S-train as an alternative for the expansion of the Cieszyńska street in Bielsko-Biała

W artykule przedstawiono wybrane analizy dotyczące zastosowania szybkiej kolei miejskiej jako alternatywy dla rozbudowy ulicy Cieszyńskiej w Bielsku-Białej. Autorzy dokonali oceny efektywności układu drogowego dla zmodernizowanej ulicy Cieszyńskiej oraz przedstawili podstawowe założenia dla utworzenia szybkiej kolei miejskiej z wykorzystanym aktualnie wyłączonej z eksploatacji linii kolejowej.The article presents selected analyzes regarding to the use of S-train as an alternative to the extension of Cieszyńska Street in Bielsko-Biała. The authors assessed the efficiency of the road system for the modernized Cieszyńska Street and presented the basic assumptions for the creation of S-train with the used of currently excluded railway line

Vasopressin anti-idiotypic antibody staining in the rat brain: colocalization with [³⁵S] [pGlu⁴, Cyt⁶]AVP_(4-9) binding sites

Vasopressin and its fragment peptides such as [pGlu4, Cyt6]AVP(4-9) represent putative neuromodulators within central nervous homeostatic, memory and behavioural circuits. To localize their central receptor systems, the previously characterized monoclonal anti-idiotypic antibody mAb 237 was employed in immunocytological investigations of rat brain tissue sections. This antibody was raised to the monoclonal idiotypic anti-AVP antibody mAb 113 which preferentially binds to the acyclic C-terminal portion of the AVP molecule and is therefore also capable of binding the naturally occurring AVP(4-9) fragment. Immunoreactive magnocellular neurones were detected in the AVP-synthesizing supraoptic but not paraventricular nuclei. Dense staining was observed within circumventricular organs lacking a blood-brain barrier (BBB). These structures include the subfornical organ, the organum vasculosum laminae terminalis, the internal layer of the median eminence, the body of the pineal gland, the choroid plexus and the area postrema, where immunoreactivity was found on capillaries, neurones and fibres. Further staining was found in the nucleus of the solitari tract and the arcuate nucleus, endowed with a leaky BBB. Distinct cell patches in the ependymal lining of the third ventricle as well as dendritic processes of juxtaependymal neurones were labelled by the anti-idiotypic antibody mAb 237. The observed staining pattern did not parallel that obtained in autoradiographic studies performed using either radiolabelled AVP or a V1-receptor antagonist, but that found with the [35S]-labelled AVP(4-9) fragment. Using [35S]-labelled AVP(4-9) fragment, specific high density binding sites could be localized autoradiographically in structures within and outside the BBB, in complete agreement with the anti-idiotypic immunoreactivity. Since the anti-idiotypic methodology is based on transfer of complementary structures, and the epitope recognized by the corresponding idiotypic antibody resembles the sequence of AVP(4-9), the anti-idiotypic antibodies might recognize the AVP(4-9) receptor with high affinity

Lack of interaction of vasopressin with its antisense peptides: a functional and immunological study

The peptide encoded in the 5' to 3' direction by rat vasopressin complementary RNA, rat PVA (H-Ser-Ser-Trp-Ala-Val-Leu-Glu-Val-Ala- OH) and the corresponding bovine PVA (H-Ala-Pro-Trp-Ala-Val-Leu-Glu-Val-Ala-OH) were investigated with respect to their interaction with [8-arginine] vasopressin (AVP) and V2 vasopressin receptor binding and function. Rat or bovine PVA did neither affect the binding of the hormone to the V2 receptor of bovine kidney membranes and LLC-PK1 pig kidney cells nor influence the AVP-induced cAMP-production in LLC-PK1 cells. Rat PVA was further investigated by the use of vasopressin-specific polyclonal and monoclonal antibodies with different affinity and epitope specificity. Consistent with receptor binding studies no inhibition of [3H]AVP-binding in fluid- or solid-phase antibody binding tests after preincubation with PVA was found. Direct interaction of rat PVA and [3H]AVP measured on solid surface was not observed in contrast to specific binding of the hormone with NP II and antibodies. In our study no evidence for an interaction of AVP and its antisense peptides was found

Renal and central vasopressin receptors: immunocytochemical localization

Employing an anti-vasopressin monoclonal antibody for immunization, anti-idiotypic monoclonal antibodies were obtained which induced plasminogen activator production in the renal epithelial cell line LLC-PK1. The anti-idiotypic antibodies were employed to visualize vasopressin receptors on LLC-PK1 and A7r5 smooth muscle cells by immunofluorescence. All results indicated specificity of the anti-idiotypes for both V1 and V2 vasopressin receptor subtypes. These antibodies were used for immunohistochemical localization of vasopressin receptors in rat and bovine kidney preparations. In accordance with earlier physiological and biochemical observations, vasopressin receptors were detected predominantly in collecting ducts in cortex and medulla. On the cellular level, a differential staining pattern was observed. On rat brain tissue sections, dense staining was observed within various circumventricular organs. The staining pattern corresponded to that obtained in autoradiographic studies with labeled AVP(4-9) fragment peptide and differed from the distribution of binding sites for labeled vasopressin or V1 antagonists

A new tritiated oxytocin receptor radioligand—Synthesis and application for localization of central oxytocin receptors

A new tritiated oxytocin antagonist radioligand was synthesized by introducing a tritiated propionic acid residue into the free amino group of ornithine in position 8 of the parent peptide [1-(β-mercapto-β,β-cyclopentamethylene propionic acid), 2-(O-methyl)-tyrosine, 4-threonine, 8-ornithine, 9-tyrosylamide]vasotocin (OTA), that was previously described. The tritiated compound [3H][1-(β-mercapto-β,β-cyclopentamethylene propionic acid), 2-(O-methyl)-tyrosine, 4-threonine, 8-(Nδ-propionyl)-ornithine, 9-tyrosylamide]vasotocin ([3H]PrOTA) was obtained in good yield with high specific activity (100 Ci/mmol). [3H]PrOTA exhibited the same affinity (Kd = 0.8 nM) and selectivity for the myometrial oxytocin receptor as the iodinated antagonist [125I]OTA. Autoradiographic localization of oxytocin receptors in the rat brain showed specific binding sites for [3H]PrOTA within regions of the limbic system, the neocortex, and hypothalamus, which is consistent with the binding pattern obtained with [125I]OTA. The high specific activity in combination with the long half-life of tritium and its low radiotoxicity as compared to iodine-125 makes the new tritiated antagonist a valuable tool for pharmacological studies

Monoclonal antibodies against different epitopes of peptide hormones

In order to produce monoclonal antibodies directed against different epitopes of the neurohypophyseal hormone vasopressin, the hormone was coupled to carrier proteins via photoreactive groups at different positions in the vasopressin sequence: [2‐(4‐azidophenylalanine), 8‐arginine]vasopressin (peptide P1, photoreactive group at position 2) and desamino‐[8‐N6‐(4‐azidophenylamidino)lysine]vasopressin (peptide P2, photoreactive group at position 8) were conjugated to thyroglobulin by flash photolysis. Monoclonal antibodies against these conjugates bound {[3H]8‐arginine}vasopressin with dissociation constants ranging over 40–400 nM. Epitope analysis by means of competitive ELISA showed that the monoclonal antibody obtained with peptide P1 as hapten was directed against the C‐terminal acyclic tripeptide when its conformation was stabilized by interaction with the disulphide‐linked cyclic hexapeptide. In contrast, the epitope analysis of three monoclonal anti‐(peptide P2) antibodies demonstrated that they recognized antigenic determinants in the cyclic hexapeptide ring, mainly the hydrophobic surface formed by Tyr2 and Phe3. Our results suggest that monoclonal antibodies against different epitopes in small peptide hormones can be generated selectively by using photoreactive peptides in such a way that different antigenic sites are exposed in the hapten‐carrier conjugate