Analysis of ras gene mutations in childhood myeloid leukaemia

Previous studies have shown that approximately 30% of adult acute myeloid leukaemias and 20% of adult acute lymphoid leukaemias contain point mutated ras oncogenes. In order to assess whether ras oncogenes are also involved in childhood leukaemias, we have used polymerase chain reaction (PCR) amplification and synthetic oligonucleotide probes to study the nature and frequency of ras gene mutations in childhood leukaemias, concentrating largely on the acute myeloid leukaemias (AML). Thirty-four childhood presentation AML DNAs were screened for mutations in and around codons 12, 61 and 117 of N-, K- and H-ras. Eight of these samples (24%) contained ras mutations. As in the adult disease, the gene predominantly involved was N-ras (6/8), with occasional activation of K-ras (2/6). The most common base change was a G----A transition at codon 12 or 13 (4/8). Of the patients with mutant ras, 4/8 were diagnosed as AML FAB subtype M5. Five of the 34 childhood AMLs analysed displayed abnormalities of chromosome 7. However, none of these cases contained a mutant ras gene. One AML patient was studied at relapse, 14 months after initial presentation. The presentation mutation (N61p3) was not detectable, although a new mutation (N13Cys) was readily identified. This observation extends our original finding with presentation and relapse samples of adult AML, in which it was uncommon for the relapse sample to contain the same ras mutation as the presentation DNA. In addition, two out of five patients diagnosed as juvenile CML, were found to harbour mutant ras.     

T lymphocyte abnormalities in disseminated histoplasmosis

Disseminated histoplasmosis is associated with depression of T cell-mediated immunity and in some cases anergy. In this report, two patients with disseminated disease are described. Both had a depression of T cell-mediated immunity as well as other abnormalities of immune response. In one, a patient with relapse, a marked depression in the ratio of T helper to T suppressor cells was noted. Neither patient had any predisposing condition known to be associated with disseminated disease.     

Norepinephrine-induced changes in cardiac transforming growth factor-beta isoform expression pattern of female and male rats

Transforming growth factor-beta (TGF-beta) is a ubiquitous growth-regulating protein with an essential role in tissue repair and formation of extracellular matrix (ECM). To better understand the role of different isoforms of TGF-beta in the cardiac remodeling process induced by norepinephrine (NE), the expression of TGF-beta1, TGF-beta2, and TGF-beta3 was studied and compared with the expression of collagen. NE (0.1 mg/kg. h) was intravenously infused in female and male Sprague-Dawley rats for several time periods, and freshly obtained ventricular myocardium after 1 day was dissociated into myocyte and nonmyocyte fractions. Prazosin (0.1 mg/kg x h) and metoprolol (1 mg/kg. h) were used to block alpha- and beta-adrenoceptors, respectively. After NE infusion, the three isoforms of TGF-beta were differentially induced as far as the magnitude and the time course is concerned. The increased expression of TGF-beta2 started earlier with a maximum after 12 hours and was more pronounced (10-fold elevation) than that of the other two isoforms, with a clear specificity for the left ventricle in female hearts. This specificity was also seen in male rats with 16-fold elevation of TGF-beta2 after 1 day of NE-stimulation. The increase of TGF-beta2 was significant only in the myocyte fraction obtained from female as well as from male hearts. The expression of the mRNA of all TGF-beta isoforms of collagen type I and type III, and of the matrix metalloproteinase (MMP)-2 and its inhibitor TIMP-2 was reduced predominantly by alpha-adrenoceptor blockade with prazosin. The increase in TGF-beta isoforms correlated with that of the mRNA expression of collagens, MMP-2 and TIMP-2.     

Cardiac magnetic resonance imaging study for quantification of infarct size comparing directly serial versus single time-point measurements of cardiac troponin T.

OBJECTIVES: We compared single-point cardiac troponin T (cTnT) measurements with parameters from serial sampling during 96 h after acute myocardial infarction with magnetic resonance imaging measured infarct mass. BACKGROUND: Contrast-enhanced magnetic resonance imaging (CE-MRI) allows exact quantification of myocardial infarct size. Clinically, measurement of cardiac biomarkers is a more convenient alternative. METHODS: The CE-MRI infarct mass was determined 4 days after primary percutaneous coronary intervention in 31 ST-segment elevation myocardial infarction (STEMI) and 30 non-ST-segment elevation myocardial infarction (NSTEMI) patients. All single-point, peak, and integrated area under the curve (AUC) cTnT values were plotted against CE-MRI infarct mass. RESULTS: All single-point and serial cTnT values were significantly higher in STEMI than in NSTEMI (p < 0.01) patients. Except for the admission values, all single-point values on any of the first 4 days, peak cTnT and AUC cTnT were found to correlate comparably well with infarct mass. Among single-point measurements, cTnT on day 4 (cTnTD4) showed highest correlation and performed as well as peak cTnT or AUC cTnT (r = 0.66 vs. r = 0.65 vs. r = 0.69). Receiver-operator characteristic analysis demonstrated that cTnTD4 >0.84 microg/l predicted infarct mass above median as well as peak cTnT >1.57 microg/l or AUC cTnT (receiver-operator characteristic for AUC: 0.839 vs. 0.866 vs. 0.893). However, estimation of infarct mass with cTnTD4, peak cTnT, and AUC cTnT was worse in patients with NSTEMI (r = 0.36, r = 0.5, r = 0.36) than in STEMI (r = 0.75 vs. r = 0.65 vs. r = 0.76). CONCLUSIONS: All single-point cTnTs, except on admission, give a good estimation of infarct size and perform as well as peak cTnT or AUC cTnT. Infarct estimation by single-point measurements, particularly cTnTD4, may gain clinical acceptance because the measurement is easy and inexpensive.