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• W2012058802 abstract "Objectives: To determine zeta chain expression in women before pregnancy and 14 days after oocyte retrieval.Design: Women undergoing an in vitro fertilization cycle at the University of Louisville were recruited prospectively prior to the start of the cycle.Materials and Methods: A serum sample was obtained during the stimulation cycle, prior to HCG injection. Fourteen days after oocyte retrieval, another serum sample was collected when the patient returned for a pregnancy test. Serum samples were centrifuged for ten minutes and red blood cells were discarded. Jurkat cells, from a T-cell leukemia, were incubated with serum samples for a duration of four days, from 5 women, before and after pregnancy occurred. After incubation, the serum samples were centrifuged, the T-lymphocytes were solubilized, and immunoblotting was done to evaluate the presence of zeta chain, a T-lymphocyte signal transduction protein. Next T-lymphocytes were incubated with serum from 4 pregnant patients who did not express zeta chain, and from 4 patients who had also undergone oocyte retrieval 14 days earlier, but did not become pregnant. DNA was isolated and a gel electrophoresis was performed to evaluate the presence of DNA fragmentation, suggesting apoptosis as a mechanism of zeta chain loss in pregnant patients. Finally to further evaluate apoptosis as the mechanism of zeta chain loss, T-cells were incubated for 30 minutes with sera from 2 patients that were pregnant, but did not express zeta chain. Western blotting was then done to evaluate for the presence of ubiquitin.Results: In the initial experiment, 4 of 5 serum samples from pregnant women, suppressed T-cell zeta chain expression. This suggests that zeta chain degradation could be a way in which T-cell signal transduction and immune activation is inhibited during pregnancy. DNA fragmentation, used to demonstrate apoptosis, was present in all T-cells incubated with sera from pregnant patients and none of the T-cells incubated with sera from non-pregnant patients. This finding suggests that apoptosis is the mechanism of zeta chain loss. As further support of this theory, ubiquination was evaluated, since ubiquination suggests imminent apoptosis. Increased ubiquination was found in T-cells, incubated with sera from 2 pregnant women, who suppressed zeta chain. T-cells incubated with non-pregnant sera did not demonstrate this phenomenon.Conclusions: Zeta chain expression on T-cells, cultured with sera from pregnant women, is altered in early pregnancy. Loss of zeta chain on T-cells may alter signal transduction in pregnancy and prevent immune activation. The mechanism of zeta chain loss appears to be due to apoptosis. This finding can be demonstrated as early as 14 days after conception. Objectives: To determine zeta chain expression in women before pregnancy and 14 days after oocyte retrieval. Design: Women undergoing an in vitro fertilization cycle at the University of Louisville were recruited prospectively prior to the start of the cycle. Materials and Methods: A serum sample was obtained during the stimulation cycle, prior to HCG injection. Fourteen days after oocyte retrieval, another serum sample was collected when the patient returned for a pregnancy test. Serum samples were centrifuged for ten minutes and red blood cells were discarded. Jurkat cells, from a T-cell leukemia, were incubated with serum samples for a duration of four days, from 5 women, before and after pregnancy occurred. After incubation, the serum samples were centrifuged, the T-lymphocytes were solubilized, and immunoblotting was done to evaluate the presence of zeta chain, a T-lymphocyte signal transduction protein. Next T-lymphocytes were incubated with serum from 4 pregnant patients who did not express zeta chain, and from 4 patients who had also undergone oocyte retrieval 14 days earlier, but did not become pregnant. DNA was isolated and a gel electrophoresis was performed to evaluate the presence of DNA fragmentation, suggesting apoptosis as a mechanism of zeta chain loss in pregnant patients. Finally to further evaluate apoptosis as the mechanism of zeta chain loss, T-cells were incubated for 30 minutes with sera from 2 patients that were pregnant, but did not express zeta chain. Western blotting was then done to evaluate for the presence of ubiquitin. Results: In the initial experiment, 4 of 5 serum samples from pregnant women, suppressed T-cell zeta chain expression. This suggests that zeta chain degradation could be a way in which T-cell signal transduction and immune activation is inhibited during pregnancy. DNA fragmentation, used to demonstrate apoptosis, was present in all T-cells incubated with sera from pregnant patients and none of the T-cells incubated with sera from non-pregnant patients. This finding suggests that apoptosis is the mechanism of zeta chain loss. As further support of this theory, ubiquination was evaluated, since ubiquination suggests imminent apoptosis. Increased ubiquination was found in T-cells, incubated with sera from 2 pregnant women, who suppressed zeta chain. T-cells incubated with non-pregnant sera did not demonstrate this phenomenon. Conclusions: Zeta chain expression on T-cells, cultured with sera from pregnant women, is altered in early pregnancy. Loss of zeta chain on T-cells may alter signal transduction in pregnancy and prevent immune activation. The mechanism of zeta chain loss appears to be due to apoptosis. This finding can be demonstrated as early as 14 days after conception." @default.
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• W2012058802 date "2000-09-01" @default.
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• W2012058802 title "Zeta-Chain Regulation in Early Pregnancy" @default.
• W2012058802 doi "https://doi.org/10.1016/s0015-0282(00)00754-8" @default.
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