Loss of fimbrial adhesion with the addition of Vaccinum macrocarpon to the growth medium of P-fimbriated Escherichia coli.

PURPOSE: Vaccinium macrocarpon--the American cranberry--irreversibly inhibits the expression of P-fimbriae of E. coli. Further effects on the function and expression of P-fimbriae were studied by growing P-fimbriated E. coli in solid media laced with cranberry juice. METHODS: Cranberry concentrate at pH 7.0 was added to CFA medium to a final concentration of 25%. E. coli strains JR1 and DS17 were plated on this medium with a plain CFA control and incubated at 37C. Cultures were tested for ability to agglutinate P-receptor specific beads. Bacteria were washed in PBS and agglutination retested. Cultures were also replated on plain CFA agar and rechecked for their ability to agglutinate. Transmission electron micrographs were performed on positive control and test bacteria. RESULTS: For E. coli strain JR1, P-fimbrial agglutination was inhibited after the third plating. DS17 was fully inhibited after the second plating. Washing in PBS did not affect agglutination, but replating on CFA agar allowed agglutination to recur. Electron micrographic study of control populations confirmed fimbriae. Fully inhibited bacteria had a 100% reduction in expression of fimbriae. Additionally, inhibited bacteria showed cellular elongation. CONCLUSIONS: Cranberry juice irreversibly inhibits P-fimbriae. Electron micrographic evidence suggests that cranberry juice acts on the cell wall preventing proper attachment of the fimbrial subunits or as a genetic control preventing the expression of normal fimbrial subunits or both.

Optimum testicular aspiration technique for deoxyribonucleic acid flow cytometric analysis of spermatogenesis in primates (Papio anubis).

Deoxyribonucleic acid flow cytometry on testicular tissue is an established method to evaluate spermatogenesis. Needle aspiration is less invasive and permits sequential sampling as compared with open testis biopsy. Using seven primates, we examined the needle aspiration technique. There was no significant difference among locations aspirated; the Westcott needle (Bard Urological, Covington, GA) provided the most rapid analysis, and both the Biopty gun (Bard Urological) and Westcott needle were easy to use. Baseline means +/- SD for ploidy distributions were 1N: 65.73% +/- 7.1; 2N: 18.95% +/- 5.5; 4N: 11.4% +/- 2, Experimental data generated from primate models using testis aspiration and flow cytometry may elucidate human infertile conditions.

Host-parasite relationships in acute pyelonephritis.

During a 1-year prospective study, a total of 15 patients (seven children and eight adults) were observed with acute nonobstructive pyelonephritis. P-fimbriated Escherichia coli was the causative pathogen in all 15 patients. The same serotype of E coli that was P-fimbriated was isolated from the vaginal introitus of 60% and from the fecal flora of 86% of these patients. The only host abnormality was moderate vesicoureteral reflux in 20% of the patients. Uroepithelial cells were isolated from the first morning-voided urine from patients, who had recovered from pyelonephritis, and from age-matched controls. Using fluorescein-labelled type 1 and P-fimbriated reference strains of E coli and fluorescence-activated cell sorting (FACS) analysis, we evaluated their ability to adhere to these uroepithelial cells. P-fimbriated E coli was more adherent than type 1 fimbriated E coli, and more P-fimbriated E coli adhered to the patients' cells. Our data show that both colonization with P-fimbriated strains of E coli and receptor availability are important in the pathogenesis of pyelonephritis.

Experimental pyelonephritis in the monkey. VII. Ascending pyelonephritis in the absence of vesicoureteral reflux.

Six adult male nonrefluxing monkeys were experimentally infected by inoculation of P-fimbriated E. coli into the bladder. Eight control monkeys were inoculated with a non-P-fimbriated E. coli strain. Inoculation with the P-fimbriated E. coli resulted in marked leukocytosis, prolonged bacteriuria and loss of renal function with a 66 per cent incidence of pyelonephritis. Death secondary to bilateral pyelonephritis was seen in 2 monkeys inoculated with P-fimbriated E. coli. Pyelonephritis was not seen in any of the monkeys inoculated with non-P-fimbriated E. coli. The study shows that ascending pyelonephritis can occur in monkeys in the absence of vesicoureteral reflux.

Prevention of pyelonephritis by immunization with P-fimbriae.

P-fimbriae have been found to be a virulence factor in both human and nonhuman primate urinary tract infections caused by Escherichia coli. This is because of bacterial adherence to urothelial cells which is mediated by adherence of fimbriae to a specific glycolipid receptor on the cell membrane of these cells. We purified P-fimbriae for immunization of monkeys. High titers of antifimbrial antibody protected against both acute and chronic pyelonephritis after renal inoculations of P-fimbriate Escherichia coli.

Receptors for pyelonephritogenic Escherichia coli in primates.

P-fimbriated Escherichia coli adhere to uroepithelial cells of monkeys by binding to receptors which are identical or very similar to those of man. In atraumatic experimental infections of monkeys, a P-fimbriated Escherichia coli strain induced ureteral malfunction with subsequent pyelonephritis. A non-P-fimbriated Escherichia coli strain did not. Thus, P-fimbriation of Escherichia coli represents a true virulence factor in the acquisition of pyelonephritis.

Biochemistry of the nonhuman primate prostate and seminal vesicles. I. Some chemical parameters.

Normative biochemical values for cranial prostate, caudal prostate, and seminal vesicles were obtained in four macaque species and in baboons, with a view to chemical comparison of the cranial lobe with the other two tissues. The cranial prostatic lobe and the seminal vesicles were both significantly different from the caudal prostate in prostatic acid phosphatase activity. However, citric acid and lactate dehydrogenase in the cranial lobe were comparable to concentrations in the caudal lobe and significantly different from the seminal vesicles. Dihydrotestosterone was higher in content than testosterone in all tissues of each species examined. The nonhuman primate cranial lobe seems to be a link between the seminal vesicles and the caudal prostate.

Biochemistry of the nonhuman primate prostate and seminal vesicles. II. Molecular weights of soluble proteins.

The molecular weights of the soluble proteins from cranial and caudal prostate and seminal vesicle tissues of several species of nonhuman primates were separated by electrophoresis. The tissues of animals belonging to the genus Macaca were found to be similar and are compared to those of other species. The molecular weights of these proteins are discussed with regard to known concentrations of prostate and seminal vesicle proteins.

Immunology of pyelonephritis in the primate model. VI. Effect of complement depletion.

The inflammatory response has been shown to be responsible for the renal damage from bacterial infection. Phagocytic events are responsible for damage to the host as well as the pathogen. In this study we evaluated the effect of complement depletion, in an attempt to decrease chemotaxis and opsonization and thus decrease phagocytosis. Acute renal damage was decreased from both a decrease in inflammatory cells and phagocytic events in the areas of bacterial inoculation.

Immunology of pyelonephritis in the primate model. V. Effect of superoxide dismutase.

Ascending acute pyelonephritis was produced in monkeys by infusion of bacteria through a ureteral catheter to the point of intrarenal reflux. This led to a significant inflammatory response with death of renal tubular cells in the area of the tubular granulocytes and bacteria. We gave superoxide dismutase, and found that the inflammatory response was decreased and fewer tubular cells were killed. Ultrastructural change was also decreased in tubular cells adjoining phagocytosing neutrophils. This suggests that renal damage following a bacterial infection may be due to the production and release of superoxide into the tubular lumen during phagocytosis. We believe that it is the initial event which may lead to the eventual loss of renal tissue and function called chronic pyelonephritis.

Prostate epithelial cell culture. Application to a possible nonhuman primate model of prostate disease.

We successfully cultured prostatic epithelial cells from the nonhuman primate and developed an in vitro in vivo model system for possible use in the study of prostatic disease. We used light and electron microscopy studies and prostatic acid phosphatase staining to verify this system as viable secretory prostatic epithelial cells.

Seasonal changes in the thyroid hormones of the male squirrel monkey.

The serum of male squirrel monkeys was analyzed for levels of thyroxine (T4) and triiodothyronine (T3) at intervals corresponding to (1) preparations for breeding, (2) termination of breeding, and (3) well after the breeding season. Highest concentrations of hormones were found prior to breeding when body weight was also highest. Significant differences in hormone levels were found across sampling period. The interaction between gonadotropins and thyroid hormones is discussed.

The growth and development of the squirrel monkey (Saimiri Sciureus).

Squirrel monkeys of 2 subspecies, Bolivian and Colombian, were removed from their mothers on the day of birth and nursery reared for up to 2 years of age. Infants were tested weekly for 12 weeks, then monthly for 1 year, and at 2 years of age. Tests included morphology (body weight, crown-rump length, and head measurements), behavior (reflexes, activity, reaction to the surrogate), and physiology (heart rate, respiratory rate, temperature and optokinetic nystagmus). Data show some likenesses and some differences between the 2 subspecies.

Comparative normal levels of serum triiodothyronine and thyroxine in nonhuman primates.

Normative values were obtained for triiodothyronine and thyroxine from four species of Old World primate (chimpanzees, rhesus monkeys, African green monkeys and talopoin monkeys) and a single species of New World primate (squirrel monkeys) represented by two subspecies, Colombian and Bolivian. The Bolivian squirrel monkeys exhibited the lowest values for both triiodothyronine and thyroxine. Male talapoins had the highest levels of thyroxine. Significant differences were found in levels of triiodothyronine and thyroxine between males and females of the same species and between the two subspecies of squirrel monkeys. Triiodothyronine:thryroxine ratios were consistently lower in the males of all species examined.

Age differences in short-term memory and cell loss in the cortex of the rat.

Studies with human and animal subjects have indicated age declines in short-term memory and cell loss in the cortex. Cell loss has been estamated by descriptive nonautomated methods. Declines in short-term memory may be related to reduced learning, movtivation, motor capacity, or some combination of these factors. Passive-avoidance tests of memory minimize these factors. Direct correlational studies on learning and memory in relation to cell loss in the same subject are not feasible in man and they have not been reported previously in animals. The aims of this study were to examine age differences in learning and short-term passive-avoidance memory in relation to cell packing density in the visual cortex of the Fisher 344 rat. Cell counts were made with a computer-guided, automated, image-analyzing system (TAS, Leitz). The following observations were made: (1) significant age differences in 2- and 6-hour short-term passive-avoidance retention or memory between mature and senescent rats were related to nonsignificant age differences in original learning inferred from starting latencies, running time and running distance and (2) sd compared to mature rats were associated with significant differences in neuron but not glia-vascular cell density in area 17 in the presence of nonsignificant age differences in cortical depth and brain weight. Aims of further studies are to establish the role of cell loss from the hippocampus in loss of short-term spatial memory with age and to develop criteria for differential counting of small neurons, glia, endothelial cells and pericytes.