Home fortification with calcium reduces Hb response to iron among anaemic Bangladeshi infants consuming a new multi-micronutrient powder formulation.

OBJECTIVE: To investigate whether the recommended dietary intake of Ca in anaemic infants compromises the expected Hb response, via home fortification with a new Ca- and Fe-containing Sprinkles™ micronutrient powder (MNP). DESIGN: A double-blind, randomized controlled, 2-month trial was conducted in Bangladesh. Infants were randomized to one of two MNP intervention groups containing Fe and other micronutrients, with or without Ca. Hb, anthropometrics and dietary intake were measured pre- and post-intervention while family demographics were collected at baseline. SETTING: Twenty-six rural villages in the Kaliganj sub-district of Gazipur, Bangladesh. SUBJECTS: One hundred infants aged 6-11 months. RESULTS: A significant increase in Hb (MNP, 13·3 (sd 12·6) g/l v. Ca-MNP, 7·6 (sd 11·6) g/l; P < 0·0001) was noted in infants from both groups. However, infants receiving MNP without Ca had a significantly higher end-point Hb concentration (P = 0·024) and rate of anaemia recovery (P = 0·008). Infants receiving MNP with Ca were more likely to remain anaemic (OR 3·2; 95 % CI 1·4, 7·5). Groups did not differ in dietary intake or demographic and anthropometric indicators. CONCLUSIONS: Although both groups showed significant improvement in Hb status, the nutrient-nutrient interaction between Fe and Ca may have diminished the Hb response in infants receiving the Ca-containing MNP.

Magnetic resonance imaging assessment of articular disc position in temporomandibular disorder subjects with various bite registrations.

Objective: This study aimed to evaluate the validity and reliability of three bite registrations on articular disc position in temporomandibular disorder patients using magnetic resonance imaging (MRI). Materials and Methods: Fifteen clinically symptomatic and orthodontically untreated temporomandibular disorder patients within the age range of 17-40 years (mean age: 28.5 years) were examined. Each patient was subjected to three bite registrations, namely maximum intercuspation, initial contact bite and Roth power centric bite, and evaluated with MRI. Results: On the right side, the mean vertical and horizontal measurement values of the point in the most posterior aspect of the posterior band of the articular disc in relation to horizontal reference line (HRL) and vertical reference line (VRL) in the sagittal view in the Roth power centric bite were lesser (2.720 ± 1.239 mm and 2.380 ± 1.185 mm, respectively), in comparison with the other two bites, and on the left side too, it was lesser in the Roth power centric bite (2.293 ± 0.979 mm and 2.360 ± 1.078 mm, respectively), when compared to the other two bites. Statistical analysis also showed the significance of Roth power centric bite over the other two bites. Conclusions: Favourable articular disc positional changes were observed in the Roth power centric bite followed by the initial contact bite and that maximum disc recapture was observed in most patients with the Roth power centric bite rather than in initial contact bite and maximum intercuspation positions. The Roth power centric bite could be assumed to be the ideal method for articulation and fabrication of gnathological splints for treating patients with temporomandibular disorders.

Updated survival and outcomes for older adults with inoperable stage III non-small-cell lung cancer treated with cisplatin, etoposide, and concurrent chest radiation with or without consolidation docetaxel: analysis of a phase III trial from the Hoosier Oncology Group (HOG) and US Oncology.

BACKGROUND: Concurrent chemoradiation with etoposide and cisplatin (EP/XRT) is standard treatment for inoperable stage III locally advanced non-small-cell lung cancer (LA-NSCLC). Consolidation docetaxel (D; Taxotere) after EP/XRT resulted in increased toxicity but no improvement in survival compared with observation (O). We report updated survival for the entire study population and include an analysis of efficacy and tolerability of EP/XRT with or without D in patients aged ≥ 70 years. PATIENTS AND METHODS: Hoosier Oncology Group LUN 01-24 enrolled 243 patients with LA-NSCLC and randomized 166 after EP/XRT to three cycles of D versus O. the trial was terminated after an analysis of the first 203 patients demonstrated futility of D. RESULTS: Median survival time (MST) for the overall study population was 21.5 months, and 3-, 4-, and 5-year survival rates were 30.7%, 18.0%, and 13.9%, respectively. No differences in MST or 3-year survival were noted between D and O arms. Older patients had similar MST (17.1 versus 22.8 months for younger patients, P = 0.15) but higher rates of grade 3/4 toxicity and hospitalization during induction. CONCLUSIONS: Consolidation docetaxel after EP/XRT does not improve survival in LA-NSCLC. Fit older adults with LA-NSCLC benefit from concurrent chemoradiation similarly as younger patients but experience higher rates of hospitalization and toxicity.

Machine learning based congestive heart failure detection using feature importance ranking of multimodal features.

In this study, we ranked the Multimodal Features extracted from Congestive Heart Failure (CHF) and Normal Sinus Rhythm (NSR) subjects. We categorized the ranked features into 1 to 5 categories based on Empirical Receiver Operating Characteristics (EROC) values. Instead of using all multimodal features, we use high ranking features for detection of CHF and normal subjects. We employed powerful machine learning techniques such as Decision Tree (DT), Naïve Bayes (NB), SVM Gaussian, SVM RBF and SVM Polynomial. The performance was measured in terms of Sensitivity, Specificity, Positive Predictive Value (PPV), Negative Predictive Value (NPV), Accuracy, False Positive Rate (FPR), and area under the Receiver Operating characteristic Curve (AUC). The highest detection performance in terms of accuracy and AUC was obtained with all multimodal features using SVM Gaussian with Sensitivity (93.06%), Specificity (81.82%), Accuracy (88.79%) and AUC (0.95). Using the top five ranked features, the highest performance was obtained with SVM Gaussian yields accuracy (84.48%), AUC (0.86); top nine ranked features using Decision Tree and Naïve Bayes got accuracy (84.48%), AUC (0.88); last thirteen ranked features using SVM polynomial obtained accuracy (80.17%), AUC (0.84). The findings indicate that proposed approach with feature ranking can be very useful for automatic detection of congestive heart failure patients and can be very helpful for further decision making by the clinicians and physicians in order to decrease the mortality rate.

Machine learning based classification of normal, slow and fast walking by extracting multimodal features from stride interval time series.

The gait speed affects the gait patterns (biomechanical and spatiotemporal parameters) of distinct age populations. Classification of normal, slow and fast walking is fundamental for understanding the effects of gait speed on the gait patterns and for proper evaluation of alternations associated with it. In this study, we extracted multimodal features such as time domain and entropy-based complexity measures from stride interval signals of healthy subjects moving with normal, slow and fast speeds. The classification between different gait speeds was performed using machine learning classifiers such as classification and regression tree (CART), support vector machine linear (SVM-L), Naïve Bayes, neural network, and ensemble classifiers (random forest (RF), XG boost, averaged neural network (AVNET)). The performance was evaluated in term of accuracy, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), p-value, area under the receiver operating characteristic curve (AUC). To distinguish the slow and normal gait walking, the highest performance was yielded in terms of accuracy (100%), p-value (0.004), and AUC (1.00) using RF, XGB-L followed by XGB-Tree with accuracy (88%), p-value (0.04) and AUC (1.00). To classify the fast and normal walking, the highest performance was obtained with accuracy (88%), p-value (0.04) using XGB-L, XGB-Tree and AVNET. The highest AUC (0.94) was obtained using NB. To discriminate the fast and slow gait walking, the highest performance was obtained using SVM-R, NNET, RF, AVNET with accuracy (88%), p-value (0.04) and AUC (0.94) using RF and AUC (0.96) using XGB-L.

Influences for Gender Disparity in Academic Neuroradiology.

BACKGROUND AND PURPOSE: There has been extensive interest in promoting gender equality within radiology, a predominately male field. In this study, our aim was to quantify gender representation in neuroradiology faculty rankings and determine any related factors that may contribute to any such disparity. MATERIALS AND METHODS: We evaluated the academic and administrative faculty members of neuroradiology divisions for all on-line listed programs in the US and Canada. After excluding programs that did not fulfill our selection criteria, we generated a short list of 85 US and 8 Canadian programs. We found 465 faculty members who met the inclusion criteria for our study. We used Elsevier's SCOPUS for gathering the data pertaining to the publications, H-index, citations, and tenure of the productivity of each faculty member. RESULTS: Gender disparity was insignificant when analyzing academic ranks. There are more men working in neuroimaging relative to women (χ2 = 0.46; P = .79). However, gender disparity was highly significant for leadership positions in neuroradiology (χ2 = 6.76; P = .009). The median H-index was higher among male faculty members (17.5) versus female faculty members (9). Female faculty members have odds of 0.84 compared with male faculty members of having a higher H-index, adjusting for publications, citations, academic ranks, leadership ranks, and interaction between gender and publications and gender and citations (9). CONCLUSIONS: Neuroradiology faculty members follow the same male predominance seen in many other specialties of medicine. In this study, issues such as mentoring, role models, opportunities to engage in leadership/research activities, funding opportunities, and mindfulness regarding research productivity are explored.

Studying the dynamics of interbeat interval time series of healthy and congestive heart failure subjects using scale based symbolic entropy analysis.

Considerable interest has been devoted for developing a deeper understanding of the dynamics of healthy biological systems and how these dynamics are affected due to aging and disease. Entropy based complexity measures have widely been used for quantifying the dynamics of physical and biological systems. These techniques have provided valuable information leading to a fuller understanding of the dynamics of these systems and underlying stimuli that are responsible for anomalous behavior. The single scale based traditional entropy measures yielded contradictory results about the dynamics of real world time series data of healthy and pathological subjects. Recently the multiscale entropy (MSE) algorithm was introduced for precise description of the complexity of biological signals, which was used in numerous fields since its inception. The original MSE quantified the complexity of coarse-grained time series using sample entropy. The original MSE may be unreliable for short signals because the length of the coarse-grained time series decreases with increasing scaling factor τ, however, MSE works well for long signals. To overcome the drawback of original MSE, various variants of this method have been proposed for evaluating complexity efficiently. In this study, we have proposed multiscale normalized corrected Shannon entropy (MNCSE), in which instead of using sample entropy, symbolic entropy measure NCSE has been used as an entropy estimate. The results of the study are compared with traditional MSE. The effectiveness of the proposed approach is demonstrated using noise signals as well as interbeat interval signals from healthy and pathological subjects. The preliminary results of the study indicate that MNCSE values are more stable and reliable than original MSE values. The results show that MNCSE based features lead to higher classification accuracies in comparison with the MSE based features.

Prognostic factors for hyperdiploid-myeloma: effects of chromosome 13 deletions and IgH translocations.

Chromosomal hyperdiploidy is the defining genetic signature in 40-50% of myeloma (MM) patients. We characterize hyperdiploid-MM (H-MM) in terms of its clinical and prognostic features in a cohort of 220 H-MM patients entered into clinical trials. Hyperdiploid-myeloma is associated with male sex, kappa immunoglobulin subtype, symptomatic bone disease and better survival compared to nonhyperdiploid-MM (median overall survival 48 vs 35 months, log-rank P = 0.023), despite similar response to treatment. Among 108 H-MM cases with FISH studies for common genetic abnormalities, survival is negatively affected by the existence of immunoglobulin heavy chain (IgH) translocations, especially those involving unknown partners, while the presence of chromosome 13 deletion by FISH did not significantly affect survival (median overall survival 50 vs 47 months, log-rank P = 0.47). Hyperdiploid-myeloma is therefore a unique genetic subtype of MM associated with improved outcome with distinct clinical features. The existence of IgH translocations but not chromosome 13 deletion by FISH negatively impacts survival and may allow further risk stratification of this population of MM patients.

Immunophenotypic aberrancies in acute lymphoblastic leukemia from 282 Iraqi patients.

INTRODUCTION: The identification of aberrancies in leukemia-associated immunophenotype (LAIP) of acute lymphoblastic leukemia (ALL) is quite important in the assessment of minimal residual disease (MRD). This study, the first from Iraq, aimed to assess the frequency and patterns of LAIP among Iraqi patients with ALL, to establish future strategies for evaluating MRD. METHODS: A total of 282 newly diagnosed Iraqi ALL cases were analyzed with six-parameter flow cytometry using a panel of 29 monoclonal antibodies. RESULTS: Immunological subtyping revealed that 85.5% of cases were B-ALL and the remainder T-ALL. LAIP was detected in 97.1% of B-ALL, and in 26.8% of T-ALL. The asynchronous maturation-associated antigen patterns in B-ALL were CD10strong+ /TdTdim+ , CD38dim+ /CD34+ , CD10dim+ /CD34+ , CD10strong /CD20strong+ , CD20strong+ /CD34+, and CD45dim+ /CD20strong+ in 84.6%, while the cross-lineage myeloid expression was seen in 81.3% and aberrant T-cell antigen expression in 6.2%. For T-ALL, asynchronous maturation-associated antigen patterns included the following: CD1a+ /CD5+ /sCD3+ and CD34+ /sCD3+ in 12.2%. Myeloid and B-cell antigen expression were each identified in 7.3% of T-ALL. No significant differences in LAIP were found between children and adults. CONCLUSION: The high rates and the patterns of LAIP particularly in Iraqi B-ALL patients may allow the development of more cost-effective strategies for MRD monitoring.

Symbolic time series analysis of electroencephalographic (EEG) epileptic seizure and brain dynamics with eye-open and eye-closed subjects during resting states.

OBJECTIVE: Epilepsy is a neuronal disorder for which the electrical discharge in the brain is synchronized, abnormal and excessive. To detect the epileptic seizures and to analyse brain activities during different mental states, various methods in non-linear dynamics have been proposed. This study is an attempt to quantify the complexity of control and epileptic subject with and without seizure as well as to distinguish eye-open (EO) and eye-closed (EC) conditions using threshold-based symbolic entropy. METHODS: The threshold-dependent symbolic entropy was applied to distinguish the healthy and epileptic subjects with seizure and seizure-free intervals (i.e. interictal and ictal) as well as to distinguish EO and EC conditions. The original time series data was converted into symbol sequences using quantization level, and word series of symbol sequences was generated using a word length of three or more. Then, normalized corrected Shannon entropy (NCSE) was computed to quantify the complexity. The NCSE values were not following the normal distribution, and the non-parametric Mann-Whitney-Wilcoxon (MWW) test was used to find significant differences among various groups at 0.05 significance level. The values of NCSE were presented in a form of topographic maps to show significant brain regions during EC and EO conditions. The results of the study were compared to those of the multiscale entropy (MSE). RESULTS: The results indicated that the dynamics of healthy subjects are more complex compared to epileptic subjects (during seizure and seizure-free intervals) in both EO and EC conditions. The comparison of the dynamics of epileptic subjects revealed that seizure-free intervals are more complex than seizure intervals. The dynamics of healthy subjects during EO conditions are more complex compared to those during EC conditions. Further, the results clearly demonstrated that threshold-dependent symbolic entropy outperform MSE in distinguishing different physiological and pathological conditions. CONCLUSION: The threshold symbolic entropy has provided improved accuracy in quantifying the dynamics of healthy and epileptic subjects during EC an EO conditions for each electrode compared to the MSE.

Ploidy status rarely changes in myeloma patients at disease progression.

Hyperdiploid and non-hyperdiploid multiple myeloma represents distinct biological entities characterized by different patterns of genetic changes. We sought to determine whether ploidy category (non-hyperdiploid versus hyperdiploid) remains stable over time from diagnosis to progression. Of the 43 patients studied (39 by flow cytometry DNA index and 4 by a FISH-based index), only five (12%) altered their ploidy status at progression. In three of these patients, the change may possibly be attributable to technical artifacts because of the low absolute change in DNA index. For those who retain their ploidy subtypes, the DNA index change minimally (3.75+/-4.87%). It would appear that the initiating genetic events underlying hyperdiploid and non-hyperdiploid MM that marks them out as distinct entities continue to dominate and persist during disease evolution and progression.

Molecular detection and identification of Candida and Aspergillus spp. from clinical samples using real-time PCR.

This report describes the development of a real-time LightCycler assay for the detection and identification of Candida and Aspergillus spp., using the MagNa Pure LC Instrument for automated extraction of fungal DNA. The assay takes 5-6 h to perform. The oligonucleotide primers and probes used for species identification were derived from the DNA sequences of the 18S rRNA genes of various fungal pathogens. All samples were screened for Aspergillus and Candida to the genus level in the real-time PCR assay. If a sample was Candida-positive, typing to species level was performed using five species-specific probes. The assay detected and identified most of the clinically relevant Aspergillus and Candida spp. with a sensitivity of 2 CFU/mL blood. Amplification was 100% specific for all Aspergillus and Candida spp. tested. To assess clinical applicability, 1,650 consecutive samples (1,330 blood samples, 295 samples from other body fluids and 25 biopsy samples) from patients with suspected invasive fungal infections were analysed. In total, 114 (6.9%) samples were PCR-positive, 5.3% for Candida and 1.7% for Aspergillus spp. In patients with positive PCR results for Candida and Aspergillus, verification with conventional methods was possible in 83% and 50% of cases, respectively. In conclusion, the real-time PCR assay allows sensitive and specific detection and identification of fungal pathogens in vitro and in vivo.

Coexistence of aneuploid subclones within a myeloma cell line that exhibits clonal immunoglobulin gene rearrangement: clinical implications.

A new human myeloma cell line, ANBL-6, was established and characterized at the genotypic and phenotypic levels. The cells exhibit a clonally rearranged immunoglobulin gene locus and resemble plasma cells morphologically. The ANBL-6 cells also exhibited an absolute dependence on exogenous interleukin 6 for growth. Of interest, DNA ploidy analysis suggested the existence of a near-diploid as well as a near-tetraploid population in this cell line. Cytogenetic studies confirmed the existence of two aneuploid karyotypes and further revealed a clonal relationship between the two karyotypes, as evidenced by numerous shared structural abnormalities. To determine whether the near-diploid cells functioned as stem cells for the near-tetraploid population, the near-diploid population was separated via flow cytometry and recultured prior to ploidy analysis. This population was observed to remain predominantly near-diploid over time, suggesting that these cells did not function as stem cells for the near-tetraploid population. However, the near-tetraploid cells did exhibit a growth advantage in vitro. Moreover, sequential ploidy analysis performed retrospectively on fresh bone marrow cells from the patient also suggested that there was an expansion of the near-tetraploid population during clinical relapse. These results suggest that both populations are self-regenerating and reflect the consequences of clonal evolution in the myeloma tumor. The coexistence of clonally related subclones with shared chromosomal abnormalities, however, suggests that the near-tetraploid subclone was derived from the near-diploid subclone at an unknown time during tumorigenesis.

Analysis of Hurthle cell neoplasms of the thyroid by interphase fluorescence in situ hybridization.

Recent studies have indicated that numerical chromosomal abnormalities including changes in p53 and cyclin D1 may be involved in Hurthle cell tumorigenesis. We analyzed a series of Hurthle cell neoplasms of the thyroid to evaluate the diagnostic and prognostic utility of numerical anomalies by DNA fluorescent probes for cyclin D1 and p53 gene loci and chromosomes 5, 7, 11, 12, 17, and 22. Interphase fluorescence in situ hybridization (FISH) analysis was performed on paraffin-embedded tissue sections from 10 Hurthle cell adenomas, 19 Hurthle cell carcinomas, and 7 normal thyroid tissues used as controls. Directly labeled fluorescent DNA probes for the centromere region of chromosomes 7, 11, 12, and 17 and locus-specific probes for chromosomes 5 and 22, cyclin D1, and p53 were utilized for dual-probe hybridizations. Sixty percent (6 of 10) Hurthle cell adenomas and 63% (12 of 19) Hurthle cell carcinomas showed chromosome gains. Twenty percent (2 of 10) Hurthle cell adenomas and 26% (5 of 19) Hurthle cell carcinomas showed chromosome losses. Normal thyroid tissues used as controls showed no chromosomal abnormalities. Among Hurthle cell tumors with chromosomal abnormalities, adenomas averaged 2.7 gains and 0.3 losses per case, and carcinomas averaged 3.3 gains and 0.6 losses per case. The two adenomas with chromosome losses each showed loss of one chromosome, whereas the five carcinomas with losses averaged 1.8 losses per case. Chromosome 22 was the most common loss identified, occurring in three of the 11 patients who died of disease. These results indicate that chromosomal imbalances as gains are common in both benign and malignant Hurthle cell neoplasms, but Hurthle cell carcinomas tend to have more chromosome losses than adenomas. Among Hurthle cell carcinomas in this study, chromosome losses were identified only from patients who died of disease. The loss of chromosome 22 may have prognostic value in Hurthle cell carcinoma of the thyroid.

Detection of t(2;5) in anaplastic large cell lymphoma: comparison of immunohistochemical studies, FISH, and RT-PCR in paraffin-embedded tissue.

Anaplastic large cell lymphoma (ALCL) is associated with the t(2;5)(p23;q35) translocation involving the anaplastic lymphoma kinase gene (ALK) and the nucleophosmin gene (NPM), which result in expression of a novel fusion protein, NPM-ALK (p80). Clinicopathologic studies have shown that ALK expression in ALCL is associated with improved 5-year survival rates when compared with ALCL lacking ALK expression. This study used paraffin-embedded tissue to compare interphase fluorescence in situ hybridization (FISH) and reverse transcriptase-polymerase chain reaction (RT-PCR) for the detection of t(2;5) with immunohistochemical analysis for the detection of ALK protein expression in 27 patients with CD30-positive ALCLs. ALK protein expression was detected with ALK1 antibody in 14 of the 27 patients. The neoplastic cells in 13 of these 14 lymphomas reacted with the p80NPM/ALK antibody. FISH, using a two-color ALK DNA probe, correlated 100% with the immunohistochemical results: a translocation involving the ALK gene was detected in all 14 lymphomas that reacted with anti-ALK1. RT-PCR, performed on 21 lymphomas, detected NPM-ALK mRNA in five of the lymphomas, all of which reacted with anti-ALK1 and showed ALK gene rearrangement by FISH. Lymphomas showing ALK1 reactivity occurred in a younger patient population (median age, 19.5 years) and were associated with improved 5-year survival rates (84%), as compared with lymphomas lacking ALK1 reactivity (median age, 68.0 years; 5-year survival rate, 35%; p = 0.008). We conclude that immunohistochemical studies, using antibody ALK1. and FISH for ALK gene rearrangement are equally effective for identifying patients with ALCL who have a favorable clinical outcome.

Detection of newborn aneuploidy by interphase fluorescence in situ hybridization.

OBJECTIVE: To test fluorescence in situ hybridization (FISH) probes for rapid detection of aneuploidy of chromosomes 13, 18, 21, X, and Y from newborn uncultured blood samples. MATERIAL AND METHODS: Directly labeled, multicolored, commercially available FISH probes for the five aforementioned chromosomes were validated, and their hybridization efficiencies were established. In a blinded study, eight trisomic samples were tested by this FISH method. RESULTS: The hybridization efficiency based on metaphase evaluation of each of the five probes was 100%, and no cross-hybridization occurred. The mean interphase hybridization efficiencies of the probes for chromosomes 13, 18, 21, X, and Y were 97.4 %, 89.4 %, 96.1%, 94.4 %, and 100 %, respectively. The eight abnormal samples were identified as trisomy 21 (in six), trisomy 13 (in one), and trisomy 18 (in one). CONCLUSION: The screening of aneuploidy of newborns for chromosomes 13, 18, 21, X, or Y by interphase FISH is rapid, reliable, and cost-effective. The test is especially suitable for medically urgent cases as a screen, followed by a standard chromosome analysis.

Efficacy of fluorescence in situ hybridization for detecting PML/RARA gene fusion in treated and untreated acute promyelocytic leukemia.

OBJECTIVE: To test the efficacy of fluorescence in situ hybridization (FISH) for detection of fusion of the promyelocytic leukemia (PML) and retinoic acid receptor alpha (RARA) genes in patients with treated or untreated acute promyelocytic leukemia (APL). DESIGN: We conducted a retrospective blind study on a series of stored bone marrow specimens from normal subjects and patients with APL. MATERIAL AND METHODS: Conventional cytogenetic and FISH analyses were done on interphase and metaphase cells in specimens from 31 normal subjects and 19 patients with untreated or treated APL. RESULTS: From 25 of the normal specimens, we calculated a normal cutoff of 10% for interphase cells and 0% for metaphase cells. With use of these criteria, the other six specimens from normal subjects showed normal findings, and each of the seven specimens from patients with untreated APL was abnormal by FISH analysis. The specimens from four patients in clinical relapse or with residual APL were abnormal. Of the eight specimens from patients in clinical remission, three were abnormal; two of these patients had a relapse within 8 months, and the other patient had received 1 month of chemotherapy and was entering remission. Of the other five patients in remission, four had normal FISH results and have now been in remission for 2.5 to 10 years. The other patient in remission with normal FISH results had a relapse within 6 months. PML/RARA fusion was detectable in three patients with hypogranular APL and in three with a cytogenetic variant of the t(15;17). CONCLUSION: The results of this study suggest that FISH with PML and RARA probes can be used to diagnose APL and may be useful for monitoring treated patients.

DNA fluorescent probes for diagnosis of velocardiofacial and related syndromes.

OBJECTIVE: To study the usefulness of fluorescent in situ hybridization (FISH) with the DNA probe D22S75 for detecting microdeletions in chromosome 22q11.2 in metaphases from patients with features of "CATCH 22" (cardiac anomalies, abnormal facies, thymic hypoplasia or aplasia, cleft palate, and hypocalcemia). METHODS: High-resolution chromosome analysis and FISH were performed on metaphases from 10 control subjects, 42 patients with features of CATCH 22, and 6 parents of children with CATCH 22. Patients were screened for conotruncal heart defect, palatal abnormality, and facial features. We correlated the phenotype, karyotype, and deletion of a D22S75 locus. RESULTS: Specimens from nine patients with one or more features of CATCH 22 had a single hybridization signal for D22S75, indicating a deletion of chromosome 22q11.2. Four patients had all the major features of the syndrome and a chromosomal deletion. Thirteen patients had two CATCH 22 features, five of whom had a deletion. None of the 25 patients with a single CATCH 22 feature had a deletion. One patient with a deletion detected by FISH also had a deletion noted on high-resolution banding. All six parents who had blood samples studied by FISH had normal hybridization patterns. CONCLUSION: FISH is a useful adjunct to chromosome analysis for assessing patients with features of CATCH 22. Detecting a chromosomal deletion by FISH provides a definitive diagnosis and helps to ensure appropriate medical management and genetic counseling.

Prenatal detection of aneuploidy by directly labeled multicolored probes and interphase fluorescence in situ hybridization.

OBJECTIVE: To detect aneuploidy of chromosomes 13, 18, 21, X, and Y with use of new, directly labeled, multicolored, commercially available DNA probes from interphase cells of amniotic fluid (AF). MATERIAL AND METHODS: The hybridization sites of the five probes were validated by metaphase analysis. The fluorescence in situ hybridization (FISH) normal range was determined from a series of normal AF specimens and tested on a series of normal and abnormal specimens. RESULTS: The hybridization efficiencies of the five probes were 100%. The mean AF interphase disomic signal patterns for chromosomes 13, 18, 21, XX, and XY were 95.9%, 89.1%, 94.3%, 94.7%, and 98.7%, respectively. Of a total of 508 cases analyzed, 211 were aneuploid. All cases were correctly identified and no false results occurred (in comparison with karyotypic analysis), although maternal cell contamination was relatively common. CONCLUSION: Clinical screening for aneuploidy of chromosomes 13, 18, 21, X, and Y from interphase AF cells is possible with use of these probes and FISH. Cases of maternal cell contamination and mosaicism necessitate cautious interpretation. The FISH procedure is recommended for screening of common aneuploidies, followed by a complete chromosome analysis to detect anomalies.

Application of multicolor fluorescent in situ hybridization for enhanced characterization of chromosomal abnormalities in congenital disorders.

OBJECTIVE: To determine the efficacy of multicolor fluorescent in situ hybridization (M-FISH), which paints each chromosome in a unique color, for identification of congenital derivative and marker chromosomes. MATERIAL, METHODS AND CASES: Commercially available M-FISH probes were used to label each chromosome in a specific fluorescent color. Six representative cases involving derivative chromosomes, markers, and subtle anomalies were analyzed by M-FISH. RESULTS: Three familial, rather subtle derivative chromosomes were identified by M-FISH with relative ease. A small ring that was unidentifiable by banded-chromosome analysis was identified by M-FISH. A case of a subtle telomeric anomaly could not be resolved without the use of telomeric-specific probes. The M-FISH results were confirmed by individual chromosome-specific painting probes. CONCLUSION: M-FISH was helpful for identifying a wide range of congenital chromosomal anomalies. However, for subtle chromosomal abnormalities, use of locus-specific probes may be necessary.