Comparison of different substances for subureteric injection in the management of vesicoureteric reflux in children.

INTRODUCTION: Endoscopic techniques are becoming increasingly accepted for treatment of vesicoureteric reflux as alternatives to open surgical reimplantation. However, there is some debate about the ideal injectable material. Since we have accumulated experience with several substances, an opportunity existed to compare them. MATERIALS AND METHODS: From 1991 to 2003, 101 children with vesicoureteric reflux were treated by endoscopic subureteric injection either once (74) or twice (27) by either of two pediatric urologists. There were a total of 165 ureteral injections, 83 with polytetrafluoroethylene (Teflon), 73 with polydimethylsiloxane (Macroplastique), and 9 with collagen. Each child was evaluated pre-operatively and 3 months post-operatively with a nuclear cystogram and renal ultrasonography. RESULTS: The polytetrafluoroethylene and polydimethylsiloxane groups were not significantly different with respect to sex, age, indication for surgery, severity of reflux or prior surgeries. The collagen group overall did very poorly with only 3 of 9 refluxing ureters cured. The other two substances had much more success with 61% of ureters in the polytetrafluoroethylene group cured on first injection and 75% with polydimethylsiloxane, plus another 19% and 11% cured on second attempt, respectively (total 80% and 86%). CONCLUSIONS: Subureteric injections of polytetrafluoroethylene and polydimethylsiloxane are very effective at curing vesicoureteric reflux in children with little morbidity. When comparing individual cases, ureters, and all grades of reflux, polytetrafluoroethylene and polydimethylsiloxane have similar success rates. Collagen injections were less successful, and patients with neurogenic bladders had poor results.

Evidence for seeding of beta -amyloid by intracerebral infusion of Alzheimer brain extracts in beta -amyloid precursor protein-transgenic mice.

Many neurodegenerative diseases are associated with the abnormal sequestration of disease-specific proteins in the brain, but the events that initiate this process remain unclear. To determine whether the deposition of the beta-amyloid peptide (Abeta), a key pathological feature of Alzheimer's disease (AD), can be induced in vivo, we infused dilute supernatants of autopsy-derived neocortical homogenates from Alzheimer's patients unilaterally into the hippocampus and neocortex of 3-month-old beta-amyloid precursor protein (betaAPP)-transgenic mice. Up to 4 weeks after the infusion there was no Abeta-deposition in the brain; however, after 5 months, the AD-tissue-injected hemisphere of the transgenic mice had developed profuse Abeta-immunoreactive senile plaques and vascular deposits, some of which were birefringent with Congo Red. There was limited deposition of diffuse Abeta also in the brains of betaAPP-transgenic mice infused with tissue from an age-matched, non-AD brain with mild beta-amyloidosis, but none in mice receiving extract from a young control case. Abeta deposits also were not found in either vehicle-injected or uninjected transgenic mice or in any nontransgenic mice. The results show that cerebral beta-amyloid can be seeded in vivo by a single inoculation of dilute AD brain extract, demonstrating a key pathogenic commonality between beta-amyloidosis and other neurodegenerative diseases involving abnormal protein polymerization. The paradigm can be used to clarify the conditions that initiate in vivo beta-amyloidogenesis in the brain and may yield a more authentic animal model of Alzheimer's disease and other neurodegenerative disorders.

Milameline (CI-979/RU35926): a muscarinic receptor agonist with cognition-activating properties: biochemical and in vivo characterization.

Milameline (E-1,2,5,6-tetrahydro-1-methyl-3-pyridinecarboxaldehyde, O-methyloxime monohydrochloride, CI-979, PD129409, RU35926) was characterized in vitro and evaluated for effects on central and peripheral cholinergic activity in rats and rhesus monkeys. In muscarinic binding studies, milameline displayed nanomolar affinity with an agonist ligand and micromolar affinity with antagonist ligands, with approximately equal affinities determined at the five subtypes of human muscarinic receptors (hM(1)-hM(5)) with whole cells or membranes from stably transfected Chinese hamster ovary (CHO) cells. On binding, milameline stimulated phosphatidylinositol hydrolysis in hM(1) and hM(3) CHO cells and inhibited forskolin-activated cAMP accumulation in hM(2) and hM(4) CHO cells. Additionally, it decreased K(+)-stimulated release of [(3)H]acetylcholine from rat cortical slices. Responses were not caused by the inhibition of acetylcholinesterase, and there was no significant binding to approximately 30 other neurotransmitter binding sites. In rats, milameline decreased spontaneous and scopolamine-induced swimming activity, improved water-maze performance of animals impaired by basal forebrain lesions, increased cortical blood flow, decreased core body temperature, and increased gastrointestinal motility. Electroencephalogram activity in both rats and monkeys was characterized by a predominance of low-voltage desynchronized activity consistent with an increase in arousal. Milameline also reversed a scopolamine-induced impairment of attention on a continuous-performance task in monkeys. Thus, milameline possesses a pharmacological profile consistent with that of a partial muscarinic agonist, with central cholinergic actions being produced in rats and monkeys at doses slightly lower than those stimulating peripheral cholinergic receptors.

Inhibitors of V-type ATPases, bafilomycin A1 and concanamycin A, protect against beta-amyloid-mediated effects on 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction.

The functional viability of cells can be evaluated using a number of different assay determinants. One common assay involves exposing cells to 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), which is converted intracellularly to a colored formazan precipitate and often used to assess amyloid peptide-induced cytotoxic effects. The MTT assay was employed to evaluate the role of endosomal uptake and lysosomal acidification in amyloid peptide-treated differentiated PC12 cell cultures using selective vacuolar-type (V-type) ATPase inhibitors. The macrolides bafilomycin A1 (BAF) and concanamycin A (CON) block lysosomal acidification through selective inhibition of the V-type ATPase. Treating nerve growth factor-differentiated PC12 cells with nanomolar concentrations of BAF or CON provides complete protection against the effects of beta-amyloid peptides Abeta(1-42), Abeta(1-40), and Abeta(25-35) and of amylin on MTT dye conversion. These macrolides do not inhibit peptide aggregation, act as antioxidants, or inhibit Abeta uptake by cells. Measurements of lysosomal acidification reveal that the concentrations of BAF and CON effective in reversing Abeta-mediated MTT dye conversion also reverse lysosomal pH. These results suggest that lysosomal acidification is necessary for Abeta effects on MTT dye conversion.

Differential expression of group I metabotropic glutamate receptors (mGluRs) in the rat pheochromocytoma cell line PC12: role of nerve growth factor and ras.

Glutamate treatment of PC12 cells has been shown to result in the accumulation of intracellular inositol phosphates suggesting the presence of glutamate metabotropic receptors (mGluRs) positively coupled to phospholipase C. The present study examined the expression of group I mGluRs (mGluR1 and mGluR5) in PC12 cells. Undifferentiated PC12 cells were found to express both mGluR5 mRNA and receptor protein by reverse transcription polymerase chain reaction (RT-PCR) and western blot techniques. However, mGluR1 mRNA was not detected in these cells and western blot analysis showed only faint mGluR1alpha immunoreactivity suggesting a very low level of mGluR1 expression. Nerve growth factor-induced differentiation of PC12 cells resulted in the induction of mGluR1alpha and mGluR1beta mRNA and mGluR1alpha protein. PC12 cells overexpressing dominant negative ras revealed that NGF-induced mGluR1 induction, but not mGluR5 expression, is dependent on ras pathway activation in these cells. These results suggest PC12 cells may be a useful model for investigating the regulation and expression of group I mGluR isoforms and their role in neuronal processes in vitro.

Characterization of muscarinic agonists in recombinant cell lines.

Using recombinant CHO cells that express Hm1-Hm5 receptors, reference muscarinic agonists have been characterized with respect to their activity in receptor binding and second messenger assays. In whole cell [3H]-N-methyl scopolamine binding, no agonist was found to be truly subtype selective, although some showed marked differences between several of the subtypes (e.g. m1 vs. m2). As a functional index of receptor activation, phosphatidyl-inositol (PI) turnover was measured for m1, m3, and m5 receptors while inhibition of forskolin-stimulated cAMP accumulation was measured for m2 and m4 receptors. Both full and partial agonists were delineated in PI turnover, but all agonists showed similar responses on cAMP. Alkylation studies with propylbenzylcholine mustard showed that both efficacy and potency were markedly affected in the functional assays by the number of free receptors. Thus, receptor reserve appears to play a major role in the determination of subtype selectivity for agonists using functional measures. Even with these limitations, however, the use of transformed cell lines is playing a pivotal role in the discovery of selective agonists.

Synthesis and SAR of bulky 1-azabicyclo[2.2.1]-3-one oximes as muscarinic receptor subtype selective agonists.

The synthesis of a series of potent and efficacious 1-azabicyclo[2.2.1]heptan-3-one oxime muscarinic agonists is described. The oximes have extended appendages designed to span the cavity defined by the seven transmembrane helices of the muscarinic receptor. Some members of the series are selective for receptors of the m1 subtype. One such oxime, 31, shows affinity and functional selectivity for m1 over m2, m3, and m4 muscarinic receptor types.

Acute scrotal pain in children: prospective study of diagnosis and management.

Forty-eight boys were assessed for an acutely painful scrotum. Thirty-six (75%) of them underwent radionuclide scanning of the scrotum; the average age of this group was 11 years. The scan revealed epididymitis in 19 cases, spermatic cord torsion in 9, appendix testis torsion in 7 and acute hernia-hydrocele in 1. The diagnosis was confirmed at operation in all nine cases of spermatic cord torsion. Boys who had epididymitis received antibiotics only; all were available for short-term follow-up, and 16 were also assessed at a mean of 6 months after infection. Only one boy had testicular atrophy; he had undergone repair of an inguinal hernia, which could not be ruled out as a cause. Bacteriuric epididymitis occurred in three boys; two had known predisposing genitourinary anomalies, the third had no abnormalities. Boys who had nonbacteriuric epididymitis were investigated by renal and pelvic ultrasonography or voiding cystourethrography; no important abnormalities were detected. This prospective study indicates that radionuclide scanning can reliably differentiate spermatic cord torsion from other acute scrotal disease.

Acute renal failure secondary to bilateral ureteric obstruction: review of 50 cases.

The records of 50 patients with acute renal failure secondary to bilateral ureteric obstruction were reviewed. An underlying malignant disorder was the cause of the obstruction in 38 of the patients and had not previously been diagnosed in almost half of them. Carcinomas of the cervix and prostate were the most frequent malignant disorders, and aggressive management resulted in good survival rates. Similarly, the outcome for patients with benign bilateral ureteric obstruction, usually caused by retroperitoneal fibrosis, was good with proper management.

The pathogenesis of renal dysplasia. I. Quantification of hypoplasia and dysplasia.

In order to assess the relative effects of abnormal ureteric orifice position and abnormal urodynamics on the morphogenesis of hypoplasia and dysplasia in kidneys obtained from infants, we devised a method of quantifying the renal structures. The method was based on radial glomerular counts which ranged from zero to normal (seven to nine), a score for dysplastic structures, and the ratio of normal to abnormal tissues present. These three values, when plotted against each other, correlated closely. The glomerular count, with occasional minor adjustment for inconsistencies, was the best parameter of hypodysplasia. Severe to moderate grades of hypodysplasias fell in the low and middle ranges and hypoplasia through to normal in the highest range. By grading kidneys in this way, we were able to compare the effects of ureteral ectopy and abnormal urinary dynamics on the developing kidney.

The pathogenesis of renal dysplasia. III. Complete and incomplete urinary obstruction.

We graded obstructed kidneys of infants on the hypodysplasia scale to assess the influence of complete and partial obstruction on the pathogenesis of hypodysplasia. Kidneys with complete obstruction exhibited severe grades; those with partial ureteral obstruction had near normal grades. Those kidneys subjected to partial urethral obstruction ranged from mild to severe grades which correlated with degrees of lateral ectopy of the urethral office. Renal parenchymal development was impaired by complete obstruction but was tolerant to incomplete obstruction. Abnormal orifice positions associated with urethral obstructions were considered to be manifestations of ectopic ureteric buds and the hypodysplasia to be evidence of abnormal induction of abnormal renal blastema.

The pathogenesis of renal dysplasia. II. The significance of lateral and medial ectopy of the ureteric orifice.

Renal hypoplasia and dysplasia may be primary malformations linked to a panureteric bud deformity or result from damage to the developing nephrons caused by abnormal urodynamic pressures, Kidneys with misplaced ureteric orifices were graded, according to histologic criteria, on the hypodysplasia scale. With lateral ectopy of the ureteric orifices with and without congenital urethral obstruction, the grades correlated with specific orifice positions. The grades of kidneys with caudal ectopy of the ureters indicated a more general correlation. Dysgenesis of the bud and nephrogenic mesenchyme may account for the renal hypodysplasia when the ureteric orifice is found to be ectopic.