curves. Ventricular size was assessed with tomographic magnetic resonance imaging (MRI). Graded exercise testing was performed.

Results. Systematic and random differences (mean ± SD) of velocity mapping and Doppler tricuspid time to peak velocities (peak E: 1 ± 26 ms, r = 0.43; peak A: 2 ± 11 ms, r = 0.76), E/A ratios (0.04 ± 0.5, r = 0.63) and duration of pulmonary regurgitation (20 ± 35 ms, r = 0.74) were satisfactory. In 6 patients (group I), late diastolic forward pulmonary artery flow was absent; in 13 patients (group II), this flow contributed 1% to 14% to right ventricular stroke volume. Significant differences were increased deceleration time (315 ± 91 vs. 168 ± 28 ms, p < 0.001), decreased filling fraction (44 ± 11 vs. 55 ± 16%, p = 0.02) and increased peak early filling rate (378 ± 124 vs. 286 ± 112 ml/s, p = 0.018) between control subjects and group I, and increased deceleration time (230 ± 40, p = 0.03) between control subjects and group II. Pulmonary regurgitation, ventricular size and ejection fraction did not differ significantly between patient groups. Exercise function was diminished with restrictive right ventricular physiology (p < 0.001, group II vs. control subjects).

Conclusions. Impaired relaxation and restriction to filling affect diastolic right ventricular function in children with repair of tetralogy of Fallot and pulmonary regurgitation. Restrictive right ventricular physiology is associated with decreased exercise function.

(J Am Coll Cardiol 1996;28:1827–35)>     

Compliance with and acceptability of two fortified balanced energy protein supplements among pregnant women in rural Nepal

Some evidence suggests that balanced energy protein (BEP) supplements taken during pregnancy and lactation can have positive effects on birth outcomes such as small‐for‐gestational age and birthweight, but more evidence is needed on the long‐term use and acceptability of such supplements. We conducted a mixed‐methods formative research study to assess and compare compliance with and acceptability of two BEP supplements, a lipid‐based peanut paste and a biscuit, to identify BEP supplements for subsequent inclusion in an efficacy trial. We conducted an 8‐week feeding trial of daily supplementation among two groups of 40 pregnant women each in rural Nepal. Compliance data were collected and supplements distributed at the weekly visits. Sensory properties of the supplements were assessed using a 7‐point Likert scale. In addition, in‐depth interviews with women (n = 16), family members (n = 6) and health workers (n = 6) and focus group discussions (FGDs) (n = 4) were conducted to explore themes related to general use and intention of future use of the supplement. Overall self‐reported compliance was high: medians of 91.1% in the lipid‐based peanut paste group and 96.4% in the biscuit group. Both supplements were rated highly on overall likability (median score 6/7) and sensory properties. Qualitative findings showed that sustained use of the supplements was attributed to expected health benefits, favourable sensory attributes, and family support. The FGDs suggested providing the option to choose between more than one type/flavour of supplements to improve compliance. Sharing was mostly evident in the first week with higher sharing reported in the biscuit group.     

Validation of the urine column measurement as an estimation of the intra-abdominal pressure

Objective  To evaluate the efficacy of the urine column (UC) measurement compared to the intra-vesicular pressure (IVP) measurement as an estimation of intra-abdominal pressure (IAP) in patients with IAP up to 30 mmHg. Methods  Fifteen patients undergoing a laparoscopic cholecystectomy were studied. All patients were catheterized. IVP measurements were performed using a pressure transducer connected to the culture aspiration port. UC measurements were done by holding up the tubing against a measuring rod. The symphysis pubis was used as the zero-reference. IAP was raised from 0 to 30 mmHg using increments of 5 mmHg, during which first the IVP and then UC measurement series were recorded end-expiratory. Fifty and 100 ml of saline were used as a priming volume. Results  The IVP and UC measurements showed a significant correlation with IAP. Comparing IVP and UC showed a correlation of 0.91 (p < 0.001) for 50 ml and 0.87 (p < 0.001) for 100 ml of saline as a priming volume. Using 50 ml of saline, UC was 0.68 mmHg higher than IVP (95% CI −7.21 to +5.85 mmHg). For 100 ml of saline, UC was 1.23 mmHg lower than IVP (95% CI −7.41 to +9.87 mmHg). Conclusion  UC measurement shows significant correlation to IVP measurement as an estimation of the IAP. Further study needs to be done to conclude whether UC measurement is a reliable clinical alternative to IVP measurement.     

Local administration of PF-3512676 CpG-B instigates tumor-specific CD8+ T-cell reactivity in melanoma patients

PURPOSE: Impaired immune effector functions in the melanoma sentinel lymph node (SLN) may allow for early metastatic events. Local administration of PF-3512676 (formerly known as CpG 7909) has shown immunostimulatory effects of both dendritic cell and T-cell subsets in the melanoma SLN. Here, we set out to ascertain whether these PF-3512676-induced immunostimulatory effects translate into higher frequencies of melanoma-specific CD8(+) T cells. EXPERIMENTAL DESIGN: Twenty-four stage I to III melanoma patients were randomized to preoperative local administration of either PF-3512676 or saline. CD8(+) T cells from SLN and peripheral blood were tested for reactivity by IFN-gamma ELISPOT assay against several HLA-A1/A2/A3-restricted epitopes derived from various melanoma-associated antigens (MAA) in 21 of 24 enrolled patients. Frequencies of natural killer (NK) cells and frequencies and maturation state of dendritic cell subsets in the SLN were determined by flow cytometry. RESULTS: Melanoma-specific CD8(+) T-cell response rates against >1 MAA epitope in the SLN were 0 of 11 for the saline group versus 5 of 10 for the PF-3512676-administered group (P = 0.012). Of these 5 responding patients, 4 also had a measurable response to >1 MAA epitope in the blood. Increased frequencies in the SLN of both MAA-specific CD8(+) T cells and NK cells correlated to CpG-induced plasmacytoid dendritic cell maturation. CONCLUSIONS: These data show an increase in melanoma-specific CD8(+) T-cell frequencies as well as an increased effector NK cell rate after a single dose of PF-3512676 and thus support the utility of local PF-3512676 administration as adjuvant treatment in early-stage melanoma to try and halt metastatic spread.     

Parental feeding style and the inter-generational transmission of obesity risk

OBJECTIVE: This study was designed to determine whether a community sample of obese mothers with young children used different feeding styles compared with a matched sample of normal-weight mothers. Four aspects of feeding style were assessed: emotional feeding, instrumental feeding (using food as a reward), prompting/encouragement to eat, and control over eating. RESEARCH METHODS AND PROCEDURES: Participants were from 214 families with same-sex twins; 100 families in which both parents were overweight or obese and 114 in which both parents were normal weight or lean. RESULTS: We found that obese mothers were no more likely than normal-weight mothers to offer food to deal with emotional distress, use food as a form of reward, or encourage the child to eat more than was wanted. The obese and normal-weight mothers did differ on "control"; obese mothers reported significantly less control over their children's intake, and this was seen for both first-born and second-born twins. Twin analyses showed that these differences were not in response to children's genetic propensities, because monozygotic correlations were no greater than dizygotic correlations for maternal feeding style. DISCUSSION: These results suggest that the stereotype of the obese mother, who uses food in nonnutritive ways so that her child also becomes obese, is more likely to be myth than fact. However, the results raise the possibility that lack of control of food intake might contribute to the emergence of differences in weight.