ABO blood type predicts the cytolocalization of anti-P-glycoprotein monoclonal antibody reactivity in human colon and ureter.

Classic multidrug resistance is mediated by a P-glycoprotein. Using monoclonal antibody C219 (MAb C219) in an immunohistochemical study, we found high levels of putative Golgi P-glycoprotein in normal columnar and transitional epithelium in subpopulations of patients with specific blood types. For example, Golgi staining was present in blood type A patients in 46% of normal colon samples (N = 21) and 88% of normal ureter samples (N = 17). In comparison, Golgi staining was present in blood group O patients in only 6% of normal colon samples (N = 34) and in 0% of normal ureter samples (N = 19). The association of MAb C219 Golgi staining with blood type A and lack of Golgi staining with blood type O was statistically significant in normal colon (P = .001) and normal ureter (P less than .0001). Inappropriate hyperexpression of P-glycoprotein was frequently found in colon carcinomas. Additional evidence that Golgi MAb C219 reactivity represents P-glycoprotein is presented. This includes (1) immunostaining of Golgi with two anti-P-glycoprotein MAbs, C219 and JSB-1, and (2) experiments in which Mab C219 Golgi reactivity was blocked by preincubation of MAb C219 with a specific P-glycoprotein epitope-containing peptide. The high degree of association of Golgi P-glycoprotein with blood type A may suggest a role for P-glycoprotein in processing or trafficking of specific blood group antigens.

Localization of a renal kallikrein immunoreactive-like substance in rat ureter.

An antibody against rat kallikrein was produced in rabbits and its localization was studied in various organs of the rat to confirm its specificity. The distribution of immunoreactive kallikrein was studied in rat ureter by use of immunochemical techniques. Ureteral tissue was fixed in Zamboni's-glutaraldehyde fixative and immunostained with indirect immunofluorescence and the peroxidase-antiperoxidase (PAP) method for light and electron microscopy. Preabsorption of the primary polyclonal antiserum with purified rat urinary kallikrein and substitution with normal serum were used as controls. By light microscopy, kallikrein was localized in the lamina propria and in the adventitial connective tissue surrounding the entire ureter. Immunoelectron microscopy confirmed this immunolocalization. Immunoreactive kallikrein was concentrated in fibroblasts of connective tissue and was not present in collagen fibers. Immunoreactivity was associated with the Golgi complex, free polyribosomes, and rough endoplasmic reticulum. No immunostaining was observed in other subcellular components of fibroblasts.

Presence of the Dr receptor in normal human tissues and its possible role in the pathogenesis of ascending urinary tract infection.

The Dr hemagglutinin of uropathogenic Escherichia coli recognizes the Dra blood group antigen, a component of the IFC or Cromer-related blood group complex. The present report used the Dr hemagglutinin to demonstrate location of the Dr receptor in selected human tissues and to evaluate the possible use of this lectin as a tissue marker recognizing sites sensitive for bacterial colonization. It was found that the Dr receptor was expressed in different parts of the digestive, urinary, genital, and respiratory tracts, and skin. Intense staining by Dr hemagglutinin was shown in colonic, bronchial, and endometrial glands, and skin eccrine sweat glands. Structures of the urinary tract showing strong fluorescence were renal tubular basement membrane, Bowmans' capsule, and transitional epithelium. The role of Dra antigen as receptor for adhesion for Dr-positive E. coli in ascending colonization of urinary tract and the possible importance of Dra in human pathology is discussed.

Alpha- and beta-adrenergic receptors in the horse ureter.

The presence of both alpha and beta adrenergic receptors in the caudal third ureter of the horse were studied in vitro under isometric conditions using adrenergic agonist and antagonist drugs. Isoprenaline and the beta 2- stimulating agent, salbutamol, elicited relaxation of the ureter smooth muscle strips. The responses were not affected by the beta 1- blocking agent, practolol, but were totally abolished by propranolol and the beta 2- blocking agent, butoxamine. The stimulation of alpha-adrenergic receptors with noradrenaline and phenylephrine evoked a contractile effect which was totally inhibited by phenoxybenzamine and the alpha 1- blocking agent, prazosin. It is concluded that in the horse ureter the alpha receptors are dominant and belong to alpha 1 subtype while the beta receptors are recessive and belong to beta 2- adrenoceptor subtype.

Expression of the c-erbB-2 protein in normal and transformed cells.

Two synthetic peptides from the predicted sequence of the human c-erbB-2 protein were synthesized and used to raise antisera in rabbits. The sequences chosen were identical to those in the homologous rat c-neu protein. The antibodies produced reacted with the immunizing peptides in ELISA but showed little or no cross-reaction with the partly homologous peptides found in equivalent positions in the human EGF receptor. Both antipeptide antibodies, and a monoclonal antibody (MAb) specific for the rat neu protein, immunoprecipitated a 185-kDa protein from 35S-methionine-labelled lysates prepared from a rat cell line known to express high levels of the c-neu protein. The antipeptide antibodies also recognized a protein of the same size in Western blots. In addition, both antipeptide antibodies immunoprecipitated a 190-kDa protein from labelled cell lysates prepared from human and monkey cells. Antibodies to one of the peptides, which showed no detectable cross-reaction with human, rat or monkey EGF receptor, were used to examine the expression of the c-erbB-2 protein on a variety of cultured cell types. Eleven transformed, I non-established and 2 immortalized cell types were examined by immunoprecipitation for their level of expression of the c-erbB-2 protein and of the EGF receptor. The numbers of EGF receptors varied widely between different cell lines, whereas the level of the c-erbB-2 protein, which was found on all of the cell types examined, was more constant. The number of c-erbB-2 molecules present was estimated by autoradiography to be about 100,000 per cell. The antibodies were then used to examine the location and level of expression of the human c-erbB-2 and rat c-neu proteins in normal tissues. Immunohistochemical staining showed that the c-erbB-2 protein was highly expressed in rat kidney proximal tubules and loop of Henle. The c-erbB-2 protein was also present on normal human epithelial cells but in some cases with a different distribution to that of the EGF receptor.

Thomsen-Friedenreich-related antigen in non-neoplastic ureter urothelium and transitional cell tumours of the urinary bladder. An immunohistochemical study employing the monoclonal antibody 49H.8.

This study, which is part of a larger immunohistochemical investigation of blood-group antigens in non-neoplastic urothelium and bladder cancer, reports our findings on the expression of an antigen related to the cryptic Thomsen-Friedenreich antigen (beta Gal 1-3 GalNAc) of erythrocytes. De-waxed sections of 19 ureters and of 93 transitional cell tumours, either untreated or pretreated with neuraminidase, were subjected to an indirect immunoperoxidase staining, employing the monoclonal antibody 49H.8. Staining results were compared to Lewis-secretor types, morphology, and in tumours to the clinical course as regards recurrence rate and the development of either stroma invasive recurrence or papillomatosis as well. Ureters not subjected to neuraminidase were unstained, whereas urothelium in 12 of 19 ureters subjected to neuraminidase showed staining. Serial dilution of antibody disclosed quantitative differences related to the Lewis-secretor types. Lea+b- urothelium, i.e., non-secretor urothelium, had the highest end-point titers. Endothelium was unstained. Thirty-six of the 93 tumours showed staining without prior neuraminidase treatment, 31 showed staining after neuraminidase treatment only, while 26 were unstained. Staining correlated with the pathological stage and grade (p less than 0.05), but not with the clinical course (p greater than 0.05). The results do not support previous observations on the prognostic value of Thomsen-Friedenreich antigen determination in superficial bladder cancer.

Laminar elution of tubular organs for bioluminescence analysis of the interior cell layer.

Cells lining the interior of tubular organs are of considerable interest from physiological and pathological aspects but are very difficult to prepare for biochemical analyses. The contents of such cells can be extracted by infusion of a suitable detergent serving as a membrane destroyer. The tiny ureter of the rat has been used as experimental model. Time governed elution with saponin, using a Hamilton programmable microlab for the infusion results in an effluent pattern which can be determined by sensitive bioluminescence assays. The time course of the outflux of nucleotides and enzymes showed two maxima in agreement with the presence of two epithelial layers in the ureter.

Plasma levels and renal handling of endogenous amino acids in snakes: a comparative study.

Plasma levels of 22 endogenous amino acids were measured by ion-exchange chromatography in four species of snakes: Thamnophis sirtalis, T. radix, Aipysurus laevis, and Python molurus. Despite considerable interspecific variation in the amino acid composition, all species showed relatively high plasma concentrations of histidine, a feature apparently unique to reptiles. The renal handling of these amino acids was studied by renal clearance methods. As in other vertebrates, net tubular absorption of filtered amino acids predominated. However, net tubular secretion of taurine, cysteic acid and/or phosphoserine and beta-alanine was observed, with taurine being the predominant amino acid secreted. The percentage reabsorption of the total amino acids filtered by the snake kidneys ranged from 79 to 95%. Evidence for the postrenal absorption of amino acids in these reptiles is presented. In species that normally undergo hibernation (Thamnophis spp.), the ability of the kidney to reabsorb amino acids was depressed by cold acclimation. Cold acclimation significantly decreased plasma levels of all amino acids except taurine, whose concentration increased. The increase in plasma taurine level may have resulted from cellular osmoregulation. Under these conditions, renal excretion of taurine increased concomitantly with the increase in plasma taurine concentration.

Altered glycosylation of membrane glycoproteins in human uroepithelial cell lines.

Total cellular glycopeptides of 7 human uroepithelial cell lines that differ in the grade of transformation (TGr) were analysed by gel filtration and affinity chromatography on immobilized lectins. The 4 cell lines that are tumorigenic in nude mice and invasive in vitro (TGr III) possess more highly branched, tri- and tetraantennary N-acetyllactosaminic glycans, with less biantennary glycans than the 2 non-tumorigenic, noninvasive (TGr II) cell lines examined. The only exception to this general pattern is the third cell line, which is classified as TGr II. The cellular glycopeptide distribution pattern in this cell line is similar to that of the TGr III cells. The possible relationship between altered glycosylation of membrane glycoproteins and the expression of a malignant phenotype is discussed.

The specific glycosphingolipid composition of human ureteral epithelial cells.

Total non-acid and acid glycolipid fractions were isolated from epithelial cell scrapings and the non-epithelial residue of a human upper ureter. The glycolipid fractions were structurally characterized as total mixtures by thin-layer chromatography, mass spectrometry, and proton NMR spectroscopy. Selected structural information was also obtained on binding of monoclonal antibodies and bacteria to the thin-layer chromatograms. The major epithelial cell glycolipids were Glc beta 1-1ceramide (75%), dihexosylceramide (10%) and NeuAcLacceramide (10%). In addition, 8 minor glycolipids belonging to the blood group P, Lewis and ABO systems were identified. The major glycolipids of the non-epithelial residues were mono- and dihexosylceramides together with globotriaosyl- and globotetraosylceramides. The epithelial mono- and diglycosylceramide compounds had an unusual ceramide composition with mainly C18 and C20 trihydroxy long chain bases in combination with C22-C24 hydroxy fatty acids in contrast to the non-epithelial glycolipids which contained mainly C18 dihydroxy long chain bases in combination with C16-C24 non-hydroxy fatty acids.

Immunohistochemical distribution of S-100 antigen in the urinary system of man and rat.

The immunohistochemical distribution of S-100, a protein originally isolated from the brain, has been investigated at the light and electron microscopic levels in rat and man urinary systems. In both species the antigen essentially exhibited the same location, restricted, with different degrees of staining, to certain cells in the kidney, i.e. collecting tubules, thin limbs of Henle's loop and renal papillae.

Tumor promoter 12-O-tetradecanoylphorbol-13-acetate receptors in normal human transitional epithelial cells.

As a prelude to study the promotion with TPA of in vitro transformation of human urothelial cells (HUC) in culture, we characterized tumor promoter TPA receptors in primary cultures of HUC. [3H]TPA bound specifically to intact living HUC; maximum specific binding was attained in approximately 30 min at 37 degrees C. [3H]TPA bound to HUC in a saturable and competitive manner. Scatchard analysis of specific binding to intact cells displayed a single slope corresponding to an equilibrium dissociation constant (Kd) of 0.56 nM; at saturation TPA-binding capacity was 2.37 pmol/10(6) HUC (1.43 X 10(6) sites per cell). [3H]TPA bound specifically and with high affinity to the particulate fractions of HUC; binding was both saturable and reversible. Saturation of the specific binding of [3H]TPA occurred at approximately 1 nM at 4 degrees C. Scatchard analysis of specific binding to the particulate fraction displayed a single slope corresponding to a Kd of 1.08 nM; at saturation TPA-binding capacity was 2.05 pmol/mg protein (750 000 molecules per HUC). [3H]TPA binding was inhibited by the biologically active phorbol ester, phorbol didecanoate, whereas inactive phorbol did not compete for TPA binding. Binding was not affected by sodium saccharin, epidermal growth factor, retinoic acid or dexamethasone. [3H]TPA bound specifically to the HUC cytosolic fraction but only in the presence of calcium and phosphatidylserine. Calcium-activated and phospholipid-sensitive protein kinase activity was detected in HUC fractions. These results indicate the presence of high-affinity specific receptors for TPA in HUC.

[Histamine in the urogenital tract]. / Histamin im Urogenitaltrakt.

Tissue histamine determination in the urogenital tract showed that there are significant differences concerning tissue histamine content between intraoperative and postmortem tissue samples. The difference appeared to be most significant in the case of renal pelvis samples, where the decrease was about 90% in the autopsie material compared to intraoperative tissues. In comparing tissue histamine content in special organs of different animal species we discovered considerable differences; e.g. in the ureter of the dog we measured 30 micrograms/g tissue - where as against the values in man amounted to only 6 micrograms/g tissue. The physiological meaning of the different tissue histamine levels is unknown.

Human ureter, a source of tissue plasminogen activator.

In organ cultures of human ureters, plasminogen activators were released into the medium. Unlike most tissue cultures that release mainly urokinase (UK), the activators released by ureter cultures were predominantly of the tissue-type (t-PA), as shown by radioimmunoassay. Using affinity chromatography it was possible to distinguish two activities. One minor fraction was quenched by antibodies against UK, but not by antibodies against t-PA. The other activity was quenched by antibodies against t-PA, but not by those against UK, and required the presence of fibrin to activate plasminogen. Because of the large amount of t-PA, cells derived from human ureter might well prove to be a useful source of t-PA.

Decreased fibrinolytic activity in vessels and ureters of rejecting human kidneys.

Plasminogen activator (PA) activity was determined with a histochemical fibrin slide technique in renal arteries, veins and ureters in 62 transplanted kidneys and again at transplantectomy in 12 transplants who later underwent irreversible rejection. At transplantation PA activity was the same whether the grafts later rejected or not. There was a significantly decreased PA activity in the vessels and ureters in rejected transplants. The role of the fibrinolytic system as a pathogenetic factor in rejected kidneys is discussed.