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W. H. Wollaston, who first described cystine stones, as well as the most outstanding contemporary figures and their contribution to the understanding of this uncommon type of lithiasis are described.

We report the luminescence and spectral properties of a phospholipid analogue containing a long-lifetime luminescent rhenium metal-ligand complex (MLC) covalently linked to the amino group of phosphatidyl ethanolamine. When incorporated into synthetic membranes, this lipid probe displays intensity decay times near 3 microseconds. Importantly, the probe displays highly polarized emission with a maximal fundamental anisotropy of 0.33. This probe is expected to have numerous applications for studies of microsecond diffusion and dynamics of membranes.

Background: Cdc42, a GTP-binding protein of the Rho family, controls actin cytoskeletal organization and helps to generate actin-based protruding structures, such as filopodia. In vitro, Cdc42 regulates actin polymerization by facilitating the creation of free barbed ends - the more rapidly growing ends of actin filaments - and subsequent elongation at these ends. The Wiskott- Aldrich syndrome protein, WASP, which has a pleckstrin-homology domain and a Cdc42/Rac-binding motif, has been implicated in cell signaling and cytoskeleton reorganization. We have investigated the consequences of local recruitment of activated Cdc42 or WASP to the plasma membrane., Results: We used an activated Cdc42 protein that could be recruited to an engineered membrane receptor by adding rapamycin as a bridge, and added antibody-coupled beads to aggregate these receptors. Inducible recruitment of Cdc42 to clusters of receptors stimulated actin polymerization, resulting in the formation of membrane protrusions. Cdc42-induced protrusions were enriched in the vasodilator-stimulated phosphoprotein VASP and the focal-adhesion-associated proteins zyxin and ezrin. The Cdc42 effector WASP could also induce the formation of protrusions, albeit of different morphology., Conclusions: This is the first demonstration that the local recruitment of activated Cdc42 or its downstream effector, WASP, to a membrane receptor in whole cells is sufficient to trigger actin polymerization that results in the formation of membrane protrusions. Our data suggest that Cdc42-induced actin-based protrusions result from the local and serial recruitment of cytoskeletal proteins including zyxin, VASP, and ezrin.

Chemotaxis of Escherichia coli toward phosphotransferase systems (PTSs)-carbohydrates requires phosphoenolpyruvate-dependent PTSs as well as the chemotaxis response regulator CheY and its kinase, CheA. Responses initiated by flash photorelease of a PTS substrates D-glucose and its nonmetabolizable analog methyl alpha-D-glucopyranoside were measured with 33-ms time resolution using computer-assisted motion analysis. This, together with chemotactic mutants, has allowed us to map out and characterize the PTS chemotactic signal pathway. The responses were absent in mutants lacking the general PTS enzymes EI or HPr, elevated in PTS transport mutants, retarded in mutants lacking CheZ, a catalyst of CheY autodephosphorylation, and severely reduced in mutants with impaired methyl-accepting chemotaxis protein (MCP) signaling activity. Response kinetics were comparable to those triggered by MCP attractant ligands over most of the response range, the most rapid being 11.7 +/- 3.1 s-1. The response threshold was <10 nM for glucose. Responses to methyl alpha-D-glucopyranoside had a higher threshold, commensurate with a lower PTS affinity, but were otherwise kinetically indistinguishable. These facts provide evidence for a single pathway in which the PTS chemotactic signal is relayed rapidly to MCP-CheW-CheA signaling complexes that effect subsequent amplification and slower CheY dephosphorylation. The high sensitivity indicates that this signal is generated by transport-induced dephosphorylation of the PTS rather than phosphoenolpyruvate consumption.

We describe a glucose sensor based on a mutant glucose/galactose binding protein (GGBP) and phase-modulation fluorometry. The GGBP from Escherichia coli was mutated to contain a single cysteine residue at position 26. When labeled with a sulfhydryl-reactive probe 2-(4'-iodoacetamidoanilino)naphthalene-6-sulfonic acid, the labeled protein displayed a twofold decrease in intensity in response to glucose, with a dissociation constant near 1 microM glucose. The ANS-labeled protein displayed only a modest change in lifetime, precluding lifetime-based sensing of glucose. A modulation sensor was created by combining ANS26-GGBP with a long-lifetime ruthenium (Ru) metal-ligand complex on the surface of the cuvette. Binding of glucose changed the relative intensity of ANS26-GGBP and the Ru complex, resulting in a dramatic change in modulation at a low frequency of 2.1 MHz. Modulation measurements at 2.1 MHz were shown to accurately determine the glucose concentration. These results suggest an approach to glucose sensing with simple devices., (Copyright 1999 Academic Press.)

The technology and applications of fluorescence spectroscopy are rapidly advancing. In this overview presentation we summarize some recent developments from this laboratory. Two and three-photon excitation have been observed for a wide variety of intrinsic and extrinsic fluorophores, including tryptophan, tyrosine, DNA stains, membrane probes, and even alkanes. It has been possible to observe multi-photon excitation of biopolymers without obvious photochemical or photo-thermal effects. Although not de-scribed in our lecture, another area of increasing interest is the use of engineered proteins for chemical and clinical sensing. We show results for the glucose-galactose binding protein from E. coli. The labeled protein shows spectral changes in response to micromolar concentrations of glucose. This protein was used with a novel sensing method based on the modulated emission of the labeled proteins and a long lifetime reference fluorophore. And finally, we describe a recently developed rhenium complex which displays a lifetime near 3 µs in oxygenated aqueous solution. Such long life-time probes allow detection of microsecond dynamic processes, bypassing the usual nanosecond timescale limit of fluorescence. The result of these developments in protein engineering, sensing methods, and metal-ligand probe chemistry will be the increased use of fluorescence in clinical chemistry and point-of-care analyses.

Objective: To describe a case of adult renal polycystosis causing intercostal hernia with intestinal segment. To our knowledge, no such case has been previously reported in the literature., Methods/results: Diagnosis was by abdominal CT. Treatment was by bilateral nephrectomy and surgical repair of the diaphragmatic and intercostal hernia., Conclusions: Surgery is indicated in adult renal polycystosis if kidney size causes gastrointestinal involvement and above all, if the lungs are compromised by the intercostal diaphragmatic hernia.

We describe a new approach to fluorescence sensing based on a mixture of fluorophores, one of which is sensitive to the desired analyte. If a long-lifetime analyte-insensitive fluorophore is mixed with a short-lifetime analyte-sensitive fluorophore, the modulation of the emission at conveniently low frequencies becomes equal to the fractional fluorescence intensity of the sensing fluorophore. Under these conditions, the modulation can be used to determine the analyte concentration. This can be used with any fluorophore that changes intensity in response to analyte and does not require the sensing fluorophore to display a change in lifetime. The feasibility of modulation-based sensing was demonstrated using mixtures of 6-carboxyfluorescein and [Ru 2,2'-(bipyridyl)3]2+ as a pH sensor and of the calcium probe Fluo-3 and [Ru 2,2'-(bipyridyl)3]2+ as a calcium sensor.

Rab GTPases are post-translationally modified by addition of geranylgeranyl moieties to carboxyl-terminal cysteine residues. For Rab proteins ending with xxCC xCxC and CCxx motifs this modification is catalysed by geranylgeranyltransferase type II (GGTaseII), and is entirely dependent on the Rab substrate being bound to Rab escort protein (REP). Several Rab proteins contain carboxyl-terminal CaaL prenylation motifs typical of members of the Rho family, which are modified in a REP-independent manner by geranylgeranyltransferase type I (GGTaseI). The present studies show that one such Rab protein (Rab8), which ends with a CVLL motif, is uniquely able to serve as a substrate for either REP/GGTaseII or GGTaseI in cell-free assays. The modification of Rab8 by GGTaseI did not require REP, indicating that a REP-induced conformational change is not essential for exposure of the Rab carboxyl-terminal cysteine prenylation site. To determine whether one enzyme plays a predominant role in Rab8 prenylation in vivo, the incorporation of [3H]mevalonate into Rab8 was measured in human embryonal kidney 293 cells under conditions where the activity of GGTaseI, but not GGTaseII, was blocked by the peptidomimetic inhibitor GGTI-298. The GGTaseI inhibitor did not prevent prenylation of either overexpressed Myc-tagged Rab8 or endogenous Rab8, whereas prenylation of a known GGTaseI substrate with the same carboxyl-terminal motif, Cdc42Hs, was completely blocked. To rule out the possibility that the apparent prenylation of Rab8 by GGTaseII occurs only when GGTaseI activity is eliminated, metabolic labelling studies were carried out in the absence of the GGTaseI inhibitor, using a REP-binding-deficient Rab8 construct (Y78D) that cannot serve as a substrate for GGTaseII, but is indistinguishable from wild-type Rab8 as a substrate for GGTaseI. Prenylation of the Y78D mutant was reduced by 60-70% in intact cells, consistent with the conclusion that the majority of Rab8 is prenylated by the REP/GGTaseII system in vivo.

The introduction of tacrolimus as rescue therapy represents a significant advance in the prevention of late graft failure and second liver transplantation. The authors report the blood level monitoring of tacrolimus as a rescue therapy in 21 children who underwent liver transplantation, and they report the dose-concentration relationship in the presence or absence of hepatitis C virus (HCV) in these patients. This was a retrospective study conducted from May 1993 to January 1997. Indication for the conversion from cyclosporine (CsA) to tacrolimus were acute rejection (62%), chronic rejection (33%), and CsA toxicity (5%). Mean daily dose in the first month was 0.32 mg/kg, whereas at the end of the follow-up period it was 0.14 mg/kg. Tacrolimus mean whole blood concentration levels were between 7.1 ng/ml and 9.4 ng/ml, without significant differences over time. Mean daily doses in HCV+ and HCV- patients were 0.08 and 0.24 mg/kg, respectively (p < 0.01), and mean concentrations were 8.3 and 8.4 ng/ml (NS). HCV+ children required a mean dose three times lower than the dose used in HCV- children to obtain the same tacrolimus trough blood level. Therefore, doses in HCV+ children must be decreased to achieve levels within the therapeutic range.

We describe the synthesis and characterization of two asymmetrical ruthenium(II) complexes, [Ru(dpp)2(dcbpy)]2+ and [Ru(dpp)2(mcbpy)]2+, as well as the water soluble sulfonated derivatives [Ru(dpp(SO3Na)2)2(dcbpy)]2+ and [Ru(dpp(SO3Na)2)2(mcbpy)]2+ (dpp is 4,7-diphenyl-1,10-phenanthroline, dcbpy is 4,4'-dicarboxylic acid-2,2'-bipyridine, mcbpy is 4-methyl,4'-carboxylic acid-2,2'-bipyridine, and dpp(SO3Na)2 is the disulfonated derivative of dpp) as probes for the measurement of the rotational motions of proteins. The spectral (absorption, emission, and anisotropy) and photophysical (time-resolved intensity and anisotropy decays) properties of these metal-ligand complexes were determined in solution, in both the presence and absence of human serum albumin (HSA). These complexes display lifetimes ranging from 345 ns to 3.8 microseconds in deoxygenated aqueous solutions under a variety of conditions. The carboxylic acid groups on these complexes were activated to form N-hydroxysuccinimide (NHS) esters which were used to covalently lable HSA, and were characterized spectroscopically in the same manner as above. Time-resolved anisotropy measurements were performed to demonstrate the utility of these complexes in measuring long rotational correlation times of bioconjugates between HSA and antibody to HSA. The potential usefulness of these probes in fluorescence polarization immunoassays was demonstrated by an association assay of the Ru(II)-labeled HSA with polyclonal antibody.

We developed a water-soluble luminescent probe for dissolved oxygen. This probe is based on (Ru[dpp(SO3Na)2]3) cl2, which is a sulfonated analogue of the well-known oxygen probe (Ru[dpp]3)cl2. The compound dpp is 4,7-diphenyl-1,10-phenanthroline and dpp(SO3Na)2 is a disulfonated derivative of the same ligand. In aqueous solution in the absence of oxygen (Ru[dpp(SO3Na)2]3)cl2 displays a lifetime of 3.7 microseconds that decreases to 930 ns on equilibrium with air and 227 ns on equilibrium with 100% oxygen. The Stern-Volmer quenching constant is 11,330 M-1. This high oxygen-quenching constant means that the photoluminescence of Ru(dpp[SO3Na]2)3cl2 is 10% quenched at an oxygen concentration of 8.8 x 10(-6) M, or equilibration with 5.4 torr of oxygen. The oxygen probe dissolved in water displays minimal interactions with lipid vesicles composed of dipalmityl-L-alpha-phosphatidyl glycerol but does appear to interact with human serum albumin. The absorption maximum near 480 nm, long lifetime and large Stokes' shift allow this probe to be used with simple instrumentation based on a light-emitting diode light source, allowing low-cost oxygen sensing in aqueous solutions. To the best of our knowledge this is the first practical water-soluble oxygen sensor.

We describe a new class of fluorescence polarization immunoassays based on the luminescence from a Re(I) metal-ligand complex. Re(I) complexes are extremely photostable and possess useful photophysical properties including long lifetimes, high quantum yields, and high emission polarization in the absence of rotational diffusion. In the present study, a conjugatable, highly luminescent Re(I) metal-ligand complex, [Re(bcp)(CO)3(4-COOHPy)](ClO4), where bcp is 2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline and 4-COOHPy is isonicotinic acid, has been evaluated for use in fluorescence polarization immunoassays (FPIs) with high-molecular-weight antigens. This Re(I) complex (Re) displays highly polarized emission (with a maximum anisotropy near 0.3) in the absence of rotational diffusion and a long average lifetime (2.7 microseconds) when bound to human serum albumin (HSA) in oxygenated aqueous solution. The emission polarization of the Re-HSA conjugate is sensitive to the binding of anti-HSA, resulting in a significant increase in anisotropy. The labeled HSA was also used in a competition immunoassay where unlabeled HSA was also used as an antigen. These experimental results, combined with theoretical predictions, demonstrate the potential of this Re(I) metal-ligand complex as a luminescence probe in FPIs of high-molecular-weight analytes (10(5)-10(8) Da).

We report the synthesis and spectral properties of two long-lifetime highly luminescent Ru(II) complexes containing either a sulfhydryl reactive iodoacetamido group or a less reactive choloroacetamido group, [Ru(bpy)2(5-iodoacetamido-1,10-phenanthroline)] (PF6)2 and [Ru(bpy)2(5-chloroacetamido-1,10-phenanthroline)](PF6) 2, respectively, where bpy is 2,2'-bipyridine. Ru(bpy)2(phen-IA)](PF6)2 was covalently linked to human serum albumin (HSA) and human immunoglobulin G (IgG). The photoluminescence lifetime of protein-bound probes approaches 1 microsecond under ambient conditions. In the absence of rotational motions, this probe displayed an anisotropy of 0.18 for excitation at 472 nm. Anisotropy decay data were used to determine the overall rotational correlation times of HSA and IgG. These long-lifetime sulfhydryl-reactive probes can be used to recover microsecond rotational motions and/or domain motions of proteins and/or macromolecular complexes.

A highly luminescent rhenium (I) metal-ligand complex [Re(bcp)(CO)3(4-COOHPy)](ClO4), where bcp is 2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline and 4-COOHPy is isonicotinic acid, has been synthesized and characterized. High quantum yields (> 0.5) and long excited-state lifetimes (0.3-10 micronseconds) in fluid solutions at room temperature were found for this complex, with remarkable emission sensitivity to microenvironment. This compound also displays highly polarized emission with a maximum anisotropy near 0.3 in the absence of rotational diffusion. This Re complex was conjugated to several biomolecules, including the proteins human serum albumin and bovine immunoglobulin G, as well as an amine-containing lipid. When bound to a protein or lipid, the decay time is near 3 microseconds and the quantum yield is approximately 0.12 in aqueous oxygenated solution at room temperature. This compound's unique spectral properties along with its conjugatability allowed us to utilize it as biomolecular probe in a variety of environments.

Objective: The fact that for an equivalent oversaturation of solutes, urine of lithiasic patients has a greater tendency to form crystals, indicate that urine of stone-formers may contain substances that promote crystallization., Methods: A renal calculus was analyzed by infra-red spectrophotometry, x-ray dispersion energy spectrophotometry, ultraviolet absorption spectrophotometry, acid hydrolysis cationic exchange chromatography, SDA-Page gel electropheresis with Coomassie Blue and amino acid sequence., Results: It was a pure protein calculus formed by a beta 2-microglobulin anomaly. It was a residue that had lost its first 19 amino acids and phenylalanine replaced serine in position 19, Conclusions: Changes in the amino acid sequence of urinary proteins can cause these to behave like urinary calculus-promoting substances.

Objectives: To review the diagnosis and treatment of ureteral stenosis arising from endometriosis and describe a case of mixed (intrinsic and extrinsic) ureteral endometriosis., Methods: The diagnostic and therapeutic aspects of ureteral stenosis due to endometriosis are reviewed. The case of a patient with a nonfunctioning right kidney and hematuria is described. The patient had previously undergone hysterectomy and double adnexectomy for uterine leiomyofibromas and a right ureteral lesion that warranted cuff ureterocystoneostomy leaving the extreme distal third of the pelvic ureter. CT evaluation disclosed a right retrovesical mass. The therapeutic strategy consisted in performing percutaneous nephrostomy, ureteroscopy of nonfunctioning ureter and determining the nature of the retrovesical mass by fine needle punction aspiration biopsy., Results: Percutaneous nephrostomy achieved functional recovery of the renal unit. Endoscopic incision of the stricture with intravesical invagination of the compromised segment was performed with the combined antegrade and retrograde approach. The anatomopathological findings of ureteroscopic biopsy of an intraureteral lesion in the nonfunctioning ureter disclosed ureteral endometriosis. This finding obviated fine needle punction aspiration biopsy and the patient was treated with danazol for 6 months., Conclusions: Diagnosis by ureteroscopy should be considered in intrinsic and mixed forms of ureteral endometriosis. Endoscopic incision of the ureteral stricture combined with hormone therapy is a valid therapeutic option.

Objectives: A new form of minimally invasive treatment of ureteral stricture with the Acucise catheter is described herein., Methods: The method for catheter use for the ureteral incision using radiological control alone is described., Results: The technique is easy to perform. There were no early postoperative complications in strictures at the juxtavesical or upper lumbar ureter., Conclusions: The availability of balloon dilatation catheters equipped with an electrocautery device permits incision of the ureteral stricture using radiological control alone, preferably those in the juxtavesical or upper lumbar ureter. Those localized to other levels of the ureter are not free from vascular or visceral lesions.

The present study evaluated a total of 350 patients who underwent placement of double-J catheters as a result of their lithiasic condition. The work includes a review of the patients' urological background, indication for the double-J placement, tolerance to catheter, time of permanence, reason for withdrawal, presence of encrustations in the catheter and relationship between all these parameters. A significant increase of adverse symptoms was observed when permanence of the double-J catheter lasted longer than 6 weeks. Likewise, there was a significant increase in the number of encrustations in catheters retained longer than 6 weeks, as well as when the lithiasic mass was meaningful and urine cultures were positive. The authors consider that double-J catheters effectively prevent the complications of the lithiasic condition but an excessively long permanence, more than 6 weeks, increases occurrence of side-effects significantly.

Objectives: Ureterointestinal stricture represents a serious problem whose management has traditionally been by open surgery. Endourological treatment is an alternative that is generally free from the complications of open surgery, but with a lower success rate. The efficacy of endourological treatment is analyzed in 14 cases of ureteroileal stricture (12 patients) and the different treatment options are discussed., Methods: Ureteroileal stricture was managed endourologically in 9 of 13 cases (69%) by dilatation (6 cases), incision+dilatation (2 cases) and stenting (1 case)., Results: Of the 6 cases submitted to dilatation, 3 recurred at 6 months, 2 remained patent at 18 and 24 months, respectively, and 1 was lost to follow up because the patient underwent reimplantation (40% success rate). The two cases submitted to incision are patent at 18 and 19 months, respectively., Conclusions: The overall success rate for endourological treatment of ureteroileal stricture is 28% (4/14 cases) at 19 months mean follow up.

The relationship between the Ras-related GTP-binding protein, Rab1B, and intracellular transport of nascent low density lipoprotein (LDL) receptor was studied in cultured human embryonic kidney cells (line 293) cotransfected with plasmids encoding the LDL-receptor and either wild-type Rab1B or a Rab1B mutant (N121I) known to act as a dominant suppressor of endogenous Rab1B function. [35S]Methionine pulse-chase analysis of immunoprecipitated LDL-receptor indicated that coexpression with Rab1BN121I, but not Rab1BWT, impaired its conversion from the Endo-H-sensitive 120-125 kDa form to the O-glycosylated 160-170 kDa form, consistent with a block in ER-->Golgi trafficking of the nascent receptor. In cells expressing Rab1BN121I, the newly synthesized LDL-receptor was unable to reach the cell surface as evidenced by its inaccessibility to sulfo-NHS-biotin added to the cultures. These observations provide a direct demonstration of Rab protein involvement in LDL receptor trafficking and lend support to the concept of Rab1B as a universal mediator of ER-->Golgi transport of membrane glycoproteins in mammalian cells.

Objective: The present study analyzed the hydrodynamic behaviour of the double-J stent., Methods: Thirty-two double-J stents of different sizes and biomaterials (16 new and 16 removed from patients) were hydrodynamically evaluated using two models: a) nephroureterocystectomy surgical blocks harvested from pigs and b) an in vitro experimental model of the upper urinary tract. The following parameters were measured: initial and final weights, encrusted material weight, inside and outside diameters, number, size and distribution of sideholes, wall thickness, drainage area and length of time the catheter was indwelling in the patient., Results: Double-J stent placement in a normal upper urinary tract constitutes a partial obstruction that is proportional to the caliber of the catheter. Ureteral-flow reduction is 83%, 61% and 58% for 5FR, 6FR, and 7FR catheters, respectively. Extraluminal obstruction of the catheter reduces ureteral flow more than intraluminal obstruction (74%, 43% and 25% for extraluminal versus 83%, 66%, 57% for intraluminal for 5FR, 6FR and 7FR, respectively). Vesicoureteral and vesicorenal reflux pressures were 6.2 +/- 0.3 cm H2O and 35.2 +/- 2 cm H2O, respectively., Conclusion: Selection of a double-J stent must be individualized on the basis of indication, time indwelling, sex and route of insertion.

Objective: This paper describes the endoscopic incision of uretero-ileal anastomotic strictures by a combined approach (percutaneous antegrade and endoscopic retrograde) using the intraluminal invagination technique over a balloon dilatation catheter., Methods: The different steps of the surgical procedure of incision of the stenotic segment of different techniques of ureteroileal diversion (Bricker, Camey, ureterosigmoidostomy) are described in detail. Endoscopic images and those from the image intensifier are shown., Results: Of the 6 cases described herein, incision was achieved in 5 and in 1 case stenting of the stenotic segment could not be achieved., Conclusions: Intraluminal invagination of the stenotic segment considerably facilitates endoscopic incision.

Objectives: Herein we describe the technique of endoscopic incision in the treatment of ureterointestinal stricture., Methods: A teflon sheathed stent is placed in the stenotic ureterointestinal segment utilizing a combined approach (percutaneous antegrade endoscopic transrectal) and a balloon is passed over the stent retrogradely. The balloon is then inflated and is pulled retrogradely to invaginate the anastomosis into the rectal lumen. Thereafter, endoscopic incision of all the layers and the entire length of the stricture is performed by electrocautery., Results: The radiological evaluation three months postoperatively show marked morphological and functional improvement of the compromised renal unit., Conclusions: In patients with a ureterointestinal urinary diversion, endoscopic incision of the stricture at the site of the anastomosis is a valid therapeutic technique, after discarding the presence of urinary infection or tumor at this site. Furthermore, open surgery can be done in the event this technique fails to achieve the desired results.

We report 4 cases of primary congenital paraurethral diverticula or cavities (3 diverticula and 1 cyst of Cowper's glands) that were treated by endoscopic surgery. All the foregoing cases were diagnosed as primary in the absence of coexisting pathologies or associated disorders. The wall between the diverticulum and the urethral lumen was sectioned endoscopically. All patients recovered normal clinical and morphological status.

We report three cases of retrocaval ureter with a clear indication for surgery: one patient with severe hydronephrosis and two cases with mild dilatation of the renal pelvis, calyces and upper ureter, both complicated by renal stones. The initial diagnosis was based on the ultrasound and the intravenous urogram findings, and was confirmed by a retrograde pyelogram in combination with cavography and CT. All patients were submitted to surgery (1 nephrectomy and 2 uretero-ureteric anastomosis). Renal stone was resolved at the same time in one case, and by ESWL after surgery in the other.

There is scant literature on bone lithiasis, a disease entity whose diagnosis depends fundamentally upon the anatomopathological findings. The data gleaned from animal experiments have been shown to overlap with those observed in the clinical setting. Conservative treatment should include resection of the urothelial bony metaplasia to avoid recurrence. The importance of making a correct diagnosis cannot be overemphasized since this condition may be associated with a tumoral lesion and a close follow-up is therefore warranted.

Treatment with mevinolin, a competitive inhibitor of HMGCoAR, the key enzyme of isoprenoid metabolism, causes the arrest of proliferation and the differentiation of a neuroblastoma cell line (N18TG2). Mevalonate and high density lipoproteins partially restore growth. Cholesterol synthesis in the presence of mevinolin remains active, because in these cells the key enzyme HMG-CoA reductase is not completely inhibited by this drug. The fact that cell growth is reduced, while cholesterogenesis remains active, suggests that mevinolin acts by interfering with the synthesis of some unknown compound, other than cholesterol, which is necessary for proliferation.

Chemotactic excitation responses to caged ligand photorelease of rapidly swimming bacteria that reverse (Vibrio alginolyticus) or tumble (Escherichia coli and Salmonella typhimurium) have been measured by computer. Mutants were used to assess the effects of abnormal motility behavior upon signal processing times and test feasibility of kinetic analyses of the signaling pathway in intact bacteria. N-1-(2-Nitrophenyl)ethoxycarbonyl-L-serine and 2-hydroxyphenyl 1-(2-nitrophenyl) ethyl phosphate were synthesized. These compounds are a 'caged' serine and a 'caged' proton and on flash photolysis release serine and protons and attractant and repellent ligands, respectively, for Tsr, the serine receptor. The product quantum yield for serine was 0.65 (+/- 0.05) and the rate of serine release was proportional to [H+] near-neutrality with a rate constant of 17 s-1 at pH 7.0 and 21 degrees C. The product quantum yield for protons was calculated to be 0.095 on 308-nm irradiation but 0.29 (+/- 0.02) on 300-350-nm irradiation, with proton release occurring at > 10(5) s-1. The pH jumps produced were estimated using pH indicators, the pH-dependent decay of the chromophoric aci-nitro intermediate and bioassays. Receptor deletion mutants did not respond to photorelease of the caged ligands. Population responses occurred without measurable latency. Response times increased with decreased stimulus strength. Physiological or genetic perturbation of motor rotation bias leading to increased tumbling reduced response sensitivity but did not affect response times. Exceptions were found. A CheR-CheB mutant strain had normal motility, but reduced response. A CheZ mutant had tumbly motility, reduced sensitivity, and increased response time to attractant, but a normal repellent response. These observations are consistent with current ideas that motor interactions with a single parameter, namely phosphorylated CheY protein, dictate motor response to both attractant and repellent stimuli. Inverse motility motor mutants with extreme rotation bias exhibited the greatest reduction in response sensitivity but, nevertheless, had normal attractant response times. This implies that control of CheY phosphate concentration rather than motor reactions limits responses to attractants.

Objective: Antiphospholipid antibodies (aPL) are noted with increased frequency in patients with systemic lupus erythematosus (SLE). The main manifestations found to be associated with aPL are arterial and venous thrombotic events, thrombocytopenia, and recurrent pregnancy loss. This study is an attempt to define the incidence of aPL in patients with childhood-onset SLE and in their relatives and to correlate their presence with clinical manifestations, and especially, to evaluate the risk of thrombosis in aPL-positive subjects., Methodology: We studied 37 unrelated patients and 107 of their first-degree relatives. VDRL, IgG and IgM anticardiolipin, and IgG antiphosphatidylethanolamine antibodies were studied in all probands during periods of clinical remission and in first-degree relatives at the time of interview. Lupus anticoagulant had only been studied in probands during an SLE flare-up., Results: Thirty-eight percent of probands and 19% of relatives were positive for at least one aPL, with little overlap between the different aPL studied. -No aPL-negative proband developed thrombosis. Two of the aPL-positive probands had thrombotic events before testing, and a third one showed thrombosis after testing. Only two probands had high levels of IgG aCL and showed thrombosis. The occurrence of aPL positivity in relatives was not always related to its presence in probands. None of the aPL-positive relatives had had thrombosis, but recurrent fetal loss was noted in one aPL-positive mother with SLE. Although there was a high frequency of SLE, SLE-like disease, auto-immune disorders or positive serological findings for lupus in first-degree relatives, many of these relatives did not test positive for aPL., Conclusion: The high levels of IgG aCL may be considered a risk factor for thrombosis. Findings in relatives suggest a multifactorial origin for autoimmune disease and antibody production.

After partial hepatectomy in the rat, there is substantial lipid accumulation in the liver. No information is available on the possible role of receptor-mediated endocytosis in this process. Since the low density lipoprotein (LDL) receptor is stimulated as a part of an early growth response in cell culture (Ellsworth et al.: Biochem. J. 279:175-187, 1991), the metabolism of this receptor during liver regeneration was studied. The mRNA and membrane protein level of the receptor were measured in the liver and in the adrenal glands at different times after partial hepatectomy, corresponding to different phases of the cell cycle. A discontinuous pattern of receptor expression is detectable in the regenerating liver; a large increase of mRNA and membrane protein occurs at an early time (2-4 h), suggesting that there is induction of LDL receptor gene transcription shortly after partial hepatectomy. This response seems specific for the liver following injury since the adrenal receptor does not show a different pattern in partially hepatectomized rats and sham-operated controls. After returning to control levels, the LDL receptor again increases slightly above control at 24 h, a time when cell replication begins.

Solutions of porcine kidney diamine oxidase, PKDAO, and bovine plasma amine oxidase, BPAO, were saturated with the spin-trapping agent alpha-phenyl-N-t-butylnitrone, PBN, and incubated with cadaverine or benzylamine substrate, respectively, under aerobic conditions. EPR spectra due to trapped hydroxyl radicals were obtained for both enzymes with no evidence of superoxide formation. Under anaerobic conditions, hydroxyl radicals were formed only when H2O2 was present as well as substrate. Catalase prevented hydroxyl radical formation by PKDAO but not BPAO. The results indicate that hydroxyl radical is produced in the reaction of the product H2O2 with the reduced enzymes and therefore may be important in turnover-related enzyme degradation, but is not a true reaction intermediate.

Six cases of vascular lesion associated with percutaneous removal of renal calculi are described. The lesions had been caused during percutaneous access or ultrasound fragmentation of the stone. Those that had resolved spontaneously have not been included. When vascular injury is suspected during the procedure, the appropriate measures must be taken to resolve the different complications, such as arteriovenous, arteriocaliceal or venocaliceal fistula. Since there is little reference to this subject in the Spanish literature, the experience described herein may be of interest to the urologist who has only recently begun to perform this technique.

Small aliquots of rat high-density lipoproteins (HDL) (388 +/- 67 nmol lipoprotein cholesterol) were labeled with [14C]cholesterol and administered as a bolus to perfused rat livers. Bile and perfusate samples were collected for 2 hours at 30-minute intervals. After perfusion, both the microsomes and lipid extracts were prepared from the livers. Lipid composition was examined in both liver and microsomes, and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity was evaluated in microsomes. Basal values of bile flow, lipid composition, and enzyme activity were evaluated using livers in which perfusion was discontinued before injecting the lipoprotein. In some experiments, the effect of perfusion per se was assessed by infusing saline instead of lipoprotein. After 10 minutes of lipoprotein perfusion, 50% of cholesterol administered was taken up by the perfused liver. During infusion, transient but significant increases in both bile flow and bile steroid secretion were observed. Cholesterol administration, even if rapid, represented less than 0.4% of total liver cholesterol content. However, this was enough to significantly increase the cholesterol to phospholipid (CH/PL) molar ratio in liver microsomes and at the same time decrease HMG-CoA reductase activity. In conclusion, the main response of the perfused liver to HDL cholesterol infusion is a reduced activity of the rate-limiting enzyme in cholesterol biosynthesis, due to the shift in the microsomal CH/PL molar ratio. A small proportion of the infused cholesterol enters bile as cholesterol and bile salts.

Unlike the other exploratory techniques (US, arteriography, phlebography, CT and MRI), laparoscopy constitutes the only reliable diagnostic method for absent testis, which has a high incidence in children with non palpable testes. The finding of the blind-ending spermatic veins and deferent duct confirms the diagnosis of absent testis.

Presentation of 31 patients with suspected diagnosis of upper urothelium tumour seen between 1986 and 1991. In agreement with our study protocol and the endourological treatment followed in this type of tumours, there were eight false positive, which were excluded. Five cases were treated by retrograde ureterorenoscopy while in the remaining 18 cases the anterograde percutaneous approach was used. A review of both the aspects of endoeurological treatment and the evolution of these patients during that period is included.

1. The isoprenoid metabolism of the green frog has been studied, taking into consideration the transport and uptake mechanisms of plasma lipoproteins. 2. The lipoprotein complexes separated on KBr gradient showed six discrete peaks in both sexes. 3. The mechanisms of cellular uptake have been studied by immunological procedures. A molecule homologous to rat LDL receptor, and sharing its ability to bind only specific lipoproteins, has been shown. 4. Homology at mRNA level has also been demonstrated by Northern blot analysis and two different messengers have been shown in both male and female frog.

Presentation of 31 patients with suspected diagnosis of upper urothelium tumour seen between 1986 and 1991. In agreement with our study protocol and the endourological treatment followed in this type of tumours, there were eight false positive, which were excluded. Five cases were treated by retrograde ureterorenoscopy while in the remaining 18 cases the anterograde percutaneous approach was used. A review of both the aspects of endourological treatment and the evolution of these patients during that period is included.

The effects of HDL1 lipoprotein infusion on biliary lipid secretion were studied in the in vitro model of rat perfused liver. A strong increase in bile flow was observed during and after lipoprotein infusion. This caused a significant rise in cholesterol, phospholipid and bile salt secretions. However, only the percentage of cholesterol increased with respect to the other bile lipids. The changes observed in the cholesterol/phospholipid molar ratio values of liver membrane subfractions (i.e., liver plasma membrane, mitochondria plus lysosomes and microsomes) isolated from the perfused rat liver after HDL1 administration were not significant.

In female frogs (Rana Esculenta) during gametogenesis the cholesterol synthesized in the liver by 3-hydroxy-3-methylglutaryl coenzyme A reductase is mostly exported into the blood and taken up by the oocytes. In order to understand the fate of the neosynthesized cholesterol, female and male frogs and estrogenized male controls were injected with the labelled precursor 14C mevalonate. In females and in estrogenized controls, mevalonate-derived radioactivity is found in a plasmatic lipoprotein that has been identified as vitellogenin by immunological detection. The increased 3-hydroxy-3-methylglutaryl coenzyme A reductase activity present in females in Fall is likely to be committed to provide cholesterol for the lipidation of this cholesterol-rich protein.

Sixteen hours after partial hepatectomy in the rat, the synthesis of mevalonate (MVA) is not committed to produce cholesterol and only partially utilized for dolichol formation. In order to investigate the fate of MVA in this replicative system, slices of the remaining liver were incubated with 5-3H-MVA. Labeled proteins from whole liver and purified nuclei were analyzed after extensive delipidation and separation by SDS-PAGE. Many MVA-derived proteins were identified at 16 hours, while only two labelled peptides were detectable at 24 hours. The radioactivity was localized in covalently bound lipid moieties. A highly labeled 26 kD peptide was detectable in the nucleus at 16 hours, suggesting its involvement in the cell cycle progression.

Friedreich's ataxia (FA) is a progressive degenerative disease involving both central and peripheral nervous system. It is an autosomal recessive hereditary disorder, which begins around puberty and has an unknown genetic basis and biochemical defect. The recent mapping of FA locus in human chromosome 9 by means of the analysis of the molecular genetic linkage has permitted to evaluate FA genetics with polymorphic genetic markers (RFLPs) that are secreted linked with the FA gene. The normal and mutant allele secretion of FA was evaluated in ten Spanish families with one or two members with FA by means of several cloned probes (MCT112, DR47, D9S1 and HHH220), localized in chromosome 9 and strongly linked to FA gene, with the aim of achieving a predictive diagnosis of relatives in the pediatric age and to detect healthy carriers. In 9 out of 10 families some totally or partially informative RFLP were found. In 5 of 6 relatives in pediatric age the future development of the disease could be ruled out. By contrast, the carrier status could only be identified in three relatives. In a family with two affected children a genetic recombinant for D9S1 was found. Remarkably, one of them had a better clinical evolution and preserved tendon reflexes in lower limbs.

We analyze the results achieved by extracorporeal piezoelectric lithotripsy (EPL) in 40 renal units with the following anatomic anomalies: solitary kidney (10), chronic pyelonephritis (8), medulospongiosis (6), caliceal diverticula (5), intrarenal cysts (5), pyeloureteric duplicity (3), megacaliosis (1), horseshoe kidney (1), and ureterocele (1). Adequate stone fragmentation was achieved in 80% of the patients with solitary kidney; 20% developed obstructive complications (1 patient with a 15 mm.-stone and 1 patient with obstruction of double-J catheter). At 6 months, stone remnants persist in 71.4% (5/7) after stone fragmentation in those cases with pyelonephrosis. The rates for fragmentation, elimination of remaining stone fragments, and disappearance of pain are 80%, 50% and 100% for diverticular caliceal calculi, and 67%, 50% and 100% for those in kidney with medulospongiosis. We can conclude that placement of a double-J catheter is useful before EPL in patients with solitary kidney and calculi greater than 10 mm. EPL is the first therapeutic approach in symptomatic lithiasis in caliceal diverticula or precaliceal cystic dilatation (medulospongiosis).

C4 genotyping was performed in 38 unrelated patients with systemic lupus erythematosus (SLE) aged 2-16 years at onset. Null alleles were found in 68% of patients. Ten patients had one null allele at the C4A locus and 11 others had one null allele at the C4B locus. One patient was homozygous for C4A*Q0, 2 homozygous for C4B*Q0 and 1 heterozygous C4A*Q0/B*Q0. The last patient was C4A*Q0B*Q0/B*Q0. Three patients, being heterozygous C2 deficient, had thus a combined C2 and C4 deficiency. A significant increase in C4 null alleles in SLE patients has been reported by different authors who have suggested that the C4 null alleles confer susceptibility to SLE. However, when considering only the 20 patients of French descent, no differences in gene frequencies were found between this group and the French population. Disease patterns were compared in patients with or without null C4. Renal involvement was more frequent in the C4A*Q0 or C4A*Q0/C4A*Q0 patients than in the patients without null C4 (9/11 vs 3/12). These data suggest that differences between these results and those of others might be due to clinical heterogeneity of the disease, as exemplified by the frequency of renal involvement in the different groups. In susceptible individuals the absence of one C4A gene product may predispose the patient to renal involvement.