Transport of amino acid aryl amides by the intestinal H+/peptide cotransport system, PEPT1

The oxygen tension (PO2) in the renal cortex and outer renal medulla in 26 rats was studied by use of oxygen microelectrodes before and after injection of x-ray contrast media (CM). The CM, iopromide, ioxaglate and iotrolan were administrated intravenously in iodine equivalent doses (1,600 mg iodine/kg body wt). Ringer's solution was used as the control. In the outer medulla, all three CM induced a decrease in PO2: iopromide (N = 6) from 30 +/- 3 to 18 +/- 4 mm Hg; ioxaglate (N = 7) from 32 +/- 6 to 15 +/- 4 mm Hg; and iotrolan (N = 6) from 36 +/- 3 to 14 +/- 4 mm Hg. All these decreases were significant. After the injection of Ringer's (N = 7) there was an increase from 34 +/- 3 to 35 +/- 3 mm Hg. In the cortex a slight decrease was noted for injection of CM, but this was significant only after injection of iotrolan. All tested contrast media decrease PO2 in the outer renal medulla, which may partly explain contrast medium-induced acute renal failure.     

Effects of iodinated contrast media on pulmonary airway resistance in anesthetized guinea pigs

RATIONALE AND OBJECTIVES: Bronchospasm is occasionally observed following iodinated X-ray contrast medium administration. We performed an in vivo study in guinea pigs to investigate the effects of a number of iodinated contrast media on pulmonary airway resistance and the mechanisms underlying the potential bronchoconstrictor effect. METHODS: The contrast media studied were the pharmaceutical formulations of iomeprol (400 mg I/ml), iopamidol (370 mg I/ml), and iohexol (350 mg I/ml), which are nonionic, triiodinated contrast media; diatrizoate (370 mg I/ml), an ionic, triiodinated contrast medium; iotrolan (300 mg I/ml), a nonionic, hexaiodinated contrast medium; and iocarmate (280 mg I/ml) and ioxaglate (320 mg I/ml), which are both hexaiodinated and ionic contrast media. Each contrast medium was administered intravenously at 2 g I/kg. Changes in pulmonary airway resistance were evaluated by measuring intratracheal pressure at the moment of maximum insufflation, or maximal insufflation pressure (MIP), in anesthetized guinea pigs submitted to forced ventilation. RESULTS: All contrast media except ioxaglate caused mean increases of MIP of no more than 20%. By contrast, ioxaglate caused a marked bronchoconstrictor effect, increasing MIP by 242% +/- 46%. Of the drugs tested for antagonistic action on this increase in MIP, salbutamol inhibited almost completely the increase in MIP for the first 40 min posttreatment. Similarly, lysine acetylsalicylate and indomethacin consistently reduced MIP after contrast media administration to levels only 30% and 14% above those of baseline precontrast media, respectively. Promethazine had only a minor inhibitory effect, and the response to prednisolone varied. CONCLUSION: There was no apparent relationship between the size of the increase in airway resistance and the charge or molecular weight of the contrast agent molecule or the pharmaceutical formulation. The increase induced by ioxaglate must be attributed to inherent molecular toxicity mediated through a direct action on the production of bradykinin and/or the prostanoid products of the cyclooxygenase pathway, rather than through a direct action on the release of histamine.     

Selective coronary artery perfusion in vitro: a method to study cardiac effects of contrast media

RATIONALE AND OBJECTIVES: The authors evaluated selective perfusion of the coronary arteries in the isolated rat heart as a model for studying contrast medium-induced cardiac effects and compared the effects of iodixanol, iotrolan, and ioxaglate with this model. MATERIALS AND METHODS: Isolated, spontaneously beating rat hearts were used. Control hearts were perfused in the Langendorff or the selective perfusion mode receiving Krebs Henseleit buffer. Contrast media were injected selectively into the left coronary artery. Left ventricular pressure and electrocardiographic parameters were monitored continuously throughout the experiments. RESULTS: The stability of the selective perfusion preparation was similar to that of the conventional Langendorff preparation. Ioxaglate (0.3 g iodine per kilogram body weight) significantly (P < .05) depressed left ventricular contractility and decreased (P < .05) left ventricular pressure. Iodixanol and iotrolan had minor cardiac effects. CONCLUSION: Selective coronary artery perfusion seems to be a suitable model for studying direct cardiac effects of contrast media. The nonionic dimers, iodixanol and iotrolan, induce only minor changes in cardiac function.     

A comparison between the platelet activating properties of different contrast media used in radiology and MRI

Previous studies have demonstrated that some intravascular radiographic contrast media (CM) used in angiography, especially non-ionic monomers, may cause platelet activation. This study was designed to elucidate which properties of the CM were responsible for this activity. Platelet activation engendered by CM was studied using flow cytometry to detect platelet degranulation (as CD62 expression) and fibrinogen binding. In order to elucidate the relevant characteristics of the CM responsible, contrast agents of differing structures, properties, formulations and osmolalities were studied; ionic and non-ionic, monomeric and dimeric. Gadolinium chelate MR enhancing agents and saline solutions of differing osmolalities were also investigated. Ionic dimeric sodium meglumine ioxaglate, non-ionic dimeric iodixanol and non-ionic dimeric iotrolan did not produce increased degranulation compared with saline controls. However, all agents produced a mild increase in bound fibrinogen. Experiments using saline solutions demonstrated that these effects are not attributable to the high osmolality of some CM. The broad comparison facilitated by this study shows that previous generalizations regarding platelet activation by CM, based on an ionic-non-ionic division, are not valid. We presume that some chemical structural property of the compounds is responsible and it is of note that the chemically distinct gadolinium chelates, gadolinium DTPA and gadolinium DTPA-BMA, also caused platelet activation to a similar degree. CD62 expression correlated with fibrinogen binding suggesting that at least one common pathway of platelet activation is involved.     

Contrast media-induced renal morphologic lesions in diabetic rats

Diabetes mellitus was induced in rats with streptozotocin and after 3 months the animals (n = 48) received an i.v. injection of 1 or 3 g I/kg in the form of high-osmolar diatrizoate, low-osmolar iopromide or iohexol, or of 0.6 g I/kg of high-osmolar Gd-DTPA. The controls were given an i.v. injection of physiologic saline. After 2 hours the kidneys were fixed by perfusion and the renal morphologic changes were semiquantitatively analyzed by two independent observers unaware of the agent administered. The contrast media (CM) induced pronounced cytoplasmic vacuolization in the proximal convoluted tubular cells. Such a lysosomal alteration may indicate CM uptake into the cell, and the ultrastructural evaluation revealed intracellular injuries related to the process. The alterations were most marked following administration of iohexol, but diatrizoate also induced a statistically highly significant vacuolization (p < 0.001). The lysosomal alterations following iopromide administration were not as striking, and Gd-DTPA induced only minor changes.     

Liver toxicity of diatrizoate and a non-ionic contrast medium (C29). Coeliac angiography in the rabbit

Toxic effect on the liver of diatrizoate and a new non-ionic contrast medium (C29) were investigated using coeliac angiography in rabbits. The contrast media were injected in doses of 5 ml/kg rabbit with a concentration of 370 mg I/ml. Serum levels of ALAT, ASAT, ALP and bilirubin did not change significantly after the injections.     

The prevalence of insulin resistance and other cardiovascular disease risk factors in healthy elderly southwestern Nigerians

A derivative of coumarin, 8-nitro-7-hydroxycoumarin (8-NO2-7-OHC), was synthesised, purified and characterised. The cytostatic and cytotoxic nature of this compound was determined using both human and animal cell lines grown in vitro for 96 h in the presence of drug (0-500 microM, equivalent to 0-104 microg/ml). 8-NO2-7-OHC was shown to be cytotoxic to three cell lines, but cytostatic to all cell lines tested. With K562 and HL-60 cells, cell death was found to occur by apoptosis. This cytotoxic effect was found to be irreversible, with cell death continuing to occur following a 96 h recovery period. The cytostatic effects were found to be irreversible in four of the five cell lines tested. 8-NO2-7-OHC demonstrated its cytostatic effects within 24 or 48 h, while its cytotoxic effects appeared more gradually. The IC50 of 8-NO2-7-OHC was 475-880 microM, depending on the cell line tested. It was shown to exert its cytostatic effect through an alteration of cell cycle. It also inhibited DNA synthesis. The toxicity of 8-NO2-7-OHC does not appear to be mediated through the multi-drug resistance (MDR) protein since it caused significant cytostatic and cytotoxic effects to CHrC5 cells, which have an increased expression of this protein. This compound was shown to be non-mutagenic in a standard Ames test, both with or without a mammalian enzyme activation system. The applications and mode of actions of coumarins are discussed.     

Cyclooxygenase-2-selective antagonists do not inhibit growth of colorectal carcinoma cell lines

Cytogenetic effects of two ionic contrast media (CM), Urografin 76% a sodium-meglumine diatrizoate, and Telebrix 38, a sodium-meglumine ioxythalamate, were tested on lymphocytes of patients undergoing brain CT Scan. Both compounds have approximately similar iodine concentrations. Chromosomal aberrations were scored in peripheral lymphocytes obtained from 15 patients undergoing brain CT with either urografin 76% or telebrix 38 before and after examination. Results showed no difference in aberration frequency for patients who underwent brain CT without contrast materials compared to controls. However, injection of CM resulted in a high frequency of chromosomal aberrations which significantly differed from controls (P < 0.05). The effect of urografin 76% appeared to be similar to telebrix 38. Therefore, both CM exhibited clastogenic effects on peripheral lymphocytes in vivo. An increase in chromosomal aberrations due to CM used in this study were similar to that reported for other ionic and non-ionic compounds.     

Head pain associated with sixth-nerve palsy: spontaneous dissection of the internal carotid artery

BACKGROUND: Iodixanol (Visipaque, Nycomed Imaging AS, Oslo, Norway) is a new non-ionic and isotonic X-ray contrast medium. OBJECTIVE: To assess its safety and efficacy for paediatric excretory urography. MATERIALS AND METHODS: A three-centre trial in which 72 patients were randomised into three parallel groups: iodixanol 270 mgI/ml, iodixanol 320 mgI/ml and iohexol 300 mgI/ml (Omnipaque, Nycomed Imaging, Oslo, Norway). Doses ranging from 1 to 3 ml/kg never exceeded 50 ml. Pulse rate and blood pressure were recorded before, during, and after the examination. Adverse events, including injection associated discomfort, were recorded during and up to 24 h after the examination. The diagnostic quality of the urograms was assessed on a four-level scale. RESULTS: No serious adverse event occurred in any of the three groups. One patient who was given iodixanol 270 mgI/ml, three who received iodixanol 320 mgI/ml, and one who received iohexol 300 mgI/ml experienced transient adverse events. More than 80 % of the urograms in all three groups were rated "good" or "excellent". CONCLUSION: Iodixanol, either 270 mgI/ml or 320 mgI/ml, is well tolerated and efficacious for excretory urography in children.     

Diagnostic quality in hysterosalpingography. Comparison between iodixanol and iotrolan

PURPOSE: To compare the diagnostic quality and adverse events of the water-soluble, non-ionic, iso-osmolar, dimeric contrast media iodixanol and iotrolan in hysterosalpingography (HSG). MATERIAL AND METHODS: One hundred and twenty patients participated in a double-blind randomized parallel-group phase-III study. Questionnaires and patient interviews were used. Diagnostic quality was assessed using a visual analogue scale and the quality in specific regions was rated using a categorical scale. RESULTS: Overall diagnostic quality was equal for the two groups. There was no statistically significant difference between the two groups in diagnostic quality in the specific regions of interest. No differences between the contrast media were found with regard to adverse events. CONCLUSION: Both contrast media provided images of high diagnostic quality in HSG.     

Flavopiridol: a cytotoxic flavone that induces cell death in noncycling A549 human lung carcinoma cells

Flavopiridol (NSC 649890, L86-8275), a potent inhibitor of cyclin-dependent kinase 1/p34cdc2 phosphorylation and kinase activity, is currently undergoing Phase I clinical testing as a potential antineoplastic agent. Previous studies have suggested that flavopiridol is cytostatic but not cytotoxic when applied to exponentially growing cells. In the present study, various human tumor cell lines were assayed for trypan blue exclusion and ability to form colonies after exposure to flavopiridol under a variety of growth conditions. When log phase A549 non-small cell lung cancer cells were examined 72 h after the start of a 24-h flavopiridol exposure, as many as 90% of the cells accumulated trypan blue. A 24-h exposure to 250-300 nM resulted in trypan blue uptake in 50% of A549 cells at 72 h and a 50% reduction in colony formation. Similar results were observed in HCT8 ileocecal adenocarcinoma, T98G glioblastoma, MCF-7 breast adenocarcinoma, and HL-60 leukemia cells. With A549 cells, identical results were obtained in actively growing logarithmic phase cells and growth-arrested confluent cells. Treatment with the DNA synthesis inhibitor aphidicolin only minimally affected the cytotoxicity of flavopiridol. In contrast, the RNA synthesis inhibitor 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole or the protein synthesis inhibitor cycloheximide reduced the cytotoxicity of flavopiridol. These results suggest that: (a) flavopiridol is not only cytostatic, but also cytotoxic to a variety of human tumor cell lines; (b) flavopiridol is equally active against cycling and noncycling A549 cells; and (c) RNA and protein synthesis appear to play a role in flavopiridol-induced cytotoxicity.     

The additive effects of glucose and insulin on the proliferation of infragenicular vascular smooth muscle cells

PURPOSE: Peripheral vascular disease involving the infragenicular arterial tree is common in patients with diabetes mellitus (DM). Accelerated proliferation of vascular smooth muscle cells (VSMCs) plays an important role in the development of atherosclerosis. Insulin and glucose stimulate VSMC proliferation and are elevated in patients with non-insulin-dependent DM. We have previously described the mitogenic effect of insulin on VSMCs in vitro; the effects of insulin and glucose separately and in combination on the proliferation of VSMCs grown in serum-free media were studied. METHODS: Human infragenicular VSMCs isolated from diabetic patients with end-stage peripheral vascular disease undergoing below-knee amputation were used. Cells from passages 3 to 5 were grown in serum-free media with varying glucose (0.05%, 0.1%, 0.2%, 0.4%, 0.6%, and 0.8%) and insulin (no added insulin, 100 ng/mL, and 1000 ng/mL) concentrations for 6 days. RESULTS: Insulin stimulated VSMC growth at glucose concentrations more than 0.2% (0.4% glucose with no added insulin resulted in 13,073 +/- 336 cells/mL, 0.4% glucose with 100 ng/mL insulin resulted in 16,536 +/- 1175 cells/mL, 0.4% glucose with 1000 ng/mL insulin resulted in 17,500 +/- 808 cells/mL, 0.6% glucose with no added insulin resulted in 14,167 +/- 1062 cells/mL, 0.6% glucose with 100 ng/mL insulin resulted in 18,984 +/- 1265 cells/mL, 0.6% glucose with 1000 ng/mL insulin resulted in 20,450 +/- 1523 cells/mL, 0.8% glucose with no added insulin resulted in 15, 853 +/- 1650 cells/mL, 0.8% glucose with 1000 ng/mL insulin resulted in 26,302 +/- 1919 cells/mL; P <.05 compared with glucose with no added insulin). Glucose stimulated VSMC proliferation up to a concentration of 0.2% (42% and 117% higher growth at 0.1% and 0.2% glucose, respectively, compared with the baseline, P <.05), regardless of the insulin concentration in the media. The greatest growth (26,302 +/- 1919 cells/mL) occurred in the group with the highest concentration of both insulin (1000 ng/mL) and glucose (0.8% glucose; P <.05). CONCLUSION: Both insulin and glucose stimulate the growth of diabetic infragenicular VSMCs. The mitogenic effects of insulin and glucose are additive and may contribute to the development of atherosclerosis in patients with DM.     

Cytostatic and cytotoxic properties of the marine product bistratene A and analysis of the role of protein kinase C in its mode of action

Bistratene A is a polyether which was isolated from the marine ascidian Lissoclinum bistratum Sluiter. The hypothesis has been tested that the cytostatic effect of bistratene A is mediated by modulation of protein kinase C (PKC). Human-derived A549 lung and MCF-7 breast adenocarcinoma cells are extremely sensitive to growth inhibition induced by activators of PKC. Therefore, the effect of bistratene A on these cell lines was compared with that of the known PKC activator 12-O-tetradecanoylphorbol-13-acetate (TPA). The ability of bistratene A to modulate PKC activity in cellular cytosol was assessed to determine the involvement of PKC in the induction of cytostasis. Bistratene A inhibited the growth of both cell lines and initial seeding density determined its cytostatic potency. IC50 values were between 1.0 and 2.9 nM. Bistratene A also had a profound effect on the colony forming ability of A549 cells, preventing clonal growth at 5 nM. Using the incorporation of [3H]thymidine into cells to assess DNA synthetic activity and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay to define cytotoxicity, the compound was found to have both cytostatic and cytotoxic properties. Bistratene A decomposed by 50% after only 2.8 hr in cell culture medium. TPA induced rapid motility and the formation of a network of branched colonies in both cell lines grown on Matrigel, whereas bistratene A did not cause the same effect. Cell cytosol was analysed for phorbol ester binding sites after treatment with bistratene A or TPA. Incubation with TPA (10 nM) caused a reduction in binding sites to 57% of binding in control cells after 30 min and to 35% after 24 hr. Bistratene A did not cause a significant change in binding sites. Assays of PKC activity in cellular cytosol revealed that bistratene A was unable to activate or inhibit the enzyme at concentrations of up to 10 microM. The results suggest that bistratene A is an exquisitely potent cytostatic agent in the two cell lines studied, but modulation of PKC is not involved in the mode of action by which it elicits this effect.     

Local and systemic effects of water-soluble contrast media and barium in rats with chronic small bowel obstruction

RATIONALE AND OBJECTIVES: The local effects on the small intestine and systemic changes produced by different contrast media in small bowel obstruction, with time courses of 4 days, were evaluated. MATERIALS AND METHODS: Four groups, each with 10 normal rats and another four groups (also each with 10 rats) that had ligation of the terminal ileum (obstructed rats) for 4 days were given 3 mL of barium, meglumine sodium diatrizoate, iohexol, or saline (control animals). Radiographs were taken immediately, 1 and 4 hours after administration of contrast media. Immediately before sacrifice, blood samples were taken to determine the hematocrit (Hct), hemoglobin (Hb), white blood cell count (WBC), red blood cell count (RBC), and serum sodium, and potassium and chloride concentrations. Specimens of small bowel were taken for histologic and morphometric analysis. RESULTS: In obstructed rats, the image quality with iohexol improved on final radiographs despite being diluted in the great intestinal contents. There was an improvement in the serum electrolyte concentrations in the obstructed animals that were given any one of the contrast media, the best improvement being in the iohexol groups. A shortening of the length of epithelial cells when any one of the contrast media was administered was observed, as was an increase in the lymphatic space area in the diatrizoate group in normal rats. In the bowel proximal to the obstruction, the lymphatic space area was increased in the diatrizoate group and the size of the epithelial cells was higher in the diatrizoate and iohexol groups compared to the barium and saline groups. CONCLUSION: Our results suggest that iohexol offers good radiologic efficacy and excellent systemic and local tolerance in small bowel obstruction.     

Aspirin and fraxiparine in the prevention of laser induced thrombosis in the presence of iodinated contrast media

The purpose of this study was to investigate the thromboembolic properties of ionic and nonionic contrast media in rats pretreated with aspirin and/or fraxiparine using an experimental model of laser induced thrombosis in the mesenteric microvessels of 17 groups of five male Wistar rats each. Two ionic (ioxaglate and diatrizoate) and two nonionic contrast media (iopamidol and iohexol), alone or associated with antithrombotic drugs (aspirin and/or fraxiparine) were studied. To evaluate the effects of these substances in this model, the number of laser beams needed to induce platelet thrombus formation, the number of emboli detached from the thrombus and the duration of embolization were quantified. Platelet aggregation induced by ADP, induced hemorrhagic time (IHT) and haemoglobin loss level were also determined. Both contrast media injected at 3 ml/kg caused a significant increase in the number of emboli and the duration of embolization (p<0.05). Pretreatment with aspirin and/or fraxiparine in the presence of ionic contrast media showed antithrombotic activities equal to those obtained when they were tested alone (p<0.05), while in the presence of nonionic contrast media, these drugs only neutralised the prothrombotic effects. There were no differences with the NaCl treated group (p>0.05). The ionic contrast media, and to a lesser extent the nonionic contrast medium: iohexol, inhibited platelet aggregation, while iopamidol behaved as an activator. The antithrombotic drugs tested in this study prevent the prothrombotic activities of contrast media therefore suggesting their use before radiographic procedures.     

Ozone Degradation of Iodinated Pharmaceutical Compounds

This study investigates the aqueous degradation of four iodinated x-ray contrast media (ICM) compounds (diatrizoate, iomeprol, iopromide, and iopamidol) by ozone and combined ozone and hydrogen peroxide. In laboratory scale experiments, second-order kinetic rate constants for the reactions of the ICM compounds with molecular ozone and hydroxyl radicals, and overall at pH 7.5, were determined. For the four ICM compounds the degradation rate constants with molecular ozone were low and in the range of 1–20?M?1?s?1, whereas the rate constants with hydroxyl radicals were in the range of 1×109–3×109?M?1?s?1. Diatrizoate had the lowest rate constant of the four compounds with respect to molecular ozone reactions. At pH 7.5, the extent of compound degradation was proportional to the applied ozone dose and inversely related to the initial compound concentration at a given ozone dose. At this pH approximately 90% of the degradation could be attributed to hydroxyl radical reactions. Enhancement of the radical mechanism by the addition of hydrogen peroxide during ozonation led to complete removal of the nonionic compounds, and >80% removal of diatrizoate, at relatively low oxidant mass ratios (H2O2/O3<0.25). A similar enhancement in compound degradation was evident with the presence of small concentrations of humic substances ( ～ 4–5?mg?L?1). Ozone oxidation led to major cleavage of the ICM compounds and the release of inorganic iodine; the proportion of iodine release was similar among the nonionic ICM compounds but much greater for diatrizoate.     

Bunazosin hydrochloride inhibits exaggerated growth of vascular smooth muscle cells from spontaneously hypertensive rats by suppressing the response to growth factors

Selective alpha1-adrenoreceptor blockers were recently reported to have an in vivo antiproliferative effect on hypertensive cardiovascular organs. Cultured vascular smooth-muscle cells (VSMCs) from spontaneously hypertensive rats (SHRs) show exaggerated growth compared with cells from Wistar-Kyoto (WKY) rats. We investigated the effects of an alpha1-adrenoreceptor blocker, bunazosin hydrochloride (HCl), on the growth of VSMCs from SHRs. In the absence of serum, bunazosin HCl significantly inhibited basal DNA synthesis by VSMCs from SHRs, but not by cells from WKY rats. In the presence of serum, bunazosin HCl significantly inhibited DNA synthesis by VSMCs from both rat strains. Angiotensin (Ang) II, platelet-derived growth factor (PDGF)-AA, and epidermal growth factor (EGF) dose-dependently increased DNA synthesis by VSMCs from SHRs, but not by VSMCs from WKY rats. Bunazosin HCl significantly suppressed the response of DNA synthesis to PDGF-AA and EGF, but not to Ang II, in VSMCs from SHRs. Expression of basic fibroblast growth factor (bFGF), transforming growth factor-beta1 (TGFbeta1), and PDGF messenger RNA (mRNA) was markedly greater in VSMCs from SHRs than in cells from WKY rats. Bunazosin HCl significantly inhibited the expression of bFGF and TGFbeta1 mRNA in VSMCs from SHRs, but not in cells from WKY rats. These findings suggest that the inhibition of growth factor hyperresponsiveness and inhibition of the expression of growth factors in VSMCs from SHRs are associated with the antiproliferative effect of bunazosin.     

The haemodynamic effects of iodinated water soluble radiographic contrast media: a review

All classes of iodinated water-soluble radiographic contrast media (RCM) are vasoactive with the iso-osmolar dimers inducing the least changes in the vascular tone. The mechanisms responsible for RCM-induced changes in the vascular tone are not fully understood and could be multifactorial. A direct effect on the vascular smooth muscle cells causing alterations in the ion exchanges across the cell membrane is thought to be an important factor in RCM-induced vasodilatation. The release of the endogenous vasoactive mediators adenosine and endothelin may also play a crucial role in the haemodynamic effects of RCM particularly in the kidney. In addition, the effects of RCM on blood rheology can cause a reduction in the blood flow in the microcirculation. The purpose of this review is to discuss the pathophysiology of the haemodynamic effects of RCM and to offer some insight into the biology of the endothelium and vascular smooth muscle cells as well as the pharmacology of the important vasoactive mediators endothelin and adenosine.     

Insulin-like growth factor I activates the invasion suppressor function of E-cadherin in MCF-7 human mammary carcinoma cells in vitro

The calcium-dependent cell-cell adhesion molecule E-cadherin has been shown to counteract invasion of epithelial neoplastic cells. Using three monoclonal antibodies, we have demonstrated the presence of E-cadherin at the surface of human MCF-7/6 mammary carcinoma cells by indirect immunofluorescence coupled to flow cytometry and by immunocytochemistry. Nevertheless, MCF-7/6 cells failed to aggregate in a medium containing 1.25 mM CaCl2, and they were invasive after confrontation with embryonic chick heart fragments in organ culture. Treatment of MCF-7/6 cells with 0.5 microgram ml-1 insulin-like growth factor I (IGF-I) led to homotypic aggregation within 5 to 10 min and inhibited invasion in vitro during at least 8 days. The effect of IGF-I on cellular aggregation was insensitive to cycloheximide. However, monoclonal antibodies that interfered with the function of either the IGF-I receptor (alpha IR3) or E-cadherin (HECD-1, MB2) blocked the effect of IGF-I on aggregation. The effects of IGF-I on aggregation and on invasion could be mimicked by 1 microgram ml-1 insulin, but not by 0.5 microgram ml-1 IGF-II. The insulin effects were presumably not mediated by the IGF-I receptor, since they could not be blocked by an antibody against this receptor (alpha IR3). Our results indicate that IGF-I activates the invasion suppressor role of E-cadherin in MCF-7/6 cells.     

Gallium-67 radiotoxicity in human U937 lymphoma cells

Promising clinical results have been obtained with radiolabeled antibodies in lymphoma patients. The higher uptake by lymphomas of 67Gallium (67Ga) compared with monoclonal antibodies makes selective radiotherapy by the widely available 67Ga appealing. However, the gamma radiation of 67Ga used in scintigraphy is considered to be almost non-toxic to lymphoma cells. However, in addition to photon radiation 67Ga emits low energy Auger electrons and 80-90 keV conversion electrons which could be cytotoxic. The objective of the present study was the assessment of radiotoxicity of 67Ga on a lymphoid cell line: U937. Proliferation (MTT-assay) and clonogenic capacity (CFU-assay) were measured after 3 and 6 days incubation with 10, 20 and 40 microCi ml-1 67Ga. Growth inhibition was 36% after 3 days incubation and 63% after 6 days incubation with 40 microCi 67Ga ml-1. Clonogenic capacity was reduced by 51% after 3 days and 72% after 6 days incubation with 40 microCi ml-1 67Ga. A survival curve showed an initial shoulder and became steeper beyond 200-250 pCi cell-1 (low linear energy transfer type). Iso-effect doses of 67Ga and 90Yttrium (90Y) were determined. The iso-effect dose of 40 microCi 67Ga ml-1 (cumulative dose of conversion electrons 306 cGy) was 2.5 microCi 90Y ml-1 (cumulative dose 494 cGy) and the iso-effect dose of 80 microCi 67Ga ml-1 was 5.0 microCi 90Y/ml. The main cytotoxic effect of 67Ga seems to be induced by the 80 keV conversion electrons. We conclude that the conversion electrons of 67Ga have a cytotoxic effect on U937 cells and that in our experiments a 16-fold higher microCi-dose of 67Ga than of 90Y was needed for the same cytotoxic effect. We believe that 67Ga holds promise for therapeutic use.