Effects of cyclic AMP on expression of LFA-1, Mac-1, and VLA-4 and eosinophilic differentiation of a human leukemia cell line,EoL-1

Abstract: We examined the effect of dibutyryl cAMP (dbcAMP) on the expression of LFA-1 (CD1 la/CD18), Mac-1 (CD11b/CD18), and VLA-4 (CD49/CD29) and on eosinophilic differentiation of a human leukemia cell line, EoL-1. Dibutyryl cAMP induced eosinophilic differentiation of EoL-1 cells from 6–9 days after the start of culture with down-regulation of CD 11a, CD 18, and CD49 expression and up-regulation of CD11b expression. Changes in integrin expression, except for CD18, were seen predominantly in the fraction containing eosinophilic granule-positive cells, suggesting that the changes were dependent on eosinophilic differentiation. On the other hand, dbcAMP-induced changes of integrin expression were reversible and were not seen on day 9 when dbcAMP was removed on day 3, whereas eosinophilic differentiation was still present. A combination of G-CSF and TNFα, which also induced eosinophilic differentiation of EoL-1 cells, increased CD11b expression slightly but had no significant effect on the expression of the other integrins. Butyrate and PMA up-regulated CD11b expression without eosinophilic differentiation. The results collectively suggest that the regulation of integrin expression on EoL-1 cells is partly dependent and partly not dependent on eosinophilic differentiation. The possible involvement of protein kinase A and protein kinase C in these changes is suggested.     

Ectopic ACTH-producing adenocarcinoma of the stomach.

A 73-year-old female was admitted to our hospital because of weight loss and pretibial edema. Plasma levels of adenocorticotropic hormone (ACTH) and cortisol were elevated, and neither hormone showed circadian rhythm. Dexamethasone (2 mg for 2 days) failed to reduce the urinary excretion of 17-hydroxycorticosteroids and the plasma cortisol level. The stomach biopsy specimens showed a moderately-differentiated papillo-tubular adenocarcinoma. Computed tomography of the abdomen showed multiple metastases to the liver. Immunohistochemical staining of the autopsy specimens showed immunoreactive ACTH in the primary tumor cells of the stomach as well as the metastatic tumor cells of the liver. On the basis of the clinical, histological and immunohistochemical findings, we diagnosed this patient as having ectopic ACTH syndrome caused by adenocarcinoma of the stomach.     

Three elderly cases with Brugada-type electrocardiographic pattern unmasked by pilsicainide at the toxic level

We describe three elderly patients with Brugada-type electrocardiographic abnormalities unmasked by a high plasma concentration of the Class IC antiarrhythmic drug pilsicainide, caused by renal dysfunction. The electrocardiograms showed right bundle branch block pattern and ST segment elevation in the precordial leads in patients 1 and 2, and a polymorphic wide QRS wave in patient 3. The serum concentrations of pilsicainide were 2.85, 2.50, and 4.18 microg/ml, which were much higher than the therapeutic range (0.16-0.24 microg/ml). Withdrawal of the drug reversed the electrocardiographic changes. Careful observation is necessary to avoid life-threatening arrhythmias whenever pilsicainide is administered to elderly patients.     

Clinical significance of the calcification of coronary arteries in patients with angiographically normal coronary arteries.

In order to clarify the clinical significance of coronary calcification in patients with angiographically normal coronary arteries, exercise electrocardiography was used and left ventricular function was examined noninvasively and invasively. The patient groups were as follows: (1) patients with coronary artery calcification on only the left anterior descending artery but no narrowing lesion on any other arteries (calcified group), (2) patients with a significant stenosis on only the left anterior descending artery (stenotic group), and (3) the control group. The left ventricular function in the calcified group, as indicated by systolic time intervals and invasive parameters such as ejection fraction and mean systolic ejection rate, showed a depression similar to that in the stenotic group, compared with the control group. The incidence of electrocardiographically ischemic responses to exercise testing was significantly higher in the calcified group (75%, p less than 0.01) and the stenotic group (68%, p less than 0.01) than in the control group (25%). Exercise tolerance time and the maximum double product were markedly smaller in the calcified and the stenotic groups as compared with the control group. These results indicate that the left ventricular function and coronary reserve in the calcified group were reduced and almost identical with those in the stenotic group. The authors conclude that a calcified coronary artery, even if patent, cannot supply an adequate blood flow for the myocardium, resulting in impaired left ventricular function.     

Association between diabetic nephropathy and left ventricular longitudinal myocardial function in type 1 diabetes mellitus patients with preserved ejection fraction

The International Journal of Cardiovascular Imaging - Left ventricular (LV) longitudinal myocardial dysfunction can be observed even in type 2 diabetes mellitus (DM) (T2DM) patients with preserved...     

Serum creatine kinase MM isoforms in hypertrophic cardiomyopathy.

1. To determine whether a persistent release of creatine kinase from the myocardium occurs in patients with hypertrophic cardiomyopathy, the activities of serum creatine kinase MM isoforms were measured in 22 patients with hypertrophic cardiomyopathy and in 14 normal control subjects. 2. Serum creatine kinase MB activity was significantly higher in patients with hypertrophic cardiomyopathy (7.8 +/- 3.8 i.u./l) than in normal control subjects (0.4 +/- 0.8 i.u./l; P less than 0.01). 3. Serum MMa, MMb and MMc activities in patients with hypertrophic cardiomyopathy were 19.4 +/- 4.1%, 26.7 +/- 2.5% and 33.5 +/- 7.0% of the total creatine kinase MM activity, respectively. These values for each isoform were significantly different from those in normal control subjects (11.3 +/- 3.0%, 21.5 +/- 4.4% and 40.7 +/- 7.0%, respectively). The MMa/MMc activity ratio was significantly higher in patients with hypertrophic cardiomyopathy (0.61 +/- 0.25) than in normal control subjects (0.30 +/- 0.10; P less than 0.01). 4. Our results indicate that a small amount of the myocardial tissue isoform of creatine kinase MM (MMa) is constantly released in many patients with hypertrophic cardiomyopathy.     

Induction of eosinophilic granules,nonspecific esterase activity and CD14 expression in the human eosinophilic leukemia cell line,EoL-1

We examined the expression of eosinophilic granules, esterase activity and CD14 in a human eosinophilic cell line, EoL-1. Unstimulated EoL-1 cells were weakly positive for nonspecific esterase, but negative for surface CD14, and contained a few eosinophilic granule-positive cells. A combination of G-CSF and TNF-α increased the eosinophilic granule-containing cells, but failed to increase esterase activity or CD14 expression. IFN-γ alone or in combination with TNF-α enhanced nonspecific esterase activity but failed to induce CD14 expression or increase eosinophilic granule-containing cells. dbcAMP increased eosinophilic granule-containing cells, nonspecific esterase activity and CD14 expression. Specific esterase activity was not detected in any circumstances. EoL-1 cells fractionated by density gradients or CD14 expression showed nonspecific esterase activity and CD14 expression in both the eosinophilic granule-positive and negative cell populations. Forskolin and butyrate had a synergistic effect on CD14 induction and protein kinase A was suggested to play a role in dbcAMP-induced CD14 expression. A protein kinase C activator, phorbol 12-myristate 13-acetate, did not increase eosinophilic granules, nonspecific esterase activity or CD14 experession in EoL-1 cells. The results show that EoL-1 cells can express nonspecific esterase and CD14, but the expression is not necessarily restricted to cells which have differentiated into the monocyte/macrophage lineage.     

Establishment of a myelodysplastic syndrome (MDS)/secondary AML-derived T lymphoid cell line K2-MDS

We have established a T lymphoid cell line, K2-MDS, from the peripheral blood mononuclear cells (PBMC) of a patient with acute myeloblastic leukemia (AML) transformed myelodysplastic syndrome (MDS). K2-MDS cells are positive for the expression of CD4, CD5, CD13, CD25, CD71, CD95, HLA-DR and cytoplasmic CD3. Southern blotting analysis shows T cell receptor (TCR) beta chain genes rearrangements, whereas immunoglobulin heavy chain (IgH) genes are not rearranged. Further, the patient PBMC contains TCR beta chain genes rearrangements in the same manner as K2-MDS cells. The data indicate that K2-MDS is a T lymphoid cell line derived from a myelodysplastic clone in the patient PBMC. This new MDS-derived cell line K2-MDS may be a useful in vitro model for studies on the pathogenetic mechanisms leading to MDS.     

Accuracy of measurement of radiochromic film density for high-energy X-rays

Radiochromic film (RC-film) is of great interest as a film-type dosimeter for radiation oncology applications. We present a two-dimensional image-based evaluation of the measurement accuracy of a commercial RC-film product (Gafchromic MD-55-2 film, ISP TECHNOLOGIES, Inc.) by using a commercial Laser Densitometer (Model 1710, Computerized Medical Systems, Inc.) as an optical density imaging system. The coefficient of variation of the density (pixel-value) in one sample was approximately 3% to 11% at 3 Gy or less, and 3% or less at 4 to 60 Gy. Although the coefficient of variation between three samples at the same dose was about 14% at 1 Gy, it decreased as the dose increased, reaching several percent. In 1 to 6 Gy samples, geometric imaging artifacts [interference (moire) patterns] were observed, and it was found that scan-sampling pitch influenced the accuracy of measurement of the density of the sample. To improve the accuracy of density measurement, sufficient knowledge about characteristic features of the density measuring system is essential.