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Preeclampsia, an important cause of maternal and fetal morbidity and mortality, is associated with increased sFLT1 levels and with structural and functional damage to the glycocalyx contributing to endothelial dysfunction. We investigated glycocalyx components in relation to preeclampsia in human samples. While soluble syndecan-1 and heparan sulphate were similar in plasma of preeclamptic and normotensive pregnant women, dermatan sulphate was increased and keratan sulphate decreased in preeclamptic women. Dermatan sulphate was correlated with soluble syndecan-1, and inversely correlated with blood pressure and activated partial thromboplastin time. To determine if syndecan-1 was a prerequisite for the sFlt1 induced increase in blood pressure in mice we studied the effect of sFlt1 on blood pressure and vascular contractile responses in syndecan-1 deficient and wild type male mice. The classical sFlt1 induced rise in blood pressure was absent in syndecan-1 deficient mice indicating that syndecan-1 is a prerequisite for sFlt1 induced increase in blood pressure central to preeclampsia. The results show that an interplay between syndecan-1 and dermatan sulphate contributes to sFlt1 induced blood pressure elevation in pre-eclampsia.

Major depressive disorder is a common and devastating psychiatric disorder. While pharmacotherapy and psychotherapy can be effective, a significant proportion of patients remain treatment resistant. Traditional antidepressants need to be taken for several weeks or months before the therapeutic effects become evident. For treatment-resistant patients, electroconvulsive therapy (ECT) is still the most effective treatment. Postictal slowing of electroencephalogram (EEG) activity has been associated with the therapeutic effects of ECT, but the mechanistic basis of this remains poorly studied. For decades this has encouraged researchers to investigate the antidepressant effects of isoflurane anesthesia with promising, but inconsistent, results. More recently, evidence of the rapid-acting antidepressant effects of subanesthetic doses of ketamine, an N-methyl-D-aspartate receptor (NMDAR) antagonist and a dissociative anesthetic, has sparked a renewed interest in the development of novel antidepressant therapies. Another treatment to show positive results is nitrous oxide (N2O), a gaseous anesthetic with NMDAR antagonist properties. One of the proposed mechanisms of ketamine’s action is related to its ability to increase glutamatergic signaling, leading to further changes in synaptic potentiation and in the function of neuronal networks. These changes have been suggested to involve the actions of brain-derived neurotrophic factor (BDNF) signaling via its receptor TrkB. Downstream of TrkB, the inhibition of glycogen synthase kinase 3β (GSK3β), the induction of mammalian target of rapamycin (mTOR) mediated protein synthesis, and the consolidation of synaptic changes have been implicated in ketamine´s actions. The first aim of this study is to investigate the molecular changes induced by isoflurane anesthesia in the adult mouse hippocampus using phosphoproteomics in the absence of a priori information. We find that brief isoflurane anesthesia induces 318 phosphorylation changes in a total of 237 proteins. While confirming the phosphorylation alterations on selected proteins, we also discover that various anesthetics, including urethane and ketamine, regulate these targets in a similar manner. In the second part, we investigate the effects of N2O on molecular signatures implicated in ketamine’s action. Findings reveal that N2O produces cortical excitation, followed by the rebound emergence of slow EEG activity following gas cessation, which coincide with the phosphorylation of TrkB, GSK3β and p70S6k (a kinase downstream of mTor). Moreover, we demonstrate that these pathways become regulated during the postictal period after flurothyl-induced seizures or during slow EEG activity induced by hypnotic agent medetomidine. Notably, medetomidine is not effective in the learned helplessness test. Finally, we investigate the dose-dependent changes induced by ketamine in TrkB signaling. An acute administration of sedative-anesthetic doses of ketamine, accompanied by increases in slow EEG activity, is found to increase the phosphorylation of the investigated pathways. These changes appear independent of ketamine’s metabolite hydroxynorketamine, an agent shown to have antidepressant-like behavioral effects in rodents. Masennus on yleinen psykiatrinen sairaus, joka aiheuttaa mittavaa inhimillistä kärsimystä ja merkittäviä yhteiskunnallisia kustannuksia. Masennusta hoidetaan farmakoterapialla, mutta moni potilaista ei saa merkittävää hyötyä lääkkeistä useiden viikkojen tai kuukausienkaan käytön jälkeen. Näille hoitoresistenteille masennuspotilaille psykiatrinen sähköhoito on yhä tehokkain hoitomuoto. Yhdeksi sähköhoidon tehoa ennustavaksi tekijäksi on esitetty kouristuksen jälkeisen aivojen hidasaaltotoiminnan lisääntymistä aivosähkökäyrässä. Ajatus kannusti tutkijoita selvittämään isofluraanilla aikaansaadun anestesian masennuslääkevaikutuksia jo vuosikymmeniä sitten, mutta tulokset jäivät epäselviksi. Tuoreemmissa tutkimuksissa on puolestaan toistettavasti havaittu N-metyyli-D-aspartaatti (NMDA) -reseptoreja salpaavan ketamiinin nopea masennusoireita lievittävä vaikutus. Hiljattain kliinisessä tutkimuksessa on havaittu myös NMDA-reseptoreja salpaavan typpioksiduulin (N2O) masennusoireita nopeasti lievittävä vaikutus. Isofluraanin ja typpioksiduulin vaikutusten taustalla olevat neurobiologiset mekanismit ovat kuitenkin yhä pääosin tuntemattomia. Ketamiinin vaikutusmekanismiksi on sen sijaan esitetty glutamatergisen hermovälityksen ja aivokuoren ärtyvyyden lisääntymistä, jonka on ajateltu johtavan muutoksiin synapsien ja hermosolujen toiminnassa. Näiden vaikutusten on esitetty aiheutuvan aivoperäisen hermokasvutekijän (BDNF) kohteena toimivan TrkB-reseptorin aktivaatiosta ja sitä seuraavista muutoksista glykogeenisyntaasikinaasi 3β:n toiminnassa ja rapamysiinin mekaanisen kohteen (mTOR) ohjaamassa proteiinisynteesissä. Ensimmäisessä tutkimuksessa keskityimme selvittämään isofluraanianestesian aiheuttamia molekulaarisia muutoksia hiiren hippokampuksessa, jonka tutkimisessa hyödynsimme fosfoproteomiikkaa. Havaitsimme 318 fosforylaatiomuutosta 237 eri proteiinissa. Vahvistaessamme havaittuja muutoksia valikoiduissa proteiineissa havaitsimme monien eri anesteettien, kuten uretaanin ja ketamiinin, aiheuttavan samankaltaisia muutoksia. Tutkimuksen toisessa osassa selvitimme N2O:n vaikutuksia ketamiinin säätelemiin solusignalointimekanismeihin hiiren etuaivokuorella. Ilokaasualtistuksen aikana havaittiin hermoston aktiivisuuteen liitettyjen merkkiaineiden lisääntymistä. Annostelun päätyttyä aivosähkökäyrässä havaittiin voimistunutta hidasaaltotoimintaa, jonka aikana kerätyissä aivonäytteissä havaittiin TrkB-välitteisen signaloinnin lisääntyneen. TrkB-signalointi lisääntyi myös näytteissä, jotka kerättiin flurotyylin aiheuttamien kouristuksien tai hidasaaltotoimintaa suoraan lisäävän medetomidiinin annostelun jälkeen. Medetomidiini ei kuitenkaan aiheuttanut masennuslääkkeille tyypillisiä käyttäytymisen muutoksia ns. opittu avuttomuus -mallissa. Lopuksi selvitimme ketamiinin annosriippuvaisia vaikutuksia TrkB-signalointiin hiiressä. Anesteettiset ketamiiniannokset lisäsivät aivosähkökäyrän hidasaaltotoimintaa, jonka aikana kerätyissä näytteissä havaittiin TrkB-signalointipolun aktivoituneen. Ketamiinin aiheuttamat muutokset näihin solusignalointimekanismeihin eivät näyttäisi olevan riippuvaisia sen metaboliatuotteesta hydroksinorketamiinista, jolla on esitetty olevan masennuslääkevaikutuksia koe-eläimiin. Tutkimustuloksemme viittaavat siihen, että hidasoskillaatioilla ja ketamiinin vaikutuksiin liitetyillä solusignalointimuutoksilla olisi yhteys. Lisäksi tulokset kannustavat selvittämään, ovatko aivokuoren ärtyvyys ja sitä seuraava hidasaaltotoiminnan lisääntyminen nopeavaikutteisten masennuslääkkeiden yhteinen ominaisuus.

Objective— Endothelial sphingosine-1-phosphate (S1P) receptor-1 (S1P 1 ) affects different vascular functions, including blood vessel maturation and permeability. Here, we characterized the role of the zS1P 1 ortholog in vascular development in zebrafish. Methods and Results— zS1P 1 is expressed in dorsal aorta and posterior cardinal vein of zebrafish embryos at 24 to 30 hours postfertilization. zS1P 1 downregulation by antisense morpholino oligonucleotide injection causes early pericardial edema, lack of blood circulation, alterations of posterior cardinal vein structure, and late generalized edema. Also, zS1P 1 morphants are characterized by downregulation of vascular endothelial cadherin ( VE-cadherin ) and Eph receptor EphB4a expression and by disorganization of zonula occludens 1 junctions in posterior cardinal vein endothelium, with no alterations of dorsal aorta endothelium. VE-cadherin knockdown results in similar vascular alterations, whereas VE-cadherin overexpression is sufficient to rescue venous vascular integrity defects and EphB4a downregulation in zS1P 1 morphants. Finally, S1P 1 small interfering RNA transfection and the S1P 1 antagonist (R)-3-amino-(3-hexylphenylamino)-4-oxobutylphosphonic acid (W146) cause EPHB4 receptor down-modulation in human umbilical vein endothelial cells and the assembly of zonula occludens 1 intercellular contacts is prevented by the EPHB4 antagonist TNYL-RAW peptide in these cells. Conclusion— The data demonstrate a nonredundant role of zS1P 1 in the regulation of venous endothelial barrier in zebrafish and identify a S1P 1 / VE-cadherin / EphB4a genetic pathway that controls venous vascular integrity.

The sphingosine-1-phosphate type 1 (S1P(1)) receptor is a new target in the treatment of auto-immune diseases as evidenced by the recent approval of FTY720 (Fingolimod). The ligand-binding pocket of the S1P(1) receptor has been generally characterised but detailed insight into ligand-specific differences is still lacking. The aim of the current study is to determine differences in ligand-induced S1P(1) receptor activation using an in silico guided site-directed mutagenesis strategy. S1P(1) mutant receptors (modifications of residues Y98(2.57), R120(3.28), F125(3.33)) were probed with a chemically diverse set of S1P(1) agonists (S1P, dihydro-S1P (dhS1P), R-, S- and racemic FTY720-P, VPC24191, SEW2871). Mutation of the R(3.28) residue generally results in a reduction of the potency of all ligands although the synthetic ligands including FTY720-P are less sensitive to these mutations. The Y(2.57)F mutation does not affect the potency of any of the ligands tested, but for all ligands except FTY720-P a significant decrease in potency is observed at the Y(2.57)A mutant. The F(3.33)A mutation significantly decreased the potency of FTY720-P and is detrimental for SEW2871 and VPC24191. The non-aromatic endogenous ligands S1P and dhS1P are less affected by this mutation. Our in silico guided mutagenesis studies identified new molecular determinants of ligand-induced S1P(1) receptor activation: 1) the flexibility of the polar head of the agonist to maintain a tight H-bond network with R(3.28) and 2) the ability of the agonist to make aromatic π-stacking interactions with F(3.33). Interestingly, FTY720-P has both chemical properties and is the only ligand that can efficiently activate the Y(2.57)A mutant.

Context: Individuals differ in their sensitivity to drug treatment, including that with muscarinic receptor antagonists used in the treatment of overactive bladder (OAB), due to a combination of pharmacodynamic and pharmacokinetic reasons. Objective: To discuss the variability in drug response among individual patients, the concept of the dose-response curve, and the selection of drug dosage as well as how these factors are integrated when optimising OAB treatment using the muscarinic receptor antagonist fesoterodine as an example. Evidence acquisition: Data sources were identified in 2010 using a nonsystematic search and included articles and abstracts selected using expert opinion of their relevance to drug response in OAB. Evidence synthesis: A given drug dose is unlikely to yield the same quantitative response in all patients, and undertreatment (too little efficacy) or overtreatment (too many side effects) may occur. The position and shape of the dose-response curve for a drug may differ between patients and within a patient for desired and adverse effects. The availability of two or more drug doses allows for titration (flexible dosing) to find the dose exhibiting the optimal clinical efficacy that is tolerable for an individual patient. Optimally, a patient would have symptom resolution with no adverse events (AEs). Realistically, an efficacy-to-tolerability ratio (therapeutic index) exists for each of the combinations of selected efficacy metrics and AE reports. Conclusions: Dose titration is important for selection of an effective dose of a treatment with minimal side effects. (C) 2011 European Association of Urology. Published by Elsevier B. V. All rights reserved

Nebivolol is a selective β 1 -adrenoceptor antagonist which, in addition, displays endothelium-dependent vasodilating properties in humans and other species. β 3 -Adrenoceptors have been proposed to be a molecular target of nebivolol-induced vasodilatation. Therefore, we have investigated possible β 3 -adrenoceptor agonism by nebivolol for relaxation of the human and rat urinary bladder (prototypical β 3 -adrenoceptor-mediated responses) as well as for cAMP accumulation in Chinese hamster ovary cells stably transfected with the human β-adrenoceptor subtypes. Nebivolol concentration-dependently relaxed both human and rat isolated urinary bladder strips but with low potency, similar to that reported for vasodilatation. However, nebivolol-induced bladder relaxation in either species was not inhibited by the β 3 -adrenoceptor antagonist SR 59,230A (10 μM), although this compound inhibited the isoprenaline-induced relaxation with the expected potency. In radioligand binding studies nebivolol had lower affinity for human β 3 -adrenoceptors than the other two β-adrenoceptor subtypes, but this low affinity was in line with its potency to relax the bladder or isolated blood vessels. In functional studies nebivolol even in high concentrations did not stimulate cAMP formation via any of the three cloned human β-adrenoceptors or in rat bladder smooth muscle cells. Taken together these data demonstrate that nebivolol can relax not only vascular but also urinary bladder smooth muscle. However, they do not support the hypothesis that nebivolol is an agonist at cloned human β 3 -adrenoceptors or in rat or human urinary bladder.

Purpose Comparison of efficacy of propiverine extended release (ER) 30 mg o.d. in the treatment of male OAB administered as monotherapy (MT) or add-on to α-blockers (combination treatment, CT) in relation to maximum urinary flow (Qmax) in a non-interventional study. Methods Men ≥40 years with OAB symptoms, Qmax ≥10 ml/s, prostate volume

The widespread non-neuronal synthesis of acetylcholine (ACh) has changed the paradigm of ACh acting solely as a neurotransmitter. Indeed, the presence of ACh in many types of proliferating cells suggests a role for this neurotransmitter in the control of cell division. The parasympathetic system is a major pathway regulating micturition, but ACh-mediated control plays a more complex role than previously described, acting not only in the detrusor muscle, but also influencing detrusor function through the activity of urothelial muscarinic receptors. Here we investigated the role of muscarinic receptors in mediating cell proliferation in the human UROtsa cell line, which is a widely used experimental model to study urothelium physiology and pathophysiology. Our results demonstrate that UROtsa cells express the machinery for ACh synthesis and that muscarinic receptors, with the rank order of M3 > M2 > M5 > M1 = M4, are present and functionally linked to their known second messengers. Indeed, the cholinergic receptor agonist carbachol (CCh) (1–100 μM) concentration-dependently raised IP3 levels, reaching 66 ± 5% over basal. The forskolin-mediated adenylyl cyclase activation was reduced by CCh exposure (forskolin: 1.4 ± 0.14 pmol/ml; forskolin + 100 μM CCh: 0.84 ± 0.12 pmol/ml). CCh (1–100 μM) concentration-dependently increased UROtsa cell proliferation and this effect was inhibited by the non-selective antagonist atropine and the M3-selective antagonists darifenacin and J104129. Finally, CCh-induced cell proliferation was blocked by selective PI-3 kinase and ERK activation inhibitors, strongly suggesting that these intracellular pathways mediate, at least in part, the muscarinic receptor-mediated cell proliferation.

β-Adrenoceptors are important mediators of smooth muscle relaxation in the urinary bladder, but the concomitant presence of a muscarinic agonist, e.g., carbachol, can attenuate relaxation responses by reducing potency and/or efficacy of β-adrenoceptor agonists such as isoprenaline. Therefore, the present study was designed to explore the subtypes and signalling pathways of muscarinic receptors involved in the attenuation of isoprenaline-induced isolated rat detrusor preparations using novel subtype-selective receptor ligands. In radioligand binding studies, we characterized BZI to be a M(3)-sparing muscarinic agonist, providing selective M(2) stimulation in rat bladder, and THRX-182087 as a highly M(2)-selective antagonist. The use of BZI and of THRX-182087 in the presence of carbachol enabled experimental conditions with a selective stimulation of only M(2) or M(3) receptors, respectively. Confirming previous findings, carbachol attenuated isoprenaline-induced detrusor relaxation. M(2)-selective stimulation partly mimicked this attenuation, indicating that both M(2) and M(3) receptors are involved. During M(3)-selective stimulation, the attenuation of isoprenaline responses was reduced by the phospholipase C inhibitor U 73,122 but not by the protein kinase C inhibitor chelerythrine. We conclude that both M(2) and M(3) receptors contribute to attenuation of β-adrenoceptor-mediated relaxation of rat urinary bladder; the signal transduction pathway involved in the M(3) component of this attenuation differs from that mediating direct contractile effects of M(3) receptors.

PURPOSE: The known importance of testosterone for the development of benign prostatic hyperplasia (BPH) prompted us to test the hypothesis whether polymorphisms of two genes (CYP19A1 and CYP3A4) involved in testosterone metabolism are associated with clinical BPH-parameters. METHODS: A random sample of the population-based Herne lower urinary tract symptoms cohort was analysed. All these men underwent a detailed urological work-up. Two polymorphisms in the CYP19A1 gene [rs700518 in exon 4 (A57G); rs10046 at the 3'UTR(C268T)] and one in the 3'UTR of CYP3A4 [rs2740574 (A392G)] were determined by TaqMan assay from genomic DNA of peripheral blood. These polymorphisms were correlated to clinical and laboratory BPH-parameters. RESULTS: A total of 392 men (65.4 +/- 7.0 years; 52-79 years) were analysed. Mean International Prostate Symptom Score (IPSS; 7.5), Q (max) (15.4 ml/s), prostate volume (31 ml) and prostate specific antigen (PSA) (1.8 ng/ml) indicated a typical elderly population. Both polymorphisms in the CYP19A1 gene were not correlated to age, IPSS, Q (max), prostate volume and post-void residual volume. Serum PSA was higher in men carrying the heterozygous rs10046 genotype (2.0 +/- 0.1 ng/ml) than in those with the CC-genotype (1.7 +/- 0.2 ng/ml, P = 0.012). Men carrying one a mutated allele of the CYP3A4 gene had smaller prostates (27.0 +/- 2.0 vs. 32 +/- 0.8 ml, P = 0.02) and lower PSA levels (1.6 +/- 0.3 vs. 1.9 +/- 0.1 ng/ml). CONCLUSIONS: The inconsistent associations observed herein and for other gene polymorphisms warrant further studies. In general, the data regarding the association of gene polymorphism to BPH-parameters suggest that this disease is caused by multiple rather than a single genetic variant. A rigorous patient selection based on anatomo-pathological and hormonal profile may possible reduce the number of confounders for future studies thus enabling a more detailed assessment of the association between genetic factors and BPH-parameters

Top of pageAbstract The adipocytokine leptin has distinct functions regulating vascular tone, inflammation, and collateral artery growth. Arteriogenesis is an inflammatory process and provides a mechanism to overcome the effects of vascular obstruction. We, therefore, tested the effects of leptin in hypoperfused rat brain (three-vessel occlusion). Systemic leptin administration for 1 week after occlusion surgery increased cerebral hemodynamic reserve similar to granulocyte–macrophage colony-stimulating factor (GM-CSF), as indicated by improved CO2 reactivity (vehicle 0.53%±0.26% versus leptin 1.05%±0.6% per mm Hg arterial pCO2, P

Aims Structural adaptation of the vessel wall in response to sustained alterations in haemodynamic forces is known as vascular remodelling. Detailed knowledge on the mechanism underlying this vascular response is limited, and we aimed to study the function of Nur77 in smooth muscle cells (SMCs) in arterial remodelling. Methods and results Carotid artery ligation in mice results in flow-induced, outward remodelling of the contralateral carotid artery, and we observed enhanced Nur77 expression during this process. Transgenic mice that express Nur77 or its dominant-negative variant, denoted as ‘ΔTA’ in arterial SMCs, were exposed to carotid artery ligation, and after 4 weeks pressure–diameter relationships were measured. Structural outward remodelling is inhibited in Nur77-transgenic mice when compared with wild-type and ΔTA-transgenic mice. The key determinants of remodelling vascular tone and macrophage accumulation were studied. No difference in contractile and relaxant responses was detected in isolated aorta, carotid, and mesenteric artery segments between transgenic and wild-type mice. SMC-specific overexpression of Nur77 in transgenic mice reduced macrophage accumulation and repressed matrix metalloproteinase (MMP)1 and MMP9 expression at early time points. MMP2 protein expression was reduced in Nur77-transgenic mice, whereas in ΔTA-transgenic mice MMP2 expression was increased. Conclusion Nur77 is induced during outward remodelling and inhibits this vascular adaptation in mice. Nur77-mediated inhibition of arterial remodelling involves a reduction in both macrophage accumulation and MMP expression levels.

Fesoterodine is an antimuscarinic agent indicated for the treatment of overactive bladder (OAB) symptoms. The objective of this study was to evaluate the efficacy of fesoterodine versus placebo over selected intervals during a 24-hour period in subjects with OAB. In a post hoc analysis, data were analyzed from two randomized, double-blind, placebo-controlled 12-week phase III trials in which subjects with a history of OAB symptoms for 6 months were treated with morning doses of fesoterodine 4 mg, fesoterodine 8 mg, or placebo. These trials are registered at ClinicalTrials.gov (NCT00220363 and NCT00138723). Changes were evaluated in number of micturitions, urgency episodes, urgency urinary incontinence (UUI) episodes, and mean voided volume (MVV) divided into three 8-hour intervals: 08:00-15:59 (daytime), 16:00-23:59 (evening), and 00:00-07:59 (nighttime). Comparisons with placebo were made using analysis of covariance (for least squares mean changes) and Wilcoxon rank sum test (for median percent changes); differences were considered significant at p

Study Type – Symptom prevalence (retrospective cohort) Level of Evidence 2b OBJECTIVES To determine the role of treatment-associated improvement in nocturia in health-related quality of life (HRQL) in patients with lower urinary tract symptoms (LUTS) suggestive of benign prostatic hyperplasia, and secondarily to confirm the role of nocturia in HRQL at baseline and to compare the effects of watchful waiting, transurethral microwave treatment (TUMT) and transurethral resection of the prostate (TURP) to those of α1-adrenoceptor antagonists (α-blockers) on nocturia. PATIENTS AND METHODS We retrospectively analysed using multiple regression a large single-centre database of patients receiving routine care for treatment-associated alterations of symptoms and HRQL (assessed at baseline, 2611 men) and 6–12 months after initiation of treatment (1258 men). RESULTS Among the symptoms assessed using the International Prostate Symptom Score, nocturia (together with urgency and weak stream) had the strongest correlation with HRQL at baseline and after treatment. Watchful waiting, α-blockers, TUMT and TURP reduced nocturia episodes by a mean (sd) of 7 (53)%, 17 (40)%, 32 (47)% and 75 (23)%, respectively. The treatment-associated improvements in nocturia (together with those of weak stream) had the strongest association with those of HRQL. CONCLUSIONS We conclude that among all LUTS assessed in the IPSS, nocturia has one of the strongest associations with HRQL, and that treatment-associated improvements in nocturia contribute considerably to overall improvements in HRQL.

We investigated the contribution of the intracellular calcium (Ca (i) (2+) ) transient to acetylcholine (ACh)-mediated reduction of pacemaker frequency and cAMP content in rabbit sinoatrial nodal (SAN) cells. Action potentials (whole cell perforated patch clamp) and Ca (i) (2+) transients (Indo-1 fluorescence) were recorded from single isolated rabbit SAN cells, whereas intracellular cAMP content was measured in SAN cell suspensions using a cAMP assay (LANCE((R))). Our data show that the Ca (i) (2+) transient, like the hyperpolarization-activated "funny current" (I (f)) and the ACh-sensitive potassium current (I (K,ACh)), is an important determinant of ACh-mediated pacemaker slowing. When I (f) and I (K,ACh) were both inhibited, by cesium (2 mM) and tertiapin (100 nM), respectively, 1 micro M ACh was still able to reduce pacemaker frequency by 72%. In these I (f) and I (K,ACh)-inhibited SAN cells, good correlations were found between the ACh-mediated change in interbeat interval and the ACh-mediated change in Ca (i) (2+) transient decay (r (2) = 0.98) and slow diastolic Ca (i) (2+) rise (r (2) = 0.73). Inhibition of the Ca (i) (2+) transient by ryanodine (3 microM) or BAPTA-AM (5 microM) facilitated ACh-mediated pacemaker slowing. Furthermore, ACh depressed the Ca (i) (2+) transient and reduced the sarcoplasmic reticulum (SR) Ca(2+) content, all in a concentration-dependent fashion. At 1 microM ACh, the spontaneous activity and Ca (i) (2+) transient were abolished, but completely recovered when cAMP production was stimulated by forskolin (10 microM) and I (K,ACh) was inhibited by tertiapin (100 nM). Also, inhibition of the Ca (i) (2+) transient by ryanodine (3 microM) or BAPTA-AM (25 microM) exaggerated the ACh-mediated inhibition of cAMP content, indicating that Ca (i) (2+) affects cAMP production in SAN cells. In conclusion, muscarinic receptor stimulation inhibits the Ca (i) (2+) transient via a cAMP-dependent signaling pathway. Inhibition of the Ca (i) (2+) transient contributes to pacemaker slowing and inhibits Ca (i) (2+) -stimulated cAMP production. Thus, we provide functional evidence for the contribution of the Ca (i) (2+) transient to ACh-induced inhibition of pacemaker activity and cAMP content in rabbit SAN cells.

Purpose Previous studies on associations between voiding dysfunction and quality of life (QoL) have largely been limited to baseline data. Therefore, we have explored associations between Q max and voiding and storage sub-scores of the International Prostate Symptom Score (IPSS) before and after treatment with QoL. Methods Analysis of a single-center database of 2,316 men with voiding dysfunction attributed to benign prostatic hyperplasia undergoing various medical and surgical treatment forms. Results Q max exhibited little correlation with QoL before or after treatment. IPSS inversely correlated with QoL at baseline and after treatment, and IPSS improvements correlated with those of QoL. The associations applied to both the voiding and storage sub-score of the IPSS, with the latter consistently exhibiting somewhat tighter associations. Conclusions Our post-treatment data support the idea of a cause–effect relationship between voiding symptoms and QoL irrespective of treatment form. While both voiding and storage symptoms contribute to this relationship, storage symptoms play a somewhat greater role.

Doxazosin and related, quinazoline-based alpha(1)-adrenoceptor antagonists can induce apoptosis in prostate and various other normal, benign, smooth muscle, endothelial and malignant cells. Such apoptosis-inducing effects occur independently of alpha(1)-adrenoceptor antagonism and typically require much high concentrations than those required for receptor occupancy. Several studies have invested efforts towards the elucidation of the molecular mechanisms underlying doxazosin-induced apoptosis. These include various tumor cells, cardiomyocytes, endothelial cells and bladder smooth muscle cells. While the high concentrations of doxazosin required to induce apoptosis challenge the use of this and related drugs for clinical optimization of apoptosis induction, such quinazoline structure may represent chemical starting points to develop more potent apoptosis-inducing agents free of alpha(1)-adrenoceptor antagonistic action and suitable for cancer treatment with minimal and well-tolerated side effects.

Objectives: To explore associations between the number of vaginal deliveries (primary aim) or gender (secon- dary aim) and overactive bladder (OAB) symptoms as well as their response to treatment with a muscarinic antagonist. Methods: Preplanned secondary analysis of an observational study of solifenacin in OAB patients. Episode frequencies of OAB symptoms, pad use and scores on OAB rating scales were documented in 4450 patients before and after a 12–14 week treatment period with solifenacin 5 or 10 mg. Results: Women without, and with one, two or more than two vaginal deliveries and men were similar in their baseline characteristics. All groups also exhibited rather similar reductions in symptoms and improvements in rating scales upon treatment. Conclusion: These data indicate that solifenacin, and perhaps other muscarinic receptor antagonists, are similarly suitable for the treatment of OAB symptoms in both genders, irrespective of previous vaginal deliveries.

Anticholinergic therapy is the first-line therapy for overactive bladder (OAB) syndrome. Especially in the last years, the number of available substances has increased because of the launch of solifenacin, darifenacin, and fesoterodine. Additionally, slow-release and transdermal formulations have led to a large variety of available treatment options. The efficacy of all substances has been proven in randomised, double-blind studies, and reviews and meta-analyses have also underlined the efficacy of all available anticholinergics and have been updated regularly. All available drugs are efficacious for OAB treatment, and clinically relevant differences among them have not been proven consistently. Moreover, age, gender, and the type of OAB (dry vs. wet) seem to lack clinically relevant impact on the efficacy of OAB treatment. The various drugs are similar in tolerability, with the exception of more dry mouth and central nervous effects with slow-release oxybutynin. Knowledge of pharmacokinetic properties of the individual substances is important in order to choose the right therapy for each patient

BACKGROUND: In major depressive disorder, selective serotonin reuptake inhibitors target the serotonin transporter (SERT). Their response rates (30-50%) are modified by SERT promotor polymorphisms (5-HTTLPR). OBJECTIVES: To quantify the relationship between SERT occupancy and response, and whether 5-HTTLPR is a modifier. METHODS: Drug-free depressed outpatients (n=49; both sexes; aged 25-55 years), received paroxetine (20 mg/day). We quantified SERT occupancy with iodine-123-labeled 2beta-carbomethoxy-3beta-(4-iodophenyl)-tropane single-photon emission computed tomography imaging at baseline and after 6 weeks; we genotyped 5-HTTLPR (S, LG, LA). Primary outcomes: percentage decrease in 17-item Hamilton Depression Rating Scale and response (>/=50% decrease of 17-item Hamilton Depression Rating Scale). RESULTS: A significant positive relationship between SERT occupancy and clinical response existed only in the LA/LA genotype (P

Sphingosine-1-phosphate (S1P) signalling via G protein-coupled receptors is important for the regulation of cell function and differentiation. Specific Regulators of G protein Signalling (RGS) proteins modulate the function of these receptors in many cell types including vascular smooth muscle cells (VSMCs). Therefore, we investigated the role of altered expression levels of RGS proteins in S1P receptor function in VSMCs and transfected CHO cells. The mRNA expression of the S1P 1 receptor, RGS4 and RGS16 were down-regulated in VSMCs during phenotypic modulation induced by culturing, whereas mRNA levels of RGS2, RGS3, S1P 2 and S1P 3 receptors were unchanged. Interestingly, the expression level of RGS5 was transiently up-regulated. Despite major alterations in RGS levels, S1P-induced calcium elevation in VSMCs was not altered. Co-transfection of RGS2, RGS3, RGS4, RGS5 and RGS16 into CHO-Flp-In cells stably expressing the S1P 1 or S1P 3 receptor did not modify S1P-induced inhibition of cAMP accumulation to a major extent. Similar results were obtained with SEW2871, a selective S1P 1 receptor agonist. However, the inhibition of cAMP accumulation by the agonist FTY720-P via the S1P 1 receptor was significantly decreased by co-transfection with RGS5. These results indicate that mRNA of the S1P 1 receptor, RGS4, RGS5 and RGS16 is differentially regulated during phenotypic modulation. However, major alterations in RGS protein expression have only limited effect on S1P receptor function.

Two groups of drugs, alpha blockers and 5-alpha-reductase inhibitors (5ARI), are currently widely used for the medical treatment of benign prostatic syndrome (BPS). Alpha blockers are characterized by a rapid onset of efficacy. If given at an adequate dose, all alpha blockers have a similar efficacy, yet quantitative differences regarding side effects exist. The onset of clinical efficacy of 5ARIs is delayed and dependent on prostate volume. Symptom improvement is generally less pronounced than with alpha blockers, yet this difference declines with time. 5ARI, in contrast to alpha blockers, reduce prostate volume and the risk of long-term BPS complications such as prostate surgery or acute urinary retention. The combination therapy of alpha blockers and 5ARI is superior to either monotherapy; however, this superiority becomes evident only after prolonged (>1 year) therapy. Because of additive side effects, this combination should be reserved for BPS patients with a high risk of progression. Regarding plant extracts, no definitive recommendation can be given because of a limited number of high-quality clinical trials. The use of antimuscarinics in men with BPS with a dominance of storage symptoms and without significant obstruction is promising, although further trials, particularly with a longer study duration, are required.

The new version of the German guidelines for assessing benign prostatic hyperplasia (BPH) was written in 2006 and 2007 by experts of the BPH working group on behalf of the German Association of Urology and Federation of German Urologists. The full version is expected to be published in 2008. Recommendations for assessing BPH were modified and updated and include patient history, symptom questionnaires, physical examination, urine analysis, prostate-specific antigen, uroflowmetry, ultrasound examination of the urinary bladder (including postvoid residual urine), and ultrasound examination of the upper urinary tract or creatinine measurement. Optional tests are voiding diary, pressure-flow studies, ultrasound measurement of detrusor wall thickness, urethrocystography, and urethrocystoscopy. For the first time, tests for the differential diagnosis of bladder symptoms and BPH are described. Furthermore, the latest knowledge was added on disease progression, indications for urodynamic assessment, and ultrasound measurement of detrusor wall thickness for evaluating bladder outlet obstruction. International quality standards were applied in order to increase the guidelines' value; all tests were judged using the levels of evidence and grades of recommendation of the U.S. Department of Health and Human Services classification

Urinary bladder function is known to mature during fetal and postnatal development, including changes in neurotransmitter regulation of detrusor contraction. However, only few experimental data are available about muscarinic receptor antagonist function in the urinary bladder from young animals. In the present study, we compare the muscarinic receptor-mediated contractions in juvenile and adult porcine detrusor and the effects of antimuscarinic compounds. Urinary bladders from young (8–12 weeks; 12- to 35-kg body weight) and mature pigs (>40 weeks; >100 kg) were compared. Muscarinic receptor expression was assessed by real time polymerase chain reaction and radioligand binding. Muscle contraction was measured with a force transducer; L-type Ca2+ currents (I Ca,L) of isolated detrusor myocytes were recorded with standard voltage clamp technique. Juvenile and adult detrusor expressed similar quantities of the messenger RNA of M2 and M3 receptors. The number of [3H]QNB-binding sites and their affinity for the radioligand were also similar between juvenile and adult detrusor. In contrast, maximum contractile responses to the muscarinic receptor agonist carbachol were slightly larger in juvenile than adult bladders. On the other hand, carbachol was slightly less potent in juvenile than in adult tissue. The M3 antagonist DAU 5884 and the spasmolytic drug propiverine inhibited contractile responses with comparable efficacies and potencies in juvenile and adult tissue. I Ca,L was somewhat smaller in juvenile than in adult cells. Taken together, these data suggest that expression and function of M2 and M3 receptors are similar in the detrusor of juvenile and mature pigs. Therefore, similar responses to antimuscarinic compounds could be expected in young and adult patients.

The normal physiological contraction of the urinary bladder, which is required for voiding, is predominantly mediated by muscarinic receptors, primarily the M-3 subtype, with the M-2 subtype providing a secondary backup role. Bladder relaxation, which is required for urine storage, is mediated by beta-adrenoceptors, in most species involving a strong beta(3)-component. An excessive stimulation of contraction or a reduced relaxation of the detrusor smooth muscle during the storage phase of the micturition cycle may contribute to bladder dysfunction known as the overactive bladder. Therefore, interference with the signal transduction of these receptors may be a viable approach to develop drugs for the treatment of overactive bladder. The prototypical signaling pathway of M-3 receptors is activation of phospholipase C (PLC), and this pathway is also activated in the bladder. Nevertheless, PLC apparently contributes only in a very minor way to bladder contraction. Rather, muscarinic-receptor-mediated bladder contraction involves voltage-operated Ca2+ channels and Rho kinase. The prototypical signaling pathway of beta-adrenoceptors is an activation of adenylyl cyclase with the subsequent formation of cAMP. Nevertheless, cAMP apparently contributes in a minor way only to beta-adrenoceptor-mediated bladder relaxation. BKCa channels may play a greater role in beta-adrenoceptor-mediated bladder relaxation. We conclude that apart from muscarinic receptor antagonists and beta-adrenoceptor agonists, inhibitors of Rho kinase and activators of BKCa channels may have potential to treat an overactive bladder

Conditions associated with hypertrophy of the urinary bladder have repeatedly been associated with an increased urinary excretion of transforming growth factor ( TGF)beta in both rats and patients. Because TGF beta can have both growth- promoting and - inhibiting effects, we have studied its effects on cell growth and death in primary cultures of rat bladder smooth muscle cells. TGF beta 1, TGF beta 2, or TGF beta 3 did not cause apoptosis, but all three isoforms inhibited DNA synthesis with similar potency (EC50 of approximately 0.1 ng/ ml) and efficacy. Such inhibition was antagonized by a specific TGF beta receptor antagonist and independent of the presence of serum. Mitogen- activated protein kinases ( MAPKs) are involved in the control of cell growth, and all three TGF beta isoforms inhibited activation of the extracellular signal- regulated kinase, c- Jun NH 2- terminal kinase, and p38 MAPK subfamilies. Nevertheless, the inhibitory effects of the TGF beta isoforms on DNA synthesis were not affected by presence of inhibitors of the three MAPK pathways. TGF beta did not alter cell size as measured by flow cytometry or mitochondrial activity, an integrated measure of cell size and number. We conclude that our data do not support the hypothesis that TGF beta is a mediator of rat bladder hypertrophy.

Agonist exposure can cause internalisation of G-protein coupled receptors (GPCRs), which may be a part of desensitisation but also of cellular signaling. Previous methods to study internalisation have been tedious or only poorly quantitative. Therefore, we have developed and validated a quantitative method using a sphingosine-1-phosphate (S1P) receptor as a model. Because of a lack of suitable binding studies, it has been difficult to study S1P receptor internalisation. Using a N-terminal HisG-tag, S1P(1) receptors on the cell membrane can be visualised via immunocytochemistry with a specific anti-HisG antibody. S1P-induced internalisation was concentration dependent and was quantified using a microplate reader, detecting either absorbance, a fluorescent or luminescent signal, depending on the antibodies used. Among those, the fluorescence detection method was the most convenient to use. The relative ease of this method makes it suitable to measure a large number of data points, e.g. to compare the potency and efficacy of receptor ligands.

Summary 1 The present study was designed to investigate the relationship between innervation density and contractile responses to field stimulation and exogenous agonists at early time points after induction of bladder outlet obstruction (BOO) in rats. 2 When compared with sham-operated animals, 1, 3 and 7 days of BOO were associated with a 75%, 80% and 90% increase of bladder weight. Field stimulation caused a frequency-dependent increase in force of contraction. The force of contraction was reduced at each frequency in BOO rats with the greatest decrease after 1 day and a gradual but incomplete recovery thereafter. In contrast, contractile responses to ATP, carbachol and KCl were markedly reduced after 1 day of BOO and fully recovered after 7 days. The neurofilament staining was not altered by 1 day of BOO, but gradually decreased with increasing duration of BOO reaching the lowest levels after 7 days. 3 We conclude that impaired cellular contractility seems to underlie the early reductions of field stimulation-induced contraction, possibly reflecting surgical trauma of the tissue. However, at later time points a reduced nerve density, possibly reflecting a partial denervation, appears to be the main reason for impaired contractile response to field stimulation.

Both hypertension and congestive heart failure are characterized by a reduced vasodilatory capacity. In both conditions, the impairment of endothelial function is mainly the result of a reduced nitric oxide availability. The highly beta1-selective third-generation adrenoceptor blocker nebivolol displays additional endothelium-dependent vasodilating actions in humans as well as in animal models. In this study, we investigated whether these vasodilating properties of nebivolol are preserved in conditions with endothelial dysfunction. The vasodilatory effects of nebivolol were compared with those of the muscarinic agonist methacholine in isolated aortic rings obtained from spontaneous hypertensive rats and rabbits with experimental heart failure. The methacholine-induced responses were attenuated in aortic rings from both spontaneous hypertensive rats and congestive heart failture rabbits (42+/-6% and 25+/-3% vs. 89+/-3% and 54+/-7% for controls, respectively; P 0.05, n=6-13). These results implicate that the favorable hemodynamic profile of nebivolol may be preserved in patients with hypertension or congestive heart failure despite an impaired endothelial function

Muscarinic acetylcholine receptors, particularly M 3 receptors, are physiologically the most important mechanism to induced urinary bladder smooth muscle contraction. Their prototypical signaling response is a stimulation of phospholipase C (PLC), and this also has been shown in the urinary bladder. Nevertheless, it has remained controversial whether PLC signaling mediates bladder contraction induced by muscarinic receptor agonists. Studies in favor and against a role for PLC differed in their experimental protocol (single versus repeated concentration-response curves within a single preparation) and in the PLC inhibitors that have been used. We have now tested whether previous differential conclusions regarding a role for PLC are related to inhibitors and/or experimental protocols. In a single curve protocol, U-73,122 [1-[6-[((17β)-3-methoxyestra-1,3,5[10]-trien-17-yl)amino]hexyl]-1 H -pyrrole-2,5-dione] did not attenuate carbachol responses. In a repeated curve protocol, ET-18-OCH 3 (1- O -octadecyl-2- O -methyl- sn -glycero-3-phosphorylcholine) lacked significant inhibition relative to vehicle time controls. In contrast, D609 ( O -tricyclo[5.2.1.02,6]dec-9-yl dithiocarbonate potassium salt) depressed maximal carbachol effects but also nonspecifically inhibited contraction induced by KCl. Neomycin did not affect the carbachol-induced rat urinary bladder contraction. We conclude that previously reported differences relate to the use of inhibitors rather than experimental protocols and that the overall data do not support a role for PLC in M 3 muscarinic receptor-mediated rat bladder contraction.

We have compared the ability of three radioligands, [(125)I]-cyanopindolol, [(3)H]-CGP 12,177 and [(3)H]-dihydroalprenolol, to label the three human beta-adrenoceptor subtypes. Saturation and competition binding experiments were performed using membrane preparations from Chinese hamster ovary cells stably transfected with the three subtypes. While [(3)H]-CGP 12,177 had very similar affinity for beta(1)- and beta(2)-adrenoceptors (about 40 pM), [(125)I]-cyanopindolol and [(3)H]-dihydroalprenolol had 4- to 6-fold higher affinity for beta(2)- as compared to beta(1)-adrenoceptors (10 vs 45 and 187 vs 1,021 pM, respectively). The affinity of [(125)I]-cyanopindolol at beta(3)-adrenoceptors was considerably lower (440 pM) than at the other two subtypes. The beta(3)-adrenoceptor affinity of [(3)H]-CGP 12,177 and [(3)H]-dihydroalprenolol was so low that it could not be estimated within the tested range of radioligand concentrations (up to 4,000 pM and 30,000 pM for [(3)H]-CGP 12,177 and [(3)H]-dihydroalprenolol, respectively). We conclude that all three radioligands are ill-suited to label beta(3)-adrenoceptors, particularly in preparations co-expressing multiple subtypes. In the absence of alternatives, [(125)I]-cyanopindolol appears the least unsuitable to label beta(3)-adrenoceptors. There is a need for high-affinity radioligands which are either selective for beta(3)-adrenoceptors or reasonably non-selective among all three beta-adrenoceptor subtypes.

Background The temporal relations between the onset of echocardiographic changes and clinical diagnosis of right ventricular (RV) failure are unresolved. We have characterized such relations in a rat monocrotaline (MCT) model of RV failure. Methods Eight-week-old male Wistar rats were injected with MCT (60 mg/kg) or vehicle and underwent serial echocardiography. RV free-wall thickness (RVWT), pulmonary artery acceleration time normalized to cycle length (PAAT/CL), RV end-diastolic diameter (RVEDD), and tricuspid annular plane systolic excursion (TAPSE) were measured. Results Significant differences in echocardiographic parameters between MCT-treated and control rats were found as early as 14 days before RV failure for RVWT, 10 days for PAAT/CL, and 7 days for RVEDD and TAPSE. The time intervals between the onset of changes in RVWT, PAAT/CL, RVEDD, and TAPSE and diagnosis of RV failure were 11.3 ± 0.8, 10.9 ± 0.7, 6.5 ± 0.5, and 5.4 ± 0.7 days, respectively. The sequence of echocardiographic changes was consistent in all animals during development of RV failure. Conclusions Pulmonary hypertension (assessed by PAAT/CL) and RV free-wall thickening (characterized by RVWT) precede RV dilation and RV systolic dysfunction (measured by RVEDD and TAPSE, respectively). Echocardiographic analysis permits accurate determination of the stage of disease development in MCT-induced RV failure.

alpha(1)-Adrenoceptor antagonists such as alfuzosin, doxazosin, tamsulosin and terazosin are first-line agents for the treatment of lower urinary tract symptoms suggestive of benign prostatic hyperplasia (BPH), but are only second-line agents (doxazosin and terazosin only) for the treatment of arterial hypertension. Sexual function is complex and includes multiple domains such as sexual desire (libido), erectile function and ejaculatory function. Erectile and ejaculatory functions are frequently reduced in patients with BPH and can impact on their quality of life. Therefore, the treatment of BPH should aim to maintain or even restore sexual function.alpha(1)-Adrenoceptor antagonists lack major effects on sexual desire in placebo-controlled studies. Reports on erectile function are inconsistent, with both beneficial and adverse effects being reported, but impotence can occur in some patients without clear differences between drugs. Ejaculatory dysfunction during treatment may represent (relative) an ejaculation. It occurs more frequently with tamsulosin than with other drugs of this class, but the differences are not big enough to be consistently detectable in directly comparative studies. We propose that such differences between drugs should be weighed against differences in cardiovascular tolerability when choosing the optimal treatment for each patient.

Anticholinergics act in the treatment of overactive bladder by blocking muscarinic receptors of which five subtypes exist. Their desired effects occur via M(3) receptors, but a role for M(2) receptors is being discussed. Adverse effects such as dry mouth and constipation occur also via M(3) receptors, but M(2) and M(1) receptors can mediate side effects in the heart or on cognitive function, respectively. Therefore, an M(3)-selective drug such as darifenacin could theoretically be less effective but also have fewer cardiac or central nervous side effects. However, the limited available clinical data do not support a smaller efficacy or better general tolerability. The lack of adverse effects on cognitive function is well documented for darifenacin, but it cannot yet be determined definitively whether this discriminates it from other modern anticholinergics

We describe the development and validation of analytical methodology for the determination of sphingosine 1-phosphate (S1P) in plasma. It uses solid-phase extraction (SPE) followed by an automated reversed-phase gradient HPLC column-switching system with a pre-column derivatization with o-phthalaldehyde (OPA) and fluorescence detection. The limit of quantification was determined at 100 ng/ml exogenous sphingosine 1-phosphate with a relative standard deviation for precision and accuracy

We compared the density and function of M-2 and M-3 muscarinic acetylcholine receptor subtypes in the urinary bladder of young adult (3 months) and old (23 months) male Wistar rats. Old rats had a reduced density of muscarinic receptors (96 +/- 10 vs. 156 +/- 21 fmol/mg protein), but competition experiments with the M-3-selective darifenacin did not indicate alterations in the relative roles of M-2 and M-3 receptors, with the former being more abundant. The amount of immunodetectable alpha-subunits of various G-proteins potentially linked to muscarinic receptor function was unchanged. The potency of carbachol to contract bladder strips was also unaltered; its maximum effects as well as those of a single KCl concentration were unchanged if raw data or those corrected for strip length were analysed, but somewhat reduced when those corrected for strip weight were analysed. Antagonistic effects of atropine, the M-2-selective Ro 320-6206 and the M-3-selective darifenacin were unchanged. Agonistic effects of the M-3-sparing agonist 4-(2-oxo-2,3-dihydro-benzoimidazol-1-yl)-[1,4']bipiperidinyl-1'-carboxyl ic acid ethyl ester were similarly poor in young and old rats. Additional experiments were concomitantly performed in submandibular glands from the same animals. While total muscarinic receptor density in submandibular glands was not significantly affected by age (56 +/- 5 vs. 61 +/- 4 fmol/mg protein), the relative contribution of M-3 receptors significantly declined from 68 +/- 3% to 57 +/- 2% based upon darifenacin competition curves. We conclude that aged Wistar rats express fewer muscarinic receptors in their urinary bladder, but there is no change in the relative abundance of M-2 and M-3 receptors; this is accompanied by only minor if any alterations in receptor responsiveness. In contrast, submandibular gland expresses similar receptor numbers in young and old rats, but slightly fewer M-3 receptors in old animals

Muscarinic receptor antagonists such as oxybutynin and tolterodine are the most widely prescribed forms of treatment of the syndrome of overactive bladder and urge urinary incontinence. Recently, three new muscarinic antagonists, that is, darifenacin (Enablex®), solifenacin (Vesicare®) and trospium (Sanctura®) have been introduced in the US. We review and compare their efficacy and tolerability with an emphasis on the data from the studies performed for the US registration purposes.

Die Pravalenz von Harninkontinenz ist bei beiden Geschlechtern hoch. Frauen sind haufiger von einer Stress- und Manner haufiger von einer Dranginkontinenz betroffen, andere Inkontinenzarten sind demgegenuber selten. Pathophysiologisch liegt bei Stressinkontinenz ein insuffizienter Harnrohrenverschluss und bei Dranginkontinenz unwillkurliche Detrusorkontraktionen vor. Medikamente sind ein Teil im Gesamtkonzept der Inkontinenztherapie. Bei Stressinkontinenz ist es mit dem neuen Wirkstoff Duloxetin, einem Serotonin- und Noradrenalinwiederaufnahmehemmer, moglich, den urethralen Verschlussdruck zu erhohen und Inkontinenzepisoden um bis zu 64% zu reduzieren. Bei Dranginkontinenz bewirken Muskarinrezeptorantagonisten eine Relaxierung des Detrusors, was eine signifikante Abnahme der Miktionsfrequenz und Inkontinenzepisoden sowie eine Zunahme der Blasenkapazitat zur Folge hat. Trospiumchlorid, Tolterodin, Solifenacin, Oxybutynin und Propiverin sind in Deutschland zugelassene antimuskarinerge Medikamente. Bei geringen Unterschieden in der Effektivitat weisen diese Medikamente z. T. grose Unterschiede bei Nebenwirkungen auf. Mundtrockenheit ist haufig ein therapielimitierender Faktor. Keines der zugelassenen Medikamente hat allerdings bei allen Patienten eine ausreichende Wirkung, weshalb sich zurzeit neue Medikamente mit anderen Wirkmechanismen in der Erprobung befinden.

Cyclic AMP is the prototypical second messenger of beta-adrenergic receptors, but recent findings have questioned its role in mediating smooth muscle relaxation upon beta-adrenergic receptor stimulation. We have investigated the signaling mechanisms underlying beta-adrenergic receptor-mediated relaxation of rat urinary bladder. Concentration-response curves for isoproterenol-induced bladder relaxation were generated in the presence or absence of inhibitors, with concomitant experiments using passive tension and KCl-induced precontraction. The adenylyl cyclase inhibitor 9-(tetrahydro-2-furanyl)-9H-purin-6-amine (SQ 22,536; 1 microM), the protein kinase A inhibitors 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7; 10 microM), N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H89; 1 microM), and Rp-adenosine 3',5'-cyclic monophosphorothioate (Rp-cAMPS; 30 microM), and the guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (ODQ; 3 microM) produced only minor if any inhibition of relaxation against passive tension or KCl-induced precontraction. Among various potassium channel inhibitors, BaCl2 (10 microM), tetraethylammonium (3 microM), apamin (300 nM), and glibenclamide (10 microM) did not inhibit isoproterenol-induced relaxation. Some inhibition of the isoproterenol effects against KCl-induced tone but not against passive tension was seen with inhibitors of calcium-dependent potassium channels such as charybdotoxin and iberiotoxin (30 nM each). A combination of SQ 22,536 and ODQ significantly inhibited relaxation against passive tension by about half, but not that against KCl-induced tone. Moreover, the combination failed to enhance inhibition by charybdotoxin against KCl-induced tone. We conclude that cAMP and cGMP each play a minor role in beta-adrenergic receptor-mediated relaxation against passive tension, and calcium-dependent potassium channels play a minor role against active tension.

Purpose: Two thirds of patients with overactive bladder (OAB) are continent, but our knowledge on the treatment of the syndrome is largely based on studies with incontinent patients. Therefore, we have explored baseline symptoms and treatment responses to tolterodine in continent relative to incontinent OAB patients. Materials and Methods: Data from an open-label, observational study involving 3 824 patients with OAB symptoms were analyzed for baseline symptoms and alterations thereof upon a 9 months treatment with 4 mg q.d. tolterodine ER. Results: Baseline symptoms (number of urgency episodes, total urination frequency, daytime frequency, nocturia, and three scales of OAB severity) were similar in 1147 continent and 2571 incontinent patients, the latter having 4.8 +/- 3.7 incontinence episodes per 24 h. Tolterodine ER-induced reduction of OAB symptoms was very similar in both groups of patients. Conclusions: The severity of OAB in continent patients can be similar to that in incontinent ones. Symptom improvement upon treatment with tolterodine ER is very similar in both groups. We propose that continent patients may similarly deserve treatment with a muscarinic receptor antagonist as incontinent ones. (c) 2004 Elsevier B.V. All rights reserved

Urine storage and micturition are tightly controlled by the peripheral and central nervous systems. The parasympathetic pelvic nerves mediate signals for voiding, whereas the sympathetic hypogastric and the somatic pudendal nerves mediate signals for urine storage. Glutamate is a key transmitter of the descending neuronal pathways during the storage phase of the micturition cycle. Its effects on the activity of the hypogastric and pudendal nerves are modulated by serotonin and/or noradrenaline. Duloxetine inhibits the neuronal uptake of serotonin and noradrenaline and thereby enhances their effects in the central nervous system including the spinal nuclei responsible for urine storage. Thus, in a bladder irritation model in anaesthetised cats duloxetine dose-dependently restored bladder capacity and enhanced activity of the external urethral sphincter/rhabdosphincter. Such effects should prove beneficial in the treatment of patients with stress urinary incontinence. (C) 2004 Elsevier B.V. All rights reserved

The beta-blocker nebivolol is a racemic mixture of D- and L- enantiomers that displays negative inotropic as well as direct vasorelaxant activity. In addition, it has been proposed that nebivolol exerts endothelium-protective effects caused by its antioxidant properties. In the present study we investigated the effect of D-, L-, and d/l-nebivolol on reactive oxygen species (ROS)-induced endothelial damage and compared it with carvedilol and metoprolol. Isolated rat aortic rings were exposed to ROS by electrolysis of the organ bath medium. Before and after electrolysis, endothelial function was measured by preconstricting the vessels with phenylephrine followed by the addition of methacholine. Carvedilol and nebivolol protected against ROS-induced endothelial damage, whereas metoprolol did not. The protective effect of nebivolol proved not to be stereoselective. Furthermore, we attempted to determine whether nebivolol acts a scavenger itself or whether another mechanism is involved. By means of HPLC measurements it was shown that nebivolol concentrations were decreased after exposure to electrolysis-induced ROS, thus indicating that nebivolol is degraded by its reaction with ROS. Functional experiments, in the rat aorta, demonstrated that exposure of nebivolol to ROS also affects its vasodilator activity. In conclusion, the present study demonstrates that nebivolol alleviates ROS-induced impairment of endothelium-dependent vasorelaxation. This protective effect is very likely the result of a direct ROS-scavenging action by the nebivolol molecule itself.

In recent years it has been shown that angiotensin II (Ang II) stimulates formation of reactive oxygen species (ROS), presumably by activation of NAD(P)H oxidase. This ROS formation has been primarily associated with cellular growth regulation by Ang II. The objective of the present study was to investigate whether these ROS contribute to Ang II-induced vasoconstriction. Experiments were performed in isolated rat thoracic aorta. Concentration response curves were constructed for Ang II in the absence and presence of the NAD(P)H oxidase inhibitor DPI, and ROS scavengers catalase and EUK-8. Inhibition of NAD(P)H oxidase as well as scavenging of ROS, decreased the contractile response to Ang II. Administration of NADPH, a substrate for NAD(P)H oxidase, produced vasoconstriction that proved to be sensitive for DPI, catalase, and EUK-8. Exposure of the vessels to exogenous ROS, induced by electrolysis of the organ bath medium, also resulted in a contractile response that was decreased by ROS scavenging. The results suggest that ROS play a role in Ang II-induced vasoconstriction via the activation of NAD(P)H oxidase.

Having been founded in 1873 as Archiv fur experimentelle Pathologie und Pharmakologie, Naunyn-Schmiedeberg’s Archives of Pharmacology is the oldest existing pharmacology journal. Historically, it has been a flagship journal for German language pharmacology but following publication of papers in English since 1973, it has increasingly become international (Starke 1998). In the past, most published primary research in this and many other leading pharmacology journals came from Western Europe, Northern America and Japan, but in recent years, other parts of the world increasingly contribute to the advancement of pharmacological knowledge. To explore how this is reflected in the pharmacological literature, we have compared the countries of origin of published papers in Naunyn-Schmiedeberg’s Archives of Pharmacology in 2001 and 2010.

β(3)-adrenoceptor agonists are currently in clinical development for the treatment of overactive bladder and considered for several other indications. This Perspective discusses desirable properties of such drugs mainly based on the example of overactive bladder, but at least partly they should also be applicable to other indications of β(3)-adrenoceptor agonists or other drug classes and therapeutic areas. These include degree of selectivity for the molecular target in terms of affinity, intrinsic efficacy and ligand-directed signaling. The ability to cause agonist-induced desensitization and the potential impact of gene polymorphisms also need to be considered. Depending on intended indication, specific pharmacokinetic considerations may also apply. These findings challenge the usefulness of high-throughput screening assays based upon a single molecular response in an artificial system and emphasize the need for early use of in vivo testing in species considered to be predictive for the human situation

Serum activity of the enzyme secretory sphingomyelinase (sSM) is higher in patients with congestive heart failure (CHF) than in those with arterial hypertension or in healthy controls; more importantly, this elevation is not related to the aetiology of CHF, but rather to its severity, and well correlated with peak oxygen uptake, cytokine activation, skeletal muscle strength, and peripheral vasodilator capacity.1 In a proportional hazard analysis, serum sSM activity was related to survival independent of age, NYHA class, or blood pressure. As sSM is largely derived from the endothelium, these findings highlight the emerging role of endogenous sphingolipids as regulators of cardiovascular function and the role of the endothelium in such regulation. Against this background, we will highlight some recent findings on the complex interplay between the local formation of sphingomyelin metabolites and the endothelium. Various stressful stimuli can activate different isoforms of sphingomyelinase, which catalyses the hydrolysis of sphingomyelin to ceramide. Ceramidase can metabolize ceramide further into sphingosine, which in turn can be phosphorylated by sphingosine kinase (SphK) to yield sphingosine-1-phosphate (S1P). Other enzymes allow for a reversal of such reactions and/or can form other biologically active sphingomyelin metabolites such as sphingosylphosphorylcholine. S1P is a ligand for at least five subtypes of G-protein coupled receptors, designated S1P1–5, which were originally described as endothelial differentiation genes. S1P1–3 are the major S1P receptor subtypes expressed in the cardiovascular system, in both endothelium and vascular smooth muscle cells; at least at the mRNA level, S1P1 appears to be the most abundantly expressed subtype in the endothelium.2 In many cases, ceramide and sphingosine on the one and S1P on the other hand have opposite effects on cellular function, e.g. by stimulating cell death and apoptosis vs. cell growth and differentiation, respectively. Accordingly, sphingomyelinases … *Corresponding author. Tel: +31-20-566-6762; fax: +31-20-696-5976. E-mail address : m.c.michel{at}amc.uva.nl