Novel protective mechanism for interleukin-33 at the mucosal barrier during influenza-associated bacterial superinfection.

Influenza A is a highly contagious respiratory virus that causes seasonal epidemics and occasional worldwide pandemics. The primary cause of influenza-related mortality is bacterial superinfection. There are numerous mechanisms by which preceding influenza infection attenuates host defense, allowing for increased susceptibility to bacterial pneumonia. Herein, we demonstrate that influenza inhibits Staphylococcus aureus-induced production of interleukin-33 (IL-33). Restoration of IL-33 during influenza A and methicillin-resistant S. aureus superinfection enhanced bacterial clearance and improved mortality. Innate lymphoid Type 2 cells and alternatively activated macrophages are not required for IL-33-mediated protection during superinfection. We show that IL-33 treatment resulted in neutrophil recruitment to the lung, associated with improved bacterial clearance. These findings identify a novel role for IL-33 in antibacterial host defense at the mucosal barrier.

Granulocyte serology: current concepts and clinical signifcance.

Applying serologic procedures to the detection of RBC and lymphocyte antigens has facilitated the identification of granulocyte antigens with established clinical significance, which are now classified in the human neutrophil antigen system. Granulocyte alloantibodies and autoantibodies have been implicated in a variety of clinical conditions including alloimmune neutropenia, autoimmune neutropenia, febrile and severe pulmonary transfusion reactions, drug-induced neutropenia, refractoriness to granulocyte transfusions, and immune neutropenia after hematopoietic stem cell transplantation. Although the intrinsically fragile nature of granulocytes contributes to the inherent challenges of granulocyte serology, several advances in laboratory procedures have improved detection of granulocyte antibodies. This review will provide a current perspective about the importance and use of granulocyte serology for detection of granulocyte antibodies that have significant medical effects.

The effect of dosing regimen on the pharmacokinetics of risedronate.

AIMS: To examine the effect of timing of a risedronate dose relative to food intake on the rate and extent of risedronate absorption following single-dose, oral administration to healthy male and female volunteers. METHODS: A single-dose, randomized, parallel study design was conducted with volunteers assigned to four treatment groups (31 or 32 subjects per group, 127 subjects total). Each subject was orally administered 30 mg risedronate. Group 1 was fasted for 10 h prior to and 4 h after dosing (fasted group); Groups 2 and 3 were fasted for 10 h and were dosed 1 and 0.5 h, respectively, before a high-fat breakfast; and Group 4 was dosed 2 h after a standard dinner. Blood and urine samples were collected for 168 h after dosing. Pharmacokinetic parameters were estimated by simultaneous analysis of risedronate serum concentration and urinary excretion rate-time data. RESULTS: Extent of risedronate absorption (AUC and Ae ) was comparable (P=0.4) in subjects dosed 2 h after dinner and 0.5 h before breakfast; however, a significantly greater extent of absorption occurred when risedronate was given 1 or 4 h prior to a meal (1.4- to 2.3-fold greater). Administration 0.5, 1, or 4 h prior to a meal resulted in a significantly greater rate of absorption (Cmax 2.8-, 3.5-, and 4.1-fold greater, respectively) when compared with 2 h after dinner. CONCLUSIONS: The comparable extent of risedronate absorption when administered either 0.5-1 h before breakfast or 2 h after an evening meal support previous clinical studies where risedronate was found to have similar effectiveness using these dosing regimens. This flexibility in the timing of risedronate administration may provide patients an alternative means to achieve the desired efficacy while maintaining their normal daily routine.

Effect of heat on stored red cells during non-flow conditions in a blood-warming device.

BACKGROUND AND OBJECTIVES: While blood is flowing within a transfusion-warming device, the blood temperature is usually less than that applied externally. If the flow is temporarily stopped, the temperature can rise above 37 degrees C in some warming devices. We sought to determine whether temperatures near 45 degrees C achieved during prolonged non-flow conditions in a blood warmer are harmful to red cell integrity. MATERIALS AND METHODS: After 42 days of storage at 4 degrees C, red cells were exposed to 44.7 degrees C for 30 min while stationary in a blood warming device (Augustine Medical, Inc., 241 Fluid Warming Set) and examined for cell counts, hemolysis and osmotic fragility. RESULTS: Red cell, white cell and platelet counts, hemoglobin, PCV and potassium were unchanged following heat treatment. Plasma hemoglobin was 508+/-132 mg/dl following heat treatment compared to 396+/-188 for the control (p>0. 05). In the osmotic fragility test, hemolysis remained within normal limits when tested at 0.60 and 0.65% sodium chloride (NaCl) and was unchanged at the 0.5% NaCl level. At the 0.75% NaCl level, there was 16+/-5.1% hemolysis of heated 42-day-old red cells compared to 11+/-3.4% for the control (both of which being above the 9% upper limit for fresh control red cells). CONCLUSIONS: We conclude that elevated temperatures achieved during temporary cessation of flow in the Augustine Medical, Inc., 241 Fluid Warming Set for as long as 30 min do not cause notable hemolysis or other damage to red cells.

Collection of two peripheral blood stem cell concentrates from healthy donors.

When peripheral blood stem cell (PBSC) concentrates are used for allogeneic transplants, two or more apheresis procedures must often be performed. To determine how many cells could be collected from healthy people by two back-to-back apheresis procedures and what effect these collections would have on donors, we gave 19 healthy people 5 micrograms kg-1 day-1 and 21 people 10 micrograms kg-1 day-1 of granulocyte colony stimulating factor, filgrastim, for 5 days. We then collected two PBSC concentrates, one on day 5 and one on day 6. A third group of six people was given filgrastim 10 micrograms kg-1 day-1 for 5 days but had no PBSC concentrates collected. PBSC concentrate cell counts and donor cell counts, symptoms, and blood chemistries were assessed for up to 1 year. On day 5, three times more CD34+ cells were collected from donors given 10 micrograms kg-1 day-1 than those given 5 micrograms kg-1 day-1 (P = 0.009) but on day 6 the quantity of cells collected was the same (P = 0.23). The total number of CD34+ cells collected was two times greater in donors given the higher dose of filgrastim (median = 579 x 10(6); range = 174-1639 x 10(6) compared to 237 x 10(6); 103-1670 x 10(6); P = 0.061). Platelet counts fell after each PBSC concentrate collection, but there were no differences between the two groups of donors in platelet counts measured immediately after each collection. The platelet counts also fell in people who did not donate PBSC concentrates. The lowest counts in all three groups of people also occurred on day 10. In PBSC donors given 10 micrograms kg-1 day-1 of filgrastim the absolute neutrophil count (ANC) fell below premobilization counts on day 14. In donors given 5 micrograms kg-1 day-1 the ANC fell below premobilization counts on days 21, 28 and 49, CD34+ cell counts were significantly lower than premobilization counts on days 14 and 28 in donors given 10 micrograms kg-1 day-1 of filgrastim and on day 14 in those given 5 micrograms kg-1 day-1. No decrease in neutrophil or CD34+ cell counts occurred after filgrastim was given in the people who did not donate PBSC concentrates. The incidence of symptoms was similar in both groups of PBSC concentrate donors, except that those given 10 micrograms kg-1 day-1 were more than twice as likely to experience myalgias as those receiving the lower dose (P = 0.029). Several blood chemistries changed. Levels of alkaline phosphatase, LDH, SGPT, SGOT, uric acid and sodium increased. Levels of bilirubin, total protein, potassium, calcium and chloride decreased. In conclusion, twice as many CD34+ cells were collected from donors given 10 micrograms kg-1 day-1 of filgrastim. Platelet, neutrophil and CD34+ cell counts fell after the PBSC concentrate collections. The fall in platelet counts was due to both the collection and the administration of filgrastim. The falls in neutrophil and CD34+ cell counts were due to the loss of haematopoietic progenitor cells in the PBSC concentrates. Allogeneic PBSC concentrate donors should be given 10 micrograms kg-1 day-1 of filgrastim, and if possible only one component should be collected in order to avoid thrombocytopenia.

Expression of neutrophil antigens after 10 days of granulocyte-colony-stimulating factor.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) is becoming the standard agent for mobilizing granulocytes. Most granulocyte donors are given a single dose of G-CSF, but in some cases they are given G-CSF for several days, and multiple granulocyte concentrates are collected. The administration of a single dose of G-CSF induces several changes in the expression of neutrophil antigens, but the effects of multiple daily doses of G-CSF are not known. STUDY DESIGN AND METHODS: Seven healthy people received 5 microg per kg of G-CSF for 10 days. Their expression of several neutrophil antigens before, during, and after the administration of G-CSF was analyzed through the use of flow cytometry. RESULTS: The expression of L-selectin (CD62L), Fcgamma receptor (FcgammaR) III (FcgammaRIII, CD16), and the leukocyte function antigen (CD11a) decreased throughout the course of G-CSF administration, while the expression of FgammaR I (FcgammaRI, CD64) and lipopolysaccharide-binding protein receptor (CD14) increased. The expression of FcgammaR II (FcgammaRII, CD32) also increased, but not until the fourth day of G-CSF administration. The expression of amino peptidase N (CD13), C3bi receptor (CD11b), and the neutrophil beta2 integrin unit (CD18) did not change during the administration of G-CSF, but that of both CD13 and CD18 increased 3 days after the last dose. The expression of neutrophil-specific antigen NB1 initially increased, returned to pre-G-CSF levels after 4 days, and then increased again after 10 days of G-CSF administration. CONCLUSION: Changes in the expression of several neutrophil antigens occurred throughout a 10-day course of G-CSF Most of the changes occurred after one dose, but additional changes occurred later in the 10-day course and after its completion. These changes may affect the function of G-CSF-mobilized granulocytes.

Risedronate gastrointestinal absorption is independent of site and rate of administration.

PURPOSE: Two studies were conducted to compare the absorption of risedronate administered as a solution to three different gastrointestinal sites (study A) and to determine the extent of absorption of risedronate solution administered by rapid and slow infusion to the second part of the duodenum (study B). METHODS: Each study was designed as a single-dose, crossover (three periods, study A; two periods, study B) trial in healthy male subjects, with a 14-day washout period between dosing. Subjects fasted overnight before drug administration and for 4 hours after drug administration. In study A, a risedronate solution of 40 mg in 30 mL of water was administered directly into the stomach, the second part of the duodenum, or the terminal ileum over 1 minute via a nasoenteral tube in a three-period crossover design. In study B, a risedronate solution of 40 mg in 30 mL of water was administered directly into the second part of the duodenum over 1 minute and over 1 hour in a randomized, two-period crossover design. Serum and urine samples were obtained for 48 hours after dosing for risedronate analysis. RESULTS: Eight subjects completed each study. No statistically significant site-specific differences in any pharmacokinetic parameter were observed (study A). Based on the area under the serum concentration-time profile and the amount of drug excreted in the urine unchanged, the extent of risedronate absorption did not differ significantly following a rapid or a slow infusion (study B). Only minor symptomatic complaints were reported by subjects, such as headaches and body aches. CONCLUSIONS: These studies indicate that the rate and extent of risedronate absorption are independent of the site of administration along the gastrointestinal tract, and that the extent of absorption is not affected by the rate of administration.

Radioactive contamination incidents involving protective clothing.

The study focuses on incidents at Department of Energy facilities involving the migration of radioactive contaminants through protective clothing. The authors analyzed 68 occurrence reports for the following factors: (1) type of work; (2) working conditions; (3) type of anti-contamination material; (4) area of body or clothing contaminated; and (5) nature of spread of contamination. A majority of reports identified strenuous work activities such as maintenance, construction, or decontamination and decommissioning projects. The reports also indicated adverse working conditions that included hot and humid or cramped work environments. The type of anti-contamination clothing most often identified was cotton or water-resistant disposable clothing. Most of the reports also indicated contaminants migrating through perspiration-soaked areas, typically in the knees and forearms. On the basis of their survey, the authors recommend the use of improved engineering controls and resilient, breathable, waterproof protective clothing for work in hot, humid, or damp areas where the possibility of prolonged contact with contamination cannot be easily avoided or controlled.

Donor attitudes about exporting and importing blood.

BACKGROUND: The movement of blood among different areas of the United States and the collection of more blood than is needed locally in some areas are increasing. Little is known of donors' attitudes about this blood resource sharing. STUDY DESIGN AND METHODS: One thousand donors from five regions of the American Red Cross Blood Services were surveyed by telephone. Demographic information about the donors and the regions was obtained, and the donors were asked to describe their attitudes about blood resource sharing as well as other blood donation-related issues. RESULTS: Donors are not very knowledgeable about whether their community is self-sufficient in its blood supply. In regions that import blood, 29 to 43 percent of donors believed that enough blood was collected to meet all local needs, and, in regions that export blood, only 22 to 24 percent of donors believed that more than enough blood was collected. About three-fourths of the donors believed it acceptable to send their blood to other communities if it is needed there. However, this attitude was based on the premise that all local needs would be met first. Only 4 percent of donors would be less willing to donate if their blood was being sent to another community. CONCLUSION: Donors are not very aware of blood resource sharing but are willing, under certain circumstances, to donate blood for use outside their local communities.

Comparison of two blood cell separators in collecting peripheral blood stem cell components.

To ensure that a sufficient number of CD34+ cells are collected for an allogeneic blood progenitor cell transplant, the most effective blood cell separator should be used to collect peripheral blood stem cell (PBSC) components. We compared the effectiveness of two blood cell separators. We gave 29 healthy people 7.5 or 10 micrograms kg-1 of granulocyte colony stimulating factor (G-CSF) daily for 5 days and collected one PBSC component with either a Fenwal CS3000 (n = 15) or a Cobe Spectra (n = 14) blood cell separator. The volume of blood processed was the same for each machine (8.4 +/- 1.0 L; range = 4.9-9.4 L for the CS3000 and 8.9 +/- 1.0 L; range 6.7-10.9 L; P = 0.71). The components collected with the CS3000 contained more mononuclear cells (39.6 +/- 21.9 x 10(9) compared with 26.9 +/- 5.6 x 10(9), P = 0.02) and fewer neutrophils (1.38 +/- 1.88 x 10(9) compared with 5.53 +/- 8.71 x 10(9), P = 0.001). The total number of CD34+ cells collected with the two instruments was the same (470 +/- 353 x 10(6) for the CS3000 and 419 +/- 351 x 10(6) for the Spectra; P = 0.64) as was the number of CD34+ cells collected per litre of whole blood processed (55.9 +/- 42.0 x 10(6) L-1 compared with 45.9 +/- 37.9 x 10(6) L-1; P = 0.59). The mononuclear cell collection efficiency was greater for the CS3000 (82.4 +/- 54.9% compared with 53.3 +/- 14.1; P = 0.04) but the CD34+ cell collection efficiencies were the same (87.4 +/- 61.1% for the CS3000 compared with 56.3 +/- 23.5% for the Spectra; P = 0.07). In conclusion, both blood cell separators collected components which contained large numbers of CD34+ cells, but those collected with the CS3000 contained fewer neutrophils and the CS3000 was more efficient at collecting mononuclear cells.

Composition of peripheral blood progenitor cell components collected from healthy donors.

BACKGROUND: Peripheral blood progenitor cell (PBPC) components are being collected from healthy donors for allogeneic transplantation, but the quantity, quality, composition, and variability of PBPCs collected from healthy people given granulocyte-colony-stimulating factor (G-CSF) have not been evaluated. STUDY DESIGN AND METHODS: PBPC components were collected from 150 healthy people who were given G-CSF (5, 7.5, or 10 microg/kg/day) for 5 days. The components were evaluated for white cell (WBC), mononuclear cell, CD34+ cell, neutrophil, platelet, and red cell (RBC) composition. RESULTS: The quantities collected were: WBCs, 35.0 +/- 16.4 x 10(9) (range, 11.9-163.3 x 10(9)); mononuclear cells, 33.3 +/- 14.4 x 10(9) (range, 11.9-139.6 x 10(9)); CD34+ cells, 412 +/- 287 x 10(6) (range, 70-1658 x 10(6)); neutrophils, 1.71 +/- 3.59 x 10(9) (range, 0-27.6 x 10(9)); RBCs, 7.2 +/- 4.0 mL (range, 0-22.1 mL); and platelets, 480 +/- 110 x 10(9) (range, 250-920 x 10(9)). PBPC components collected from people given G-CSF at 7.5 or 10 microg per kg per day contained significantly more CD34+ cells (respectively, 428 +/- 300 x 10(6); range, 70-1658 x 10(6) and 452 +/- 294 x 10(6); range, 78-1380 x 10(6)) than those from people given G-CSF at 5 microg per kg per day (276 +/- 186 x 10(6); range, 91-767 x 10(6)) (p = 0.007 and p = 0.002). When 10 microg per kg per day of G-CSF was given, 50 percent of the components contained enough CD34+ cells for transplantation to a 75-kg recipient (375 x 10(6) CD34+ cells), but 10.6 percent of the components contained less than 150 x 10(6) CD34+ cells and thus would provide a transplantable dose only for a 30-kg patient. CONCLUSION: One PBPC component collected from a healthy donor given 7.5 or 10 microg per kg per day of G-CSF should contain 70 to 1660 x 10(6) CD34+ cells, with 0 to 22 mL of RBCs. Because of the great variability in the number of CD34+ cells collected, the quantity of CD34+ cells in each component should be measured after each procedure to ensure that sufficient quantities of cells are present for a successful transplant.

The kinetics of G-CSF mobilization of CD34+ cells in healthy people.

When healthy people are given granulocyte colony stimulating factor (G-CSF) for 10 days the number of CD34+ cells in the peripheral blood begins to increase on the fourth day, reaches a maximum on the sixth day and then decreases. In this study, we further define the time and variability of peak mobilization of CD34+ cells. Twenty-two healthy people were given G-CSF (7.5 or 10 micrograms kg-1 day-1) subcutaneously each morning for 5 days and peripheral blood CD34+ cell counts were analysed immediately prior to the fourth (day 4) and fifth (day 5) G-CSF injection and 24 h after the fifth injection (Day 6). White blood cell (WBC) and neutrophil counts were greatest on day 6 [WBC = 43.8 +/- 13.9 x 10(9) L-1 (mean +/- 1 SD) and neutrophils = 36.6 +/- 12.8 x 10(9) L-1]. In contrast the CD34+ cell counts on day 6 (107 +/- 104 x 10(6) L-1) were less than on day 5 (128 +/- 136 x 10(6) L-1) (P = 0.048) but still greater than on day 4 (60.7 +/- 40.2 x 10(6) L-1) (P < 0.0001). The CD34+ cell counts of 10 donors were measured 2, 4 and 6 h after the fifth injection to determine if the counts increased further between days 5 and 6. The number of CD34+ cells in the blood on day 5 2 h after the fifth injection (193 +/- 277 x 10(6) L-1) was greater than the number prior to the injection (158 +/- 190 x 10(6) L-1), 4 h post-injection (139 +/- 158 x 10(6) L-1) and 6 h post-injection (170 +/- 236 x 10(6) L-1), but the differences were not significant (P = 0.29, 0.25 and 0.45). The number of CD34+ cells in the blood of 12 people were measured before and after the fourth G-CSF dose. Prior to the day 4 injection the CD34+ count was 61 +/- 40 x 10(6) L-1. At 2, 4 and 6 h the counts were 60 +/- 40, 61 +/- 29 and 64 +/- 30 x 10(6) L-1, respectively, and the differences were not significant (P = 0.99, P = 0.98, and P = 0.73). In conclusion, when healthy volunteers are given daily G-CSF injections, the number of mobilized CD34+ cells was the greatest on day 5, slightly less on day 6 and the least on day 4. If only one PBSC component is needed, PBSCs can be collected on day 5 after only 4 days of G-CSF. If PBSC components are collected on both days 5 and 6, the fifth dose can be given either before or after the collection of the first PBSC component.

Blood counts in healthy donors 1 year after the collection of granulocyte-colony-stimulating factor-mobilized progenitor cells and the results of a second mobilization and collection.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF)-mobilized blood cells are being used for allogeneic transplants, but the long-term effects of G-CSF on healthy individuals are not known. Furthermore, it is not certain how many CD34+ cells can be collected in a second mobilization and collection procedure. STUDY DESIGN AND METHODS: Nineteen people were given 2, 5, 7.5, or 10 micrograms of G-CSF per kg per day for 5 days, and blood progenitor cells were collected by apheresis on the sixth day; this was done on two occasions separated by at least 12 months. Blood counts obtained before and after each course of G-CSF and the quantity of cells collected were compared. RESULTS: There were no differences in white cell (WBC), platelet, red cell, and WBC differential counts measured before each course of G-CSF, and all the values were in the normal range. In a subset of 12 people who received 7.5 or 10 micrograms of G-CSF per kg per day for both courses, the numbers of neutrophils, mononuclear cells, and CD34+ cells in the blood after each course were similar (34.1 +/- 7.31 x 10(9)/L vs. 36.4 +/- 12.3 x 10(9)/L, p = 0.24; 6.59 +/- 2.28 x 10(9)/L vs. 5.63 +/- 2.11 x 10(9)/L, p = 0.24; and 92.0 +/- 55.6 x 10(5)/L vs. 119.2 +/- 104.6 x 10(6)/L; p = 0.48, respectively), as were the quantities of mononuclear cells (31.0 +/- 8.4 x 10(9) vs. 31.0 +/- 6.1 x 10(9); p = 0.64) and CD34+ cells (417 +/- 353 x 10(6) vs. 449 +/- 286 x 10(6); p = 0.53) collected in the two apheresis procedures. Furthermore, there was a positive correlation between the quantity of CD34+ cells collected from each of the 12 people per liter of whole blood processed in the two procedures (r2 = 0.86, p < 0.001). CONCLUSION: One year after the administration of G-CSF to healthy people, their blood counts were normal and unchanged from pretreatment counts. If healthy people donate blood progenitor cells after a second G-CSF course the quantity of CD34+ cells collected will be similar to that obtained in the first collection.

Systemic chemotherapy for the treatment of metastatic melanoma.

Several new treatment regimens for patients with metastatic melanoma have shown improved overall response rates when compared with the historical results with dacarbazine as a single agent. These regimens have included the addition of tamoxifen to cisplatin-containing regimens or the combination of chemotherapeutic agents with biologicals. Despite the seeming improved overall response rates, the complete remission rates remain less than 20%. Additionally, relatively few new agents have shown significant activity against melanoma cell lines, offering promise for future treatments. In the midst of all this pessimism, however, we should recognize that we have learned important information from these studies that will provide us with the basis for the next generation of clinical trials. We should continue our aggressive search for new agents or combinations of agents that will improve the outlook for our patients while continuing the laboratory studies that may provide insight into new mechanisms for attacking malignant cells.

Changes in blood counts after the administration of granulocyte-colony-stimulating factor and the collection of peripheral blood stem cells from healthy donors.

BACKGROUND: After the collection of granulocyte-colony-stimulating factor (G-CSF)-mobilized peripheral blood stem cells from healthy donors, the donor platelet counts fall. However, the magnitude and duration of this decrease are not known. STUDY DESIGN AND METHODS: Sixty healthy people were given G-CSF (5, 7.5, or 10 micrograms/kg/day) for 5 days (Days 1-5), and 1 peripheral blood stem cell component was collected on Day 6. The platelet count, white cell count, absolute neutrophil count, hematocrit, and red cell count were measured before administration of G-CSF (Day 0), before collection of peripheral blood stem cells on Day 6, and on Days 8, 10, 13, 16, and 20. RESULTS: The platelet count fell from 261 +/- 47 x 10(9) cells per L on Day 0 to 159 +/- 30 x 10(9) cells per L on Day 8 (p < 0.0001) and reached its lowest level on Day 10 (146 +/- 30 x 10(9)/L; p < 0.001). Compared to Day 0 levels, the platelet count was lower on Day 13 (185 +/- 49 x 10(9)/L, p < 0.001), was the same on Day 16 (270 +/- 53 x 10(9)/L), and was greater on Day 20 (333 +/- 60 x 10(9)/L, p < 0.0001). The white cell count returned to pretreatment values on Day 13, and the absolute neutrophil count returned to pretreatment values on Day 10 (Day 0 white cell count = 6.05 +/- 1.59 x 10(9)/L and Day 0 absolute neutrophil count = 3.97 +/- 1.52 x 10(9)/L). On Day 20, both were less than pretreatment values (white cell count = 5.14 +/- 1.24 x 10(9)/L, p = 0.0007 and absolute neutrophil count = 3.20 +/- 1.24 x 10(9)/L, p = 0.0036). The red cell counts on Day 16 (4.52 +/- 0.41 x 10(12)/L) and Day 20 (4.42 +/- 0.39 x 10(12)/L) were less than Day 0 values (4.73 +/- 0.43 x 10(12)/L, p = 0.008 and p < 0.0001, respectively). The hematocrit on Day 20 (39.2 +/- 3.2%) was also less than that on Day 0 (41.2 +/- 4.8%; p = 0.01). The changes in these blood counts were not affected by the dose of the G-CSF. CONCLUSION: After stimulation with granulocyte-colony-stimulating factor and the collection or peripheral blood stem cells, the platelet counts in normal donors were decreased for at least 7 days (Days 6-13). Two weeks after collection of peripheral blood stem cells (Day 20), platelet production was increased, but the production of neutrophils and red cells was decreased. If two or more peripheral blood stem cell components are collected, then the platelet count should be measured after the second and subsequent collections. Further studies on the long-term effect of G-CSF on blood counts are needed.

Treatment of normal individuals with granulocyte-colony-stimulating factor: donor experiences and the effects on peripheral blood CD34+ cell counts and on the collection of peripheral blood stem cells.

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) has been used in patients to increase the level of circulating hematopoietic progenitors. Although G-CSF has been administered to some healthy individuals, the kinetics of mobilization of peripheral blood stem cells (PBSCs), the optimum dose schedule and the incidence and nature of adverse reactions in normal individuals are not completely defined. STUDY DESIGN AND METHODS: Normal individuals (n = 102) who received G-CSF for 5 or 10 days at doses of 2, 5, 7.5, or 10 micrograms per kg per day were studied. The subjects were observed for symptoms and physical changes, and blood samples were obtained for a variety of laboratory tests. After 5 or 10 days of G-CSF treatment, PBSCs were collected by apheresis and analyzed. RESULTS: Overall, 89 percent of the individuals completed the 5-day treatment protocol and 88 percent completed the 10-day protocol without modification of the dose of G-CSF administered. Ninety percent of donors experienced some side effect of G-CSF. The most frequent effects noted were bone pain (83%), headache (39%), body aches (23%), fatigue (14%), and nausea and/or vomiting (12%). The dose of G-CSF administered directly affected the proportion of people with bone pain (p = 0.025) or body aches (p = 0.045) or who were feeling hot or having night sweats (p = 0.02) or taking analgesics (p = 0.01). With the 5-day dose schedule, several changes in serum chemistries occurred, including increases in alkaline phosphatase (p = 0.001), alanine aminotransferase (p = 0.0013), lactate dehydrogenase (p = 0.0001), and sodium (p = 0.0001). Decreases occurred in glucose (p = 0.045), potassium (p = 0.0004), bilirubin (p = 0.001), and blood urea nitrogen (p = 0.0017). In donors who received G-CSF for 5 days, the absolute neutrophil count was increased after one G-CSF dose, and it reached a maximum on Day 6, as did the number of CD34+ cells (64.6 +/- 55.9 x 10(6) cells/L). In those same donors, the platelet count after apheresis on Day 6 was 32 +/- 13 percent lower than pretreatment values (250 +/- 42 x 10(9) cells/L). In donors receiving G-CSF for 10 days, the neutrophil count reached a maximum on Day 8, but the number of CD34+ cells peaked on Day 6 (58.3 +/- 52.1 x 10(5) cells/L) and then declined. The platelet count decreased from pretreatment values by 28 +/- 12 percent prior to apheresis on Day 11. When individuals were treated for 5 days with G-CSF, the quantity of CD34+ cells collected was directly related to the G-CSF dose. When 5 micrograms per kg per day was given, 2.80 +/- 1.81 x 10(8) cells were collected, compared with collection of 4.67 +/- 3.11 x 10(8) cells when 10 micrograms per kg per day was given (p = 0.04). More important, PBSCs collected after 10 days of G-CSF administration (5 micrograms/kg/day) had significantly fewer CD34+ cells (0.82 +/- 0.37 x 10(8) cells, p = 0.01) than did PBSCs collected after 5 days of G-CSF (5 micrograms/kg/day). CONCLUSION: Most normal donors receiving G-CSF experience side effects, but these are mild to moderate in degree. Some alterations in blood chemistries occur, but none were clinically serious. Because of the symptoms associated with G-CSF, these individuals must be monitored closely. The treatment of normal donors with G-CSF for more than 5 days significantly decreased the number of circulating CD34+ cells and the quantity collected by apheresis.

Assessment of personnel protective equipment use at two Department of Energy facilities.

Research facilities under the jurisdiction of the United States Department of Energy are required to formally report significant radioactive contamination incidents to the Department of Energy under Department of Energy Order 5000.3. Through the reporting process, the Department of Energy is given a detailed incident description through a notification report, periodically apprised of subsequent facility response through update reports, and informed of the corrective actions that the facility will implement to prevent incident recurrence through a final report. For tracking and trending purposes, the notification, update, and final reports are entered into the Department of Energy Occurrence Reporting and Processing System database. This paper details an analysis that was performed on the radioactive contamination incident reports that were entered into the Department of Energy Occurrence Reporting and Processing System database by two similar research facilities at Los Alamos National Laboratory. The analysis focused on personnel protective equipment requirements at the two facilities to determine whether personal skin and clothing contamination incidents could be better avoided.

Determination of ABO glycosyltransferase genotypes by use of polymerase chain reaction and restriction enzymes.

BACKGROUND: The molecular basis of red cell ABO group antigens has been determined. The genes encoding the group A and B glycosyltransferases and a nonfunctional group O transferase have been cloned and sequenced. All three genes were similar. When compared to the nucleotide sequence of the A gene, the O gene has a one-base deletion that leads to a frame shift and results in a nonfunctional protein. The B gene differs from the A gene at seven nucleotides. STUDY DESIGN AND METHODS: Techniques using polymerase chain reaction and restriction enzymes to determine ABO transferase genotypes from white cell DNA were modified. Nucleotide sequence differences within the genes were analyzed by the application of selected restriction enzymes. Restriction enzymes Asp718 and BstEII were used to analyze the genes at nucleotide 258, and BssHII and Kas I were used to analyze the genes at nucleotide 523. ABO red cell phenotypes were compared in 60 unrelated individuals with ABO transferase genotypes. The ABO phenotypes and genotypes of individuals from two different families were also analyzed to determine if this method could distinguish individuals who were homozygous for A or B transferase genes from those who were heterozygous. RESULTS: The phenotypes and genotypes were consistent for all unrelated individuals, and within the families, heterozygous individuals could be distinguished from homozygous individuals. Nevertheless, two individuals from one family were found to have a group A red cell phenotype, but when the transferase genes were analyzed at nucleotide 523 with enzymes BssHII and Kas I, both A and B transferase genes were detected. Further analysis of the transferase genes at nucleotide 700 by using restriction enzymes Alu I and Hpa II and those at nucleotide 793 by using enzyme BstNI found that both transferase genes in the two individuals were similar to the A transferase gene. CONCLUSION: An A allele of the group A glycosyltransferase was detected that had the same sequence as the B gene at nucleotide 523 but was identical to the A gene at positions 700 and 793. The identification of this variant gene makes genotyping at nucleotide 523 unreliable. However, analysis of the genes at other sites of nucleotide variation may accurately identify phenotypes.

Identification of a new white cell antigen.

BACKGROUND: Antibodies to white cell antigens can cause alloimmune neonatal neutropenia, autoimmune neutropenia, and transfusion reactions. CASE REPORT: A full-term male infant developed a skin infection and was found to be neutropenic on his fourth day of life. He had a transient increase in his neutrophil count after treatment with intravenous immunoglobulin, but his neutrophil count was not consistently normal until he was 6 weeks old. Serum from the baby's mother reacted in a granulocyte immunofluorescence assay but not in a granulocyte agglutination assay. The mother's serum was tested in the granulocyte immunofluorescence assay against neutrophils from 103 healthy, unrelated people, and it reacted with cells from 66 percent of those people. The expression of SL correlated weakly with the expression of NA1 (r = 0.23; p = 0.02) and 5a (r = 0.20; p = 0.05) antigens. SL antigen expression on neutrophils was not associated with the expression of NA2, NB1, NB2, NC1, 5b, 9a, or Mart. The expression of SL on neutrophils from members of an extended family was analyzed, and the antigen was found to be inherited in an autosomal-dominant manner. Anti-SL also reacted with T-lymphocytes in a flow cytometry assay but did not react with red cells or platelets. No lymphocytotoxic antibodies were detected in the mother's sera. The anti-SL was tested against neutrophils in an immunoprecipitation and immunoblotting assay, but no molecules were identified. The neutrophil-specific antigens NA are located on Fc gamma receptor III (CD16). To determine if the SL antigen was also located on Fc gamma receptor III, anti-SL was also tested in a monoclonal antibody immobilization of granulocyte antigens assay. Anti-SL did not react with molecules recognized by CD16 monoclonal antibodies. CONCLUSION: A new white cell antigen SL, with a frequency of 66 percent, was identified on neutrophils and T-lymphocytes as a result of the evaluation of a case of neonatal alloimmune neutropenia. The molecule bearing the SL antigen was not identified in immunoblotting, immunoprecipitation, or monoclonal antibody immobilization of granulocyte antigens assays.