Small Number Of Donors Research Articles

Transcriptional signatures of environmentally relevant exposures in normal human mammary epithelial cells: benzo[a]pyrene

Changes in gene expression in a panel of primary normal human mammary epithelial cell strains, developed from healthy breast tissue obtained at reduction mammoplasty from different donors, in response to benzo[a]pyrene exposure have been investigated. It was expected that both gene expression changes common to cell strains derived from different donors as well as inter-individual variation would be observed. Therefore, the strategy that has been adopted is to identify potentially important changes, or useful changes from a biomonitoring perspective, using gene-array technology and a small number of donors; then investigate selected transcription responses using a large number of tissue donors and a cheaper method of transcript detection (real-time polymerase chain reaction). Here we report results from four primary normal human mammary epithelial cell strains that were treated with benzo[a]pyrene in vitro for either 6 or 24 h. Transcription was monitored using high-density oligonucleotide arrays (Affymetrix HuGeneFL). Total RNA was used for the preparation of labeled targets that were hybridized to microarrays containing probes representing more than 6800 human genes and expressed sequence tags. Gene expression data were analyzed using the GeneChip® software (MAS 5.0). Altered gene expression patterns were observed in response to benzo[a]pyrene in human mammary epithelial cell strains from different donors. Specifically, the dioxin inducible cytochrome P450 CYP1B1 was consistently induced in response to 6 and 24 h exposure to benzo[a]pyrene in cell strains from all four donors. Two other genes that were relatively consistently induced were IL1β and MMP1. Less consistent changes in other metabolism genes ( CYP1A1, CYP11B2, and NQO1) and certain cell cycle control genes GOS2 and AF1Q were also induced, while EGR1 was suppressed. Although no change in p53 transcription was observed, an accumulation of p53 protein was detected using antibodies. A similar accumulation of Waf1 ( p21) was also observed using immunohistochemistry, this was expected since p53 is p21's transcription factor. Significant inter-individual variations in both the levels and patterns of gene expression were observed, in response to benzo[a]pyrene exposure. These studies provide a complementary approach to molecular epidemiology for the investigation of differential susceptibility to chemical carcinogens, and specifically polycyclic aromatic hydrocarbons.

A randomized, double‐blind comparison of donor tolerance of 400 mL, 200 mL, and sham red cell donation

Volume replacement could allow the safe collection of twice the normal amount of red cells in a standard donation. Studies in small numbers of donors have shown that a temporary decrease in red cell mass is well tolerated when donors give twice the usual amount (170-225 mL) of red cells in a standard 405- to 495-mL donation. Sham-donation control groups have not been included in previous studies of increased red cell donation, and perceptions of donation effects could have been biased. In the study reported here, 17 male and 13 female volunteers were randomly assigned to make a sham donation, 1-unit donation, or 2-unit donation on an automated blood cell separator. Donor tolerance was assessed by ambulatory heart rate monitoring and by a poststudy interview. Hemoglobin, hematocrit, ferritin, serum iron, total iron-binding capacity, red cell 2,3 DPG, and serum erythropoietin were measured before and after donation for comparison of the erythropoietic responses in the three study groups. Red cells collected totaled 206 +/- 10 mL in the 1-unit group and 414 +/- 21 mL in the 2-unit group. Changes in heart rate, systolic blood pressure, and diastolic blood pressure with donation and changes in heart rate recorded by ambulatory monitoring did not differ for the experimental groups. Postdonation changes from baseline values were evaluated on Days 2, 7, and 14. Changes in hemoglobin were significantly different between groups (p < 0.017) in all postdonation tests. There were differences between groups in erythropoietin response, red cell 2,3 DPG, ferritin levels, and hemoglobin synthesis. Hemoglobin synthesis and mean changes in 2,3 DPG, erythropoietin, ferritin, and postdonation hemoglobin were greater in the 2-unit group than in the 1-unit group. Donor tolerance of red cell donations of 414 +/- 21 mL, a volume of red cells twice that in a standard 450-mL blood donation, does not differ from donor tolerance of standard or sham donations. Physiologic adjustments and the hematopoietic response to reduced red cell mass were greater in the 2-unit group, but the donation of 1 unit or 2 units did not cause detectable symptoms of reduced oxygen-carrying capacity.

Living related liver transplantation in children: a report of the first 58 recipients at the University of Chicago.

From November 1989 58 living donor liver transplants were performed in 56 children ranging in age from 1 month to 13 years. Donors were adults (> 18 years of age) with a close relationship to the recipient. ABO compatibility and normal donor health were required. Liver segments two and three were transplanted in 53 cases, and segments two, three and four in 5 cases. Actuarial patient survival at 2 years was 89%; graft survival was 76%. Six recipients died: four secondary to sepsis and two because of post-transplant lymphoproliferative disease. The main cause of graft loss was arterial thrombosis, occurring in six patients (10%). Since refinement of the technique, there have been few donor complications, but these have included a biliary tract injury and a hepatic artery thrombosis. Both donors are well, without long-term adverse sequelae. Overall, the outcome of living donor transplantation is excellent; morbidity has been encountered in a small number of donors.

Quality of life in Norwegian kidney donors

The Norwegian kidney transplant programme is based on more extensive use of living donors than in other countries. Re-examination of the physical health of kidney donors has in the majority of studies revealed no long-term medical risk. The quality of life after donation has only been described in very few studies, and with a small number of donors. This report concerns a cross-sectional study of 494 donors with a mean observation time from donation of 6.7 years. A standardized questionnaire containing 19 items related to quality of life was used. The donors were compared to the general adult population in a county in mid-Norway. Out of the 19 quality of life items, the donors scored significantly better in 13. Donors who had experienced that the corresponding recipient had died have been tested separately. The results in this subgroup were equal to the population, but inferior compared to the rest of the donors. We conclude that the quality of life of Norwegian kidney donors is better than the general population. We will continue our policy of an extensive use of living kidney donors.

Donation of factor VIII by plasma exchange after DDAVP administration

We report our experience with a new factor VIII product, the potency and safety of which are at least equivalent to industrial products. Carefully screened, highly motivated donors are informed about requirements for the treatment of hemophilia and committed to give donations for factor VIII monthly. 2.5 L of plasma is obtained by plasma exchange after i.v. infusion of DDAVP, which triples the level of factor VIII and doubles the level of Von Willebrand Factor. Plasma replacement is with autologous supernatant from previous donations or from manual plasmapheresis. An exchange can usually be completed in 2 h. Cryoprecipitate is obtained in minibags after freezing and fast thawing. The factor VIII yield has averaged 2764 ± 922 IU per donation. The concentration of factor VIII in cryoprecipitate has averaged 23.8 ± 7.7 IU/mL and the mean specific activity was 1.22 ± 0.48 IU/mg protein. Cost of production was estimated at 1.3 Fr per IU. After 6 months quarantine, during which the donors are checked repeatedly for infectious diseases, the concentrate may be released. Thus, plasma exchange and use of DDAVP in a small number of donors allow us to obtain a “clean” factor VIII concentrate with a lower cost than industrial concentrates. This strategy might be of special interest for countries that lack industrial fractionation capabilities.

Donor variables affecting survival of autologous platelets.

While platelet survival studies have been carried out by numerous laboratories for many years, general agreement has not been reached on a single method or for the value of this test. Available data indicate that in spite of certain difficulties, reliable comparisons can be obtained using relatively small numbers of donors. Indeed, a difference in platelet survival of as little as 10 percent can be detected by using eight donors, whereas only four donors are required if one wishes to detect a 20 percent change. Should studies be done using donors preselected according to strict criteria of age, sex, health, etc.? It might be that less scatter among the data points would be observed and perhaps even fewer donors would be required to obtain significant values. In surveying the available information, it is clear that many investigators do have data on normal controls which could be extracted and analyzed to provide an answer to this question. Such information would be of great value in determining the criteria for selection of donors for autologous platelet survival studies.

Fertilization rate of bovine sires

Improvement of the superovulation technique will help to enhance the efficiency of embryo and animal production. Blocking inhibin using inhibin antiserum (IAS) is known to promote follicular development by increasing the level of FSH. Previously, we reported that coadministration of IAS and eCG produced more than 100 oocytes from a single female C57BL/6 mouse at 4 weeks old. The oocytes derived from the IAS + eCG (IASe) treatment were able to fertilize and develop normally into offspring. In this study, we examined the effect of IASe treatment on the numbers of ovulated oocytes in major inbred (A/J, BALB/cByJ, C3HeJ, DBA/2J, and FVB/NJ) and outbred (CD1) mice strains at 4 weeks old. We confirmed the fertilization and developmental ability of the IASe-derived oocytes. IASe treatment ovulated 1.5 to 3.2 times higher numbers of oocytes than eCG treatment alone. The fertilization rate of IASe-derived oocytes was similar to that of eCG-derived oocytes. In vitro and in vivo developmental rates of the embryos derived from IASe were similar to the rates of embryos derived from eCG. We have shown that superovulation by IASe is very effective in obtaining high numbers of ovulated oocytes from small numbers of oocyte donor in a number of mice strains. The superovulation technique will contribute to the archiving of cryopreserved embryos of genetically engineered mice using small numbers of donors and has the potential to produce more live animals for rederivation of the archived mouse lines in mouse repositories.

Variable Motility Recovery of Spermatozoa Following Freeze Preservation

Semen from a random group of potentially fertile men and a small number of donors who had established an artificial insemination pregnancy was mixed with a cryoprotective agent and frozen by two different methods. After 1 week under liquid nitrogen (-196 degrees C), all samples were thawed and their motilities determined. The post-thaw motilities were compared with the prefreeze motilities. The mean motility recovery rate in the group of semen samples with high counts was better than that in the group with counts in the low-normal range. The recovery rate with the slow-freeze method was slightly higher than that with the quick-freeze technique. However, the wide range of post-thaw motility among the random donors (0 to 60%) and the donors of proven fertility (15 to 45%) demonstrated that prefreeze motility and/or paternity history was no guarantee of good motility recovery following freezing. At a time when semen banking is being contemplated for "fertility insurance" prior to vasectomy, it is apparent that a trial freeze is necessary before recommending freeze-preservation of his semen to the man contemplating permanent surgical sterilization.