Comparative sequence analysis of incomplete DJH and TCR gene rearrangements in children with relapses of T-ALL.
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Child↗
Biomedical subjects
G Germano
Publications and source records attributed to G Germano.
Clonality profile in relapsed precursor-B-ALL children by GeneScan and sequencing analyses. Consequences on minimal residual disease monitoring.
Detection of minimal residual disease (MRD), using immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements as clone-specific targets, represents the most recent development in diagnosis and treatment of acute lymphoblastic leukaemia (ALL). Nevertheless, risk of false-negative results, due to secondary or ongoing rearrangements of Ig/TCR genes during the disease course, might hamper MRD detection. Therefore, to gain extensive information on clonal stability, we performed PCR-GeneScan analysis of Ig/TCR gene rearrangements at diagnosis and subsequent relapse in bone marrow samples from 53 childhood precursor-B-ALL patients. In addition, sequencing analysis of junctional regions at diagnosis and relapse provided a detailed insight in the stability and changes of Ig/TCR gene rearrangements during the disease course. At least one stable clonal Ig/TCR target was found in 94% of patients. In three patients complete differences in Ig/TCR rearrangements between diagnosis and relapse were observed, suggesting relapse with a new clone. At relapse, 71% of diagnostic clonal PCR targets was conserved. Since the comparison of Ig/TCR gene rearrangements at diagnosis and relapse in our precursor-B-ALL patients did not show significant difference in the stability of different clonal PCR targets (IGH, 70%; IGK, 71%; TCRD, 67%; TCRG, 75%), we conclude that there is no 'preferential' clone-specific target for MRD monitoring.
Adolescent↗
Helicobacter pylori virulence genes and host IL-1RN and IL-1beta genes interplay in favouring the development of peptic ulcer and intestinal metaplasia.
Helicobacter pylori infection outcome might depend on genotypic polymorphisms of both the bacterium and the host. We ascertained: (1) the functionality of H. pylori oipA gene; (2) the polymorphism of the hostinterleukin (IL-1beta) gene (-31 C/T) and of the IL-1RN gene (intron 2 VNTR); (3) the association between the above genes and the histological and pathological outcome of H. pylori infection. One hundred and sixty-five H. pylori positive and 137 H. pylori negative subjects (23 gastric adenocarcinoma, 58 peptic ulcer, 221 gastritis) were studied. oipA was sequenced, IL-1beta was RFLP analysed. Antral and body mucosal biopsies were histologically evaluated. Functional oipA genes were correlated with cagA gene; both genes were significantly associated with gastritis activity, peptic ulcer and gastric adenocarcinoma. In these patients heterozygousIL-1RN 1/2 and IL-1beta C/T genotypes were more frequent than in gastritis patients. Intestinal metaplasia was associated with cagA, functional oipA and IL-1RN 2 allele. In conclusion, peptic ulcer and the preneoplastic intestinal metaplasia are associated with H. pylori virulence genes and with IL-1RN 2 host allele. An interplay between bacterial virulence factors and cytokines genotypes, is probably the main route causing H. pylori infection to lead to benign mild disease, benign severe disease or preneoplastic lesions.
Adenocarcinoma↗
Pituitary adenylate cyclase activating peptide (PACAP) immunoreactivity and mRNA expression in the duck gastrointestinal tract.
The presence and distribution of pituitary adenylate cyclase activating peptide (PACAP) immunoreactivity were studied in the duck gastrointestinal tract using immunohistochemistry and radioimmunoassays. Expression and distribution of PACAP mRNA were also studied using reverse transcriptase polymerase chain reaction (RT-PCR) and hybridization techniques. In addition, a partial coding sequence (cds) of the duck growth hormone-releasing hormone (GRF)/PACAP gene was identified. The presence of both PACAP-38 and PACAP-27 was demonstrated, the former being the predominant form. PACAP immunoreactivity was found in neurons and fibers of the enteric nervous system (ENS), in endocrine cells and in the gut associated lymphoid tissue (GALT). Double immunostaining showed that PACAP is almost completely colocalized with vasoactive intestinal peptide (VIP) in the ENS. Moreover, PACAP was also found in nitric oxide synthase (NOS)-containing neurons and nerve fibers. Radioimmunoassay (RIA) performed on denervated gut showed that more than one-half of the duodenal PACAP is extrinsic in origin. RT-PCR, Northern blot analysis and in situ hybridization confirmed the immunohistochemical data. The findings of the present study suggest that, in birds, PACAP may have multiple roles in regulating gastrointestinal functions.
Animals↗
Nuclear cardiology and electron-beam computed tomography: competitive or complementary?
Electron-beam computed tomography (EBCT) and nuclear cardiology techniques are both valuable in the noninvasive assessment of patients with suspected coronary artery disease. The techniques, however, are different in the information they provide about the patient. EBCT provides anatomic information on coronary atherosclerosis, whereas myocardial perfusion single-photon emission computed tomography assesses the physiologic significance of coronary stenosis. Because of these differences, the techniques are highly complementary. In considering the complementary nature of these methods, it is important to clarify the issues being raised. An important question in the consideration of a patient with known or suspected coronary artery disease is, What is the risk in an individual patient of developing clinical coronary artery disease? The answer to this question will determine who needs aggressive medical management. A second question in a suspected coronary artery disease patient is, What is the risk of cardiac death? As will be discussed, this risk, in general, determines the need to consider coronary revascularization. In the former question, EBCT testing and clinical assessment alone is usually sufficient, and in some cases nuclear testing can be of additional value. In answering the second question, on the basis of currently available data, the EBCT and nuclear cardiology studies appear to be operating in a complementary fashion.
Cholesterol↗
Computer simulation of topological defects around a colloidal particle or droplet dispersed in a nematic host.
We use molecular dynamics to study the ordering of a nematic liquid crystal around a spherical particle or droplet. Homeotropic boundary conditions and strong anchoring create a hedgehog (radial point defect) director configuration on the particle surface and in its vicinity; this topological defect is canceled by nearby defect structures in the surrounding liquid crystal, so as to give a uniform director field at large distances. We observe three defect structures for different particle sizes: a quadrupolar one with a ring defect surrounding the particle in the equatorial plane; a dipolar one with a satellite defect at the north or south pole; and a transitional, nonequatorial, ring defect. These observations are broadly consistent with the predictions of the simplest elastic theory. By studying density and order-parameter maps, we are able to examine behavior near the particle surface, and in the disclination core region, where the elastic theory is inapplicable. Despite the relatively small scale of the inhomogeneities in our systems, the simple theory gives reasonably accurate predictions of the variation of defect position with particle size.
Journal Article↗
Pituitary adenylate cyclase activating peptide (PACAP) immunoreactivity in the ureter of the duck.
The presence, distribution and colocalisation of pituitary adenylate cyclase activating peptide (PACAP) immunoreactivity have been studied in the duck ureter by using Western blot analysis, radioimmunoassays (RIA) and immunohistochemistry. The presence of both PACAP-38 and PACAP-27 was demonstrated, PACAP-38 being the predominant form. PACAP-immunoreactive fibres and neurons were found in all the ureteral layers. Double immunostaining showed that PACAP was almost completely colocalised with vasoactive intestinal peptide (VIP). Moreover, PACAP was found in substance P (SP)-containing ureteral nerve fibres and in SP-containing dorsal root ganglion neurons. RIA performed on denervated ureters demonstrated that almost half of the ureteral PACAP was extrinsic in origin. These findings suggest that, in birds, PACAP has a role in diverse nerve-mediated ureteral functions.
Animals↗
When to stress patients after coronary artery bypass surgery? Risk stratification in patients early and late post-CABG using stress myocardial perfusion SPECT: implications of appropriate clinical strategies.
OBJECTIVES: The study compared the prognostic significance of myocardial perfusion single-photon emission computed tomography (SPECT) (MPS) in patients early and late after coronary artery bypass graft surgery (CABG). BACKGROUND: The long-term effectiveness of CABG is limited by graft stenosis. The greatest incidence of graft occlusion occurs between five and eight years after surgery. However, little is known regarding the appropriate time to stress patients post-CABG with respect to risk stratification. METHODS: We identified 1,765 patients, who underwent MPS 7.1 +/- 5.0 years post-CABG. All patients underwent rest T1-201/stress Tc-99m sestamibi MPS and were followed up > or =1 year after testing. Patients with early CABG or PTCA (<60 days after MPS) were censored. The prognostic population consisted of 1,544 patients. A semiquantitative visual analysis employing a 20-segment model was used to define summed stress score (SSS), summed rest score (SRS), summed difference score (SDS), and the number of nonreversible segments (NRS). RESULTS: During follow-up, 53 cardiac deaths (CD) occurred. There was a significant increase in annual CD rates as a function of SSS. A multivariate analysis identified age, ischemia (SDS), and infarct size (NRS) as independent predictors of CD. Nuclear variables added incremental value to prescan information. The annual CD rate was relatively low (1.3%) in patients < or =5 years post-CABG. In this subgroup only age and infarct size (NRS) were predictive of CD. CONCLUSION: MPS is strongly predictive of subsequent CD in post-CABG patients and adds incremental value over clinical and treadmill test information. Our data suggest that symptomatic patients < or =5 years and all patients >5 years post-CABG may benefit from testing.
Aged↗
Recent advances in myocardial perfusion imaging.
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Coronary Disease↗
Rapid detection of clonality in patients with acute lymphoblastic leukemia.
BACKGROUND AND OBJECTIVES: Polymerase chain reaction (PCR) detection of clonal T-cell receptor (TCR) gamma and delta gene rearrangements is widely used in clonality assessment of lymphoid leukemias and lymphomas and for detection of minimal residual disease of acute lymphoblastic leukemia (ALL). Standard analyses for clonality assessment include Southern blotting or PCR-based detection of clonal TCR gene rearrangements. The latter consist of heteroduplex PCR analysis by separation of PCR products on non-denaturing polyacrylamide gel (PAGE). We describe a rapid and sensitive method to identify specific clonal rearrangements in PCR fragments obtained by amplification of TCRgamma and TCRdelta genes. DESIGN AND METHODS: We applied a semi-automated electrophoretic technique (PhastSystem , Amersham Pharmacia Biotech) and compared it with standard homo-heteroduplex analysis in 21 cases of childhood acute lymphoblastic leukemia (ALL). RESULTS: The results obtained for each sample analyzed by standard homo-heteroduplex detection were completely reproduced by the PhastSystem approach. INTERPRETATION AND CONCLUSIONS: We conclude that heteroduplex analysis of TCR gene rearrangements using the semi-automated PhastSystem is a simple, rapid, cheap and highly reproducible method which can be used as an alternative to traditional analysis for detection of clonality.
Child↗
Quantitative gated SPECT.
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Gated Blood-Pool Imaging↗
Quantitative assessment of motion artifacts and validation of a new motion-correction program for myocardial perfusion SPECT.
UNLABELLED: Patient motion during myocardial perfusion SPECT can produce images that show artifactual perfusion defects. The relationship between the degree of motion and the extent of artifactual perfusion defects is not clear for either single- or double-head detectors. Using both single- and double-head detectors and quantitative perfusion SPECT (QPS) software, we studied the pattern and extent of defects induced by simulated motion and validated a new automatic motion-correction program for myocardial perfusion SPECT. METHODS: Vertical motion was simulated by upward shifting of the raw projection datasets in a returning pattern (bounce) and in a nonreturning pattern at 3 different phases of the SPECT acquisition (early, middle, and late), whereas upward creep was simulated by uniform shifting throughout the acquisition. Lateral motion was similarly simulated by left shifting of the raw projection datasets in a returning pattern and in a nonreturning pattern. Simulations were performed using single- and double-head detectors, and simulated motion was applied to projection images from 8 patients who had normal 99mTc-sestamibi SPECT findings. Additionally, images from 130 patients with actual clinical motion were assessed before and after motion correction. The extent of perfusion defects was assessed by QPS, and a 20-segment, 5-point scoring system was used to assess the effect of motion on the presence and extent of perfusion defects. RESULTS: Of 12 bounce simulations, the bouncing motion failed to produce significant (>3%) perfusion defects with either the single- or the double-head detector. With the single-head detector, early shifting created the largest defect, whereas with the double-head detector, shifting during the middle of the acquisition created the largest defect. With regard to upward creep, defects were of larger extent with the double- than the single-head detector. With the single-head detector, 8 of 20 simulated motion patterns yielded significant perfusion defects of the left ventricle, 7 (88%) of which were significantly improved after motion correction. With the double-head detector, 12 of 20 patterns yielded significant defects, all of which improved significantly after correction. Of 2,600 segments in the 130 patients with actual clinical motion, only 1.3% (30/2,259) of segments that were considered normal (score = 0 or 1) changed to abnormal (score = 2-4) after motion correction, whereas 27% (92/341) of abnormal segments were reclassified as normal after motion correction. CONCLUSION: Artifactual perfusion defects created by simulated motion are a function of the time, degree, and type of motion and the number of camera detectors. Application of an automatic motion-correction algorithm effectively decreases motion artifacts on myocardial perfusion SPECT images.
Aged↗
Prediction of myocardial infarction versus cardiac death by gated myocardial perfusion SPECT: risk stratification by the amount of stress-induced ischemia and the poststress ejection fraction.
UNLABELLED: The combination of myocardial perfusion and poststress ejection fraction (EF) provides incremental prognostic information. This study assessed predictors of nonfatal myocardial infarction (MI) versus cardiac death (CD) by gated myocardial SPECT and examined the value of integrating the amount of ischemia and poststress EF data in risk stratification. METHODS: We identified 2,686 patients who underwent resting (201)Tl/stress (99m)Tc-sestamibi gated SPECT and were monitored for >1 y. Patients who underwent revascularization < or = 60 d after the nuclear test were censored from the prognostic analysis. Visual scoring of perfusion images used 20 segments and a scale of 0--4. Poststress EF was automatically generated. RESULTS: Cox regression analysis showed that after adjusting for prescan data, the most powerful predictor of CD was poststress EF, whereas the best predictor of MI was the amount of ischemia (summed difference score [SDS]). Integration of the EF and SDS yielded effective stratification of patients into low-, intermediate-, and high-risk subgroups. Patients with EF >50% and a large amount of ischemia were at intermediate risk (2%--3%), whereas those with mild or moderate ischemia were at low risk of CD (<1%/y). Patients with EF between 30% and 50% were at intermediate risk even in the presence of only mild or moderate ischemia. In patients with EF <30%, the CD rate was high (>4%/y) irrespective of the amount of ischemia. CONCLUSION: Poststress EF is the best predictor of CD, whereas the amount of ischemia is the best predictor of nonfatal MI. Integration of perfusion and function data improves stratification of patients into low, intermediate, and high risk of CD.
Adenosine↗
Evidence-based risk assessment in noninvasive imaging.
UNLABELLED: Assessment of important clinical and economic outcomes has become central to the evaluation of patient care. Outcome research is deeply rooted in epidemiology, including the use of multivariable, risk-adjusted regression analysis. In our current health care environment, these methods are increasingly being used to assess the quality of care and to profile physicians and laboratories. Nuclear medicine physicians therefore need to better understand outcome methodologies in order to evaluate patient outcomes, develop guidelines, and decide on patient management. METHODS: This review describes the methods of assessing the diagnostic and prognostic value of nuclear medicine techniques and, briefly, the methodologic limitations of sample size, frequency and type of events, and follow-up periods and the incremental value of imaging. Also described are logistic regression and Cox proportional hazards modeling. Models for risk assessment are designed to identify whether patients require conservative (i.e., low-risk) or aggressive (i.e., high-risk) treatment. Treatment selection is currently based on risk assessment and the formation of an integrated, empiric risk stratification algorithm of care. This review also includes the methods of assessing economic effectiveness and quality-of-life issues for patients examined with nuclear medicine techniques. CONCLUSION: In this era of constrained resources, low-cost outpatient-based care may be of increasing importance. High-quality evidence of the clinical and economic outcome of nuclear imaging is essential for helping health care providers and payers assess its value.
Cardiovascular Diseases↗
Technical aspects of myocardial SPECT imaging.
Myocardial perfusion SPECT imaging accounts for well over 90% of all myocardial perfusion imaging performed in the United States today. Although clearly superior to the traditional planar technique in terms of image contrast and consequent diagnostic and prognostic yield, the SPECT approach also involves additional acquisition and processing steps. This article concisely describes the entire technical sequence of myocardial SPECT imaging, from the acquisition of projection images to their filtration and tomographic reconstruction and concluding with reorientation of the tomographic transaxial images. A simplified explanation of the frequency, or Fourier, domain and the operation of digital filters is also presented to help the reader intuitively understand these important concepts.
Artifacts↗
Quantitative analysis of regional motion and thickening by gated myocardial perfusion SPECT: normal heterogeneity and criteria for abnormality.
UNLABELLED: Quantitation of regional myocardial function is valuable in patients with coronary artery disease. This study assessed normal heterogeneity and developed and validated normal limits for quantitative regional motion and thickening by gated myocardial perfusion SPECT. METHODS: Patients underwent rest (201)Tl/exercise (99m)Tc-sestamibi gated SPECT. Reference values of motion and thickening for 20 myocardial segments were obtained in 105 patients with <5% likelihood of coronary disease (low-likelihood group). Criteria for abnormality of motion and thickening were defined for each segment, using receiver operator characteristic analysis, in 101 patients with coronary disease (training group). Semiquantitative visual interpretation was used as the gold standard. These criteria were prospectively validated in 100 patients (validation group). Criteria for grading motion and thickening abnormalities by severity levels were also defined and validated. RESULTS: Normal thickening decreased substantially along the longitudinal axis of the left ventricle, from 69% +/- 13% at the apex to 25% +/- 11% at the basal segments, whereas normal motion varied within the same ventricular plane. Validation of the criteria for abnormality yielded high accuracy in the detection of motion abnormalities (sensitivity, 88%; specificity, 92%) and thickening abnormalities (sensitivity, 87%; specificity, 89%). Quantitative motion and thickening segmental scores showed good agreement with visual scores. CONCLUSION: Normal regional myocardial contraction by gated myocardial perfusion SPECT is characterized by a substantial apex-to-base decline in thickening and by circumferential heterogeneity in endocardial motion. The assignment of segment-specific threshold values for defining motion and thickening abnormalities provided reasonably accurate identification and grading of regional myocardial dysfunction.
Aged↗
Comparison of gated single-photon emission computed tomography with magnetic resonance imaging for evaluation of left ventricular function in ischemic cardiomyopathy.
To perform a head-to-head comparison between magnetic resonance imaging (MRI) and gated single-photon emission computed tomography (SPECT) for the evaluation of left ventricular (LV) function (LV ejection fraction [LVEF], LV volumes, and regional wall motion) in patients with ischemic cardiomyopathy, we studied 22 patients with chronic coronary artery disease and LV dysfunction. Multislice, multiphase echoplanar MRI was performed with Philips Gyroscan ACS-NT15. Image analysis was performed using the MASS software package to determine LV end-systolic volume, LV end-diastolic volume, and LVEF. The same parameters were calculated using quantitative gated SPECT software (QGS, Cedars-Sinai Medical Center). The different parameters were compared using linear regression, and correlation coefficients were calculated. Regional wall motion was also determined from both techniques, according to a 13-segment model and a 3-point scoring system (from 1 = normokinesia to 3 = akinesia or dyskinesia). A summed wall motion score was also calculated for MRI and gated SPECT. Good correlations were found between MRI and gated SPECT for all parameters: (1) summed wall motion scoreMRI versus summed wall motion scoreSPECT: y = 0.74x + 8.0, r = 0.88, p <0.01; (2) LV end-systolic volumeMRI versus LV end-systolic volumeSPECT: y = 0.94x - 12.3, r = 0.87, p <0.01; (3) LV end-diastolic volumeMRI versus LV end-diastolic volumeSPECT: y = 0.93x - 18.4, r = 0.84, p <0.01; and (4) LVEFMRI versus LVEFSPECT: y = 0.97x + 0.68, r = 0.90, p <0.01. For regional wall motion, an exact agreement of 83% was found, with a kappa statistic of 0.77 (95% confidence intervals 0.71 to 0.83, SE 0.03), indicating essentially excellent agreement. Thus, close and significant correlations were observed for assessment of LVEF, LV volumes, and regional wall motion by MRI and gated SPECT in patients with ischemic cardiomyopathy.
Aged↗
Identification of severe and extensive coronary artery disease by postexercise regional wall motion abnormalities in Tc-99m sestamibi gated single-photon emission computed tomography.
Postexercise wall motion abnormality (WMA) in patients with normal resting myocardial perfusion may represent prolonged postischemic stunning, and may be related to the presence of severe angiographic coronary artery disease (CAD). This study assesses the diagnostic value of postexercise WMA by technetium-99m (Tc-99m) sestamibi gated single-photon emission computed tomography (SPECT) in patients with normal resting perfusion. Ninety-nine patients underwent exercise gated Tc-99m sestamibi/resting thallium-201 SPECT and coronary angiography within 90 days of nuclear testing. All patients had normal perfusion at rest. Multivariate logistic regression analysis demonstrated an incremental value of wall motion and perfusion over perfusion data alone in identifying severe and extensive CAD. Sensitivity for identifying any severely stenosed coronary artery by WMA was significantly higher than by severe perfusion defect (78% vs 49%, p <0.0001). Overall specificities of severe perfusion defect and WMA were 91% and 85%, respectively (p = NS). Thus, postexercise WMA detected by gated Tc-99m sestamibi SPECT in patients with normal resting perfusion is a sensitive marker of severe and extensive CAD.
Aged↗