Distribution of organotin compounds in the bivalves of the Aegean Sea, Greece.

Five bivalve species--Mytilus galloprovinciallis (Mediterranean mussels), Venus gallina (stripped venus), Modiola barbatus L. (bearded horse mussels), Pecten jacobeus (scallops) and Callista chione (hard clams)--were collected from seven areas in Aegean Sea, Greece, between August 2001 and January 2003 and analyzed for organotins (OTs). The concentrations (as geometric means) found were 17.1 ng g-1 for tributyltin (TBT), 18.8 ng g-1 for dibutytltin (DBT), 7.8 ng g-1 for monobutyltin (MBT) and 13.0 ng g-1 for triphenyltin (TPhT) (wet weight), which are at similar or lower levels than those reported worldwide. Studying OTs distribution between different bivalve species, lower concentrations were observed in mediterranean mussels, possibly due to their growth in water column (grown on sea net pens in mussel farms), in contrast to the free-ranging species, collected from fishing grounds. Concentrations of the OTs in the examined bivalves varied seasonally.

Improved glucuronide hydrolysis in the determination of urinary 1-hydroxypyrene.

The efficiency of the glucuronide hydrolysis in the determination of urinary 1-hydroxypyrene was investigated as a function of the reaction conditions. A significant improvement could be obtained by increasing the enzyme concentration described in the literature.

Barnidipine block of L-type Ca(2+) channel currents in rat ventricular cardiomyocytes.

The effects of barnidipine and nifedipine on L-type Ca(2+) current (I(Ca(L))) were investigated in ventricular cardiomyocytes from rats. Both barnidipine and nifedipine reduced I(Ca(L)) in a concentration and voltage dependent manner; the EC(50) were 80 and 130 nM at a holding potential of -80 mV, respectively, and 18 and 6 nM at -40 mV, respectively. Both drugs induced a leftward shift of the steady-state inactivation curve of I(Ca(L)). Using a twin pulse protocol, the relationships between the amount of block of I(Ca(L)) by either drug, seen during the second pulse, and the length of the first pulse were described by monoexponential functions reflecting onset of block, dependent on drug concentration. The onset of block by barnidipine was three times faster than that by nifedipine. With both drugs, recovery of I(Ca(L)) was 50 times slower than under control conditions and described by monoexponential functions reflecting offset of block (independent of drug concentration). The offset of block with barnidipine was three times slower than that with nifedipine. The time constants of block and unblock of I(Ca(L)) by both drugs were used to calculate binding and unbinding and to predict their effects at two frequencies. It is suggested that barnidipine exhibits a higher affinity to the inactivated Ca(2+) channel state as compared to nifedipine.

Effects of nitric oxide donors on cardiac contractility in wild-type and myoglobin-deficient mice.

1. The effects of the nitric oxide (NO) donors S-nitroso-N-acetylpenicillamine (SNAP), sodium(Z)-1-(N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA-NONOate), and (Z)-1-[N-(2-Aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA-NONOate) on force of contraction (F(c)) were studied in atrial and ventricular muscle strips obtained from wild-type (WT) and myoglobin-deficient (myo(-/-)) mice. 2. SNAP slightly reduced F(c) in preparations from WT mice at concentrations above 100 microM; this effect was more pronounced in myo(-/-) mice. 3. DEA-NONOate reduced F(c) in preparations from myo(-/-) mice to a larger extent than those from WT mice. 4. DETA-NONOate reduced F(c) in preparations from myo(-/-) but not from WT mice. 5. Pre-incubation with an inhibitor of the soluble guanylyl cyclase (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one; 100 microM) prevented the effects of SNAP, DEA-NONOate and DETA-NONOate on F(c) in myo(-/-) mice. 6. It is suggested that, in physiological conditions, myoglobin acts as intracellular scavenger preventing NO from reaching its intracellular receptors in cardiomyocytes, whereas, in myoglobin-deficient conditions, NO is able to reduce contractility via activation of the soluble guanylyl cyclase/cyclic GMP pathway.

Activation of soluble guanylyl cyclase by YC-1 in aortic smooth muscle but not in ventricular myocardium from rat.

1. The effects of YC-1 (3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole), an activator of soluble guanylyl cyclase, on tension, levels of cyclic GMP and cyclic AMP, and cardiac L-type Ca2+-current (I[Ca(L)]) were investigated in aortic smooth muscle and ventricular heart muscle from rat. 2. YC-1 (0.1-30 microM) induced a concentration-dependent relaxation in aortic rings precontracted with phenylephrine (3 microM). The relaxant effects of YC-1 were reversed by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (30 microM; ODQ), potentiated by zaprinast (10 microM) and antagonized by Rp-8-Br-cGMPS (100 microM). 3. In ventricular heart muscle strips, YC-1 (30 microM) exhibited no effects on force of contraction (Fc) in the absence or presence of either zaprinast (10 microM) or 3-isobutyl-1-methylxanthine (30 microM). Fc was slightly increased by YC-1 (30 microM) in the presence of isoprenaline (100 nM), but this effect was not influenced by ODQ (30 microM). 4. Cardiac I[Ca(L)] was not significantly affected by YC-1 (30 microM), either in the absence or presence of isoprenaline (30 nM). 5. In aortic rings, cyclic GMP levels were increased almost 3 fold by YC-1 (30 microM); this effect was abolished by ODQ (30 microM). In isolated ventricular cardiomyocytes, cyclic GMP levels were not affected by YC-1 (30 microM) but almost doubled by activation of particular guanylyl cyclase with atriopeptin II (100 nM). 6. YC-1 (30 microM) did not increase cyclic AMP levels either in aortic rings or in ventricular cardiomyocytes. In contrast, isoprenaline (3 microM) increased cyclic AMP levels about two fold in both tissues. In cardiomyocytes, the effect of isoprenaline (3 microM) was slightly enhanced by YC-1 (30 microM). 7. It is concluded that relaxation of smooth muscle preparations by YC-1 is mediated mainly by activation of soluble guanylyl cyclase and subsequent increase in cyclic GMP levels. The failure of YC-1 to affect cardiac Fc, levels of cyclic GMP, and I[Ca(L)] suggests that soluble guanylyl cyclase is not influenced by YC-1 in rat heart muscle or only barely present in this tissue.

Failure of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) to inhibit soluble guanylyl cyclase in rat ventricular cardiomyocytes.

1. The effects of 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), an inhibitor of soluble guanylyl cyclase (sGC), were investigated in aortic rings and ventricular cardiomyocytes from rats. The production of cyclic GMP was stimulated by NO.-donors or carbachol. Additionally, the effects of ODQ were studied in cytosolic extracts from both tissues in which the cyclic GMP production was stimulated by S-nitroso-N-acetylpenicillamine (SNAP). 2. In endothelium-intact aortic rings, SNAP (100 microM), 2,2'-(hydroxynitrosohydrazino)bis-ethana-mine (DETA NONOate; 100 microM), or carbachol (10 microM) increased cyclic GMP levels about 4 fold. These effects were abolished by ODQ (50 microM). 3. In cardiomyocytes, SNAP (100 microM), DETA NONOate (100 microM), or carbachol (10 microM) increased cyclic GMP levels about 2 fold. These effects were not affected by ODQ (50 microM). 4. In cytosolic extracts from aortic rings and cardiomyocytes, SNAP (100 microM) induced about 50 fold increases in cyclic GMP levels. ODQ (50 microM) reduced these effects by about 50%. 5. In extracts from cardiomyocytes, increases by SNAP (100 microM) of cyclic GMP levels were attenuated by myoglobin dependent on concentration: at 300 microM myoglobin, SNAP (100 microM) increased cyclic GMP levels only 3 fold. Inhibitory effects of ODQ (50 microM) were abolished by 300 microM myoglobin. 6. It is suggested that both NO. and ODQ can bind to myoglobin which, at high concentrations. can diminish their effects on sGC. Such a scavenger function of myoglobin could explain why NO. and ODQ exert only minor effects in cardiomyocytes (with high myoglobin content) but strong effects in aortic tissue (virtually devoid of myoglobin).

Differential effects of isoliquiritigenin and YC-1 in rat aortic smooth muscle.

We investigated the effects of isoliquiritigenin and YC-1 (3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole) on tension in endothelial-free rat aortic rings precontracted with phenylephrine (3 microM). Both compounds induced a concentration-dependent relaxation (EC50 of YC-1 1.9 microM and of isoliquiritigenin 9.4 microM). The effects developed faster with YC-1 than with isoliquiritigenin, and the effects of YC-1 were potentiated by isoliquiritigenin (10 microM). 1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one (30 microM) inhibited the effect of YC-1, but not of isoliquiritigenin. These results suggest that the effects of YC-1 are due to stimulation of soluble guanylyl cyclase activity, whereas the effects of isoliquiritigenin are rather related to inhibition of phosphodiesterase activity.

Cardiac effects of isoliquiritigenin.

The effects of isoliquiritigenin on force of contraction (Fc), L-type Ca2+ current (I(Ca)) and intracellular Ca2+ concentration ([Ca2+]i) were investigated in rat ventricular heart muscle. Isoliquiritigenin increased Fc and I(Ca) and, after longer exposure times, resting tension and [Ca2+]i. The effect of isoliquiritigenin (100 microM) on I(Ca) was diminished by Rp-cAMPS (30 microM). 1H-[1,2,4]oxa- diazolo[4,3-a]quinoxalin-1-one (50 microM) did not influence the effects of isoliquiritigenin on Fc and I(Ca). The positive inotropic effects of isoprenaline and forskolin, but not of 3-isobutyl-1-methylxanthine, were potentiated by isoliquiritigenin (100 microM). In the presence of milrinone (10 microM), no further effects of isoliquiritigenin (100 microM) on Fc and I(Ca) were observed. It is suggested that the increase in Fc and I(Ca) by isoliquiritigenin is due to an accumulation of cyclic AMP. These effects are probably unrelated to an effect of the drug on soluble guanylyl cyclase, as reported for smooth muscle, but rather due to a direct inhibition of phosphodiesterase III activity.

Alkalinisation does not modify the effect of verapamil on myocardial Ca2+ current.

The effect of verapamil on L-type Ca2+ current (ICa) was compared at external pH 7.4 and 8.5 in rat ventricular myocytes. Alkalinisation increases the fraction of uncharged molecules of verapamil (pK 8.75) and thereby facilitates membrane permeation of the drug. Verapamil (1 microM) reduced the amplitude of ICa (ICa(peak)) by 36 +/- 4% at pH 7.4 and by 40 +/- 6% at pH 8.5, whereas alkalinisation from pH 7.4 to 8.5, without drug, increased ICa(peak) by 12 +/- 3%. It is suggested that the efficiency of verapamil is not influenced by the amount of protonation or membrane permeation.

Differences in the nitric oxide/soluble guanylyl cyclase signalling pathway in the myocardium of neonatal and adult rats.

The effects of a nitric oxide-donor, S-nitroso-N-acetylpenicillamine, and a direct activator of soluble guanylyl cyclase, 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1), on force of contraction (F(c)) and L-type Ca(2+) currents (I(Ca(L))) were investigated in myocardial preparations from neonatal and adult rats. Since hearts from adult and neonatal animals contained 160 and 47 mg/100 g wet weight myoglobin, respectively, its possible interaction with both drugs was also investigated. Both S-nitroso-N-acetylpenicillamine (100 microM) and YC-1 (30 microM) were ineffective in myocardial preparations from adult rats but reduced the magnitude of I(Ca(L)) and F(c) in preparations from neonatal rats. The latter effects were antagonised by 1H-[1,2, 4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ; 50 microM) and attenuated by myoglobin (30-300 microM), which also attenuated the effects of both drugs on pre-contracted aortic rings. The differential effects of S-nitroso-N-acetylpenicillamine and YC-1 in the myocardium from adult and neonatal rats may result from developmental changes in the content of myoglobin and/or in the NO/soluble guanylyl cyclase signal pathway.

Involvement of K(+) channels in the relaxant effects of YC-1 in vascular smooth muscle.

This study addresses the question whether K(+) channels are involved in the vasorelaxant effects of 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl-indazole (YC-1 ). In rat aorta, guinea pig aorta, and guinea pig a. carotis, YC-1 inhibited contractions induced by phenylephrine (3 microM) more potently than those induced by K(+)(48 mM). In rat aorta, tetraethylammonium (10 mM), charybdotoxin (0.2 microM), and iberiotoxin (0.1 microM), but not glibenclamide (10 microM), attenuated the relaxant effects of YC-1. In guinea pig a. carotis, YC-1 (30 microM) induced a hyperpolarisation which was antagonised by 1H-[1,2,4]oxadiazolo[4, 3-a]quinoxalin-1-one (ODQ; 50 microM). In rat aorta, YC-1 (30 microM) increased the rate constant of 86Rb-efflux. The effect of YC-1 was potentiated by zaprinast (10 microM), but inhibited by ODQ (50 microM) or charybdotoxin (0.2 microM). In smooth muscle cells from rat aorta, YC-1 (10 microM) increased BK(Ca) channel activity. It is suggested that YC-1-induced vasorelaxation is partially mediated by the activation of K(+) channels.

Kinetics and state-dependent effects of verapamil on cardiac L-type calcium channels.

The voltage dependence and the kinetics of block by verapamil of L-type calcium current (ICa) were investigated in ventricular myocytes from rat hearts using the whole-cell patch-clamp technique. ICa was elicited repetitively in response to depolarizing voltage pulses from -80 mV to 0 mV at different pulse intervals and durations. Verapamil reduced the magnitude of ICa in a frequency-dependent manner without tonic component. The time course of ICa remained unchanged suggesting that not open but inactivated channels were affected by the drug. The interaction of verapamil with inactivated channels was investigated by the application of twin pulses. In the presence of verapamil, the duration of the first pulse significantly determined the magnitude of ICa during the second pulse. Variation of the duration of the first pulse between 12 and 3000 ms, followed by a pulse interval of 100 ms, resulted into a gradual decrease of ICa during the second pulse (180 ms), described by concentration-dependent monoexponential decay curves (tau = 1060 +/- 138 ms at 0.3 microM (n = 3); tau = 310 +/- 24 ms at 1 microM (n = 6), and tau = 125 +/- 7 ms at 10 microM (n = 5); means +/- SEM). Under control conditions, the changes in ICa were comparably negligible. The recovery of ICa from block was analyzed by the application of a twin pulse protocol in which two depolarising voltage pulses at fixed length (1. pulse at 3 s and 2. pulse at 180 ms) were interrupted by variable pulse intervals (6 ms-60 s). Under control conditions, recovery from inactivation was fast (tau = 11 +/- 0.7 ms; means +/- SEM; n = 3). In the presence of verapamil, recovery from block was about 500 times slower than under control conditions, independent of the drug concentration (tau = 5.05 +/- 0.44 s at 0.3 microM (n = 3), tau = 6.7 +/- 0.69 s at 1 microM (n = 4), and tau = 6.02 +/- 0.9 s at 10 microM (n = 5); means +/- SEM). Since development of block was dependent on the concentration of verapamil, whereas recovery from block was independent from the drug concentration, it is assumed that the described time constants for block and unblock reflect voltage-dependent net binding (tau on) and unbinding (tau off), respectively, of verapamil at its receptor sites. A computer simulation, including the time constants of block development at 0 mV and of recovery from block at -80 mV, predicted reasonably well the observed frequency-dependent block of ICa by verapamil. The development of either measured or calculated block of ICa, using 180 ms depolarising voltage pulses from -80 mV to 0 mV, was fitted by identical monoexponential association curves (tau = 7 s each at 0.2 Hz and tau = 1.7 s each at 1 Hz). When Ba2+ was used as the charge carrier, which removes the calcium-dependent inactivation of the current, verapamil (3 microM) was less efficient: ICa was decreased by 57 +/- 6% (means +/- SEM; n = 6), whereas IBa was decreased by 24 +/- 4% (means +/- SEM; n = 5). It is proposed that verapamil binds to calcium channels in their inactivated state at more positive potentials and dissociates from the channels in the resting state at more negative potentials. In the proposed scheme of periodical drug binding and unbinding, dependent on the state of the channels, the development of frequency-dependent block of ICa by verapamil is adequately predicted by the construction of cumulative association/ dissociation curves which include the experimentally determined time constants of development and recovery from block at 0 mV and -80 mV, respectively.

Action of tertiary phenylalkylamines on cardiac transient outward current from outside the cell membrane.

The effects of the phenylalkylamines verapamil (V), gallopamil (G), and devapamil (D) and their corresponding quaternary derivatives on the transient outward current (Ito) were examined in rat ventricular cardiomyocytes using the whole-cell patch-clamp technique. The question was addressed, whether phenylalkylamines act on Ito from the inside or the outside or from both sides of the cell membrane. To this end, the myocytes were either superfused extracellularly or perfused intracellularly with drug-containing solutions. In addition, the effects of verapamil were investigated at different pH-values. V, G, and D (30 microM each), applied extracellularly, reduced the steady state current of Ito, Ito(150 ms), to 34 +/- 3.3, 33 +/- 6, and 30 +/- 5, respectively (% of control; means +/- SEM). The effects of V (30 microM) on Ito were similar at various external pH-values (reduction of Ito(150 ms) by 69 +/- 6 at pH 6.5, by 66 +/- 4 at pH 7.4, by 68 +/- 8 at pH 8.5, and by 58 +/- 10 at pH 9.5; % of control; means +/- SEM). In contrast, the effect of 4-aminopyridine (300 microM) on Ito was enhanced after alkalinisation: the peak current of Ito was reduced to 49 +/- 5 at pH 7.4 and to 5 +/- 2 at pH 9.2 (% of control; means +/- SEM). V, G, and D (300 microM) failed to produce any effect on Ito, when applied intracellularly (values of Ito(150 ms): 97 +/- 6, 105 +/- 4, and 94 +/- 4, respectively; % of control; means +/- SEM). In contrast, 4-aminopyridine (3 mM) depressed the peak current of Ito to 69 +/- 6% of control (mean +/- SEM), when applied intracellularly. The permanently charged quaternary derivatives of the phenylalkylamines q-V, q-G, and q-D (300 microM) did not significantly affect Ito, when applied extracellularly (values of Ito(150 ms): 94 +/- 2, 90 +/- 3, and 94 +/- 3, respectively; % of control; means +/- SEM) but diminished Ito, when applied intracellularly (reduction of Ito(150 ms) to 43 +/- 5, 56 +/- 7, and 63 +/- 4, respectively; % of control; means +/- SEM). Intracellularly applied V (300 microM) did not reduce Ito at pH 6.5 at which V is protonated to 99.4%. It is suggested that tertiary phenylalkylamines act on Ito by binding to a membrane site accessible from the outside, whereas their quaternary derivatives affect Ito by binding to a membrane site located at the inside of the cell membrane. In contrast, 4-aminopyridine is supposed to act on Ito from the inside of the cell membrane.

Extracellular site of action of phenylalkylamines on L-type calcium current in rat ventricular myocytes.

The effects of the phenylalkylamines verapamil, gallopamil, and devapamil on L-type calcium currents (ICa) were studied in ventricular myocytes from rat hearts using the whole-cell patch-clamp technique. In particular, the question was addressed, whether the pharmacological binding sites for these drugs were located at the inner and/or at the outer surface of the cell membrane. Therefore, tertiary verapamil, gallopamil, and devapamil and their corresponding quaternary derivatives were applied either from the outside or the inside of the cell membrane. Extracellular application of verapamil, gallopamil and devapamil (each at 3 microM) reduced ICa to 16.1 +/- 8.6%, 11 +/- 8.9%, and 9.3 +/- 6% of control, respectively. Intracellular application of the same substances, via the patch pipette filled with 30 microM of either verapamil, gallopamil, or devapamil, failed to depress ICa. The quaternary derivatives of the phenylalkylamines (30 microM) were ineffective both when applied extracellularly or intracellularly. It is suggested that phenylalkylamines block ICa in ventricular myocytes by acting on a binding site of the calcium channel molecule located at the outer surface of the cell membrane.

Active biomonitoring of polycyclic aromatic hydrocarbons by means of mosses.

Spherical bags, packed with 20 g of peat moss (Sphagnum spp.), were exposed to ambient air at a distance of 1 km from a plant manufacturing electrodes for the production of aluminium, near Rotterdam, The Netherlands. In these bags, the concentrations of six polycyclic aromatic hydrocarbons were determined, and compared with the concentrations in moss bags that had been exposed in relatively clean areas. From the results it can be concluded that, in addition to their useful application for biomonitoring of heavy metals, mosses can be applied in active biomonitoring of polycyclic aromatic hydrocarbons in ambient air.

Retention of neutral and basic heteroaromatic hydrocarbons in RPLC systems and its use in predictive studies. I. Concentration of the organic modifier.

The retention of polynuclear heteroaromatic hydrocarbons in reversed-phase liquid chromatographic systems was studied using methanol-water or acetonitrile-water as the mobile phase. The relation between the concentration of the modifier and the capacity factor was investigated, and capacity factors in pure water were obtained by means of extrapolation of this relation and were used in quantitative chromatographic retention-activity relationships. It was shown that the general equation for the relationship between the n-octanol/water partition coefficient (log Kow) and retention of apolar compounds in octadecyl-silica/methanol-water systems, first presented by Tomlinson and Hafkenscheid, is valid for neutral heteroaromatic hydrocarbons. For basic heteroaromatic compounds, this relationship is not valid. Both log Kow and the logarithm of the bioconcentration factor in the guppy (Poecilia reticulata) of neutral heteroaromatic hydrocarbons can be predicted satisfactorily from capacity factors extrapolated to pure water.

Prediction of environmental fate and effects of heteroatomic polycyclic aromatics by QSARs: the position of n-octanol/water partition coefficients.

The HPLC and TLC retention, n-octanol/water partition coefficients (log Kow), bioconcentration factors, and acute toxicity data of 29 heteroatomic polycyclic aromatic hydrocarbons and 7 parent polycyclic aromatics were determined experimentally. For the same set of compounds, molecular weights, fragmental log Kow values, and molecular connectivities were calculated. Quantitation of the mathematical relationships between the variables was used to validate the predictive potential of various parameters. The importance of log Kow in predictive studies is highlighted. It is concluded that the internal concentration of a pollutant in the organism should be used as a parameter in future QSAR work.

Increased cardiac alpha-1-adrenoceptor density in rats following treatment with amiodarone.

This study was undertaken to investigate the interaction between amiodarone and alpha-1-adrenoceptors in rat cardiac cells. The level (Bmax) and affinity (Kd) of alpha-1-adrenoceptors in heart cells were determined by [3H]prazosin radioligand binding following amiodarone treatment. In cultured intact cardiocytes treated for 48 h with 10 microM amiodarone, [3H]prazosin binding increased by 31% compared with the control cells (p<0.05). The increase was both dose and time dependent and was found to be specific because no significant change occurred in creatine kinase activity. Additionally, under the same conditions, an increase in [3H]prazosin binding to cultured cardiocyte cell membranes was also obtained. Oral gavage of amiodarone to rats for 8 d resulted in a 25% increase in [3H]prazosin binding to isolated ventricle membranes compared with control rats (p<0.05). We conclude that amiodarone treatment can increase the response to alpha-1-adrenoceptors agonist in the heart due to an increase in the density of alpha-1-adrenoceptors.

United Nations Environment Programme Capacity Building Pilot Project--training and interlaboratory study on persistent organic pollutant analysis under the Stockholm Convention.

Within the framework of a United Nations Environment Programme (UNEP) Capacity Building Project for training of laboratory staff in developing countries on persistent organic pollutant (POP) analysis, an interlaboratory study was organised following an initial evaluation of the performance of laboratories (reality check) and a series of training sessions. The target compounds were polychlorinated biphenyls (PCB) and organochlorine pesticides (OCP). Seven laboratories from five countries (Ecuador, Uruguay, Kenya, Moldova, and Fiji) participated. Most of the laboratories had no experience in determining PCBs. Although chromatograms improved considerably after the training and installation of new gas chromatographic (GC) columns at participating laboratories, the level of performance in the interlaboratory study was essentially on par with the moderate performance level achieved by European POP laboratories in the 1980s. Only some individual results were within +/-20% of the target values. The relative standard deviations (R.S.D.s) in POP concentrations determined by laboratories in a sediment sample were >200% in a number of cases. The results for a certified herring sample were better with at least some R.S.D. values below 50% and most below 100%. Clean up was as one of the main sources of error. After inspection it was ascertained that training of laboratory staff and investments in simple consumables such as glassware and GC columns would help to improve the quality of the analysis more than major investments in expensive instrumentation. Creating an effective network of POP laboratories at different continents together with a series of interlaboratory studies and workshops is suggested to improve the measurements of POPs in these countries.

Inhibition of L-type Cav1.2 Ca2+ channels by 2,(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and 2-[1-(3-dimethyl-aminopropyl)-5-methoxyindol-3-yl]-3-(1H-indol-3-yl) maleimide (Go6983).

Phosphatidylinositol 3-kinase (PI3-K) is involved in physiological processes of cellular proliferation and inflammation and, as postulated recently, in the regulation of L-type Ca(2+) channels. The latter conclusion arose in part from the inhibitory action of the compound 2,(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002), which has been established as a selective PI3-K inhibitor (IC(50) = 1.4 microM). Herein we show, however, that LY294002 and an inhibitor of protein kinase C (PKC), 2-[1-(3-dimethylaminopropyl)-5-methoxyindol-3-yl]-3-(1H-indol-3-yl) maleimide (Go6983), act as direct Ca(2+)-channel inhibitors, with IC(50) values of approximately 20 and 10 microM, respectively. Because both drugs are commonly used at concentrations of approximately 10 microM or higher, the interpretation of such experiments is questionable with respect to a regulatory action of PI3-K or PKC on L-type Ca(2+) channels.