Immunologic Studies of Prothrombin in Newborns

Abstract

SummaryProthrombin of newborns was investigated by means of the quantitative immunoelectrophoresis and the two-dimensional immunoelectrophoresis. The plasma of 10 of 24 healthy term newborns and of 5 preterm infants taken immediately after birth showed one prothrombin precipitate in the a-2-fi-globulin position when a buffer containing 25 mM calcium lactate was used. No differences in the mobility and form of the precipitate could be observed as compared to healthy adults. The plasma of the other 14 term newborns and one small-for-date infant showed two precipitates, one in the a-2-&globulin position and a second in the a-1-globulin position. Twenty-four hr after the injection of vitamin K, the precipitate in a-1-globulin position was no longer evident.When a buffer without calcium was used in the two-dimensional immunoelectrophoresis, only one precipitate in the a-1-globulin position was found in all newborns. In the quantitative immunologic determination, the prothrombin was found to be decreased as compared to that of adults. The quantitative immunologic determination and the activity assay gave equal results in those infants who had only one precipitate in the two dimensional immunoelectrophoresis. In those newborns, who had two precipitates, the quantitative immunologic determination gave higher results than the measurement of the activity assay.From this study, no evidence of differences between the prothrombin of newborns and that of adults could be derived. Vitamin K deficiency, as judged from the appearance of a second peak in a-1-globulin position in the two dimensional immunoelectrophoresis was a frequent finding in term newborns, even on the 1st day of life. small for date. Five were prematures with a gestational age of 3 1-35 wk. None of these five prematures had perinatal problems, in spite of their prematurity. All but two children received 1 mg/kg vitamin K (Konakion) im shortly after birth.Immediately after birth, blood was drawn from the umbilical vein and again 24 hr afterwards by puncturing a peripheral vein. Blood samples were collected in plastic tubes containing 1/10 volume of 0.1 M trisodium citrate. The plasma was stored at -20' C until the determinations were carried out.Prothrombin activity was determined by the one stage assay (20) using a test kit of the Behring Co.Prothrombin was quantified immunologically according to Laurel] (1 I), using a monospecific rabbit antiserum against prothrombin (Behring Co): 1% agarose, barbital buffer of a pH of 8.6, 6-7 V/cm.Calibration curves for the activity and for the quantitative immunologic determination were done with a pooled plasma of 20 healthy adults and the results were expressed in percent of this pooled piasma. Two-dimensional immunoelectrophoresis (4, 5) was done, using rabbit antiserum against human prothrombin 1) with a barbital buffer of a pH of 8.6 and 2) with a barbital buffer of a pH of 8.6 and 25 mM calcium lactate. Agarose, 1%, first dimension 6-7 V/ cm for 4 hr, second dimension 2-3 V/cm for 12-14 hr.Plasma of healthy adults a...

Prothrombin Expression in the Adult and Fetal Rabbit Liver

“…The sp act of plasma prothrombin in these roform as described by Chomezynski and Sacchi (12). The RNA infants' plasma is the same as in adults (2,(5)(6)(7)(8).…”

“…The sp act of plasma prothrombin in these roform as described by Chomezynski and Sacchi (12). The RNA infants' plasma is the same as in adults (2,(5)(6)(7)(8).was precipitated with sodium acetate to remove glycogen (1 3). The reason for the lower levels of prothrombin in infants' mRNA was selected from total RNA using oligo (Dt)-cellulose plasma has not been elucidated.…”

Improvement of Vitamin K Status of Breastfeeding Infants with Maternal Supplement of Vitamin K2 (MK40)

“…It is well-known that PIVKA-II is a sensitive and specific marker of the vitamin K status. 14 This precursor molecule of prothrombin can be detected by several methods, including crossed immunoelectrophoresis (CIE), 15 ratio method, 16 absorption method, 17 and specific antibody method. 18 In this study, ECLIA, a newly developed assay system possessing higher sensitivity than previous methods, was used.…”

Variation of Des-Gamma-Carboxyprothrombin (PIVKA-II) Levels in Cord Blood Throughout Gestation

“…It is well known that PIVKA-II is a sensitive and specific marker of vitamin K status. 7,8 This precursor molecule of prothrombin can be detected by several methods, including crossed immunoelectrophoresis (CIE), 9 ratio method, 9,10 absorption method, 11 and specific antibody method. 12 A new method, electrochemiluminescence immunoassay (ECLIA), has been developed.…”