Detection of pap, sfa and afa adhesin-encoding operons in uropathogenic Escherichia coli strains: relationship with expression of adhesins and production of toxins

A total of 243 Escherichia coli strains isolated from patients with urinary tract infections (UTI) were investigated for the presence of pap, sfa and afa adhesin-encoding operons by using the polymerase chain reaction. It was found that 54%, 53% and 2% of the strains exhibited the pap, sfa and afa genotypes, respectively. Pap+ and/or sfa+ strains were more frequent in cases of acute pyelonephritis (94%) than in cases of cystitis (67%) (P < 0.001) and asymptomatic bacteriuria (57%) (P < 0.001). The pap and/or sfa operons were found in 90% of strains expressing mannose-resistant haemagglutination (MRHA) versus 37% of MRHA-negative strains (P < 0.001). The presence of pap and sfa operons was especially significant in strains belonging to MRHA types III (100%) (without P adhesins) and IVa (97%) (expressing the specific Gal-Gal binding typical of P adhesins). Both pap and sfa operons were closely associated with toxigenic E. coli producing alpha-haemolysin (Hly+) and/or the cytotoxic necrotizing factor type 1. There was an apparent correlation between the pap and sfa operons and the O serogroups of the strains. Thus, 93% of strains belonging to O1, O2, O4, O6, O7, O14, O15, O18, O22, O75 and O83 possessed pap and/or sfa operons, versus only 32% of strains belonging to other serogroups (P < 0.001). The results obtained in this study confirm the usefulness of our MRHA typing system for presumptive identification of pathogenic E. coli exhibiting different virulence factors. Thus, 85% of strains that possessed both pap and sfa adhesin-encoding operons showed MRHA types III or IVa previously associated with virulence of E. coli strains that cause UTI and bacteraemia.     

Epitopes of the P-fimbrial adhesin of E. coli cause different urinary tract infections

PURPOSE: This is a study of the interaction of the tip protein of P-fimbriae E. coli, its specific urothelial adhesin, and urothelial receptors for the adhesin. This tip protein has several epitopes that adhere to different isoreceptors containing the urothelial alpha-gal-1-4 beta-gal disaccharide. Renal tubular cells of our monkey model contain the globoside isoreceptor, and thus ureteral inoculation of E. coli with the class II tip protein leads to pyelonephritis. The class III tip protein adheres to the Forssman antigen and causes cystitis in humans. MATERIALS AND METHODS: An E. coli strain, DS17 which originally caused pyelonephritis in a child, is P-fimbriated and contains a class II tip adhesin. A mutant was produced to contain a class III tip adhesin. Eight monkeys had a ureteral inoculation of E. coli DS17 and 4 monkeys with E. coli DS17-1. In addition, we studied in vitro adherence by these strains. RESULTS: We show that in vitro adherence by the tip protein of P-fimbriae to bladder cells of the monkey occurs by several mechanisms, adhering to specific receptors for the class II and III epitopes of the tip protein as well as by means of type 1 fimbriae. In addition, the PapE protein of the fibrillum of the P-fimbriae adheres to fibronectin. As always, electrostatic and hydrophobic interaction remain important contributions to adherence. E. coli DS17 caused pyelonephritis, but DS17-1 caused cystitis. Bacteriuria was prolonged by DS17 infection. CONCLUSION: The site of a urinary tract infection from P-fimbriated E. coli can be predicted by the epitope of the tip protein of P-fimbriae.     

Pathogenicity island sequences of pyelonephritogenic Escherichia coli CFT073 are associated with virulent uropathogenic strains

Urinary tract infection is the most frequently diagnosed kidney and urologic disease, and Escherichia coli is by far the most common etiologic agent. Defined blocks of DNA termed pathogenicity islands have been found in uropathogenic strains to carry genes not generally found in fecal strains. We have identified one of these regions of DNA within the chromosome of the highly virulent E. coli CFT073, isolated from the blood and urine of a woman with acute pyelonephritis. This strain, which is cytotoxic for cultured renal cells and causes acute pyelonephritis in transurethrally infected CBA mice, contains two distinct copies of the pap operon and is hemolytic. One pap operon was localized on a cosmid clone which was used to identify three overlapping cosmid clones. By using restriction mapping, DNA hybridization, sequencing, and PCR amplification, a region of approximately 50 kb was found to be present in this uropathogenic strain and to have no corresponding sequences in E. coli K-12. This gene block also carries hemolysin genes hlyCABD. The pathogenicity island begins 7 bp downstream of dadX (catabolic alanine racemase; 26.55 min) and ends at a position in the K-12 genome 75 bp downstream of the metV tRNA gene (62.74 min); this suggests that a chromosomal rearrangement has occurred relative to the K-12 linkage map. The junctions of the pathogenicity island were verified by PCR amplification directly from the genomic DNA of strain CFT073. DNA sequencing within the boundaries of the junctions revealed genes not previously identified in E. coli or in some cases bearing no known homologs. When used as probes for DNA hybridization, these sequences were found significantly more often in strains associated with the clinical syndromes of cystitis (82%) and acute pyelonephritis (79%) than in fecal strains (19%; P < 0.001).     

GH dependence and GH withdrawal syndrome in GH treatment of short normal children: evidence from growth and cardiac output

BACKGROUND: Cytokines and cytokine receptors are involved in the systemic and local inflammatory response in patients with urinary tract infections. METHODS: We examined urine and serum concentrations of soluble IL-6 receptor (sIL-6R), IL-10 and granulocyte colony-stimulating factor (G-CSF) in 29 women with acute pyelonephritis caused by Escherichia coli 2 weeks after the infection, during the subsequent episode of cystitis or asymptomatic bacteriuria and also later when the same patients were free from bacteriuria. Concentrations of sIL-6R, IL-10 and G-CSF were related to the expression of five virulence markers of E. coli and to glomerular filtration rate (GFR) after pyelonephritis. RESULTS: On admission because of acute pyelonephritis the serum concentration of sIL-6R was similar to that of 12 healthy controls. Two weeks after the infection when all patients had received antibiotic treatment, the serum concentration of sIL-6R was significantly higher compared to that on admission (p < 0.001) and also higher compared to healthy controls (p = 0.001). Patients with increased concentrations of sIL-6R in serum 2 weeks after infection had significantly lower GFR at follow-up (p < 0.05). Patients with acute pyelonephritis had higher concentrations of G-CSF and IL-10 in serum compared to healthy subjects (p < 0.001 and p = 0.06, respectively). G-CSF in serum was higher in patients infected by E. coli producing cytotoxic necrotizing factor (p < 0.05). Patients infected by strains producing hemolysin had lower concentrations of sIL-6R (p < 0.001). Patients with detectable levels of the anti-inflammatory cytokine IL-10 in serum had significantly higher concentrations of IL-6 and the soluble tumor necrosis factor receptors I and II in serum as compared to patients in whom IL-10 was not detectable (p < 0.001, p = 0.001 and p < 0.05, respectively. CONCLUSION: These investigations, together with our previous findings summarized in this paper, contribute to an increased understanding of the local and systemic inflammatory response arising in response to acute pyelonephritis.     

Cytotoxicity of hemolytic, cytotoxic necrotizing factor 1-positive and -negative Escherichia coli to human T24 bladder cells

Approximately one-half of Escherichia coli isolates from patients with cystitis or pyelonephritis produce the pore-forming cytotoxin hemolysin, a molecule with the capacity to lyse erythrocytes and a range of nucleated cell types. A second toxin, cytotoxic necrotizing factor 1 (CNF1), is found in approximately 70% of hemolytic, but rarely in nonhemolytic, isolates. To evaluate the potential interplay of these two toxins, we used epidemiological and molecular biologic techniques to compare the cytotoxicity of hemolytic, CNF1(+), and CNF1(-) cystitis strains toward human T24 bladder epithelial cells in vitro. A total of 29 isolates from two collections of cystitis-associated E. coli were evaluated by using methylene blue staining of bladder monolayers at 1-h intervals after inoculation with each strain. Most (20 of 29) isolates damaged or destroyed the T24 monolayer (less than 50% remaining) within 4 h after inoculation. As a group, CNF1(+) isolates from one collection (11 strains) were less cytotoxic at 4 h than the CNF1(-) strains in that collection (P = 0.009), but this pattern was not observed among isolates from the second collection (18 strains). To directly evaluate the role of CNF1 in cytotoxicity of hemolytic E. coli without the variables present in multiple clinical isolates, we constructed mutants defective in production of CNF1. Compared to the CNF1(+) parental isolates, no change in cytotoxicity was detected in these cnf1 mutants. Our results indicate that CNF1 does not have a detectable effect on the ability of hemolytic E. coli to damage human bladder cell monolayers in vitro.     

Clonal relationships among bloodstream isolates of Escherichia coli

The clonal relationships among 187 bloodstream isolates of Escherichia coli from 179 patients at Boston, Mass., Long Beach, Calif., and Nairobi, Kenya, were determined by multilocus enzyme electrophoresis (MLEE), analysis of polymorphisms associated with the ribosomal operon (ribotyping), and serotyping. MLEE based on 20 enzymes resolved 101 electrophoretic types (ETs), forming five clusters; ribotyping resolved 56 distinct patterns concordant with the analysis by MLEE. The isolates at each study site formed a genetically diverse group and demonstrated similar clonal structures, with the same small subset of lineages accounting for the majority of isolates at each site. Moreover, two ribotypes accounted for approximately 30% of the isolates at each study site. One cluster contained the majority (65%) of isolates and, by direct comparison of the ETs and ribotypes of individual isolates, was genetically indistinguishable from the largest cluster for each of two other collections of E. coli causing pyelonephritis and neonatal meningitis (R. K. Selander, T. K. Korhonen, V. V?is?nen-Rhen, P. H. Williams, P. E. Pattison, and D. A. Caugent, Infect. Immun. 52:213-222, 1986; M. Arthur, C. E. Johnson, R. H. Rubin, R. D. Arbeit, C. Campanelli, C. Kim, S. Steinbach, M. Agarwal, R. Wilkinson, and R. Goldstein, Infect. Immun. 57:303-313, 1989), thus defining a virulent set of lineages. The isolates within these virulent lineages typically carried DNA homologous to the adhesin operon pap or sfa and the hemolysin operon hly and expressed O1, O2, O4, O6, O18, O25, or O75 antigens. DNA homologous to pap was distributed among isolates of each major cluster, whereas hly was restricted to isolates of two clusters, typically detected in pap-positive strains, and sfa was restricted to isolates of one cluster, typically detected in pap- and hly-positive strains. The occurrence of pap-positive isolates in the same geographically and genetically divergent lineages suggests that this operon was acquired early in the radiation of E. coli, while hly and sfa were acquired subsequently, most likely by pap-positive and pap- and hly-positive precursors, respectively.     

Comparison of Escherichia coli strains recovered from human cystitis and pyelonephritis infections in transurethrally challenged mice

Urinary tract infection, most frequently caused by Escherichia coli, is one of the most common bacterial infections in humans. A vast amount of literature regarding the mechanisms through which E. coli induces pyelonephritis has accumulated. Although cystitis accounts for 95% of visits to physicians for symptoms of urinary tract infections, few in vivo studies have investigated possible differences between E. coli recovered from patients with clinical symptoms of cystitis and that from patients with symptoms of pyelonephritis. Epidemiological studies indicate that cystitis-associated strains appear to differ from pyelonephritis-associated strains in elaboration of some putative virulence factors. With transurethrally challenged mice we studied possible differences using three each of the most virulent pyelonephritis and cystitis E. coli strains in our collection. The results indicate that cystitis strains colonize the bladder more rapidly than do pyelonephritis strains, while the rates of kidney colonization are similar. Cystitis strains colonize the bladder in higher numbers, induce more pronounced histologic changes in the bladder, and are more rapidly eliminated from the mouse urinary tract than pyelonephritis strains. These results provide evidence that cystitis strains differ from pyelonephritis strains in this model, that this model is useful for the study of the uropathogenicity of cystitis strains, and that it would be unwise to use pyelonephritis strains to study putative virulence factors important in the development of cystitis.     

Associations between virulence factors of Shiga toxin-producing Escherichia coli and disease in humans

Associations between known or putative virulence factors of Shiga toxin-producing Escherichia coli and disease in humans were investigated. Univariate analysis and multivariate logistic regression analysis of a set of 237 isolates from 118 serotypes showed significant associations between the presence of genes for intimin (eae) and Shiga toxin 2 (stx2) and isolates from serotypes reported in humans. Similar associations were found with isolates from serotypes reported in hemorrhagic colitis and hemolytic-uremic syndrome. The enterohemorrhagic E. coli (EHEC) hemolysin gene was significantly associated with isolates from serotypes found in severe diseases in univariate analysis but not in multivariate logistic regression models. A strong association between the intimin and EHEC-hemolysin genes may explain the lack of statistical significance of EHEC hemolysin in these multivariate models, but a true lack of biological significance of the hemolysin in humans or in disease cannot be excluded. This result warrants further investigations of this topic. Multivariate analysis revealed an interaction between the eae and stx2 genes, thus supporting the hypothesis of the synergism between the adhesin intimin and Shiga toxin 2. A strong statistical association was observed between the stx2 gene and severity of disease for a set of 112 human isolates from eight major serotypes. A comparison of 77 isolates of bovine origin and 91 human isolates belonging to six major serotypes showed significant associations of the genes for Shiga toxin 1 and EspP protease with bovine isolates and an increased adherence on HEp-2 cell cultures for human isolates, particularly from diarrheic patients and healthy persons.     

Clinical evaluation of nitrite test for the detection of bacteriuria

A nitrite test for bacteriuria was compared with routine microscopic examination in 1,318 clinical urine specimens and with bacterial culture in 132. Sensitivity, specificity, positive predictive value and accuracy rate are as follows; for diagnosis of bacteriuria more than 10(4) CFU/ml, 53.4%, 88.6%, 90.4 and 65.2%, respectively; for more than 10(5) CFU/ml, 55.4%, 87.8%, 88.4% and 67.2%, respectively. The positive rate for the nitrite test was 21.4% for bacteriuria of > or = 10(4) CFU/ml in gram positive cocci and 56.9% in gram negative rods. False negative results were obtained from gram positive cocci without nitrate reductive activity or from patients with acute uncomplicated cystitis because of insufficient incubation time in urinary tract. However, this simple test is valuable in the detection of bacteriuria in clinical practice with high specificity.     

Alport syndrome with a peculiar pattern of distribution of the alpha3-alpha6 chains of type IV collagen in renal basement membrane

Chronic bacteriuria is a common occurrence among spinal-cord injury patients and others with neuropathic bladders. If bacteria are present in the urinary tract, the patient may develop symptoms of infection or remain asymptomatic. We have compared virulence properties of 28 Escherichia coli isolates from patients with symptomatic urinary tract infections (UTI) and 29 E. coli isolates from patients with asymptomatic bacteriuria (ABU). Bacteria from patients with symptomatic UTI were more likely to be hemolytic than isolates from patients with ABU (P = 0.05) or fecal isolates obtained from healthy volunteers (P < 0.001). Bacteria from patients with symptomatic UTI were also more likely than strains isolated from patients with ABU (P = 0.08) or fecal strains (P < 0.001) to exhibit D-mannose-resistant hemagglutination of human erythrocytes. The results suggest that E. coli isolates from nonimmunocompromised patients who require intermittent catheterization and who develop symptomatic UTI may be distinguished from bacteria recovered from patients who remain asymptomatic and possibly from normal fecal E. coli.     

Treatment of urinary tract infections

Urinary tract infections (UTIs) are common conditions in clinical practice. For uncomplicated UTIs, the causative organisms and their antimicrobial susceptibility profiles are generally predictable, and empiric short course (3 day) antibiotic therapy after an abbreviated laboratory workup is advocated. Acute pyelonephritis requires a 2 week antibiotic course, often with initial parenteral therapy. Women with frequent recurrences of UTIs may require intermittent self-treatment or continuous or postcoital antibiotic prophylaxis. Catheter-associated UTIs generally only require treatment if the patient shows signs of systemic infection. Treatment of asymptomatic bacteriuria is only recommended in certain circumstances. Careful consideration of the clinical circumstances, the patient's known or predicted urinary tract anatomy, and the antibiotic susceptibility of the bacterial pathogen(s) are critical factors in the choice of appropriate therapy for urinary tract infections.     

The occurrence of ambient temperature-regulated adhesins, curli, and the temperature-sensitive hemagglutinin tsh among avian Escherichia coli

Escherichia coli establishes a secondary respiratory tract infection in birds following inhalation of contaminated dust and litter particles. Escherichia coli express adhesins under conditions reflective of the ambient temperatures present in poultry houses. These microbial adhesins allow E. coli to attach to cell types that it initially encounters in the respiratory tract. Ambient temperature-regulated adhesins represent a new class of bacterial hemagglutinins that include pili and the thin, aggregative, flexible filaments known as "curli." This study examines the occurrence of the ambient temperature-regulated adhesins, curli (crl, csgA), and an avian-specific, temperature-sensitive hemagglutinin, tsh, among avian and mammalian E. coli isolates. The avian hemagglutinin gene tsh was present in approximately 46% of clinical avian E. coli isolates. This gene was not detected among commensal E. coli isolated from healthy broiler chickens. Unlike tsh, curli genes were ubiquitous among E. coli. However, curli were observed in only half of the avian E. coli examined by electron microscopy. Curli were not present among several nonavian E. coli positive for crl and csgA. Approximately 25% of avian E. coli isolates agglutinated chicken erythrocytes when bacteria were grown at room temperature. Hemagglutination was not specific to E. coli isolated from poultry. Presence of either tsh or curli genes was not indicative of an isolate's ability to agglutinate chicken red blood cells. No discernible structures were observed mediating attachment of the bacteria to chicken red blood cells. An additional avian-specific hemagglutinin appears to be present among avian E. coli.     

Hemagglutination ability and adherence to the Buffalo green monkey kidney cell line of uropathogenic Escherichia coli

The hemagglutination ability and adherence capacity to the Buffalo green monkey (BGM) kidney cell line of 160 wild-type strains of Escherichia coli isolated from bacteriuric patients were investigated. It was found that P-fimbriated E. coli strains adhered significantly better to BGM cells than did strains in which P-fimbriae were not detected, which is in accordance with the capacity of P-fimbriated strains to cause unobstructive pyelonephritis and with receptor distribution for P-fimbriae in the urinary tract. The strains which exhibited other adhesions, alone or simultaneously, showed reduced adherence to BGM cells, while non-agglutinating strains, mostly isolated from urine of patients with asymptomatic bacteriuria, did not adhere at all or adhered poorly to the utilized cell line. The BGM cells served as a good experimental model for investigation of uropathogenic E. coli adherence; because these cells originate from the upper urinary tract, they are viable and not coated with Tamm-Horsfall protein.     

Human herpesvirus-6 (HHV-6): current status

Bacteria isolated from the urine of 142 patients with chronic pyelonephritis (CPN) were examined for pathogenic properties of the strains (bacteriuria, hemolytic, proteolytic properties, urease, adhesive activity, antibiotic resistance, the ability to inactivate bactericidal activity of the serum) to control the effect of the treatment: antibiotics combined with plasmapheresis or antibiotics combined with laser radiation (intravascular, transcutaneous, or both). Combined application of intravascular and transcutaneous laser irradiation in multimodality treatment reduces the number of highly pathogenic strains as well as antibactericidal activity of the urine strains. It also promotes normalization of bacteriuria level. Plasmapheresis is inferior to laser radiation but ranks the second in efficacy of action on urinary microflora. Thus, use of efferent methods, especially transcutaneous plus intravascular laser radiation, plasmapheresis, in combined treatment of pyelonephritis decreases pathogenicity of urine strains and normalizes bacteriuria.     

papG alleles among Escherichia coli strains causing urosepsis: associations with other bacterial characteristics and host compromise

Alleles I, II, and III of the P adhesin gene papG were sought by PCR among 75 Escherichia coli blood isolates from adults with urosepsis, and the papG genotype was compared with associated bacterial characteristics and host compromise status. Allele II predominated over allele III in the total population (71% of the strains versus 7% for allele III; P < 0.01). Allele I was not encountered. In comparison with allele II, allele III was significantly associated with the presence of hly and the absence of iuc (which encode hemolysin and aerobactin biosynthesis), nonagglutination of digalactoside-coated beads, absence of aerobactin production, membership in serogroups O6 and O18, and host compromise, particularly cancer and upper urinary tract abnormalities.     

Clinical, and light and electron microscopical findings in sows with cystitis

The clinical findings, and urinary and morphological changes in the urinary bladder were investigated in 25 sows with a urinary tract infection. Eubacterium suis was isolated from 12 of the sows but not from the other 13. The clinical signs did not always correlate with the morphological changes. The only clinical sign indicating the beginning of cystitis appeared to be a significant bacteriuria. Other urinary changes occurred later when the inflammatory processes were more severe. In contrast with cystitis due to other bacteria, infection with E suis frequently resulted in a macrohaematuria and urinary pH values above 8.0. However, the light and electron microscopical findings in the bladder mucosa were similar in the sows with and without cystitis due to E suis. The transformation of goblet cells and the development of mucin cysts were probably due to the local bladder defence mechanisms. More severe lesions were observed with E suis infections, which resulted in changes in the ureterovesical junctions and in ascending renal infection and uraemia.     

Urinary immunoreactive interleukin-1 alpha and interleukin-6 in bacteriuric institutionalized elderly subjects

Urinary immunoreactive interleukin-1 alpha and interleukin-6 levels were measured in specimens obtained from elderly institutionalized subjects, including 67 asymptomatic subjects (51 of whom were bacteriuric), 34 with fever from nonurinary sources, 15 with bacteriuria and 9 with symptomatic urinary infection. For bacteriuric subjects urinary interleukin-1 alpha and interleukin-6 levels were measurable in 18 (35%) and 22 (43%) asymptomatic subjects, respectively, 9 (60%) and 8 (53%) with nonurinary sources of fever, respectively, and 6 (67%) and 7 (78%) with urinary infection, respectively. For subjects without bacteriuria 1 of 16 (6.3%) who were asymptomatic and 5 (25%) with nonurinary sources of fever had measurable urinary interleukin-1 alpha, and 2 (13%) and 1 (5.3%), respectively, had measurable interleukin-6. Presence of interleukin-1 alpha or interleukin-6 was significantly associated with bacteriuria for asymptomatic and symptomatic subjects. Interleukin-1 alpha or interleukin-6 quantitative levels were lower in subjects without than with bacteriuria. Quantitative levels of interleukin-6 tended to decrease for bacteriuric subjects with symptomatic infection between acute and convalescent specimens. These observations suggest that interleukin-1 alpha and interleukin-6 are produced in association with bacteriuria in some elderly subjects. Variation in local cytokine production with time and the clinical significance of these observations require further study.     

Neuropathic pruritus

Colonization of the urinary tract by Candida species occurs particularly in diabetic or immunocompromised patients. We report the cases of two patients presenting with pneumaturia and urinary tract infection who were initially thought to have colovesical fistulae. In both patients a diagnosis of emphysematous pyelonephritis or cystitis due to candidal infection was subsequently made. These cases serve as a reminder of this rare presentation of a not uncommon urinary tract infection.     

Allele-dropout using PCR-based diagnosis for the splicing mutation in intron-2 of the CYP21B-gene: successful amplification with a Taq/Pwo-polymerase mixture

The splicing mutation in intron 2 (nucleotide 656) of the 21-hydroxylase gene (CYP21B) is the most common mutation causing congenital adrenal hyperplasia (CAH). Homozygosity for nt656G is associated with the classical phenotype of CAH. In several studies, a number of clinically asymptomatic relatives of CAH-patients were genotyped as nt656G homozygotes. We have proposed that the putative asymptomatic nt656G/G individuals are incorrectly typed due to dropout of one allele (in most cases nt656C) during PCR amplification. Here, we report the successful amplification of all alleles at nt656 with a Taq/Pwo DNA polymerase mixture in the primary PCR reaction. The results were independent from the type of polymerase used for sequencing reactions as the second step in mutation analysis.     

Monolayer formation and DNA synthesis of the outer epithelial cells from pearl oyster mantle in coculture with amebocytes

OBJECTIVES: In the absence of specific symptomatology in children and neurogenic bladder disease patients, the early diagnosis of acute pyelonephritis is a challenge. The aim of the present study was to determine if dimercaptosuccinic acid (DMSA) lesion-positive (acute pyelonephritis) patients have elevated urinary alpha1-microglobulin (alpha1-MG) excretion (no false negatives) and if DMSA lesion-negative (cystitis) patients have normal urinary alpha1-MG excretion (no false positives). METHODS: A selected population of 62 children above 3 months of age with a proven urinary tract infection were administered a DMSA scan. A control scan was performed after the acute phase of the illness, and the diagnosis of pyelonephritis (n = 44) was made retrospectively. The urinary alpha1-MG was determined by immunonephelometry. RESULTS: The urinary alpha1-MG-creatinine ratio was highly sensitive (98%) and specific (100%) and correlated with the DMSA scintigraphy images. Only 1 of the 44 patients with pyelonephritis and all of the cystitis patients (n = 18) had a normal urinary alpha1-MG (<10 mg/g). The drop in absolute DMSA uptake correlated significantly (r = 0. 758, p < 0.001) with the urinary alpha1-MG-creatinine ratio. The urinary alpha1-MG-creatinine ratio was significantly higher (p < 0. 02) in bilateral than in unilateral pyelonephritis. CONCLUSION: DMSA lesion-positive (acute pyelonephritis) patients have elevated urinary alpha1-MG excretion and DMSA lesion-negative (cystitis) patients have normal urinary alpha1-MG excretion.