Production of interleukin-1 by bone marrow myeloma cells.

Plasma cells isolated from bone marrow (BM) aspirates of 12 patients with multiple myeloma (MM) and nine patients with monoclonal gammopathy of undetermined significance (MGUS) were analyzed for production of cytokines with bone-resorbing activity, such as interleukin-1 (IL-1), tumor necrosis factor (TNF), and lymphotoxin (LT). Culture supernatants of plasma cells from MM, but not from MGUS or normal donor, invariably contained high amounts of IL-1-beta and lower amounts of IL-1-alpha. With a single exception, TNF/LT biologic activity was not detected in the same supernatants. IL-6 was present in two of five supernatants tested. Normal B lymphocytes released both IL-1 and TNF/LT activities for four days after activation in vitro; however, production of these cytokines ceased at the final stage of plasma cell. Unexpectedly, the mRNA extracted from MM plasma cell hybridized with TNF- and LT-specific, as well as IL-1-specific probes, although the culture supernatants did not contain detectable TNF/LT biologic activity. When tested in the fetal rat long bone assay, MM plasma cell supernatants displayed a strong osteoclast-activating factor (OAF) activity, which was greatly reduced but not completely abolished by neutralizing anti-IL-1 antibodies. Anti-TNF or anti-LT antibodies were ineffective in the same test. We conclude that the IL-1 released in vivo by malignant plasma cells has a major role in pathogenesis of lytic bone lesions of human MM.

Interleukin 1 as an autocrine growth factor for acute myeloid leukemia cells.

Production of interleukin 1 (IL-1) by leukemic cells was studied in 13 cases of acute myeloid leukemia. Intracytoplasmic immunofluorescence studies showed that the cells invariably contained the cytokine. Endogenous labeling studies demonstrated that acute myeloid leukemia cells produced either only the 33-kDa propeptide or both the propeptide and the 17-kDa mature form of IL-1 beta. The 33-kDa propeptide IL-1 alpha was always produced but was less frequently released. Involvement of IL-1 in leukemic cell growth was investigated using two antibodies specific for IL-1 subtypes, which inhibited spontaneous cell proliferation in the six cases studied. After acid treatment of the cells, a surface receptor for IL-1 could be demonstrated, which mediated 125I-labeled IL-1-specific uptake by leukemic cells. Furthermore, recombinant IL-1 alpha or IL-1 beta induced significant cell proliferation in 10 of 12 cases. The above findings were uncorrelated with the cytologic type (French-American-British classification) of leukemia. Our studies suggest that IL-1 may act as an autocrine growth factor in most cases of acute myeloid leukemia.

Aminoethers of (+)-1,3,3-trimethyl-2-oxabicyclo[2.2.2]-octan-6-hydroxyimine with hypotensive activity. II.

The synthesis of a series of N-substituted 3-amino-2-hydroxypropyl ethers (III a-1) starting from (+)-1,3,3-trimethyl-2-oxabicyclo [2.2.2]-octan-6-hydroxyimine is described. Some of these compounds showed a remarkable hypotensive and a weak bradycardic activity in rats, as well as a moderate local anesthetic activity in mice. All compounds were found to be devoid of antiarrhythmic activity in rats and of beta-blocking activity in dogs.

Derivatives of 1,3,3-trimethyl-6-azabicyclo-[3.2.1]octane with hypotensive activity. III.

The synthesis of a series of alkyl and aryl thioureas (II) starting from 1,3,3-trimethyl-6-azabicyclo[3.2.1]octane is described. Some of alkyl thioureas showed a moderate hypotensive activity in rats. Effects on heart rate and hypoglycemic activity in rats, infiltration anesthesia and antiarrhythmic activity in mice are also reported.

Pharmacological properties of a new non-steroidal anti-inflammatory drug: flunoxaprofen.

The anti-inflammatory, analgesic and antipyretic activities of S-(+)-2(4-fluorophenyl)-alpha-methyl-5 benzoxazole acetic acid (flunoxaprofen: Flu), a new non-steroidal anti-inflammatory drug, were compared with those of indomethacin and other non-steroidal anti-inflammatory drugs (NSAIDs) in experimental animals. Flu showed strong inhibitory activity on acute and subacute inflammation tests in rats, such as carrageenin hind paw oedema (oral: 6-25 mg/kg; rectal: 50-100 mg/kg); pellet-induced granuloma formation (5-20 mg/kg/day) and adjuvant-induced arthritis (10 mg/kg/day). Its potency was comparable with that of indomethacin (I) and higher than that of acetyl salicylic acid (ASA), ibuprofen (IBU) or phenylbutazone (P). The analgesic activity of Flu, evaluated by the hot plate method and tail pinching in mice, was slightly lower than that of I but higher than that of ASA and IBU. In pyretic rabbits Flu showed an antipyretic activity higher than that of ASA and IBU. The ability of Flu to affect platelet aggregation, mucopolysaccharide synthesis by fibroblasts and the proteolytic action of trypsin was also investigated.

Nucleotide deficit and functional platelet alterations in patients on regular dialysis treatment (RDT).

In RDT patients hemocoagulative changes are repeatedly found; of these the most important are platelets' functional defects. Biochemical and biophysical modifications responsible for this pathology have not been completely clarified. In 20 non-thrombocytopenic patients, dialyzed 3 times weekly for over 1 year, we evaluated, using standard methodology, platelet adhesivity and aggregation induced by collagen and ADP at varying dosages. All blood samples were collected after the longest interdialytic period and just before dialysis. At the same time we evaluated the basic metabolic pool and, after collagen stimulation, the intraplatelet functional storage pool of ATP and ADP. The dosages were obtained using simple, reproducible and modern bioluminescence technique, which utilises microorganism light emission (bioluminescence) due to the oxidation of the bacterial substrate by catalyzing enzyme (luciferase) (1251 Luminometer LKB). We compared this data with that obtained from 40 healthy subjects. In the patients examined, adhesivity and aggregation result altered. The intraplatelet content of all nucleotides in both pools is significantly reduced if compared to the control group. The ATP and ADP concentration of both metabolic and functional pools cannot be correlated to the following: serum creatinine, BUN, calcemia, PTH, Hb. On the contrary we found that basal metabolic ATP values are inversely related (p less than 0.01) to serum phosphate levels. An analysis of results of this preliminary study leads us to hypothesize that hyperphosphatemia could interfere with intraplatelet glycolysis inducing a reduction of intracellular ATP. As all platelet functions are ATP and ADP dependent, we could consider the nucleotide deficit a cause of "uraemic platelet" disfunction, not modifiable with RDT but perhaps only through an appropriate control of phosphate levels.

[Cytochemical studies of bone marrow plasma cells in monoclonal gammapathies (author's transl)]. / Etude cyto-enzymologique des plasmocytes médullaires des dysglobulinémies monoclonales myélomateuses et non myélomateuses.

Three plasma cell enzymatic reactions--ATPase, acid phosphatase (AP), alpha-naphthyl acetate esterase (alpha-NAE)--were tested in a large number of bone marrow samples from patients with multiple myeloma (MM), benign monoclonal gammapathy (BMG)--idiopathic (iBMG) and secondary (sBMG)--, polyclonal hypergammaglobulinemia (PH), Waldenström's macroglobulinemia (WM) and from normoglobulinemic subjects (NS). The purpose of the present study was to evaluate the significance of these reactions in differential diagnosis of BMG and initial or atypical MM. For each reaction results were expressed as scores and normal limits were statistically established. Our findings in MM, HP and NS are consistent with previous reports (similar enzymatic pattern in NS and PH; depressed ATPase activity, raised AP and alpha-NAE activities in most MM cases). In the BMG class, ATPase activity was normal in about 40% of iBMG, while it was depressed in other cases; most sBMG had diminished ATPase activity. AP patterns were normal for both BMG groups. alpha-NAE activity was normal in 50% of iBMG, while it was raised in the other cases. All the reactions were normal in WM. In a bayesian analysis of cytochemical patterns of NS and MM subjects ATPase showed the highest diagnostic significance, followed by alpha-NAE and AP. ATPase, therefore, seems to be the most useful criterion for the diagnosis of initial MM; low ATPase score BMG patients should therefore be carefully followed-up.

A comparison of two staging systems for myeloma.

The data were evaluated for 43 myeloma patients who had had complete follow-up. A comparison was made of the prognostic significance of the staging systems proposed by Durie and Salmon (DS) and by Merlini, Waldenström, and Jayakar (MWJ) for multiple myeloma. Both of the three-stage systems show good statistical correlation with survival in our global analysis, although a higher log-rank P value was found for DS. Moreover, only stage I and III survival curves are really heterogeneous in MWJ, while each curve is significantly different from the other in DS.

Bone marrow percentage of plasma cells in the staging of monoclonal gammopathies.

In the present study, the relationships between percentage of bone marrow plasma cells (BMP%), tumor cell mass, stage, and survival are statistically analyzed in a large number (90) of monoclonal gammopathies (MG). Though BMP% has not been included among the criteria for the staging of multiple myeloma, our results indicate that the above mentioned parameters correlate at highly significant degrees. BMP% is therefore proposed as a new, useful parameter in the staging of MG.

[The nature of crystals in plasma cells present in a case of apparently benign biclonal gammapathy]. / Sulla natura dei cristalli endoplasmacellulari presenti in un caso di gammapatia biclonale apparentemente benigna.

A 77 years old white man presents, in his blood serum, a moderate gammapathy with 2 monoclonal peaks, IgAk and IgGlambda, and in his urine a little quantity of BJk, without any clinical or radiological manifestation. The sternal bone marrow showed 3-5% plasmacells, mostly containing many needlelike inclusions in the cytoplasm. These inclusions, red-violet at the May-Grünwald-Giemsa, were PAS and Sudan negative; positive at the Danielli reaction (for proteins). The alpha-naphtil-acetate-esterase and the acid phosphatase were present in one outer layer of the inclusions; ATP-ase was absent. At the electron microscopy, the inclusions were localized outside the rough endoplasmic reticulum; they exhibited a crystalline structure and were surrounded by an envelope which reminded the lysosomes. Basing on the morphological pattern, as well as on the presence of some lysosomal enzymes and on the lack of staining of crystals with fluorescinated anti-Ig sera, the hypothesis is stressed of an abnormal lysosomal hyperactivity, possibly leading to crystallization of the enzymatic proteins in the lysosomes.