Acid phosphatase and non-specific alpha-naphthol acetate esterase activities of monocytes in patients with cancer of the digestive tract during surgical treatment

In 22 patients with cancer of the alimentary tract the activities of acid phosphatase and non-specific alpha-naphthol acetate esterase in monocytes were tested. The enzyme activity was tested in the peripheral blood before surgical intervention, in blood from vessels draining the tumour before its excision and in the peripheral blood before surgical intervention, in blood from vessels draining the tumour before its excision and in the peripheral blood 2--3 weeks after tumour excision. In parallel tests the enzyme activity was estimated in the peripheral blood of 22 healthy individuals. The study indicates that the non-specific alpha-naphthol acetate esterase activity of monocytes derived from patients with cancer and control group did not show a marked difference. The acid phosphatase activity in monocytes derived from a tumour efferent vessel was found to be higher in majority of the cases than the activity of this enzyme in monocytes derived from the peripheral blood. After removing the tumour the acid phosphatase activity of monocytes was elevated in half of the cases. It seems possible that the increase of acid phosphatase activity in monocytes derived from cancer patients may be due to the activation of monocytes in contact with cancer antigens or antigen-antibody complexes.     

The depressing effect of tumour on bactericidal capacity of plasma and leukocytes.

Leukocyte--plasma suspensions and plasma from tumour patients have been tested in vitro against staphylococcus aureus for bactericidal capacity. Blood was collected during surgical intervention from peripheral vein and from the nearest point of vein to tumour. Second collection of peripheral blood was done 2--3 weeks later after surgery. Decrease of bactericidal activity of plasma has been found in blood collected from vein near the tumour. We can suggest that tumours release some factors disturbing the bactericidal capacity or that the tumour absorbs some factors of plasma or changes the functions of granulocytes.     

Nitroblue tetrazolium test in patients with breast cancer undergoing mastectomy and radiotherapy

The reduction of nitrotetrazolium blue with stimulated and unstimulated granulocytes was performed in 40 patients with breast cancer. Blood was collected from peripheral vein before surgical intervention, during surgery from vein draining of tumor, two weeks after surgery and three months after surgery when radiotherapy was finished. In parallel experiments the NBT test was made in the peripheral blood of 20 healthy individuals. We have observed decreased reduction of nitrotetrazolium blue in stimulated granulocytes, which may indicate that granulocytes from these patients are unable for efficient stimulation. No evident effect concerning the advancement of disease and applied therapy could be found. The importance of determining the NBT test in cancer patients is briefly discussed.     

Changes in leukocyte enzyme activity during experimental amyloidogenesis and in patients with amyloidosis]

The activity of hydrolytic and redox myeloperoxidase enzymes was determined in the neutrophils and lymphocytes of the peripheral blood of patients with secondary amyloidosis and also of the animals with amyloidogenesis and amyloidosis (caseine model). During the amyloidogenesis the activity of hyocrolytic enzymes and of myeloperoxidase in the neutrophils was found to decrease; this was particularly marked at the stage of the initial amyloid deposition. Changes in the enzyme activity in the animals against the background of already developed amyloidosis coincided with such in the blood cells of patients with secondary amyloidosis. The results obtained are discussed from the aspects of resorption theory.     

Changes in the myeloperoxidase and acid phosphatase activities in human peripheral blood neutrophils with the cells stimulated in vitro]

In four series of experiments human peripheral blood neutrophils were found capable of synthetizing the active forms of such enzymes as myeloperoxidase and acid phosphatase after stimulation with opsonized zymosan, and the optimum conditions for testing the synthetizing activity of neutrophils were established. The examination of 39 practically healthy donors revealed an approximate equilibrium between the synthesis of the active forms of the enzyme and its excretion from the cell after this cell was activated. Considerable changes in the enzymatic activity of peripheral blood neutrophils were found in patients with yersiniosis, chronic herpes, chronic bronchitis and AIDS-associated complex. The possibility of a deficient enzymatic activity in some patients was suggested.     

Ectopic production of methionine enkephalin and beta-endorphin.

Immunoreactive methionine enkephalin and beta-endorphin were sought by serial dilution of tissue extracts and assay of chromatographic fractions in non-endocrine tumour tissue from three patients with the ectopic adrenocorticotrophin syndrome associated with carcinoid tumours and in normal lung tissue and thymic tissue from a patient with myasthenia gravis. In all cases serial dilution of extracts showed parellelism to standard radioimmunoassay curves. The two peptides were found in high concentration in the three tumours but were undetectable in the control tissues. In a single case tested the methionine enkephalin concentration in a vein draining the tumour was twice that in a peripheral vein. In view of their profound effect on behaviour in animals and potent analgesic activity in animals and man the ectopic secretion of methionine enkephalin and beta-endorphin may modify the clinical features of a wide variety of tumours and produce some of the diverse clinical syndromes associated with malignancy.     

Biological consequences of occupational exposure to elementary mercury in light of hematologic tests

Principal haematological tests in peripheral blood, non-specific phosphatases and myeloperoxidase in granulocytes were determined in a group of 89 males occupationally exposed to the elementary mercury during manufacture of chlorine. Examined males were employed for 2-26 years. Blood mercury levels did not exceed 50 micrograms per l-1. The obtained results indicate that low concentrations of the elementary mercury in the environment do not produce significant abnormalities in both quantitative and qualitative picture of peripheral blood but markedly inhibit neutrophil non-specific phosphatases activity.     

Influence of age on brain vascular and cardiovascular monoamine oxidase activity in the rat

Monoamine oxidase (MAO) activity in brain microvessels and cardiovascular tissues was examined in rats of different age. MAO activity continued to increase with age in the heart, but in contrast, reached maximum activity in three weeks in the aorta, mesenteric artery and mesenteric vein. Between 7 and 60 weeks, there was a small decline in the MAO activity in the testicular artery. The highest MAO activity was found in the cerebral microvessels and increased with age. The half-life of MAO was estimated in the heart and peripheral blood vessels in young and old animals. The half-life of cardiac MAO was increased with age whereas that of the mesenteric vein, mesenteric artery and aorta remained constant between 7 and 112 weeks. Thus an explanation for this increased cardiac MAO activity in old rats was a reduced rate of degredation of this enzyme. The high activity of the enzyme in the brain microvessels suggests that it may participate in regulating the influx and efflux of monoamines in the central nervous system.     

Granulocyte and plasma cytokine activity in acute cadmium intoxication in rats.

Changes in the number and ex vivo function of peripheral blood neutrophils were investigated following intraperitoneal administration of cadmium-chloride in rats. Besides a dose-dependent increase in the number of peripheral blood neutrophils, changes were found in the functional state of isolated polymorphonuclear leukocytes (PMNs). Increased spontaneous adhesion and activation, and TNF activity in a conditioned medium were observed in cultures of granulocytes in comparison to granulocytes from control (saline-treated) animals. Increased levels of plasma activity of inflammatory cytokines, tumor necrosis factor (TNF) and interleukin-6 (IL-6) were noted following cadmium administration. Cytological signs of pulmonary inflammation were revealed histologically and the majority of neutrophils recovered from the lungs by enzyme digestion exhibited a capacity of nitroblue tetrazolium (NBT) reduction. Our data demonstrate that acute cadmium intoxication leads to a systemic inflammatory response characterized by numerical and functional changes in the granulocyte compartment and to increased levels of inflammation-related cytokine activity in the circulation. Correlations between the increased number of peripheral blood neutrophils and IL-6 plasma activity (r=0.776, p<0.00001) and the number of neutrophils recovered from the lung tissue (r=0.893, p<0.00001) suggested that systemic cadmium-induced inflammation might be involved in the pulmonary toxicity of cadmium.     

Localization of the luminol-dependent chemiluminescence reaction in human granulocytes

The granulocyte luminol-dependent chemiluminescence (CL) reaction is linked to the enzyme myeloperoxidase reacting with products of the respiratory burst activation. The results presented in this paper, show that the light generated in granulocytes originate both from intracellular and extracellular reactions; however, depending on the stimulus used the one or the other will dominate the activity measured. Furthermore, lysosomal fusion is proposed to be required for the intracellular CL reaction.     

The activity of neutrophil myeloperoxidase in patients with Hodgkin's disease.

Neutrophil myeloperoxidase activity has been studied in twenty one patients diagnosed with Hodgkin's disease. The presented data indicate no differences in total MPO activity, whereas we observed some differences in the percentage of granulocytes with different degree of scores. Changes in the intensity of reaction may indicate the possibility of exocytosis as a mechanism releasing MPO from the cell to the surrounding area. In peripheral, circulating neutrophils, such a phenomenon seems to be of no avail and may disorganise anti-cancer defence.     

Myeloperoxidases of human myeloid leukemia cells HL-60 grown in culture and in nude mice

Human myeloid leukemia HL-60 cells were grown either in suspension culture or in nude mice. The two types of myeloperoxidase, the small and the large type, in crude extracts of these cells were analyzed by sucrose density gradient centrifugation. The proportions of the small and the large myeloperoxidase varied markedly depending on the growth conditions of cells. In cells in culture, the small myeloperoxidase amounted to 80% of the total myeloperoxidase, whereas in the solid tumors it amounted to only about 30% of the total. Both in cultured cells and solid tumors, 40% of the total myeloperoxidase was found in the soluble fraction and the rest in the granule fraction. However, adult and fetal blood granulocytes contained only the large myeloperoxidase, which was mainly recovered in the granule fraction. Antiserum prepared against purified large myeloperoxidase of HL-60 solid tumors reacted with the small myeloperoxidase as well as the large enzyme of HL-60 cells in culture. The antiserum also precipitated myeloperoxidase of adult and fetal blood granulocytes. An Ouchterlony double immunodiffusion test also revealed their immunological identity.     

The effect of D-penicillamine on human myeloperoxidase, a mechanism for the efficacy of the drug in rheumatoid arthritis

We investigated the effect of D-penicillamine on the ability of myeloperoxidase, purified from human leukocytes, to catalyse the oxidation of chloride ions to hypochlorite (HOCl) in the presence of H2O2. It is shown that, due to the interaction of D-penicillamine with both myeloperoxidase itself and HOCl, the chlorinating activity of myeloperoxidase in the presence of H2O2 and chloride ions is prevented. A concentration of 100 microM D-penicillamine inhibits the chlorinating activity of myeloperoxidase completely, which Is due to the stabilization of Compound II, an inactive form of the enzyme. In addition, HOCl reacts directly with D-penicillamine. Analysis of the reaction products of D-penicillamine and HOCl showed that D-penicillamine was oxidized to penicillamine disulphide and penicillamine sulphinic acid, and eventually deaminated (indicated by the release of ammonia). Lower concentrations of D-penicillamine (10 microM) inhibited myeloperoxidase less, but still acted as effective scavengers of HOCl. In very low concentrations (1 microM), D-penicillamine did not scavenge HOCl effectively, but rather stimulated the chlorinating activity of myeloperoxidase. However, when instead of D-penicillamine a comparable amount of ascorbate was added, a similar but even larger stimulation was observed. Since the concentration of free D-penicillamine in serum from rheumatoid patients treated with this drug is about 20 microM (Saetre, R. and Rabenstein, D.L. (1978) Anal. Chem. 50, 276-280), the therapeutic effect of D-penicillamine may be due to the protection of tissues against the reactive HOCl released by activated granulocytes at inflammation sites.     

Cyclic GMP-dependent and cyclic AMP-dependent protein kinases,protein kinase modulators and phosphodieterases in arteries and veins of dogs Distribution and effects of arteriovenous fistula and arterial occlusion

Possible involvement of cyclic GMP-dependent and cyclic AMP-dependent protein kinases, protein kinase modulators and cyclic nucleotide phosphodiesterases in functions of vascular tissues were investigated in the dog. All of the above activities, localized in the smooth muscle-rich inner layer of the blood vessels, were found to be higher in the arteries than in the veins. The peripheral arteries were disproportionately richer in cyclic GMP-dependent protein kinase (as indicated by high ratios of cyclic GMP-dependent to cyclic AMP-dependent protein kinase) than were the veins, with the exception of the pulmonary artery, an atypical arterial tissue exposed to low blood pressure. Interestingly, the protein kinase ratio for the aorta, an artery with no significant role in blood pressure regulation, was not higher than that for the vena cava. Creation of femoral arteriovenous fistulae in the dogs led to preferential reductions in the cyclic GMP-dependent enzyme activity both in the proximal and distal arteries, whereas it was elevated in the stressed vein distal to the anastomotic site. The cyclic GMP-dependent enzyme was preferentially reduced in the saphenous artery distal to occlusion. Changes in the cyclic GMP-dependent enzyme activity appeared to precede gross atrophy or hypertrophy of the vessels. It is suggested that the vascular cyclic GMP-dependent protein kinase may be closely related to peripheral resistance and its regulation.     

Cyclic GMP-dependent and cyclic AMP-dependent protein kinases, protein kinase modulators and phosphodiesterases in arteries and veins of dogs. Distribution and effects of arteriovenous fistula and arterial occlusion.

Possible involvement of cyclic GMP-dependent and cyclic AMP-dependent protein kinases, protein kinase modulators and cyclic nucleotide phosphodiesterases in functions of vascular tissues were investigated in the dog. All of the above activities, localized in the smooth muscle-rich inner layer of the blood vessels, were found to be higher in the arteries than in the veins. The peripheral arteries were disproportionately richer in cyclic GMP-dependent protein kinase (as indicated by high ratios of cyclic GMP-dependent to cyclic AMP-dependent protein kinase) than were the veins, with the exception of the pulmonary artery, an atypical arterial tissue exposed to low blood pressure. Interestingly, the protein kinase ratio for the aorta, an artery with no significant role in blood pressure regulation, was not higher than that for the vena cava. Creation of femoral arteriovenous fistulae in the dogs led to preferential reductions in the cyclic GMP-dependent enzyme activity both in the proximal and distal arteries, whereas it was elevated in the stressed vein distal to the anastomotic site. The cyclic GMP-dependent enzyme was preferentially reduced in the saphenous artery distal to occlusion. Changes in the cyclic GMP-dependent enzyme activity appeared to precede gross atrophy or hypertrophy of the vessels. It is suggested that the vascular cyclic GMP-dependent protein kinase may be closely related to peripheral resistance and its regulation.     

Lipid peroxidation and the myeloperoxidase activity of neutrophilic leukocytes in shereshevsky?-Turner syndrome

The level of lipid peroxides, activity and thermostability of myeloperoxidase have been studied in neutrophil extracts from the peripheral blood of patients with the Shereshevsky-Turner syndrome and in normal subjects. A statistically significant rise in the level of lipid peroxides and respective decrease in the activity and thermostability of myeloperoxidase have been revealed in the patients as compared with the control group. These findings point to functional inferiority of neutrophils. It is probable that at the basis of the disorders seen there lie an abnormal set of sex chromosomes and upset hormonal regulation determined by the abnormality.     

Comparative analysis of function of reproductive organs of cow and female reindeer. Cellular composition of blood in vessels of reproductive organs

Cellular composition of blood was studied in vessels of reproductive organs and jugular vein in females of reindeer and cow depending on their physiological state. Physiological leukocytosis was revealed in reproductive organs of the animals, the most expressed in reindeer. In both species, during the estrous cycle, in blood of vessels of reproductive organs the content of lymphocytes and monocytes is higher than that of granulocytes. With onset of pregnancy, in vessels of ovary and uterus in reindeer and cow the number of lymphocytes and monocytes decreases, while the number of granulocytes increases due to a rise of eosinophils and basophils. The more successful reproduction of reindeer females under conditions of North seems to be owing to an increased immune reactivity of reproductive organs.     

Deficiency of beta-glucuronidase in neutrophils from patients with precancerous states of the larynx.

Activity of beta-glucuronidase (GR) and acid phosphatase (AP) has been determined in peripheral blood neutrophils from 24 men with precancerous states of the larynx that is leukoplakia papillomas and pachydermia by means cytochemical methods described by Hayashi et al., and Barka and Anderson, respectively. The results obtained were expressed in terms of absolute counts of enzyme-positive and enzyme-negative cells with regard to enzyme activity variation within the enzyme-positive neutrophil population; the enzyme activity index score has been calculated. The control group consisted of 20 healthy subjects of the same sex. No significant alterations were found so far as AP activity is concerned between the group studied. In contrast, activity of GR in patients with precancerous states exhibited significant lowering. The most striking feature was in almost complete absence from the blood of GR-positive neutrophils with high activity of the enzyme. Majority of these cells showed only traces of the GR activity. According to authors opinion the deficiency of GR in neutrophils of patients with precancerous lesions pertains to problem of neutrophil-mediated cytotoxic effect against mammalian tumour cells.     

The anlage of the duodenal lymphatic bed: morphogenetic prerequisites, structure and importance

The anlage of the duodenal lymphatic bed takes place on the 3d month of the human intrauterine life. In the intestinal villi there are chyle sinuses, that fuse into presumptive lymphatic vessels, having capillary structure, when they get out of the villi. Intraorganic vessels turn into extraorganic ones, they flow into the pancreatoduodenal lymphatic vessels. In their lumens stromal anlage of the lymphatic nodes are formed as a result of invagination of the blood vessels. Development of the lymphatic bed in the duodenum preceded with formation of intestinal villi (this contributes to absorption of the intestinal content) and deformity of the superior mesenteric vein (this probably makes difficult the organ's drainage). The situation is solved owing to the lymphatic bed anlage. It is, evidently, formed by means of switching off a part of embryonal veins from the blood stream.     

Development of an Arterio‐venous Difference Method to Study the Metabolic Physiology of the Femoral Adipose Tissue Depot

Gluteofemoral adipose tissue (AT) has interesting positive associations with metabolic health, yet little is known of its metabolic physiology. Here, we describe a technique for cannulation of a vein draining the femoral fat depot. Using ultrasound guidance, a cannula was introduced into a superficial branch of the great saphenous vein. We also obtained arterialized blood and, for comparison, blood representing drainage from forearm muscle and from subcutaneous abdominal AT. We measured appropriate biomarkers of skeletal muscle (creatinine) and AT (nonesterified fatty acids (NEFAs), glycerol, leptin) drainage. Blood obtained in this way from the saphenous vein did not show creatinine release (creatinine concentration 100.5 ± 0.4%, mean ± s.e.m., of that in arterialized blood), whereas creatinine concentrations in blood draining forearm muscle averaged 121 ± 1% of those in arterialized blood. Fatty acid release from the tissue drained was suppressed after feeding and increased during β‐adrenergic stimulation. We also demonstrated leptin secretion. These findings suggest that blood so obtained is representative of AT drainage with little apparent contribution of skeletal muscle. We believe this technique will facilitate physiological studies of a lower‐body fat depot in humans.