Skip to main content
NCBI home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1992 Sep;45(9):802–805. doi: 10.1136/jcp.45.9.802

Possibility of separating toxins from bacteria associated with sudden infant death syndrome using anion exchange chromatography.

D B Drucker 1, H A Aluyi 1, J A Morris 1, D R Telford 1, B A Oppenheim 1, B A Crawley 1
PMCID: PMC495109  PMID: 1401212

Abstract

AIMS: To develop techniques for the characterisation of toxins elaborated by a strain of Escherichia coli associated with sudden infant death syndrome (SIDS). METHODS: E coli SIDS 04, isolated from the nasopharynx of a case of SIDS, was studied. Cell-free toxin preparations were standardised, their protein measured, and analytical separation of proteins achieved using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Acetone precipitation of proteins was required prior to Coomassie blue staining of bands. Preparative separation was achieved on an anion exchange column using a programmed concentration gradient of NaCl in TRIS buffer. Fractions were tested individually or pooled for presence of lethal toxin including endotoxin. Lethal toxin was detected with the chick embryo test system. Endotoxin was measured using a chromogenic modification of the Limulus amoebocyte assay. RESULTS: Twenty one peaks were detected by chromatography. Ten individual, or pooled, fractions were assayed for endotoxin which ranged from 27-33 pg/ml. Much greater variation was found when the same fractions were assayed in chick embryos. E coli fractions varied considerably in lethal toxicity, from 0/10 to 10/10 chick embryos killed/tested. Certain E coli fractions tested individually (lethality four out of 10 to eight out of 10) proved more lethal (10 out of 10) if pooled prior to testing. CONCLUSIONS: In E coli infection associated with SIDS relatively low concentrations of extracellular protein are lethally toxigenic for the chick embryo model of SIDS. These proteins can be separated analytically by SDS-PAGE and preparatively by anion exchange chromatography. Toxicity of individual fractions is not correlated with endotoxin concentrations in samples tested.

Full text

PDF
802

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bacon C. J. Over heating in infancy. Arch Dis Child. 1983 Sep;58(9):673–674. doi: 10.1136/adc.58.9.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bettelheim K. A., Goldwater P. N., Dwyer B. W., Bourne A. J., Smith D. L. Toxigenic Escherichia coli associated with sudden infant death syndrome. Scand J Infect Dis. 1990;22(4):467–476. doi: 10.3109/00365549009027079. [DOI] [PubMed] [Google Scholar]
  3. Bohach G. A., Schlievert P. M. Detection of endotoxin by enhancement with toxic shock syndrome toxin-1 (TSST-1). Methods Enzymol. 1988;165:302–306. doi: 10.1016/s0076-6879(88)65046-4. [DOI] [PubMed] [Google Scholar]
  4. Czuprynski C. J., Noel E. J., Ortiz-Carranza O., Srikumaran S. Activation of bovine neutrophils by partially purified Pasteurella haemolytica leukotoxin. Infect Immun. 1991 Sep;59(9):3126–3133. doi: 10.1128/iai.59.9.3126-3133.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Desmond H. P., Crampton J. M., Theakston R. D. Rapid isolation and partial characterization of two phospholipases from Kenyan Echis carinatus leakeyi (Leakey's saw-scaled viper) venom. Toxicon. 1991;29(4-5):536–539. doi: 10.1016/0041-0101(91)90030-u. [DOI] [PubMed] [Google Scholar]
  6. Entrup M., Brinkmann B. Histologische Lungenbefunde beim plötzlichen Kindstod. Z Rechtsmed. 1990;103(6):425–433. doi: 10.1007/BF01263151. [DOI] [PubMed] [Google Scholar]
  7. Finck-Barbançon V., Prévost G., Piémont Y. Improved purification of leukocidin from Staphylococcus aureus and toxin distribution among hospital strains. Res Microbiol. 1991 Jan;142(1):75–85. doi: 10.1016/0923-2508(91)90099-v. [DOI] [PubMed] [Google Scholar]
  8. Fleming P. J., Gilbert R., Azaz Y., Berry P. J., Rudd P. T., Stewart A., Hall E. Interaction between bedding and sleeping position in the sudden infant death syndrome: a population based case-control study. BMJ. 1990 Jul 14;301(6743):85–89. doi: 10.1136/bmj.301.6743.85. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gordon D., Cohen R. J., Kelly D., Akselrod S., Shannon D. C. Sudden infant death syndrome: abnormalities in short term fluctuations in heart rate and respiratory activity. Pediatr Res. 1984 Oct;18(10):921–926. doi: 10.1203/00006450-198410000-00001. [DOI] [PubMed] [Google Scholar]
  10. Honda T., Shiba A., Seo S., Yamamoto J., Matsuyama J., Miwatani T. Identity of hemolysins produced by Bacillus thuringiensis and Bacillus cereus. FEMS Microbiol Lett. 1991 Apr 15;63(2-3):205–209. doi: 10.1016/0378-1097(91)90087-q. [DOI] [PubMed] [Google Scholar]
  11. Jakeman K. J., Rushton D. I., Smith H., Sweet C. Exacerbation of bacterial toxicity to infant ferrets by influenza virus: possible role in sudden infant death syndrome. J Infect Dis. 1991 Jan;163(1):35–40. doi: 10.1093/infdis/163.1.35. [DOI] [PubMed] [Google Scholar]
  12. Keane W. F., Gekker G., Schlievert P. M., Peterson P. K. Enhancement of endotoxin-induced isolated renal tubular cell injury by toxic shock syndrome toxin 1. Am J Pathol. 1986 Jan;122(1):169–176. [PMC free article] [PubMed] [Google Scholar]
  13. Lee S., Barson A. J., Drucker D. B., Morris J. A., Telford D. R. Lethal challenge of gnotobiotic weanling rats with bacterial isolates from cases of sudden infant death syndrome (SIDS). J Clin Pathol. 1987 Dec;40(12):1393–1396. doi: 10.1136/jcp.40.12.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McKendrick N., Drucker D. B., Morris J. A., Telford D. R., Barson A. J., Oppenheim B. A., Crawley B. A., Gibbs A. Bacterial toxins: a possible cause of cot death. J Clin Pathol. 1992 Jan;45(1):49–53. doi: 10.1136/jcp.45.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Morris J. A., Haran D., Smith A. Hypothesis: common bacterial toxins are a possible cause of the sudden infant death syndrome. Med Hypotheses. 1987 Feb;22(2):211–222. doi: 10.1016/0306-9877(87)90145-9. [DOI] [PubMed] [Google Scholar]
  16. Rao P. N., Dube D. A., Weightman N. C., Oppenheim B. A., Morris J. Prediction of septicemia following endourological manipulation for stones in the upper urinary tract. J Urol. 1991 Oct;146(4):955–960. doi: 10.1016/s0022-5347(17)37974-0. [DOI] [PubMed] [Google Scholar]
  17. Schlievert P. M. Enhancement of host susceptibility to lethal endotoxin shock by staphylococcal pyrogenic exotoxin type C. Infect Immun. 1982 Apr;36(1):123–128. doi: 10.1128/iai.36.1.123-128.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Shinagawa K., Mitsumori M., Matsusaka N., Sugii S. Purification of staphylococcal enterotoxins A and E by immunoaffinity chromatography using a murine monoclonal antibody with dual specificity for both of these toxins. J Immunol Methods. 1991 May 17;139(1):49–53. doi: 10.1016/0022-1759(91)90350-o. [DOI] [PubMed] [Google Scholar]
  19. Steinschneider A., Weinstein S. L., Diamond E. The sudden infant death syndrome and apnea/obstruction during neonatal sleep and feeding. Pediatrics. 1982 Dec;70(6):858–863. [PubMed] [Google Scholar]
  20. Telford D. R., Morris J. A., Hughes P., Conway A. R., Lee S., Barson A. J., Drucker D. B. The nasopharyngeal bacterial flora in the sudden infant death syndrome. J Infect. 1989 Mar;18(2):125–130. doi: 10.1016/s0163-4453(89)91094-3. [DOI] [PubMed] [Google Scholar]
  21. Watt B. Infant botulism--one cause of 'cot death'. Microbiol Sci. 1987 Feb;4(2):57–58. [PubMed] [Google Scholar]
  22. Weinstein S. A., Smith L. A. Preliminary fractionation of tiger rattlesnake (Crotalus tigris) venom. Toxicon. 1990;28(12):1447–1455. doi: 10.1016/0041-0101(90)90158-4. [DOI] [PubMed] [Google Scholar]
  23. Werthammer J., Brown E. R., Neff R. K., Taeusch H. W., Jr Sudden infant death syndrome in infants with bronchopulmonary dysplasia. Pediatrics. 1982 Mar;69(3):301–304. [PubMed] [Google Scholar]
  24. de Azavedo J. C., Lucken R. N., Arbuthnott J. P. Chick embryo assay for staphylococcal toxic shock syndrome toxin-1. Methods Enzymol. 1988;165:344–347. doi: 10.1016/s0076-6879(88)65051-8. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES