The results of EuReCa ONE highlight that OHCA is still a major public health problem accounting for a substantial number of deaths in Europe. EuReCa ONE very clearly demonstrates marked differences in the processes for data collection and reported outcomes following OHCA all over Europe. Using these data and analyses, different countries, regions, systems, and concepts can benchmark themselves and may learn from each other to further improve survival following one of our major health care events.

Staphylococcal skin colonization is a common feature of atopic dermatitis (AD) in adults. Little is known about prevalence and persistence of staphylococci in children. Forty-one AD children (mean age, 70 months) and 41 age-matched controls were studied. S. aureus was isolated from 38 AD patients (93%; 32% of controls, P less than .001) and 37% of AD patients (5% of controls, P less than .001) harbored toxigenic (enterotoxins, toxic shock syndrome toxin) S. aureus strains. No individual biotype prevailed. On follow-up (mean interval, 9 months), 70% of S. aureus strains were reisolated. Nasal and cutaneous S. aureus strains were identical in 73% of AD patients (7% of controls, P less than .001), reflecting increased self-contamination. Identical staphylococcal strains in AD children and their mothers were observed in 38% (S. aureus) and 16% (coagulase-negative strains; P less than .001). The prevalence of staphylococcal colonization in AD children is comparable to that in adults. High rates of self-contamination, transmission to contacts, and prevalence of toxigenic strains in AD children may have clinical and epidemiologic implications.

In order to investigate whether coupling to and/or activation of guanine-nucleotide-binding proteins (G proteins) is involved in agonist-induced internalization of m4 muscarinic acetylcholine receptors (mAChRs), a deletion mutant [des-(264-394)mAChRl was constructed that lacks a substantial portion of the putative third intracellular loop. The wild-type receptor and des-(264 -394)mAChR stably expressed in Chinese hamster ovary cells in essentially comparable amounts, exhibited identical antagonist-binding affinities. Consistent with the reported importance of the third cytoplasmic loop for G, protein activation, the des-(264-394)mAChR showed a drastically reduced potential to mediate agonist-induced inhibition of adenylyl cyclase. In contrast, the ability of the mutant receptor to couple to GI proteins was not impaired, as demonstrated by a similar guaninenucleotide-sensitive and pertussis-toxin-sensitive high-affinity agonist-receptor binding for both mAChRs. In contrast, des-(264-394)mAChR was hardly able to stimulate the GTPase activity of G proteins, suggesting impaired activation of G, proteins rather than ineffective coupling to GI proteins. Internalization of wild-type receptor and des-(264 -394)mAChR was observed with similar agonist concentrations and showed similar maximal values. However, des-(264-394)mAChR displayed a significantly reduced rate of receptor internalization. A similar attenuation of wild-type mAChR internalization was obtained upon pertussis toxin treatment. In conclusion, our data provide evidence that the molecular determinants of the m4 mAChR involved in G,-protein coupling and activation are not identical and that activation of, but not coupling to, G, proteins regulates m4 mAChR internalization.Muscarinic acetylcholine receptors (mAChRs) belong to the superfamily of seven transmembrane receptors which activate signal-transduction pathways through their interaction with guanine-nucleotide-binding regulatory proteins (G proteins). The mAChR family consists of five receptors of which the ml, m3 and m5 receptor subtypes are efficiently coupled to stimulation of phosphoinositide-hydrolyzing phospholipase C via pertussis toxin (PTx)-insensitive G, proteins, and m2 and m4 receptors which are preferentially coupled to the inhibition of adenylyl cyclase and to other effectors via PTxsensitive Gi and Go proteins [l]. Studies involving mutations of the ml, m2 and m3 mAChRs have led to the conlusion that third cytoplasmic loop regions are involved in G proteinspecificity. Exchanging third cytoplasmic loops among m l and m2 mAChRs [2] and m2 and m3 mAChRs [3, 41 altered the preferred second messenger pathways between the control of phospholipase C and adenylyl cyclase activities. In Correspondence to C . J. van Koppen,

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Nitric oxide (NO) is a reactive endogenous molecule with multiple functions and its cellular signaling activity is mainly mediated by activation of the soluble isoform of guanylyl cyclase, a heterodimeric (alpha/beta) hemeprotein. The expression of the NO-sensitive soluble isoform of guanylyl cyclase was studied in various cultured melanocytic cells by measuring the accumulation of guanosine 3',5'-cyclic monophosphate in the presence and absence of NO donors. Here we report that 3-morpholino-sydnonimine, a donor of NO redox species, and (Z)-1-[2- (2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate, a direct NO donor, induced a 20-fold increase in intracellular guanosine 3',5'-cyclic monophosphate in nonmetastatic melanoma cells and normal melanocytes in culture that could be related to cellular melanin content in a concentration-dependent manner. The increased intracellular guanosine 3',5'-cyclic monophosphate was due to stimulation of the activity of soluble guanylyl cyclase as such increase was completely abolished by using a specific inhibitor of soluble guanylyl cyclase. The involvement of functional soluble guanylyl cyclase was further confirmed by the presence of alpha1 and beta1 subunits in these cells at both mRNA and protein levels. In contrast, none of the NO donors induced guanosine 3',5'-cyclic monophosphate production in metastatic melanoma cells, which could be attributed to the absence of the beta1 subunit that is essential for catalytic activity of the soluble isoform of guanylyl cyclase. Metastatic melanoma cells produced higher levels of intracellular guanosine 3',5'-cyclic monophosphate in response to natriuretic peptides than other cell types, however, due to upregulation of membrane-bound guanylyl cyclase activities, but they are less pigmented or unpigmented. The present finding suggests that NO signaling in association with melanogenesis is dependent on the soluble isoform of guanylyl cyclase, whereas absence of soluble guanylyl cyclase but the presence of membrane-bound guanylyl cyclase correlates with the metastatic behavior of melanoma cells.