Novel dimeric DOTA-coupled peptidic Y1-receptor antagonists for targeting of neuropeptide Y receptor-expressing cancers

ABSTRACT:

Preclinical Evaluation of Radiolabeled DOTA-Derivatized Cyclic Minigastrin Analogs for Targeting Cholecystokinin Receptor Expressing Malignancies

Targeting of cholecystokinin receptor expressing malignancies such as medullary thyroid carcinoma is currently limited by low in vivo stability of radioligands. To increase the stability, we have developed and preclinically evaluated two cyclic 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA)-minigastrin analogs radiolabeled with (111)In and (68)Ga.

Value of the radiolabelled GLP-1 receptor antagonist exendin(9-39) for targeting of GLP-1 receptor-expressing pancreatic tissues in mice and humans

Radiolabelled glucagon-like peptide 1 (GLP-1) receptor agonists have recently been shown to successfully image benign insulinomas in patients. Moreover, it was recently reported that antagonist tracers were superior to agonist tracers for somatostatin and gastrin-releasing peptide receptor targeting of tumours. The present preclinical study determines therefore the value of an established GLP-1 receptor antagonist for the in vitro visualization of GLP-1 receptor-expressing tissues in mice and humans.

Induction of p38, tumour necrosis factor-α and RANTES by mechanical stretching of keratinocytes expressing mutant keratin 10R156H

Epidermolytic hyperkeratosis (bullous congenital ichthyosiform erythroderma), characterized by ichthyotic, rippled hyperkeratosis, erythroderma and skin blistering, is a rare autosomal dominant disease caused by mutations in keratin 1 or keratin 10 (K10) genes. A severe phenotype is caused by a missense mutation in a highly conserved arginine residue at position 156 (R156) in K10.

Psychosocial stress and cardiovascular risk : current opinion

Cardiovascular disease (CVD) is a major cause of morbidity and mortality worldwide. Epidemiologic research of the last half-century has clearly shown that psychosocial factors related to the social environment, personality characteristics, and negative affect increase the risk of incident CVD and also impact prognosis of cardiac patients. Several mechanisms may explain this link, including a genetic predisposition, poor lifestyle choices, low adherence to health recommendations, and direct pathophysiologic perturbations. The latter include alteration of the hypothalamic-pituitary adrenal axis and autonomic dysfunction resulting in endothelial dysfunction, inflammation, and a prothrombotic state further downstream. Screening for psychosocial factors seems appropriate as part of the standard history and based on the clinician's knowledge of the patient and the purpose of the visit. Psychological interventions generally alleviate distress in cardiac patients, but whether they reduce the risk of hard cardiovascular endpoints and all-cause mortality is less evident. Cardiac patients with more severe depression may particularly profit from antidepressant medications. Due to their pharmacologic properties, selective serotonin reuptake inhibitors were shown to improve cardiovascular outcome. The most effective psychosocial treatment is multicomponent therapy that combines elements of cognitive behaviour therapy ("stress management") and changes in health behaviours, including the adoption of a regular exercise regimen. Gender-specific issues should probably be considered. The field of behavioural cardiology has accumulated a wealth of epidemiological, mechanistic and clinical knowledge that undoubtedly has furthered our understanding about the important role of psychosocial risk factors in patients with a heart disease.

Intraperitoneal murine Echinococcus multilocularis infection induces differentiation of TGF-beta-expressing DCs that remain immature

Intraperitoneal larval infection (alveolar echinococcosis, AE) with Echinococcus multilocularis in mice impairs host immunity. Metacestode metabolites may modulate immunity putatively via dendritic cells. During murine AE, a relative increase of peritoneal DCs (pe-DCs) in infected mice (AE-pe-DCs; 4% of total peritoneal cells) as compared to control mice (naive pe-DCs; 2%) became apparent in our study. The differentiation of AE-pe-DCs into TGF-beta-expressing cells and the higher level of IL-4 than IFN-gamma/IL-2 mRNA expression in AE-CD4+pe-T cells indicated a Th2 orientation. Analysis of major accessory molecule expression on pe-DCs from AE-infected mice revealed that CD80 and CD86 were down-regulated on AE-pe-DCs, while ICAM-1(CD54) remained practically unchanged. Moreover, AE-pe-DCs had a weaker surface expression of MHC class II (Ia) molecules as compared to naive pe-DCs. The gene expression level of molecules involved in MHC class II (Ia) synthesis and formation of MHC class II (Ia)-peptide complexes were down-regulated. In addition, metacestodes excreted/secreted (E/S) or vesicle-fluid (V/F) antigens were found to alter MHC class II molecule expression on the surface of BMDCs. Finally, conversely to naive pe-DCs, an increasing number of AE-pe-DCs down-regulated Con A-induced proliferation of naive CD4+pe-T cells. These findings altogether suggested that TGF-beta-expressing immature AE-pe-DCs might play a significant role in the generation of a regulatory immune response within the peritoneal cavity of AE-infected mice.

CD4(+)CD25(+) T cells expressing FoxP3 in Icelandic horses affected with insect bite hypersensitivity

Insect bite hypersensitivity (IBH) is an IgE-mediated dermatitis caused by bites of midges from the genus Culicoides. We have shown previously that peripheral blood mononuclear cells (PBMC) from IBH-affected horses produce higher levels of IL-4 and lower levels of IL-10 and TGF-beta1 than those from healthy horses, suggesting that IBH is associated with a reduced regulatory immune response. FoxP3 is a crucial marker of regulatory T cells (Tregs). Here we have determined the proportion of CD4(+)CD25(+)FoxP3(+) T cells by flow cytometry in PBMC directly after isolation or after stimulation with Culicoides extract or a control antigen (Tetanus Toxoid). There were no differences between healthy and IBH horses either in the proportion of FoxP3(+)CD4(+)CD25(+) cells in freshly isolated PBMC or in the following stimulation with Tetanus Toxoid. However, upon stimulation of PBMC with the allergen, expression of FoxP3 by CD4(+)CD25(+high) and CD4(+)CD25(+dim) cells was significantly higher in healthy than in IBH horses. Addition of recombinant IL-4 to PBMC from healthy horses stimulated with the allergen significantly decreased the proportion of FoxP3 expressing cells within CD4(+)CD25(+high). These results suggest that IBH is associated with a decreased number of allergen-induced Tregs. This could be a consequence of the increased IL-4 production by PBMC of IBH-affected horses.

[(99m)Tc]Demomedin C, a radioligand based on human gastrin releasing peptide(18-27): synthesis and preclinical evaluation in gastrin releasing peptide receptor-expressing models

The synthesis and preclinical evaluation of [(99m)Tc]Demomedin C in GRPR-expressing models are reported. Demomedin C resulted by coupling a Boc-protected N(4)-chelator to neuromedin C (human GRP(18-27)), which, after (99m)Tc-labeling, afforded [(99m)Tc]Demomedin C. Demomedin C showed high affinity and selectivity for the GRPR during receptor autoradiography on human cancer samples (IC(50) in nM: GRPR, 1.4 ± 0.2; NMBR, 106 ± 18; and BB(3)R, >1000). It triggered GRPR internalization in HEK-GRPR cells and Ca(2+) release in PC-3 cells (EC(50) = 1.3 nM). [(99m)Tc]Demomedin C rapidly and specifically internalized at 37 °C in PC-3 cells and was stable in mouse plasma. [(99m)Tc]Demomedin C efficiently and specifically localized in human PC-3 implants in mice (9.84 ± 0.81%ID/g at 1 h pi; 6.36 ± 0.85%ID/g at 4 h pi, and 0.41 ± 0.07%ID/g at 4 h pi block). Thus, human GRP-based radioligands, such as [(99m)Tc]Demomedin C, can successfully target GRPR-expressing human tumors in vivo while displaying attractive biological features--e.g. higher GRPR-selectivity--vs their frog-homologues.

Central Europeans towards Integration with the European Union: A Study of Public Opinion, Social Attitudes and National Identities

Since the turbulence of 1989, the countries of Central and Eastern Europe have striven to "return to Europe". Agreements have been signed with ten post-communist countries, beginning in 1991 with Czechoslovakia (before its division), Hungary and Poland. Since that time several countries have expressed a desire to become members of the EU. In 1997 the European Commission announced its opinion on the applications for EU membership of the Czech Republic, Hungary, Poland, Slovakia, and seven other applicant countries. The Commission recommended the commencement of negotiations on accession with the Czech Republic, Estonia, Hungary, Poland, and Slovenia. Mr. Kucia's report, presented in the form of a series of manuscripts totalling 91 pages, written in English and Polish and including many pages of tables and graphs, presents the results of a study of public opinion on European integration in four countries of Central Europe (CE): the Czech Republic (CZ), Hungary (H), Poland (PL), and Slovakia (SK). The research results are primarily based on a public-opinion survey known as the Central and Eastern Eurobarometer (CEEB). CEEB has been conducted on behalf of the European Commission in the Central and Eastern European countries each year in autumn since 1990. Below is a very small selection of Mr. Kucia's research findings. Throughout the 90s people in the four countries increasingly saw their countries' future tied up with the EU, since economic and political connections to the EU were growing and prospects for EU membership were increasing. Regional co-operation within CE did not gain much popular recognition. However, initially high levels of enthusiasm for the EU were gradually superseded by a more realistic approach or even scepticism. Poland was the exception in this respect; its population was more positive about the EU in 1996 than ever before. Mr. Kucia concludes that, since the political "elites" in CE are more positive about the EU than the people they serve, they should do their best to bring people round to their beliefs, lest the project of European integration become purely the business of the elites, as Mr. Kucia claims it has been in the EU up till now. He accuses the governments of the region, the EU authorities and the media of failing to provide appropriate information, especially about the two subjects which most affect them, association with the EU and the PHARE assistance programme. Respondents were asked to rank in order the countries or regions they saw their country's future most closely tied up with. In the period 92-96 the EU received the highest ratings in all of CE. The ratings were highest in CZ in 92 and 93 (46%) and in Poland in 96 (46%). They were the lowest in Hungary (22% in 94). After the EU came "Other Western European countries (non EU)", that is Austria, Sweden and Finland (before they joined the EU in 1995), Switzerland and Norway. Mr. Kucia puts the high ratings of these countries down to historical connections and geographical proximity, particularly in the case of Austria. The USA always came second in Poland, and in Hungary too its standing has always been higher than in CZ or SK. Indeed Mr. Kucia suggests that the USA's standing is disproportionately low in especially the CZ. Germany was nominated frequently by Hungarians, though in the CZ and SK, figures have been consistently low (1-2%). "Other CE/EE countries" increased their ratings in all of CE except Poland between 92 and 96. With regard to these last figures, Mr. Kucia makes an interesting note. Assuming that for the respondents in the four countries this category covered the Visegrad 4, least support was found in Poland, whose government was the most in favour of close political co-operation within the V4, while most support was in evidence in CZ and SK, for whose governments V4 was simply not a priority. Again, there is evidence of a divide between the political elites and the people. Russia has occupied a consistently modest rank. It was the highest in PL, fairly low in H and SK and the lowest in CZ. The Slovak government's policy of closer ties with Russia is reflected in a growth in the figures from 2% in 93 to 6% in 95. Every year the spontaneous answer "we should depend on ourselves" appeared, which Mr. Kucia interprets as either a sign of isolationism and disillusionment or as a call for self-reliance. Unfortunately he regards both these tendencies as unfeasible in the uniting Europe. Moving to more general conclusions, Mr. Kucia finds that the concept "Central Europe" does not have much meaning for Central Europeans. He believes that this is probably due to the failure to establish a viable regional co-operation network. Group discussions also revealed that people thought themselves European as a consequence of being Czech or Polish etc. Thus European identity is based on national identities. Generally within the surveyed period, the numbers of those who said they often think themselves European decreased, while the numbers of those who said they never think themselves European increased from 41% in PL, 36% in CZ, and 30% in H in 1990, to 67% in CZ, 58% in PL, and 51% in H in 1995.

Epidermal caspase-3 cleavage associated with interferon-gamma-expressing lymphocytes in acute atopic dermatitis lesions

Keratinocyte apoptosis mediated by Fas/Fas ligand molecular interactions and subsequent caspase activation is believed to play an important role in the pathogenesis of atopic dermatitis (AD), in particular for the formation of spongiosis. To estimate epidermal caspase activation in normal and AD skin under in vivo conditions, we analysed caspase-3 cleavage by immunohistology. In normal skin as well as non-lesional AD skin, we detected caspase-3 cleavage in single cells of the basal layer. In contrast, in acute lesional AD skin, we not only obtained evidence for increased expression of cleaved caspase-3 in keratinocytes of the basal layer but also observed caspase-3 cleavage in one or more layers of the spinous cell layer, in particular in spongiotic areas. Short-term topical treatment of the skin lesions with tacrolimus or pimecrolimus abolished the expression of cleaved caspase-3 in the spinous layer. Moreover, epidermal caspase-3 cleavage correlated with the numbers of dermal interferon-gamma (IFN-gamma)-expressing CD4+ and CD8+ lymphocytes in skin lesions of AD patients, supporting the view that IFN-gamma is important for the activation of proapoptotic pathways in keratinocytes. This is also confirmed by the observation of increased Fas expression on keratinocytes in acute AD lesions that was markedly reduced following topical calcineurin inhibitor treatment. These data suggest that caspase-3 cleavage in the spinous layer of the epidermis is a pathologic event contributing to spongiosis formation in AD, whereas cleavage of caspase-3 in basal cells might represent a physiologic mechanism within the process of epidermal renewal.