Interdependence between Zn and Cu serum concentrations and serum immunoglobulins (IgA, IgM, IgG) in liver diseases.

The object of this study was to find out, by means of statistical-mathematical methods, whether the serum concentrations of Zn and Cu had a role in the interrelation among immunoglobulins IgA, IgM, IgG in four groups of subjects with various forms of liver diseases. It was found that the correlation structure between IgA, IgM, IgG and the serum concentration of Zn or Cu were different in the four types of diseases studied, and also that of these two metal ions, Zn was the most important, influencing the interdependence between the immunoglobulins in the active forms of disease.

Variations of zinc in acute and chronic hepatitis (preliminary data).

The variations of serum zinc concentration were studied in acute and chronic hepatitis of various etiologies. Coefficients of correlation were found with serum immunoglobulins and alanine aminotransferase (ALAT). The data obtained suggest the possible utilization of these coefficients in the estimation of prognosis in acute and chronic liver diseases.

Study of interrelationship between Zn, Ca and Mg seric concentrations in healthy subjects comparatively with diverse forms of cancer.

The interdependence between the serum Zn, Ca, Mg concentrations was studied in a group of healthy subjects comparatively with a group of patients with various form of cancer. It was found that it is significant difference between the control group and the groups of cancer patients, which shows that the carcinogenic processes disturb the concentrations of these metallic ions, not only quantitatively but also the interdependence between them.

Treatment with zincum metallicum CH5 in patients with liver cirrhosis. Preliminary study.

The zinc, an important enzymatic cofactor, is involved in many metabolic processes. Its deficiency might be due either to malabsorption or to excessive utilization. In the medical literature of the latest 10 years, zinc was considered to play a part in the immune processes. The authors of the present paper intend to study the zinc and immunoglobulin levels in various diseases, i.e., chronic progressive hepatitis, liver cirrhosis (LC), dermatitis, bronchial asthma. This preliminary investigation was carried out in 30 patients with LC in whom serum zinc values were assayed by atomic absorption spectrophotometry and the immunoglobulin levels were determined using the Mancini type simple radial immunodiffusion technique. All these patients presented considerable decrease of serum zinc concentration, the values ranging between 3.06 and 7.65 mumol/l as compared with 19.8 +/- 1.5 mumol/l in the controls, alongside with the increase of immunoglobulins G and M. In the patients treated with Zincum metallicum CH5 it was observed after about 30 days of treatment that the clinical state was considerably improved and IgG and IgM as well as serum zinc had resumed their normal values. This treatment should not be interrupted since in LC, without permanent additional supply, the serum zinc returns rapidly to the initial deficit or even lower.

Study of serum ceruloplasmin and of the copper/zinc ratio in cardiovascular diseases.

The serum copper and zinc concentration, the Cu/Zn ratio and ceruloplasmin activity have been studied in 400 subjects divided into 4 groups of study: 50 normal subjects (controls), 100 patients with ischemic heart disease, 100 subjects with myocardial infarction in the past history and 150 patients with acute myocardial infarction. It was observed that in normal subjects the Cu/Zn ratio is about 1, in acute myocardial infarction it reaches 2.5 but it returns to normal during convalescence. In the patients with ischemic heart disease the ratio decreases to 0.8 and in the subjects with infarction in the past history it is about 1.6 (almost normal). The variation of these ratio is due to the serum zinc concentration which in ischemic heart disease presents values over the upper normal limit and in acute myocardial infarction below the lower normal limit. In the subjects with acute infarction ceruloplasmin (copper depending enzyme) the activity was observed to be inversely proportional to the Cu/Zn ratio in the first 4 days after the onset of the disease. The quick normalization of one or of both parameters is a sign of positive evolution of the disease.

Possible correlation between the zinc and copper concentrations involved in the pathogenesis of various forms of anemia.

The concentrations of three trace elements (iron, copper and zinc) involved in hematopoiesis were studied in 200 patients with iron-deficit anemia, hemolytic anemia and Biermer's anemia, in comparison with a group of normal subjects. The method used was atomic absorption spectrophotometry. The results obtained showed that in iron-deficit anemia resistant to iron therapy the copper and ceruloplasmin serum concentrations are at the lowest limit of normal. This copper deficit has a negative effect on the disease by preventing hemoglobin synthesis. In iron-deficit anemias responding to iron therapy the relatively low (about 90 gamma %) zinc values have a negative effect by blocking the iron in the iron reserve. In hemolytic anemia the very high zinc serum concentrations (over 200 gamma %) may lead to a shortening of the erythrocytes life span due to its "entatic state". In Biermer's anemia zinc by its high concentration has a positive role owing to carbonic anhydrase which eliminates more rapidly carbon dioxide from the organism.

The IgD-zinc relationship in patients with allergic bronchial asthma.

In recent years, evidence was brought of the zinc ion (Zn++) interference in the mechanism of certain processes occurring at molecular level, i.e., the protein synthesis, the activity of some enzymatic complexes, the regulation of transmembranal transport or the processes of immune regulation. The present paper investigates the variations of the immunoglobulin classes in patients with bronchial asthma, particularly the relationship between IgD levels and those of the zinc ion in the blood serum.

Variations of zinc, calcium and magnesium in normal subjects and in patients with neoplasias.

The serum zinc, calcium and magnesium concentration as well as the possible relationship between these elements was determined by means of atomic absorption spectrophotometry in 150 patients with prostatic, lung, liver and breast neoplasms as well as with chronic leukemia, and in 80 normal subjects. These determinations showed that zinc concentration is increased in almost all the forms of cancer except prostatic and lung cancer in which the zinc values were below the lowest normal limits. From this study it also resulted that high zinc values favour the development of cancer. The antagonic relationship between zinc and calcium or magnesium in neoplasias proves the protective role played by the latter elements in the forms of cancer in such cases.

Study of the relationship between the granulocyte LDH, alkaline phosphatase and Zn at the level of the leukocyte in patients with chronic myeloid leukemia.

Mixed leukocyte suspensions were prepared from heparinized blood collected from healthy subjects and from patients with chronic myeloid leukemia (CML). In all the suspensions determinations were made for: zinc, by atomic absorption; granulocyte alkaline phosphatase (GAP), using the method with p-nitrophenylphosphatase; granulocyte LDH, by means of the enzymatic autoanalyser LKD 8,600. In the patients with CML, the values of zinc and of granulocyte alkaline phosphatase activity were very low while the granulocyte LDH values were higher than normal. The chromatogram of the granulocyte LDH isoenzymes on DEAE-Sephadex A50 minicolumn (0.5 X 12 cm) showed an "alpha type abnormality" revealed by the increased activity of the isoenzymes with high electrophoretic mobility LDH2 and LDH1 specific for tissues with intense oxidative phosphorylation. In the normal subjects the chromatogram of the leukocyte LDH isoenzymes showed a type M (skeletal muscle) prevalence denoting intense anaerobic glycolysis. Therefore the low zinc concentrations (0.55 micrograms mg N2 as compared with the normal 1.24 micrograms mg N2) in these patients cause the decrease of GAP activity by the lack of zinc in the active center of the enzyme and the decrease of cellular permeability thus allowing the extracellular release of granulocyte LDH.

Alkaline phosphatase activity improved by the addition of zinc in granulocytes from leukemic subjects (in vitro study).

Granulocyte alkaline phosphatase (GAP) was extracted from the leukocyte suspensions of 15 subjects with chronic myeloid leukemia (CML), determining the Zn content by atomic absorption spectrophotometry. The purified enzymatic preparation was dialyzed against a ZnCl2 solution (.01 mg%) in TRIS-HCl 0.1 M, pH 7.5, for 48 hours. After dialysis Zn concentration and GAP activity reached normal values. This suggests that the GAP molecule isolated from CML subjects is not altered as it is able to receive at most 2 Zn moles responsible for the enzymatic function.

The relationship zinc--alkaline phosphatase activity in normal and pathologic granulocytes.

To study the relationship granulocyte alkaline phosphatase (AP)--zinc (Zn) in mixed leukocyte suspensions, the enzyme from 51 mixed leukocyte suspensions was separated and purified. In the pure extract, determinations were carried out for enzymatic activity--using the autoanalyzer LKB 8600--and for Zn content--using atomic absorption spectrophotometry. From the data obtained it resulted that, in agreement with the results in the literature Zn concentration in the mixed leukocyte suspensions from chronic myeloid leukemia, (CML) patients is about half that in the normal controls (0.65 +/- 0.15 as compared with 1.24 +/- 0.15 micrograms/mg total nitrogen). Granulocyte AP separated from normal leukocyte suspensions was found to bind about 95% of the total Zn content, i.e., 1.21 +/- 0.15 micrograms/mg total nitrogen and its enzymatic activity was about 16 U/g total nitrogen. In CML patients the enzyme bound a mean Zn concentration of 0.55 +/- 0.15 micrograms/mg total nitrogen and its mean activity was 2.25 U/g total nitrogen, i.e., about 86% of the normal values found in the controls. The enzymatic activity decreased to zero after only 20 min dialysis with 8-hydroxyquinoleine-5-sulfonic acid (8HC5S) as compared with the 60-80 min in the controls. It can be concluded that the leukemic leukocyte is unable to form the metal-enzyme complex owing to the deficit of the Zn involved in the enzymatic activity.

Study of trace elements in patients with rheumatoid arthritis stage I-II, before and after treatment.

The study of the copper, zinc, iron and magnesium serum values in 35 patients with rheumatoid arthritis (RA) stage I-II, before and after therapy with indocin, gold salts and transfer factor showed a significant increase of copper to 129-340 gamma/100 ml, mean value 214.6 gamma/100 ml as compared with 65-145 gamma/100 ml, mean value 107.6 gamma/100 ml in the controls. The values of serum copper were found to evaluate with the severity of diseases returning to normal values in the patients with good response to therapy. In disagreement with some authors who found decreases of serum zinc in the course of rheumatoid arthritis, in our study the values obtained did not vary significantly as compared to normal (mean value 119 gamma/100 ml in RA as compared with 94.7 gamma/100 ml in the controls). The decrease of iron and magnesium serum concentration was also found nonsignificant in the patients with RA investigated. The results are discussed in terms of physiopathology.

The Bucharest multifactorial prevention trial of coronary heart disease. General methodology and risk factor correction after five year follow-up (1971-1977).

Both the intervention and the control group included each 5000 men aged 40-60 years, randomly selected by their home address in five districts of the city of Bucharest. The intervention group underwent an "at entry" examination for risk factor detection (high serum cholesterol, high blood pressure, overweight, diabetes, minor ECG abnormalities, family history) and subsequently a five-year multifactorial intervention aimed to reduce the risk factors. Both groups were followed up in this lapse of time for major end-points: myocardial infarction, stroke, sudden death. The qualitative analysis of the results used ten evolution indices based on a quantal counting and lead to a classification of risk factors which allowed the setting up of a strategy for their correction. The quantitative analysis showed the following decreases between the first and the last examination in the intervention group: for serum cholesterol greater than or equal to 250 mg/dl -17%; for cigarettes/day greater than or equal to 15-53%; for overweight greater than or equal to 30% - 13.57%; for high blood pressure -8%; for the overall risk computed by multiple regression -33.8%.

Separation of creatine-kinase isoenzymes on ion-exchange mini-columns for diagnostic purposes.

For diagnostic purposes, creatine-kinase (CK) isoenzymes were separated on mini-columns of DEAE Sephadex A50 from the serum of 55 patients with myocardial infarction or cerebral disturbances and from 45 normal subjects. The optimum conditions of separation were determined by separation of CK isoenzymes (CK--MM, CK--MB, CK--BB) from human organ homogenates (skeletal muscle, myocardium, brain). In the patients with myocardial infarction the CK-MB isoenzyme activity amounted to more than 6% of the total enzyme value. The chromatogram in these cases was similar to that of human myocardium homogenate. The CK-BB isoenzyme was found only in patients with severe cerebral disturbances (cerebral hemorrhage). In such cases the chromatogram was similar to that of human brain homogenate. Therefore, by associating the assay of serum CK total activity with the separation of CK isoenzymes it was proved that the CK--MB and CK--BB isoenzymes can supply a useful test in differential diagnosis.

Separation and purification of LDH and alkaline phosphatase isoenzymes for the improvement of organo-specific diagnosis.

The LDH isoenzymes from mixed granulocytic populations and from homogenates of different organs, as well as alkaline phosphatase from human liver and intestine, were separated and purified by means of chromatography using ion exchangers (DEAE-cellulose and DEAE-Sephadex A50). Comparison of purified LDH isoenzyme chromatograms with the zymograms in agar gel has allowed the determination of the isoenzymatic pattern of the respective tissues and its correlation with the predominant energetic processes. The purified extracts of alkaline phosphate have lead to the characterization of the main molecular forms of the enzyme thus enabling a more precise organo-specific diagnosis.

Circadian variations of certain blood components currently investigated.

The circadian variations of serum components such as: cholesterol, LDH, GOT, alkaline phosphatase, proteins, bilirubin, ferrous ions and creatinine were investigated. Blood collection was made two times in 24 hrs.: 5-6 p.m. and 7-8 a.m. on the next day. The data obtained allowed the classification of the subjects into two groups: stable and variable. The most interesting observations were: evening decrease of cholesterol, evening increases of LDH with a tendency of intensification of the LDH5 fraction in the zymogram and evening decreases of ferrous ion.

Coronary heart disease and its risk factors in a group of 5,000 middle aged men in urban environment.

The results obtained at entry in the subjects included in a pilot study (Institute of Internal Medicine, Bucharest) for the detection and prevention of coronary heart disease and hypertension, are presented. These data are the prevalences of the risk factors of coronary heart disease (high serum cholesterol, hypertension, smoking, overweight, diabetes, nonspecific minor ECG signs, family history), as well as the prevalences of the various forms of coronary heart disease. The study of the frequency distribution of biologic parameters likely to become risk factors showed that in middle aged subjects the upper limit of the normal should be lowered from the 95th percentile to the 76th one.