JRG, Newal DR. Relaxin as an aetiological factor in diabetic embryopathy. Lancet 1988; i: 1428-30. 4. Weiss G, O’Byme EM, Steinetz BG. Relaxin: a product of the human


corpus luteum of pregnancy. Science 1976; 194: 948-49. 5. O’Byrne EM, Carriere BT, Sorensen L, et al. Plasma relaxin in pregnant women. J Clin Endocr Metab 1978; 47: 1106-10. 6. Quagliarello J, Slachter N, Steinetz B, et al. Serial relaxin concentrations in human pregnancy. Am J Obstet Gynecol 1979; 135: 43-44. 7. Olefsky JM, Saekow M, Kroc RL. Potentiation of insulin binding and insulin action by purified porcine relaxin. Ann NY Acad Sci 1982; 380: 200-15. 8. Jarrett JC, Ballejo G, Saleem TH, Tsibris JCM, Spellacy WN. The effect of prolactin and relaxin in insulin binding by adipocytes from pregnant women. Am J Obstet Gynecol 1984; 149: 250-55. 9. Pederson J. The pregnant diabetic and her newborn, 2nd edition. Baltimore: Williams and Wilkins, 1980. 10. Vasilenko P, Adams WC, Frieden EH. Comparison of systemic and uterine effects of relaxin and insulin in alloxan-treated, hyperglycemic rats. Proc Soc Exp Biol Med 1982; 169: 376-79. 11. Stewart MO, Whittaker PG, Persson B, et al. A longitudinal study of circulating progesterone, oestradiol, hCG and hPL dunng pregnancy in type I diabetic mothers. Br J Obstet Gynaecol 1989; 96: 415-23. 12. O’Byrne EM, Steinetz BG. Radioimmunoassay (RIA) of relaxin in sera of various species using an antiserum to porcine relaxin. Proc Soc Exp Biol


3. Edwards

Med 1976; 152: 272-76. 13. Steinetz BG, Randolph C, Mahoney CJ. Serum concentrations of relaxin, chorionic gonadotropin, estradiol-17 &bgr;, and progesterone in the reproductive cycle of the chimpanzee (Pan troglodytes). Endocrinology

(in press). 14. Whittaker PG, Stewart MO, Taylor A, Howell RJS, Lind T. Insulin-like growth factor I and its binding protein I during normal and diabetic pregnancies. Obstet Gynaecol 1990; 76: 223-29. 15. Szlachter BN, Quagliarello J, Jewelwitz R, et al. Relaxin in normal and pathogenic pregnancy. Obstet Gynecol 1982; 59: 167-70. 16. Loumaye E, Tewissen B, Thomas K. Characterization of relaxin radioimmunoassay using Bolton-Hunter reagent. Gynecol Obstet Invest 1978; 9: 262-67. 17. Bell RJ, Eddie LW, Lester AR, et al. Relaxin in human pregnancy serum measured with an homologous radioimmunoassay. Obstet Gynecol

1987; 69: 585-89. 18. Witt BR, Wolf GC, Wainwright CJ, et al. Relaxin, CA-125, progesterone, estradiol, Schwangerschaft protein, and human chorionic gonadotropin as predictors of outcome in threatened and nonthreatened pregnancies. Fertil Steril 1990; 53: 1029-36.

Quagliarello J, Goldsmith L, Steinetz B, Lustig D, Weiss G. Induction of relaxin secretion in nonpregnant women by human chorionic gonadotropin. J Clin Endocr Metab 1980; 51: 74-77. Stewart DR, Celniker AC, Taylor CA Jr, Cragun JR, Overstreet JW, Lasley BL. Relaxin in the peri-implantation period. J Clin Endocr

Metab 1990; 70: 1771-73. Okokon E, Collins WP, Sharma V, Lightman SL. The effect of human chorionic gonadotropin and pregnancy on the circulating level of relaxin. J Clin Endocr Metab 1991; 72: 1042-47. 22. Ladenheim RG, Tesone M, Charreau EH. Insulin action and characterization of insulin receptors in rat luteal cells. Endocrinology 1984; 115: 752-56. 23. Toyoda N, Murata K, Sugiyama Y. Insulin binding, glucose oxidation, and methylglucose transport in isolated adipocytes from pregnant rats near term. Endocrinology 1985; 116: 998-1002. 24. Sutter-Dub M-TH, Sfaxi A, Latrille F, Sodoyez-Goffaux F, Sodoyez JC. Insulin binding and action in adipocytes of pregnant rates: evidence that insulin resistance is caused by post-receptor binding defects. J Endocr 1984; 102: 209-14. 25. Sherwood OD. Relaxin. In: Knobil E, Neill JD (eds). The physiology of reproduction. New York: Raven Press, 1988: 585-673. 26. Sutter-Dub M-TH, Leclerq R, Felix JM, Jacquot R, Sutter BChJ. Serum progesterone and immuno-reactive insulin levels in the pregnant rat. Horm Metab Res 1973; 5: 18-21. 27. Pederson O, Beck-Nielson H, Klebe JG. Insulin receptors in the pregnant diabetic and her newborn. J Clin Endocr Metab 1981; 53: 1160-66. 28. Pagano G, Cassader M, Massorbrio M, et al. Insulin binding to human adipocytes during late pregnancy in healthy, obese and diabetic state. Horm Metab Res 1980; 12: 177-81. 29. Pawlina W, Larkin LH, Ogilvie S, Frost SC. Human relaxin inhibits division but not differentiation of 3T3-L-1 cells. Molec Cell Endocr 1990; 72: 55-61. 30. Neufeld ND, Kaplan SA, Lippe BM, Scott M. Increased monocyte receptor binding of [I125] insulin in infants of gestational diabetic mothers. J Clin Endocr Metab 1978; 47: 590-95. 31. Crelin ES. The development of the bony pelvis and its changes during pregnancy and parturition. Trans NY Acad Sci Ser II 1969; 31: 1049-58. 32. Haning RV, Steinetz BG, Weiss G. Elevated serum relaxin levels in multiple pregnancy after menotropin treatment. Obstet Gynecol 1985; 66: 42-45. 33. Bell RJ, Sutton B, Eddie LW, Healy DL, Johnston PD, Tregear W. Relaxin levels in antenatal patients following in vitro fertilization. Fertil Steril 1989; 52: 85-87. 21.

Johnson MR,

SHORT REPORT We recommend that whole blood should be held at 22°C to make use of inherent bactericidal activity; leucocytes should then be removed.




the following

when the temperature is below 17oc.s,7 In most blood transfusion services world wide, donor blood is cooled to 4°C within 6 h of collection to minimise bacterial

multiplication. In Amsterdam since 1987, all whole blood units have been stored at 22°C for 16-20 h before processing into components. We have investigated the effect of this step on bactericidal activity of the donor leucocytes in whole blood. If leucocytes with internalised bacteria were then removed, bacterial growth would be reduced. A strain of Y enterocolitica 03 (IP 134, the National Institute for Public Health, Utrecht, Netherlands) was held in frozen stock; an experiment it was cultured for 24 h. Dilutions were prepared in sterile phosphate-buffered saline. Immediately after collection thirty pools of 3 bags of whole blood with identical blood groups were made and inoculated under aseptic conditions with 2 x 101-3 x 10’ colony-forming units (CFU)/ml of Yenterocolitica. (In donors without serious disorders the number of circulating Y enterocolitica is assumed to vary from 1 to 100 CFU/ml.) Each pool was split into the three original bags. One bag from each pool was processed to RBC after a 6 h hold at 4°C according to the methods of the American Association of Blood Banks (Manual 1991). The other two bags were processed to buffy-coat-depleted (BCd)-RBCs after a 20 h hold at 22OC,9 then one bag of BCd-RBC was filtered through a cellulose acetate filter column (Cellselect, NPBI, Emmercompascuum, Netherlands) within 24 h of collection to give leucocyte-depleted (Ld)-RBC.lO The filtration procedure was done in a closed system by welding the tubing of the filter and the blood bag together with a sterile connection device.lO Saline-adenine-glucose-mannitol solution was added to all types of RBC so they could be stored for 5 weeks at 4°C.° 1 unit of RBC contained 2-4 x 109 leucocytes (100% of original value), BCd-RBC 0-8 x 109 leucocytes (about 30% of original value), and Ld-RBC fewer than 0-01 x 109 leucocytes (0-05% of original value). From each type of RBC concentrate in each pool a 5 ml sample for


culture was taken at week 0 (24 h after inoculation) then at weeks 1, 2,3,4, and 5. When more than 105 CFU/ml were present, that RBC concentrate was no longer tested. From 1 0,0 1,001,andOOOl ml RBC concentrate samples, triplicate pour plates were prepared with 20 ml tryptone soya agar containing vancomycin (1 mg/1000 ml

Removal of Y enterocolitica was better in Ld- RBC than in BCd-RBC or RBC at any time point (p < 003-0001, table). Ld-RBC showed growth after longer storage times than the other preparations; BCd-RBC, in turn, showed growth after longer storage times than RBC. With contamination of less than 3 x 102 CFU/ml, reduction of storage time of RBC from 5 to 3 weeks would reduce multiplication of Y enterocolitica. However, to eliminate Y enterocolitica, leucocyte depletion after holding whole blood at 22°C was the most effective method; use of 5-week storage times of Ld-RBC could be continued. To reduce bacterial contamination in routine blood bank procedures, leucocytes should be able to function (ie, carry out phagocytosis and killing) for several hours at temperatures of 18-24°C. The time for which whole blood should be held at 22°C could be between 6 and 24 h. For logistic reasons an overnight hold for 16-20 h is convenient, and all units of blood can be processed during the next day. Subsequent removal of leucocytes by filtration is the best way to avoid growth of Y enterocolitica. Furthermore, transfusion of leucocyte-depleted blood products will reduce HLA immunisation, and prevent transmission of cytomegalovirus and human T-cell leukaemia virus. We thank Ms N. Schram-Bakker and Mr W.

Schaasberg for statistical

analysis. REFERENCES 1.

Yersinia enterocolitica bacteremia and endotoxin shock associated with red blood cell transfusions—United States, 1991. Morbid Mortal


Weekly Rep 1991; 40: 176-78. Hoppe PA. Interim measures for detection of bacterially contaminated red cell components (editorial). Transfusion 1992; 32: 199-201. 3. Goldman M, Blajchman MA. Blood product associated bacterial sepsis. Trans Med Rev 1991; V: 73-83. 4. Stossel TP. Phagocytosis recognition and ingestion. Semin Hematol 1975; 2.

12: 83-116. 5. Peterson PK, Verhoef

J, Quie PG. Influence of temperature on opsonization and phagocytosis of staphylococci. Infect Immun 1977; 15: 175-79.


Hogman CF, Gong J, Eriksson L, Hambraeus A, Johansson CS. White cells protect donor blood against bacterial contamination. Transfusion 1991; 31: 620-26. FS, Schneider DL. The activation of the human neutrophil

7. Manara

respiratory burst occurs only at temperature above 17°C: evidence that activation requires membrane fusion. Biochem Biophys Res Commun 1985; 132: 696-701. RNI, Reesink HW, Dekker WJA, Fijen FJ. Preparation of

8. Pietersz

leukocyte-poor platelet concentrates from buffy coats. I. Special inserts for centrifuge cups. Vox Sang 1987; 53: 203-07. 9. Pietersz RNI, de Korte D, Reesink HW, Dekker WJA, van den Ende A, Loos JA. Storage of whole blood for up to 24 hours at ambient temperature prior to component preparation. Vox Sang 1989; 56: 145-50. 10. Pietersz RNI, Reesink HW, de Korte D. Storage of leukocyte-poor red cell concentrates: filtration in a closed system using a sterile connection device. Vox Sang 1989; 57: 29-36. ADDRESSES. Red Cross Blood Bank, Amsterdam (R. N I. Pietersz, MD, H W. Reesink, MD, W J. A Dekker) and Department of Slotervaart Microbiology, Hospital, Amsterdam, The R I Netherlands (W. Pauw, MD, L. Buisman) Correspondence to Dr N Pietersz, Red Cross Blood Bank Amsterdam, Postbox 9137, 1006 AC Amsterdam, The Netherlands.

Prevention of Yersinia enterocolitica growth in red-blood-cell concentrates.

In response to concern about Yersinia enterocolitica contamination of blood products, we have studied the effects on Y enterocolitica growth of holdin...
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