The effect of K2Cr2O7 on the growth and morphology ofEscherichia coli

Treatment with 16 μM K₂Cr₂O₇ results is a time-dependent loss of the clonogenicity ofE. coli cells. Under the same experimental conditions, optical density values do not change significantly, since cells have the ability to replicate but not to septate. In fact, microscopic analysis of samples taken over 240 min of exposure to the toxin has revealed the formation of filaments. The filamentous response appears similar to that generated by mitomycin C.     

Characterization of a nickel resistant mouse cell line

Nickel is a potent carcinogen and, at high concentrations, is toxic to mammalian cells. The effects associated with nickel exposure are well-documented but its mechanism of action in the cell has not yet been fully described. In order to understand the metabolic fate of nickel in mammalian cells, a variant cell population has been selected that continues to grow and divide in the presence of nickel chloride concentrations that are toxic to the parental cell line (Balb/c-3T3 mouse fibroblasts). Nickel resistance is not caused by altered uptake of nickel from the medium or increased clearance from the cells and is not associated with changes in metallothionein expression. Compared to the normal cells, the nickel resistant cells have a decreased number of chromosomes and numerous centromeric fusions. The expression of some proteins and the distribution of nickel bound by various proteins are altered in the nickel resistant cells. Preliminary results indicate that the nickel resistant phenotype may be transferred by genomic DNA-mediated transfection into a recipient NIH-3T3 cell line. Current investigations are directed at identifying a gene responsible for nickel resistance.     

The in vitro transformation of a rat liver epithelial cell line with chromium

The transformation of a rat liver epithelial cell line under a wide range of doses of chromium was determined by anchorage-independent growth and tumor formation in syngeneic animals. Chronic exposure to low concentrations and brief exposure to high concentrations of hexavalent chromium (K₂CrO₄) transformed the cells, but one dose (1 mM K₂CrO₄, 2h) was clearly optimal in this regard. The cytotoxicity, effects on cell cycle, rates of chromium uptake, and mutagenic activity under the different treatment conditions were evaluated. The results showed that cells could adapt to the presence of chromium under certain treatment conditions, but this was not the case for the optimal transforming dose. Cells treated with chromium above the optimal transforming dose showed evidence of a transient G2 arrest, whereas all lower levels of treatment did not. A low level continuous exposure to chromate was mutagenic, whereas high level short exposures, including the optimal transforming dose, were not. An increase in the amount of protein complexed with isolated nucleic acids was detected in cells following treatment with the optimal transforming dose of chromate. The results indicate that the effects of chromium on this in vitro system vary with dose; and the identification of those events relevant to metal carcinogenesis will require consideration of treatment conditions.     

The involvement of heterochromatic damage in nickel-induced transformation

Nickel ions produce selective damage in heterochromatic regions of chromosomes. Male Chinese hamster embryo cells, which have heterochromatin along the entire long arm of the X-chromosome, exhibit an unusually high incidence of nickel-induced transformation compared with female cells of the same species. However, 3-methylcholanthrene, a carcinogen that produces a random distribution of chromosome damage, transforms female and male cells equally. Other species that do not have as much heterochromatin on the X-chromosome exhibit similar incidences of nickel-induced tumors in males and females. Four out of five of the male nickel-transformed Chinese hamster cell lines exhibit a deletion of the heterochromatic long arm of the X-chromosome as the only common karyotypic aberration. This result indicates that a deletion of a heterochromatic chromosomal region may be an important feature of the nickel-induced carcinogenic process. All of the male nickel transformed cells lines are able to form tumors in athymic nude mice.     

Enhancement of the kidney Cd burden by SH-containing chelating agents

The accumulation of Cd in the kidneys is enhanced markedly if chelating agents that contain SH-groups like 2,3 dimercaptopropanol (BAL) are injected immediately after the metal. This is not a transient effect but persists for more than 3 d. It is less pronounced at higher chelate doses or when the pH of urine is increased. Our experiments indicate that chelating agents, which form unstable complexes at acid pH and are able to pass through the cell membrane, will cause metal accumulation in the kidneys.     

Role of Liv-52 in protection against beryllium intoxication

An ayurvedic medicine, Liv-52, was studied as a prophylactic agent against beryllium-induced toxicity in rats. Administration of berylliumper se caused severe degenerative and necrotic changes in kidneys, liver, and uterus. Beryllium exposure also reduced glycogen content, activities of alkaline phosphatase, succinate-dehydrogenase, and adenosine-triphosphatase in these organs. On the contrary, activities of acid phosphatase and glucose-6-phosphatase showed marginal increase. Liv-52-primed rats exhibited comparatively less marked toxic effects.     

The influence of different flow velocities on tumor cell survival in micropore filters

Earlier studies have shown a lower degree of lodgement and early survival of tumor cells in muscle than in liver after infusion via the femoral artery and portal vein, respectively. A possible explanation to this difference might be that the tumor cells are mechanically destroyed, and thus die more rapidly in muscle because they enter this capillary network at a much higher flow velocity. In the present study, the effect on early tumor cell (rat fibrosarcoma) survival of a high and a low flow velocity/deformation rate was evaluated in micropore (5 μm) filters, using isotope (Cr⁵¹) technique. These experiments were combined with scanning electron microscopic (SEM) analyses. The filter experiments showed no significant differences in the rate of cell death in the filters between tumor cells subjected to high or low deformation rates, and there were no qualitative differences in tumor cell appearance in the SEM study. It is, therefore, concluded that the difference in tumor cell lodgement and survival between muscle and liver is not primarily caused by differences in the rate of cell deformation upon entry of the organ capillary network.     

Immobilized metal ion affinity chromatography

This article describes the technique of immobilized metal ion affinity chromatography (1MAC). The IMAC stationary phases are designed to chelate certain metal ions that have selectivity for specific groups in peptides and on protein surfaces. The number of stationary phases that can be synthesized for efficient chclation of metal ions is unlimited, but the critical consideration is that there is enough exposure of the metal ion to interact with the proteins, preferably in a biospecific manner. The versatility of IMAC is one of its greatest assets. An important contribution to the correct use of IMAC for protein purification is a simplified presentation of the various sample elution procedures.     

Cell orientation induced by extracellular signals

Cells like fibroblasts and osteoblasts are oriented by different extracellular guiding signals like an electric field, a bent surface, and a periodically stretched surface. An automatic controller is responsible for the cell alignment. The controller contains both a deterministic and a stochastic signal. The following machine properties were determined: (1) The angle dependence of the cellular signal transformer is cos 2(psi 0 - psi). (2) The set point of the automatic controller is psi 0 = +/- 90 degrees. The cells like to orient their long axis perpendicular to the direction of the applied guiding signal. (3) The signal transformer measures the extracellular signal in a quadratic fashion. The cells cannot register the sign of the guiding field. (4) The stochastic signal in the automatic controller can be quantified by a characteristic time (approximately 130 min for fibroblasts). (5) The extracellular signal is registered in cell-made standards (ratio of the deterministic and stochastic signal equals one): 0.3 +/- 0.05 V/mm for human fibroblasts (electric field) and 85 +/- 3 microns for human fibroblasts and osteoblasts (cyclindrically bent surface). (6) The lag-time in the signal transduction system of fibroblasts is approximately 4 min.     

A new technique for the study of erythrocyte survival by double labeling and use of a well-type Ge detector

A double label procedure with⁵⁷Co and⁵⁸Co has been developed for detailed in vivo studies of erythrocyte survival. A well-type Ge detector is used in the measurements. The activities necessary for these experiments are very low, and the associated dose received by the test persons can be neglected.     

Mechanism of comutagenesis of sodium arsenite withn-methyl-n-nitrosourea

Arsenic compounds are known carcinogens. Although many carcinogens are also mutagens, we have previously shown that sodium arsenite is not mutagenic at either the Na⁺/K⁺ ATPase orhprt locus in Chinese hamster V79 cells. It can, however, enhance UV-mutagenesis. We now confirm the nonmutagenicity of sodium arsenite in line G12, a pSV2gpt-transformed V79 (hprt ⁻) cell line, which is able to detect multilocus deletions in addition to point mutations and small deletions. The lack of arsenic mutagenicity has led to studies emphasizing its comutagenicity. Sodium arsenite at relatively nontoxic concentrations (5 μM for 24 h or 10 μM for 3 h) is comutagenic withN-methyl-N-nitrosourea (MMU) at thehprt locus in V79 cells. Using a nick translation assay, which measures DNA strand breaks by incorporating radioactive deoxyribonucleoside monophosphate at their 3′OH ends in permeabilized cells, we found that much more incorporation was seen in cells treated with MNU (4 mM, 15 min) followed by 3-h incubation with 10 μM sodium arsenite compared with cells exposed to the same MNU treatment followed by 3-h incubation without sodium arsenite. This result shows that in the presence of arsenite, strand breaks resulting from MNU or its repair accumulate over a 3-h period. We suggest that the repair of MNU-induced DNA lesions may be inhibited by arsenite either by affecting the incorporation of dNMPs into the MNU-damaged DNA template or by interfering with the ligation step.     

Determination of selenium in duplicate diets of residents of Pinhel,Portugal, by neutron activation

A rapid cyclic instrumental neutron activation analysis (CINAA) method has been used to determine the selenium content of 27 duplicate diet samples from each of the 27 districts surrounding Pinhel, Portugal. The accuracy and precision of the CINAA method have been evaluated by analyzing certified reference materials and observed to be within ±5–10% for samples containing at least 40 ppb of selenium. The detection limit has been found to vary between 26–42 ppb selenium depending on the sample composition. The average daily dietary intake has been calculated as 37 μg of selenium per day.     

Effects of etretinate on the distribution of elements in rats

We studied in the rat the effects of the drug etretinate (Tigason), given at three doses 3, 10, and 30 mg/kg body wt for 1 mo, on the concentrations of Na, K, Ca, Mg, Fe, S, P, Cu, and Zn in the plasma, brain, thymus, heart, liver, lung, kidney, testicle, muscle, and bone. The elements were simultaneously determined in tissues after nitric acid dissolution by inductively coupled plasma emission spectrometry using a JY 48 instrument. At the dose of 3 mg/kg, etretinate did not induce any statistically significant modifications of the element distribution. At the dose of 10 mg/kg, the main observed modifications were in plasma an increase of copper (+38%) and a decrease of zinc (-25%). At the highest dose of 30 mg/kg, some variations of the concentrations of elements in tissues were observed. But, on no account did retinoids induce an alteration of the mineral composition of bone, despite obvious macroscopic bone alterations.     

Effects of doxorubicin on the sensitivity of L1210 leukemia cells to deformation-associated trauma

Most of the cancer cells arrested in the microcirculation during hematogenous metastasis are rapidly killed; one major mechanism is surface-membrane rupture, associated with the mechanical deformation of cancer cells in capillaries. The feasibility of increasing the susceptibility of cancer cells to lethal, deformation-associated trauma by doxorubicin, was tested in an in vitro mechanical model system, by filtering suspensions of L1210 leukemia cells through 8-μm pore-size Nuclepore® membranes, with or without prior incubation with 10^-7M doxorubicin. The results showed that mechanically-induced loss of cancer cells immediately after filtration was increased from 18 to 55% in cells previously exposed to doxorubicin for 48 h. The results indicate the feasibility of chemotherapeutic enhancement of the mechanical killing-action of the microvasculature as a potential rate-regulator of hematogenous metastasis.     

Multielement determination in rice,wheat, and barley by instrumental neutron activation analysis

INAA has been used for the determination of Na, Mg, Al, Cl, K, Sc, Cr, Mn, Fe, Co, Cu, Zn, As, Se, Br, Rb, Sr, Mo, and W in grains of rice, wheat, and barley, which were collected from different plant fields in Iraq. Samples and standards were irradiated in the IRT-5000 reactor, at neutron fluxes of 2 × 10¹³ cm⁻²·s⁻¹ and 3.2 × 10¹¹ cm⁻²·s⁻¹. Interferences of photopeaks with each other were considered, and reaction interferences were calculated and determined experimentally. Accuracy of our method was assessed by the analysis of IAEA standards Wheat Flour and Bovine liver. A good agreement has been achieved between the present results and recommended values. The precision and detection limit were determined for all elements in all types of grain.     

Copper deficiency and hyperlipoproteinemia induced by a tetramine cupruretic agent in rabbits

The effectiveness of a cupruretic agent, N,N'-bis-(2 amino ethyl)-1,3-propanediamine HCl or 2,3,2-tetramine HCl (TETA), in the induction of copper (Cu) deficiency and the ability of a Cu-deficient diet in the maintenance of the depressed Cu status 10 wk after TETA treatment were examined in this study. In the first experiment, 42 male New Zealand White rabbits, 35 d of age, were randomly divided into three dietary treatments: a copper (Cu)-deficient (2.3 mg Cu/kg diet), a Cu-adequate (13.5 mg Cu/kg diet), and a commercial ration (21.6 mg Cu/kg diet) group. A single oral dose of 100 mg of 2,3,2-tetramine HCl TETA/kg body wt/d were administered to half of the rabbits from each treatment group for 10 d while the remaining rabbits were untreated. In the second experiment, 10 similar rabbits were assigned to three treatments: Cu-deficient plus TETA (n = 4); Cu-adequate plus TETA (n = 3); and Cu-adequate alone (n = 3). The rabbits were fed a TETA dose of 100 mg/d for three 4-d periods over 3 wk, and thereafter maintained on the diets for another 10 wk. Rabbits from the first experiment fed Cu-deficient diet and treated with TETA demonstrated cardiac hypertrophy and markedly reduced plasma and liver Cu concentrations that indicated that the animals were Cu-deficient. Significant elevations (twofold) in low density lipoprotein (LDL) protein, cholesterol, triglyceride, and apolipoprotein B (apo B) concentrations were observed in TETA treated rabbits fed Cu-deficient diet. In the second experiment, the plasma LDL protein level remained elevated, the plasma Cu level was reduced 45%, and the Cu level of the heart when expressed as microgram/g dry tissue was reduced, 10 wk post TETA treatment in rabbits maintained on Cu-deficient diet. Thus, Cu deficiency and hyperlipoproteinemia was rapidly induced by TETA and was still evident 10 wk posttreatment in rabbits maintained on a Cu-deficient diet.     

Interference of Manganese on neuroendocrinal system in exposed workers

The neurotoxic effects of Manganese (Mn) are well known, and are usually caused by high occupational exposure over long periods of time. The effects caused by low doses of this metal have been poorly evaluated. For this reason, we decided to evaluate neuroendocrinal tests in a group of 14 male workers employed in a ferrousmanganese foundry (exposed to Mn air concentrations within the TLV-TWA) and in 14 male control subjects, by analyzing FSH, LH, prolactin, and cortisol. The urinary Mn, prolactin, and cortisol levels were significantly higher in the worker’s group. The preliminary results of this research seem to show that for exposure below the TLV, Mn can somehow interfere with the neuroendocrine system. In order to confirm the existence of these effects and to verify their possible correlation with the dopaminergic control system, further studies are necessary.     

Measurement of exchange of trace metals between the body and its surroundings with the use of trace methodology

Application of general tracer theory to the problem of estimating fluxes of tracee between the gastrointestinal tract and the body proper, from observations of the movement of tracer, shows that a number of assumptions must be fulfilled. Two specific sets of assumptions are discussed and, in both cases, measurement of tracer fluxes yields information on the integrated absorption of the tracee.