Luciano Vettore

The diagnostic value of serum copper levels and other hematochemical parameters in malignancies.

An increase in the serum copper (Cu++) level has been described as a sensitive index of disease activity in several hematologic and nonhematologic malignancies. In order to explore the diagnostic value of Cu++ compared to other hematochemical parameters frequently abnormal in malignancies, Cu++, serum α2 globulin (α2), plasmatic fibrinogen (Fibr), the erythrocyte sedimentation rate (ESR), and serum iron (Fe++) have been detected and evaluated in 267 patients affected with the following diseases: Hodgkins lymphoma (HL), non-Hodgkins Lymphomas (NHL), Acute Leukemias (AL), Chronic Myeloid Leukemia (CML), Chronic Lymphocytic Leukemia (CLL), Myeloma (MM), and Breast Cancer (BC). The best correlation between Cu++ increase and disease activity has been found in HL, NHL, AL, and BC. In these diseases, when the considered parameters were compared, Cu++ and ESR showed a similar pattern, i.e., a high frequency of abnormalities in active disease. It is concluded that Cu++ represents a good complement to some other aspecific parameters in evaluating the activity and diffusion of neoplasias and the therapeutic results, particulary in HL, NHL, AL and BC.

Biosynthetic Ratio of Labelled Globin Chains in Human Reticulocytes, Determined by Electrophoresis on Cellulose Acetate

A new method for determining the biosynthetic ratio of globin chains in human reticulocytes is described.Washed red cells, enriched in reticulocytes by high speed centrifugation at 30°C, are incubated for 45 min. at 37°C with Minimum Essential Medium (Eagle) and 3H-Leucine.Globin is prepared from hemolysate by acid-acetone method. Globin chains are separated by electrophoresis on “Cellogel” at pH 6.3 in 0.04 M sodium phosphate buffer, containing urea 6 M, and stained with 0.5% Coomassie Blue. The strips of cellulose acetate containing the globin chains are dissolved in 10 ml of dioxan-naphtalene scintillation fluid, and counted in a liquid scintillation counter. The biosynthetic ratio of globin chains is calculated from the ratio of the total counts incorporated into the α and non- α chain zones.This method gives results in agreement with those obtained by macro-CMC column chromatography, but it is much simpler and faster. This electrophoretic method appears particularly suitable as a diagnostic tool in a...

Erythrocytic Proteases: Preferential Degradation of Alpha Hemoglobin Chains

Proteolytic activity against native hemoglobin polypeptide chains is demonstrated, under strictly physiological conditions, in human reticulocytes of both normal subjects and individuals suffering from a variety of pathologic conditions involving erythrocytes, including beta-thalassemia. Two thirds of the activity are found in the cytoplasm and the remainder of it is associated with the reticulocyte membrane. That this proteolytic activity is due to contamination by WBC is excluded. The activity preferentially degrades the alpha-hemoglobin chains. An increase in this substrate within the erythroid cells, as observed in beta-thalassemia, does not enhance proteolysis. Protease inhibitors produce a variable decrease in proteolysis. None inhibit completely, thus showing that several enzymes, with different specificities, are involved.

Erythrocyte cation content, globin chain synthesis and glucose metabolism in dysmyelopoietic syndromes.

Red blood cells (RBC) from 13 patients suffering from dysmyelopoietic syndromes (DMPS) have been studied. About half of these subjects showed an abnormal Na+ and K+ leakage of their RBC, which were incubated at 37 degrees C for 24 h. The mean rate of the glycolytic pathway increased significantly and the glycolysis values correlated well with the Na+ gain. Moreover, 9 out of the 13 patients showed an abnormal biosynthetic ratio of haemoglobin chains in their reticulocytes. Since the multiple defects in DMPS erythrocytes do not seem clearly linked by cause-effect relationships, they probably derive from several cooperating factors in pathological erythroid precursors, leading to RBC membrane damage.

standardization of a simple method for the estimation of haemoglobin A2

An improved method for the estimation of haemoglobin A2, at the same time precise, simple and cheap, is proposed. Haemoglobin A is separated from haemoglobin A2 by electrophoresis on Cellogel in discontinuous buffer at alkaline pH. The strips of cellulose acetate containing the haemoglobin fractions are completely dissolved in 80% acetic acid. The percentage of haemoglobin A2 present in each sample is calculated from the values for the spectrophotometric absorbance at 396 nm. The average percentage of haemoglobin A2 (+/- standard deviation) determined by this method was 2.31 +/- 0.37 in 51 normal subjects, and 4.64 +/- 0.53 in 29 subjects with heterozygous beta-thalassaemia.

Globin chain synthesis in human bone marrow.

Chalevelakis et a1 (1976) liavc rcccntly dcinoiistratcd that .//I ratios of globiii chain synthcsis in normal human bone inarrow arc lcss than unity bccausc non-liaciii protciiis, dcrivcd from wliitc cells, incorporate [3H]lcucinc and separate together with tlic /3 chain, spuriously incrcasing its spccific activity. We can confirm the Chalevelakis data through different expcriinental approaches: I. An artificial inixturc of normal reticulocytes (.IS= 0.92) with wlite cells froin chronic granulocytic leukaemia, both labelled with [3H]leucine, showed ‘a/P ratio’ of 0.41. A similar decrcasc of ‘a/S ratio’ from 0.99 to 0.50 has been obtained mixing, beforc the chain scparatioii, labclled globin from normal reticulocytes with labelled acetonic powdcr froiii white cells. 2. The ‘alp ratio’ in globin from whole cell lysate of bonc inarrow was 0.50. Thc ratio bccamc about I using the following procedure : (a) magnetic rcmoval of phagocytes previously incubated for 45 min at 37°C with 1% FcCO in niinimuin cssential incdiiuin; (b) labelling of newly synthcsized globin chains by incubation of the remaining cells with [3H]leucinc (Bank & Marks, 1966) ; (c) differential lysis of the erythroid cells in 3.07% glycerol (Gottfried & Robertson, 1974); (d) removal of unlysed granuloblasts by high speed centrifugation before preparing the globin powder from tlic supernatant haemolysate by acid-acetone method (Fanelli et al, 1958). The results suggest that a/fl production ratios of unity can be obtained in nornial bone marrow by previous removal of white cells instead of elimination of the contaminant proteins by particular chromatographic procedures. The same method gives a/P ratios ranging from 1.5 to 2.3 in bone marrow from heterozygous P thalassaeniics.

Evaluation of the scintigraphic technique for detecting bone metastases in 100 breast cancer patients.

In an attempt to evaluate the utility of bone scintigraphy for the diagnosis and follow-up of skeletal metastases, radioisotopic scan (total body) by 99mTc has been performed on 100 patients suffering from breast cancer in various stages. The radiographic control followed the scan on the basis of clinical or scintigraphic suspicion of bone lesions. Twenty-one patients showed metastases by scan, and in all but 2 the radiography was concordantly positive; in 22 subjects scintigraphy and radiology demonstrated non-malignant lesions. Both scan and radiology were used to explore 169 metastatic localizations, with concordant results in 45% of the sites; however, 7% of the sites with discordant results became positive for metastasis by radiology some months after the scan positivity. The false–negative results were more frequent in the radiographic study (27%) than with the scintigraphic scan (21%). The latter technique was unable to detect osteoblastic lesions in particular. The scintigraphic test also gave reliable results in the follow-up procedure, frequently anticipating the radiological modifications of metastatic lesions. In conclusion, bone scan often appears to be more accurate and precocious than the roentgengraphic test in the detection and the follow-up of skeletal metastatic lesions; it suggests the utility of the bone scan as a preliminary test, which allows a more rational use of the radiology.

The canine erythroid cell system A new model for study of regulatory mechanism in hemoglobin synthesis

The total methodology for a new mammalian erythroid cell system which permits direct investigation of the terminal stages of hemoglobin synthesis and assembly is described. The canine system allows quantitative separation of native heme containing alpha and beta chains which recombine to for tetrameric hemoglobin with normal functional properties (n = 2.17). These chains can be utilized for investigation in a cell-free system in which the rates of synthesis of alpha and beta chains are equal (alpha/beta = 0.96 +/- 0.05). Methodology for the quantitative separation of globin allows study of the effects of balanced and unbalanced globin chain synthesis in both the intact cells and the cell-free system.