Fibrinolysis inhibitors in plaque stability: a morphological association of PAI-1 and TAFI in advanced carotid plaque.

Essentials Fibrinolysis inhibitors are localized in advanced atheroma by immunohistology of endarterectomies. Neovascular endothelium/neocapillaries show thrombin-activatable fibrinolysis inhibitor (TAFI). Macrophage areas show free plasminogen activator inhibitor (PAI-1), notably in the vulnerable part. Free PAI-1 and TAFI stabilize active plaque area by inhibition of fibrinolysis and inflammation. SUMMARY: Background Fibrinolysis plays an important role in destabilization of atherosclerotic plaques and is tightly regulated by specific inhibitors. Objective The fibrinolysis inhibitors plasminogen activator inhibitor type-1 (PAI-1) and thrombin-activatable fibrinolysis inhibitor (TAFI) were quantified and described in the morphological context of advanced carotid plaques American Heart Association VI-VIII to elucidate their role in plaque stability. Methods Immunohistochemistry in serial sections along the longitudinal axis of endarterectomies from patients with symptomatic carotid stenosis (n = 19) were studied using an antibody specific for free PAI-1 (I205), an antibody with high affinity for TAFI/TAFIa (CP17) and established antibodies for smooth muscle cells (α-actin), endothelial cells (von Willebrand factor [VWF]), macrophages (CD68) and platelets (CD42). Results PAI-1 and TAFI show a specific distribution in these advanced plaques with a maximum corresponding to the internal carotid artery (ICA). Free PAI-1 was mainly detected in macrophages and in intravascular thrombi, and TAFI in endothelial cells (ECs) but also macrophages. The one-way ANOVA analysis with Bonferroni's correction showed a significant increase of macrophages and ECs, TAFI and PAI-1 in areas with high neovascularization in endarterectomy sections corresponding to ICA. High Spearman factors for TAFI, PAI-1 and VWF indicate neovascularization as the main source of plasma proteins, transported by platelets into the atheroma (PAI-1) or expressed by ECs (TAFI). CD68 was highly associated with VWF, PAI-1 and especially TAFI, underlining the role of macrophages in fibrinolytic activity and inflammation. Conclusion The abundance of free PAI-1 and TAFI in the plaque may inhibit plasmin generation and thereby counteract plaque destabilization by fibrinolysis, cell migration and inflammation.

Neuropeptide Y and its Y2 receptor: potential targets in neuroblastoma therapy.

Neuroblastomas are pediatric tumors that develop from sympathetic precursors and express neuronal proteins, such as neuropeptide Y (NPY). NPY is a sympathetic neurotransmitter acting via multiple receptors (Y1-Y5R). Both NPY and Y2Rs are commonly expressed in neuroblastoma cell lines and tissues. The peptide secreted from neuroblastomas stimulates tumor cell proliferation and angiogenesis. As both processes are Y2R-mediated, the aim of this study was to assess Y2R as a potential therapeutic target for neuroblastoma. In vitro, Y2R antagonist (BIIE0246) prevented activation of p44/42 mitogen-activated protein kinase (MAPK) induced by endogenous NPY, which resulted in decreased proliferation and induction of Bim-mediated apoptosis. Similar growth-inhibitory effects were achieved with NPY small interfering RNA (siRNA) and Y2R siRNA. In vivo, Y2R antagonist significantly inhibited growth of SK-N-BE(2) and SK-N-AS xenografts, which was associated with decreased activation of p44/42 MAPK, as well as reduced proliferation (Ki67) and increased apoptosis (TdT-mediated dUTP nick end labeling; TUNEL). The Y2R antagonist also exerted an antiangiogenic effect. In vitro, it reduced the proliferation of endothelial cells induced by neuroblastoma-conditioned media. Consequently, the Y2R antagonist-treated xenografts had decreased vascularization and a high degree of focal fibrosis. In human neuroblastoma tissues, the expression of Y2R was observed in both tumor and endothelial cells, while NPY was predominantly expressed in neuroblastoma cells. In summary, Y2R is a promising new target for neuroblastoma therapy affecting both cancer cells and tumor vasculature.

Diagnostic performance of routine ultrasound screening for fetal abnormalities in an unselected Swedish population in 2000-2005.

OBJECTIVES: To determine the detection rate of fetal malformations and chromosomal abnormalities and the rate of false-positive ultrasound diagnoses at routine ultrasound examinations carried out by specially trained midwives in an unselected pregnant population from 2000 to 2005, and to describe the consequences of true-positive and false-positive ultrasound diagnoses of fetal malformations. METHODS: A retrospective analysis was undertaken of all babies born in Malmö, Sweden, between January 2000 and December 2005 by mothers residing in Malmö and of all fetuses with an ultrasound diagnosis of malformation made in the same time interval at the two units performing all routine pregnancy scans in Malmö. All women underwent two routine scans, at 18 and 32 weeks, including scrutiny of the fetal anatomy. Detection rates and false-positive rates were calculated per fetus. RESULTS: The prevalence of chromosomally abnormal fetuses was 0.31% (52/16 775); that of chromosomally normal fetuses with major and minor malformations was 1.80% (302/16 775) and 1.32% (222/16 775), respectively. The detection rate of fetuses with major malformations but normal chromosomes was 68% (205/302), with a detection rate at < 22 weeks of 37% (112/302). In addition, 46% (24/52) of all chromosomally abnormal fetuses were diagnosed before birth because a malformation was detected at ultrasound imaging, 33% (17/52) being detected at < 22 gestational weeks. In all, 68 pregnancies were terminated because of an ultrasound diagnosis of fetal malformation (0.4% of all pregnancies and 47% of the pregnancies in which a fetal malformation was detected by ultrasound examination before 22 weeks). A false-positive ultrasound diagnosis of malformation was made in 0.19% (31/16 180) of the normally formed fetuses and in 20 (0.12%) fetuses the abnormal finding persisted during pregnancy. No fetus assigned a false-positive diagnosis was lost by termination of pregnancy, but most were subjected to one or more unnecessary interventions before birth (e.g. amniocentesis), at birth (e.g. Cesarean section) or after birth (e.g. electrocardiogram, X-ray, ultrasound examination or treatment with antibiotics). CONCLUSIONS: In a screening program consisting of one fetal anomaly scan at 18 weeks and another at 32 weeks the detection rate of major malformations in chromosomally normal fetuses was 68% with a detection rate of 37% at < 22 weeks. The corresponding detection rates of chromosomally abnormal fetuses were 46% and 33%. Fewer than one in 500 screened fetuses had an ultrasound diagnosis of an anomaly that was not confirmed after birth.

The role of GABA(A) receptors in the control of transient lower oesophageal sphincter relaxations in the dog.

BACKGROUND AND PURPOSE: Transient lower oesophageal sphincter relaxations (TLESRs) are triggered by activation of mechanosensitive gastric vagal afferents and are the major cause of gastroesophageal reflux and therefore an important target for therapeutic intervention in gastroesophageal reflux disease (GERD). Activation of the metabotropic GABA(B) receptor has shown to inhibit TLESRs. The aim of the present study was to assess the role of the ionotropic GABA(A) receptor in the regulation of TLESRs. EXPERIMENTAL APPROACH: TLESRs were quantified using Dentsleeve manometry in dogs, and GABA(A) agonists were given i.v. prior to gastric distension. Immunohistochemistry and RT-PCR were used to localize GABA(A) receptors in the dog nodose ganglion, the source of vagal afferents which initiate TLESRs. KEY RESULTS: The prototypical GABA(A) agonist muscimol produced a dose-dependent inhibition of TLESRs ranging from 19 to 56%. The two other GABA(A) agonists evaluated, isoguvacine and 4,5,6,7-tetrahydroisoxazolo-[5,4-c]pyridin-3-ol (THIP), as well as the GABA(A) positive allosteric modulator diazepam, had no major effects on TLESRs. Evaluation of higher doses was limited by emesis (THIP and isoguvacine) or restlessness/sedation (diazepam). Of the predominant GABA(A) receptor subunits (alpha, beta and gamma components), alpha and beta but not gamma were detected in the dog nodose ganglion by RT-PCR, while immunohistochemistry in addition demonstrated nerve fibres expressing the gamma subunit. CONCLUSIONS AND IMPLICATIONS: The present observations demonstrate that GABA(A) receptors exert an inhibitory control of TLESRs. These results warrant further studies using GABA(A) isoform-selective agonists to define the identity of receptors involved.

Characterization of the cloned atlantic cod neuropeptide Y-Yb receptor: peptide-binding requirements distinct from known mammalian Y receptors.

Five members of the neuropeptide Y (NPY) receptor family have been cloned in mammals. The recently cloned NPY receptor in the Atlantic cod seems to be distinct from the mammalian subtypes as it has only 50% identity to Y1, Y4, and y6 and only 30% to Y2 and Y5. In most of the other families of G-protein-coupled receptors, species homologues have 65-90% identity between fishes and mammals. The functional expression and detailed pharmacological characterization of this cod NPY receptor, designated Yb, is reported. Membranes of cells transiently transfected with cod Yb showed saturable [(125)I]PYY binding with a K(d) of 45 pM. The pharmacological profile is similar to those of both the zebrafish Yb and Yc receptors and distinct from those of the mammalian NPY receptors. In competition experiments the cod Yb receptor had the following rank order of potencies: porcine PYY = porcine NPY = p[Leu(31), Pro(34)]NPY > zebrafish PYY > zebrafish NPY >> NPY2-36 = NPY3-36 > NPY18-36 > bovine PP = [D-Trp(32)]NPY > BIBP3226. This is in sharp contrast to the high selectivity of BIBP3226 for the Y1 receptor from all mammalian species. Together with the low amino acid identity of cod Yb with the mammalian Y1, Y4, and y6 receptors, this is further support for the notion that fish Yb constitutes a distinct NPY receptor subtype.

Cloning of a neuropeptide Y/peptide YY receptor from the Atlantic cod: the Yb receptor.

Neuropeptide Y (NPY) belongs to a family of structurally related neuroendocrine peptides that bind to G protein-coupled receptors. Five different receptor subtypes have recently been cloned in mammals and we have found another three receptor genes in the zebrafish, called zYa, zYb, and zYc, that appear to be distinct subtypes as deduced from their widely different sequences. To elucidate the evolutionary relationships between the mammalian and zebrafish receptors, we have used the zebrafish probes to isolate genomic clones from another teleost fish, the Atlantic cod, Gadus morhua. We present here the sequence of the cod Yb gene, whose deduced protein sequence is equally identical to the zebrafish Yb (69%) and Yc proteins (66%). The two zebrafish receptors are 76% identical to each other, suggesting that they arose by gene duplication in the zebrafish lineage after divergence from the cod lineage. The five cloned mammalian NPY-family receptors and the three cloned zebrafish NPY receptors indicate that this is the largest receptor family among all peptide receptors that belong to the superfamily of G protein-coupled receptors.

Quality of life and response of negative symptoms in schizophrenia to haloperidol and the atypical antipsychotic remoxipride. The Canadian Remoxipride Group.

In a large, multicenter, double-blind study of the effect of haloperidol and the atypical antipsychotic remoxipride on improvement of negative symptoms in schizophrenia, quality of life was also assessed using a modified version of the Sickness Impact Profile (SIP). Compared with previous studies, this study had a longer duration (28 weeks), and the dose of the comparator, haloperidol, was much lower. At the end of the study, compared with the baseline, both treatment groups reported comparable improvement in negative symptoms as defined by the protocol (at least 20% improvement). Similarly, both groups showed comparable changes on global and multidimensional self-assessments of quality of life. All the subfactors of the modified version of the SIP were similar in both groups, except for the subfactor that relates to alertness behavior, which possibly reflects remoxipride's lack of any sedating properties compared with haloperidol. This study presents an approach for inclusion of quality of life as an outcome measure in the design of clinical trials of new antipsychotic medications.