Effect of a constant magnetic field and gamma radiation on the hereditary structure of somatic cells. Effect of the combination of a constant magnetic field and ionizing radiation on human lymphocytes in vitro

Samples of human whole blood were exposed in CMF (field induction, 0.3 T) for 15, 30, 45, 60, 75, 90, 120, 150, 180, 240, 300 or 360 min. 15 min following exposure, the samples were gamma-irradiated in a dose of 0.0516 C/kg (137Cs) at a dose rate of 1.95 A/kg. The following chromosome aberrations were scored: deletions dicentrics, rings, and symmetrical exchanges. Exposure of the blood in CMF for 15 to 360 min decreased radiation damage to cells as compared with unexposed irradiated samples. The extention of time from 15 to 180 min increases the effect the smallest amount of chromosome damages being scored at 150-180 min. A 2.8 - fold, 3 - fold and 3.5 - fold decrease was registered in the number of aberrant cells, deletions and dicentrics, respectively. With increasing time of exposure (240 min), the radiomodifying effect started decreasing, and with 300-360 min exposure it was the same as that observed at 15-45 min.     

The effect of static magnetic field on protein concentration in serum of guinea-pigs

The influence of the static magnetic field on magnetic induction 0.005 T--0.3 T on the protein concentration in serum of guinea-pigs with regard of twenty four hours rhythm was investigated. The range of occurrenced changes were determined by the duration of the static homogeneous magnetic field.     

The lack of the effect of a strong constant magnetic field on isolated membrane preparations of Na,K-dependent ATPase

Effect of constant magnetic field (CMF) with induction 10 T on membrane preparations of Na,K-dependent ATPase of bovine brain (lipoproteid vesicules with 300-500 A diameter) were studied. No CMF effect on the activity of Na,K-dependent ATPase was observed under different experimental conditions (three temperature points 15, 20 and 37 degrees C and great variation of Na+,K+ concentrations ratio). CMF also produced no effect on the preparations of Na,K-dependent ATPase immobilized by adsorption on millipore filters.     

The effect of static magnetic field on fibrinogen degradation products level in rabbits with thrombosis

The effect of static magnetic field of induction 0.005 T, 0.12 T and 0.3 T applied in daily rhythm (one hour every day) for the period of 2 weeks and 4 weeks produces an increase of FDP level in the serum. Especially, the effect elicited by the magnetic field applied 2 weeks prior to experimental thrombosis development. The range of changes was related to the duration of the exposure to the magnetic field. No dependence of the degree of induction of the magnetic field was established.     

Fibrinolytical processes in rabbits activated by the magnetic field

Rabbits were exposed to a constant magnetic field of 0.005 T, 0.1 T and 0.3 T induction for one hour per day each day for a period of four weeks. It was found that the magnetic field increases the rate of fibrinolytical processes. A decrease in fibrinogen concentration, an increase in the level of fibrinogen degradation products and a considerably shorter time of fibrinolysis in plasma were all noted. The magnitude of these processes was proportional to duration of exposure to the magnetic field in action. These date confirms the similar effect observed in other mammalians (guinea pigs, rats). Thus, the application of a static magnetic field of intensity as low as 0.005 T to increase a fibrinolytical processes in the thrombotic therapy seems to be justified.     

Liver and spleen morphology, ceruloplasmin activity and iron content in serum of guinea pigs exposed to the magnetic field

The chronic exposure of guinea pigs to the magnetic field of induction 0.005 T and 0.3 T cause the morphological changes of spleen and functional disturbances of liver (the histochemical analysis indicate on greater amount of glycogen in hepatocytes). Everyday 1-hour exposure determined the drop in ceruloplasmin activity and an unchanged iron content in the serum of tested animals.     

Mesodermalizing Factors in Fetal Calf Serum

The inducing activities of fetal calf serum (FCS) and fetal calf serum heated for a short time (HFCS) were tested with early gastrula presumptive ectoderm of the newt, Cynops pyrrhogaster as a reactor. Inducing activity was measured by sitting-drop culture of pieces of ectoderm before and after treatment with Ca, Mg-free solution (CMF) in culture medium contained FCS or HFCS (10–50%) as inductor. Results showed that FCS did not induce mesodermalization of ectoderm, before or after treatment with CMF, and that HFCS induced mesodermalization only of ectoderm treated with CMF. The treated ectoderm cells differentiated into mesoderm cells such as muscle and notochord. The induction increased with increase in the duration of CMF treatment and with the concentration of HFCS in the medium. This indicates that FCS changes into mesodermalizing material when heated for a short time and that mesodermal induction by HFCS depends on some effect of CMF on presumptive ectoderm. Since CMF was found to be a neuralizing factor, activation of presumptive ectoderm with a neuralizing factor is probably a prerequisite for mesodermal induction with HFCS.     

Effect of chronic exposure to static magnetic field upon the K+, Na+ and chlorides concentrations in the serum of guinea pigs

The magnetic field influence on the concentration of serum K+, Na+ and chlorides was tested. The guinea pigs were exposed to the static magnetic field for six weeks 1 hour a day, 7 days a week. Magnetic field of induction 0.005 T--0.3 T produced progressively an increase in Na+ concentration and a decrease in chlorides concentration in the serum. The range of observed changes was dependent on the duration of exposure to the magnetic field. No change in K+ serum concentration was observed following magnetic field exposure.     

Epstein-Barr virus (EBV) receptors, complement receptors, and EBV infectibility of different lymphocyte fractions of human peripheral blood. I. Complement receptor distribution and complement binding by separated lymphocyte subpopulations.

Human peripheral lymphocytes were fractionated into a variety of B-, T-, and O-cell fractions and were characterized with regard to several surface receptors. There was a strong correlation between the frequency of EAC receptor-positive cells and the percentage of complement membrane fluorescence (CMF)-stained cells following exposure to fresh human serum and subsequent staining with an anti-C3 conjugate. CMF staining did not diminish in C4-deficient or hypogammaglobulinemic serum, or in the presence of EDTA or EGTA-Mg²⁺, but was completely negative with C3-depleted normal human serum. In all likelihood, the staining is therefore due to the direct binding of C3 to preformed receptors on the lymphocyte surface. In addition to the surface Ig-positive B-cell fractions, C3 receptors were also detected on part of the O-cell population and on a proportion of the Fc receptor-positive T cells.     

Increase of IGF I binding to erythrocyte receptors during the TRH-test: correlation between IGF I binding and thyroid hormone values

The effects of TRH on insulin-like growth factor I receptors were investigated on erythrocytes from 7 GH-deficient children having plasma GH levels less than 10 ng/ml during two provocation tests. Intravenous injection of synthetic TRH (0.2 mg/m2) was followed by a marked increase of IGF I binding on erythrocytes, from 3.9% +/- 0.3% to 5.9% +/- 0.3% (P less than 0.005) after 1 hour and 7.3% +/- 0.4% (P less than 0.005) after 2 hours. The IGF I binding variations were due to an increase in both the receptor affinity and the number of sites. The levels of plasma GH, IGF I, T3, T4, free T4, TSH and prolactin having been determined during the TRH test at 0, 1 hour, and 2 hours after the injection, the increase in the IGF I binding to erythrocytes at the same time correlated with the rise of thyroid hormones: triiodothyronine T3 (P less than 0.001) and thyroxine T4 (P less than 0.005) and not with the level of the other hormones. These findings suggest that thyroid hormones play a role in the regulation of insulin-like growth factor I receptors.     

The dominant lethal effect of dietary triethylenemelamine.

Triethylenemelamine (TEM) was administered in the diet to adult male mice at doses of 0.1, 0.3, 1, 10 or 50 mg/kg body weight for 45 days or at doses of 0.1 or 0.3 mg/kg b.w. for 10 days. As a comparison, male mice were treated intraperitoneally with 5 daily doses of 0.25 or 0.5 mg TEM/kg b.w. At the end of the treatment period, males were mated sequentially with 2 untreated virgin females each for 2 or 3 weeks. Near mid-pregnancy the number of implantation sites and fetal deaths were determined. TEM, administered in the diet at 10 or 50 mg/kg b.w. for 45 dyas, was lethal to male mice. Surviving males from the 1 mg/kg level failed to impregnate any females during the two matings. TEM, given in the diet at 0.1 or 0.3 mg/kg for 10 or 45 dyas, decreased fertility and increased dominant lethal mutations in a dose and time dependent manner. These results were comparable to those obtained from males treated i.p. with TEM at 0.25 or 0.5 mg/kg b.w.     

Neural Induction by Treatment with Ca2+-free Saline Solution in the Gastrula Ectoderm of Cynops pyrrhogaster and Inhibition of Neural Induction with Fetal Calf Serum

The relation between the inducing activity and the cell-dissociation effect of Ca²⁺-free (or Ca²⁺, Mg²⁺-free) saline solution (CF or CMF) on the early gastrula ectoderm was examined. In the culture medium containing no fetal calf serum (FCS), most ectoderm cells treated with CF or CMF died within a few days and only a few differentiated into epidermal cells. However, when the culture medium contained 2% FCS, ectoderm cells treated with CF or CMF differentiated into neural crest derivatives (NCDs), such as mesenchyme cells, pigment cells, and nerve cells. The frequency of the induction depended only on the duration of CF- or CMF-treatment. FCS alone had no inducing activity on ectoderm cells. On the contrary a high concentration of FCS gave an inhibitory effect on the induction. These results indicate that CF is a neuralizing factor and that CF-treated cells require FCS, not for induction, but for survival and differentiation. With CF, the maximum induction of NCDs required a longer duration than that necessary for complete cell-dissociation. This result suggests that the induction depends on some effects of CF other than cell-dissociation.     

Effects of extremely weak alternating magnetic fields on the plant gravitropism

It was shown that the rate of gravitropic response in apical segments excised from the 4-day-old seedlings of flax (Linum bienne) may be substantially influenced by combined magnetic fields (CMF) of two different types: 1) CMF tuned to the parametric resonance for Ca2+; 2) CMF containing extremely weak alternating component with the values of magnetic density ranging from 10(-6) to 10(-10) T. Our experimental data indicate that CMF affect the gravitropic response via at least two different mechanisms. The first one corresponds to the ion parametric resonance well established earlier in studies with test-systems prepared from animals. The origin of the bioeffects induced by CMF containing extremely weak alternating component remains to be established.     

Long-term influence of adjuvant therapy on natural killer cell activity in breast cancer

Summary Unstimulated IFN- and IL2-stimulated (NK) cell activities were investigated in patients with breast cancer who had received either local radiotherapy alone or adjuvant chemotherapeutic treatment with CMF combined with radiotherapy 12 to 18 months previously. When tested against the primarily NK-sensitive K562 cell line, patients who had received adjuvant chemotherapeutic treatment with CMF were shown to have a significantly decreased unstimulated and IFN-stimulated NK cell activity, as compared to both patients after radiotherapy only (P<0.002) and P<0.005, respectively) and healthy control persons (P<0.05). The former group of patients also had a significantly decreased IFN-stimulated NK cell activity, when tested against the primarily NK-insensitive Chang hepatoma cell line, as compared to patients after radiotherapy only (P<0.005) and healthy controls (P<0.05). Moreover, patients after radiotherapy only proved to have a significantly increased unstimulated (P<0.01) and IFN-stimulated NK cell activity (K562: P<0.05; Chang hepatoma cell line: P<0.05), as compared to healthy control individuals. In contrast, no difference in IL2-stimulated NK cell activity was detected. The investigation for the expression of CD3 and/or Leu 19 antigens as phenotypic markers of cells with non-MHC restricted cytotoxicity showed a significantly lower percentage of cells with the CD3+ phenotype in patients with breast cancer, irrespective of the chosen post-operative treatment, as compared to healthy controls (P<0.01). Finally, patients with breast cancer who had received radiotherapy only had a significant trend towards an increased percentage of CD³+/Leu 19+ PMNC, as compared to both patients after CMF treatment (P<0.05) and healthy controls (P<0.025). We conclude that patients with breast cancer vary on a long-term basis in their NK activity and in the phenotype of their PMNC depending on their post-operative adjuvant management.Abbreviations NK natural killer cells - IFN interferon - IL 2 interleukin 2 - CMF cyclophosphamide, methotrexate, fluorouracil - ER oestrogen receptor - PMNC peripheral blood mononuclear cells - MHC major histocompatibility complex - CTL cytotoxic T lymphocytes     

The role for monocyte chemoattractant protein-1 in the generation and function of memory CD8+ T cells

Memory T cells are resistant to the conventional costimulatory blockade and therefore impede tolerance induction. However, their migratory, survival, and functional requirements for chemokines are not well understood. We herein examine the role for MCP-1 or CCL2 in the generation, migration, and function of memory CD8+ T cells. We found that overall generation of both central memory (TCM) and effector memory (TEM) CD8+ T cells was severely impaired in the absence of MCP-1. Importantly, the survival of TEM, but not TCM, CD8+ cells was reduced without MCP-1, whereas the homeostatic proliferation of TCM, but not TEM, CD8+ cells was weakened in MCP-1-/- mice. However, once they were generated in the absence of MCP-1, in vitro function of both subsets of memory cells remained intact as determined by their proliferation and IFN-gamma production. Interestingly, the migration of TCM, but not TEM, CD8+ cells to inflammatory sites was significantly delayed without MCP-1, whereas both subsets of memory cells underwent comparable expansion and apoptosis with or without MCP-1 during the effector phase. Moreover, the function to eliminate a graft of TCM, but not TEM, CD8+ cells was impaired without MCP-1. Thus, this study demonstrates that MCP-1 plays an important role in not only migration but also generation and survival of memory T cells. This finding provides new insight into the requirement of chemokines for the generation, survival, and function of differential subsets of memory T cells and may have clinic implications for tolerance induction.     

A secreted 80 x 10(3) Mr protein mediates sensing of cell density and the onset of development in Dictyostelium.

In submerged monolayer culture, Dictyostelium cells can differentiate into prespore and prestalk cells at high cell densities in response to cAMP but not at low cell densities. However, cells at low densities will differentiate in medium taken from developing cells starved at a high density. The putative factor in the medium was designated CMF for conditioned medium factor (Mehdy and Firtel, Molec. cell. Biology 5, 705-713, 1985). In this report, we size-fractionate conditioned medium and show that the activity that allows low density cells to differentiate can be separated into high and low Mr (relative molecular mass) fractions. Interestingly, the two fractions both have the same activity and do not need to be combined to allow differentiation. The large conditioned medium factor is a protein, as determined by trypsin sensitivity, that can be purified to a single 80 x 10(3) Mr band on a silver-stained SDS-polyacrylamide gel, and has CMF activity at a concentration of approximately 4 pM (0.3 ng ml-1). Our results suggest that CMF is a secreted factor that functions in vivo as an indicator of cell density in starved cells. At high cell densities, the concentration of CMF is sufficient to enable cells to enter the multicellular stage of the developmental cycle. When present below a threshold concentration, cells do not initiate the expression of genes required for early development. This factor plays an essential role in the regulatory pathway necessary for cells to obtain the developmental competence to induce prestalk and prespore gene expression in response to cAMP.     

Growing and starving Dictyostelium cells produce distinct density-sensing factors.

Prestarvation factor (PSF) and conditioned medium factor (CMF) are two autocrine factors produced by Dictyostelium cells. Although secreted at different times in the Dictyostelium life cycle (PSF by growing cells and CMF by starving cells), both factors are glycoproteins that are used by cells to measure their own density, and both are important in cell aggregation. To examine the relationship between PSF and CMF, a CMF antisense transformant was tested for the production of PSF during growth. Although this transformant produced extremely low levels of CMF, its production of PSF was essentially normal. We conclude that these two factors are not products of the same gene.     

A strong constant magnetic field affects muscle tension development in bullfrog neuromuscular preparations

Effects of a constant magnetic field (CMF) of 0.65 T on muscle tension over 9 h were studied in the neuromuscular preparation of the bullfrog sartorius muscle. Tension was developed every 30 min by stimulation of the sciatic nerve (nerve stimulation) or of the sartorius muscle itself (muscle stimulation). In sciatic nerve stimulation, tension decreased rapidly for the first 3-4 h at a similar rate in both test (exposed to CMF) and control muscles. However, the rate of decrease became smaller and almost leveled off after 3-4 h in the test muscles, whereas tension continued to decrease monotonically in control muscles. The slope of the decrease for these later periods was significantly different between the test and the control conditions. Accordingly, tension was larger in test than in control muscles. In muscle stimulation, tension decreased monotonically from the start of experiments in control muscles, while tension in test muscles maintained their initial values for almost 3 h. Thereafter they started to decrease with a similar rate to the control. Hence, tension was always larger in test than in control muscles. A similar pattern of temporal change was observed for the rate of rise of the maximum tension for nerve or muscle stimulation. However, a significant difference was detected only in the case of muscle stimulation. The present results showed that a strong CMF of 0.65 T had biological effects on tension development of the bullfrog sartorius muscle by stimulation of the sciatic nerve as well as by stimulation of muscle itself. The presence of a small AC magnetic field component leaves open the possibility of an AC, rather than a CMF effect.     

Regulation and processing of a secreted protein that mediates sensing of cell density in Dictyostelium.

During Dictyostelium development, the expression of some genes is dependent on cell density. This effect is mediated by soluble factors referred to as conditioned medium factors (CMFs) which the developing cells secrete at very low rates and simultaneously sense. There are at least two classes of CMFs: one is an 80 x 10(3) Mr glycoprotein and the other is a heterogeneous group of molecules, with relative molecular masses between 6.5 x 10(3) and 0.65 x 10(3). Interestingly, the two classes of molecules do not need to be combined for activity. We find that the 80 x 10(3) Mr CMF but not the small CMF is sequestered in vegetative cells. The 80 x 10(3) Mr CMF is then secreted by cells during early development, while the small CMF appears only during late development. Like the 80 x 10(3) Mr CMF, the small CMFs are trypsin-sensitive and contain N- and O-linked glycosylation. The breakdown products of a fraction containing 80 x 10(3) Mr CMF cochromatographed from a Sephadex G-50 column and a reverse-phase HPLC column with small CMFs. The specific activity of CMF increases roughtly 100-fold upon breakdown. The results suggest that, during differentiation, the slowly diffusing 80 x 10(3) Mr CMF is first produced from a precursor pool already present in vegetative cells, allowing differentiation of only those cells in the immediate vicinity of the aggregation center. The breakdown of 80 x 10(3) Mr CMF to a faster-diffusing, higher specific activity form then might enable cells farther from the aggregation center to differentiate.     

Relationships between complement activation, complement binding, and EBV absorption by human hematopoietic cell lines.

Complement consumption (C.C.) and C3 deposition on the cell membrane, visualized by membrane fluorescence (CMF), were compared in a collection of established human lymphoid lines. C.C. was independent of the presence of C3 receptors. A positive CMF reaction was seen only in lines that expressed C3 receptors, however. Trypsin treatment abolished CMF and EAC rosetting but had virtually no influence on C.C. EBV absorptive capacity correlated with both C3-receptor expression, as measured by EAC rosetting, and CMF but not with C.C. This is in line with our previous finding on the association of EBV and C3 receptors.