Histamine levels in stored human blood

The histamine content in plasma samples obtained from a group of healthy blood donors, from blood stored in citrate-phosphate-dextrose supplemented with adenine (CPDA) packs for up 28 days, and from the side arm of a transfusion line was measured by radio-enzymatic assay. Healthy donors had a mean plasma histamine content of 0.79 ng per ml. Blood stored in CPDA initially showed a similar histamine level (0.69 ng/ml on day 3 of storage), but there was a progressive rise with time, and at 28 days, the level was 20.5 ng per ml. The increase in histamine is best described by a positive exponential and may be explained by a process whereby the plasma histamine level increases the degree of histamine release from blood cells. The histamine levels in the blood infused into patients tended to be higher than those found in the stored units of the same age, if these packs were less than 7 days old. This may have been caused by the unit becoming warmer during transfusion. We speculate that the histamine levels in the older units of stored blood were high enough to cause or augment transfusion reactions and that the storage age of blood may have a bearing on the incidence of transfusion reactions.     

The determination of blood histamine in asthmatic patients with a simple and sensitive method

The histamine level of whole blood and plasma in asthmatic patients was estimated by means of a simple, sensitive and specific method, which was developed to measure low histamine levels. This method consists of the following procedures; the partial purification of histamine with a small P-cellulose column; its further purification with high performance liquid chromatography (HPLC); and fluorometric detection with precolumn o-phthaldehyde (OPT) reaction. The present assay could detect as little as 0.5 ng of histamine concentration. Blood histamine levels in patient with asthmatic attack, 57 +/- 34 ng/ml (mean +/- S.D. N = 14), were significantly different from those in symptom-free period, 37 +/- 15 ng/ml (N = 15) as well as those in normal subjects, 36 +/- 17 ng/ml (N = 12). However, there were no significant differences among plasma histamine levels in normal subjects, 1.6 +/- 1.7 ng/ml (N = 12) and, in asthmatic patients during attacks, 1.6 +/- 1.8 ng/ml (N = 14) and symptom-free periods, 1.6 +/- 1.7 ng/ml (N = 15). These results indicate that plasma histamine concentration does not increase during asthmatic attacks, even though there was significant increase of blood histamine concentration.     

Plasmin activity and complement activation during storage of citrated platelet concentrates

Platelet concentrates were studied for evidence of plasmin activity and complement activation during a 7-to-10-day storage period. When measured by an amidolytic activity assay, plasmin reached a level of 845 +/- 540 nkats/L on day 7 (n = 9). Fibrin(ogen) degradation product (FDP) levels became markedly elevated on the tenth day of storage, rising to 45 +/- 22 micrograms/ml (n = 5). Antiplasmin levels decreased in platelet concentrates by 18% +/- 6% (n = 5) over 7 days, but there was no significant decrease in stored platelet-poor plasma (-1.7%, n = 5, p = 0.5). The amount of plasminogen in platelet concentrate converted to plasmin was estimated to be less than 3% by assay of total plasminogen. Supernatant plasma from stored platelet concentrates was examined for the presence of the complement activation peptides C3a and C5a. From day 0 to day 10 of storage, mean C3a levels rose from 327 ng/ml to 6690 ng/ml. An equivalent increase in C3a levels, from 336 ng/ml at day 0 to 6866 ng/ml at day 10, was also observed in stored platelet-poor plasma. C5a was not detected (less than 10 ng/ml) at any point during the storage period; however, we noted a small decrease of borderline significance (p = 0.04) in total C5 from day 0 (117 micrograms/ml) to day 10 (108 micrograms/ml). Only trace amounts of C3 fragments were found on stored platelets, and there was no evidence of the membrane attack complex. These findings indicate the presence of plasmin activity and conversion of C3 during storage of platelet concentrates.     

Increased plasma histamine levels in uraemic pruritus

1. We determined plasma levels of histamine in uraemic patients and examined their correlation with the presence of pruritus. 2. In 27 patients with chronic renal failure, plasma histamine levels were analysed by radioimmunoassay and were compared with those of 40 healthy adult subjects. The control population showed plasma histamine concentrations of 185 +/- 33 pg/ml, which were significantly lower than those of the patients with renal insufficiency. The highest levels (552 +/- 116 pg of histamine/ml) were found in 16 patients with chronic renal failure (mean serum creatinine 5.1 +/- 1.0 mg/dl) and severe itching. 3. Twelve patients with pronounced pruritus who were on maintenance haemodialysis (serum creatinine 9.2 +/- 1.2 mg/dl) had a mean plasma histamine concentration of 515 +/- 81 pg/ml. Fifteen patients on regular haemodialysis (serum creatinine 9.0 +/- 1.5 mg/dl) and who experienced itching had plasma histamine levels (322 +/- 40 pg/ml) which were significantly lower (P less than 0.01) than those of the patients with pruritus but which were elevated compared with those of the control population (P less than 0.01). 4. No correlation could be found between increased plasma histamine levels and the type of dialysis membrane used or the method of sterilization of the membrane. 5. Haemodialysis alone did not reduce plasma histamine concentrations, although high concentrations could be detected in the ultrafiltrate. In six patients a rapid decrease in plasma histamine concentration from 565 +/- 134 pg/ml to within the normal range could be detected after 60 min of combined haemodialysis and haemoperfusion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Removal of soluble biologic response modifiers (complement and chemokines) by a bedside white cell-reduction filter

BACKGROUND: Biologic response modifiers infused with stored platelet concentrates (PCs) are believed to contribute to symptoms seen during transfusion reactions. Although prestorage white cell reduction is known to decrease the production of some biologic response modifiers during storage, the possibility that poststorage (bedside) white cell reduction could reduce the amount of biologic response modifiers already present in stored PCs during bedside filtration has not been well studied. STUDY DESIGN AND METHODS: Individual PCs were pooled on storage Days 2 and 5 and passed through a third-generation white cell- reduction filter. The results from a series of in vitro PC assays were studied, before and immediately after filtration, as were levels of C3a and interleukin 8 (n = 5). Levels of other biologic response modifiers- C5a, interleukin 1 beta, interleukin 6, tumor necrosis factor alpha, and RANTES-were also studied. Removal of interleukin 8 and RANTES was studied further by using serial filtration of units of PC. RESULTS: For the in vitro platelet assays studied, pH was unchanged after filtration from prefiltration values in units of PCs pooled on storage Day 2 or 5. A 4 log10 reduction in white cells was reliably seen after filtration in Day 2 and 5 pooled PCs. Postfiltration platelet loss was 14.8 percent for Day 2 pooled PCs and 9.6 percent for Day 5 pooled PCs. For pools of both Day 2 and Day 5 platelets, postfiltration levels of CD62 (P-selectin, CD62P) were unchanged from prefiltration levels, as were results for morphology scores. Levels of C3a decreased after filtration in both the Day 2 pooled PCs (448 ng/mL before filtration vs. 20 ng/mL after filtration) and the Day 5 pooled PCs (1976 ng/mL before filtration vs. 124 ng/mL after filtration). Levels of interleukin 8 were similarly reduced after filtration in the Day 2 pooled platelets (188 pg/mL before filtration vs. 27 pg/mL after filtration) and the Day 5 pooled platelets (2234 pg/mL before filtration vs. 799 pg/mL after filtration). Levels of interleukin 8 in other components evaluated after filtration declined similarly. However, levels of the proinflammatory cytokines interleukin 1 beta and interleukin 6 did not decline after filtration. Serial filtration studies showed that, although levels of interleukin 8 and RANTES were initially lowered by filtration, they returned to prefiltration values with increases in the volume of filtration. CONCLUSION: The third-generation bedside filter used in this study reliably reduced the level of white cell contamination to 4 log10 white cells per PC. It also lowered the levels of interleukin 8, RANTES, and C3a. The filter did not, however, remove (scavenge) the proinflammatory cytokines interleukin 1 beta and 6. The mechanism of chemokine and C3a removal by the filter is unknown, but it may be related to ionic interactions between these biologic response modifiers and the filter medium.     

H2 histamine antagonists and the hypothalamo-hypophyseal-gonadal axis: effect of short-term treatment on serum levels of prolactin, FSH, LH and testosterone in male ulcer patients

Serum prolactin (PRL ng/ml) was measured in 7 male patients on cimetidine (CMT) and in 13 on ranitidine (RNT) before therapy and 5, 10, 15 and 30 days after; at the same intervals FSH (ng/ml), LH (ng/ml) and testosterone (ng/ml) were measured in 5 patients too, in order to ascertain hypothalamic, pituitary, gonadal dysfunction caused by H2 histamine blockers. In agreement with other authors, FSH, LH and testosterone were not affected: no statistical difference was found between basal values and the times of follow-up. In RNT-treated patients PRL showed a little, but not significant increase; in CMT-treated group PRL increased with a peak on 10th day (19.41 +/- 2.14 ng/ml vs. 9.50 +/- 1.72 ng/ml; P less than 0.01); on 30th day PRL levels were still higher than basal ones (15.85 +/- 2.38 ng/ml vs. 9.50 +/- 1.72 ng/ml; P less than 0.05). Unlike other trials we observed that PRL rises very early in oral treatment with CMT, perhaps for resetting of inhibiting factors, it trends to decrease and reach basal values shortly after 30th day notwithstanding therapy. It is now widely accepted that H2 blocker's action is on central histaminergic pathways modulating PRL secretion; in our opinion, the measured differences of PRL secretion with CMT and RNT in short term observations, are produced by different crossing of blood-brain barreer.     

Storage of single donor platelet concentrates: Paired comparison of storage as single or double concentrates

Modern cell separators allow the collection of two plateletpheresis concentrates (PCs) at one session. This study evaluates the quality of PCs stored as double concentrates in standard storage containers of two manufacturers. We collected 20 PCs that contained 4.5 × 10¹¹ platelets in 375 ml plasma (10 using the COBE Spectra and 10 using the Fresenius AS.TEC 204 with 500 ml bags) that were split into one unit of 3.0 × 10¹¹ platelets in 250 ml (3.0‐PC) and one of 1.5 × 10¹¹ platelets in 125 ml (1.5‐PC). Storage of one 3.0‐PC per bag of a two‐bag system corresponded to storage conditions for double PCs and storage of one 1.5‐PC per bag to storage conditions of single PCs. Cell counts, blood gas analysis, glucose and lactate levels, platelet aggregation, and activation and plasma levels of β‐ thromboglobulin (β‐TG) and complement factor 3a (C3a) were measured before storage and again on days 3 and 5. COBE 3.0‐PCs demonstrated less pH rise, lactate production, CD 62P expression and β‐TG plasma levels, and better aggregability after storage than COBE 1.5‐PCs. Fresenius 1.5‐PCs had similar platelet quality to COBE 3.0‐PCs. Fresenius 3.0‐PCs showed a fall of pH (day 5: 6.22 ± 0.56), the highest amount of anaerobic glycolysis compared to all other storage conditions investigated, high CD 62P‐ expression and β‐TG plasma levels, and impaired aggregability on days 3 and 5. The highest C3a levels were found in COBE 1.5‐PCs. 3.0 × 10¹¹ platelets in 250 ml plasma should be stored either in one bag of the COBE system or in two 500 ml bags of the Fresenius system. The COBE two‐bag system allows the storage of two PCs without loss of platelet quality. Two PCs should not be stored in the Fresenius C4L 500 ml storage containers. J. Clin. Apheresis. 16:148‐154, 2000. © 2001 Wiley‐Liss, Inc.     

白细胞过滤对血小板保存期间炎性细胞因子水平的影响及临床效果的评估

目的　考察浓缩血小板悬液 (plateletconcentratesuspend ,PCs)在保存期内IL 1β、IL 6、IL 8和TNF α的浓度变化和过滤对其的影响 ,了解在保存前滤除PCs中的白细胞是否能有效地减少这些细胞因子的积累和降低受血者非溶血性发热性输血反应 (febrilenonhemolytictransfusionreactions,FNHTR)发生率。 方法　将 1单位PCs分成两等份 ,分别给予血小板专用白细胞滤器过滤处理和不滤除白细胞处理 ,保存 5d。在 0、3、5d测定IL 1β、IL 6、IL 8和TNF α含量及白细胞计数 ,采用配对t检验进行统计分析 ;临床观察未滤组和过滤组PCs输后FNHTR发生率。结果　PCs中的白细胞计数与保存 5d时IL 1β、IL 6、IL 8和TNF α水平之间呈正相关。未滤组PCs中有较多白细胞混入 [(35 1± 81)× 10 6/袋 ],在保存期间IL 1β、IL 6、IL 8和TNF α水平明显升高 ;过滤组的PCs残余白细胞 <1× 10 6/袋在保存期间诸细胞因子均保持在 0d水平 ;临床观察显示 ,末滤PCs与过滤PCs输注后FNHTR发生率分别为 2 0 .83%和 5 .83% ,P <0 .0 1。结论　保存前用血小板专用去白细胞滤器去除PCs中残留的白细胞能有效地防止细胞因子的积累 ,同时保留 95 %以上的血小板。输注滤除白细胞的PCs能有效地减少FNHTR发生     

Cutaneous late-phase response to allergen. Mediator release and inflammatory cell infiltration.

To better define the inflammatory infiltrates and kinetics of mediator release during the cutaneous late-phase reaction (LPR), we examined skin biopsies at 8 h, and skin chamber cell counts and mediator release for 12 h after antigen challenge. Compared with the control sites, the antigen-stimulated biopsy sites contained 14 times as many basophils (P less than 0.01) and six times as many eosinophils (P less than 0.001) with one to two fold more mononuclear cells (P less than 0.03) and neutrophils (P less than or equal to 0.01). Similar changes were found in the skin chambers. Although there were neutrophils in the control chamber, they were only twice as numerous in the antigen challenged site (P less than 0.01). Eosinophils were 35-fold (P less than or equal to 0.03) more prevalent in the antigen chamber than the control chamber for hours 8-12 and basophils were noted starting in the eighth hour and were 20-fold (P less than or equal to 0.03) more concentrated in the antigen chamber during the next 4 h. The mononuclear cells were not significantly different between antigen and control blisters. With respect to inflammatory mediators, there was an initial peak of histamine (13.2 +/- 2.9 ng/ml) in the blister fluid at 1 h. The level then fell to approximately 2 ng/ml, followed by a secondary rise starting at the eighth hour and increasing to 9.8 +/- 2.8 ng/ml by the twelfth hour. This secondary increase in histamine correlated significantly (r = 0.81, P less than 0.05) with the observed influx of basophils. PGD2 in the blister fluid rose to 371+/-25 pg/ml during the first 4 h and then slowly decreased to half this level during the last 4 h. Thus, the cutaneous LPR has been shown to manifest a secondary increase in histamine levels and a markedly specific increase in eosinophils and basophils with mediator release apparently being derived from the latter cells.     

Potentiation of vancomycin-induced histamine release by muscle relaxants and morphine in rats

The intravenous injection of vancomycin sometimes causes anaphylactoid reactions, in which histamine release may play a major role. These reactions are more frequently manifested when vancomycin is injected into anesthetized patients. We examined the vancomycin-induced histamine release and the interaction of vancomycin with muscle relaxants or opioid in rats. In an in vitro study with rat peritoneal mast cells, treatment with vancomycin at concentrations of greater than 1.25 mM produced significant histamine release. Tubocurarine, vecuronium, pancuronium, succinylcholine, and morphine up to concentrations of 0.25, 1, 5, 30, and 5 mM, respectively, produced no significant histamine release. However, the nonsignificant histamine release induced by 0.5 mM vancomycin was clearly enhanced by combining vancomycin with any of these agents. In the in vivo study, the intravenous injection of vancomycin significantly increased the plasma histamine levels in rats when vancomycin was injected at 200 mg/kg of body weight (63.2 +/- 34.0 ng/ml [mean +/- standard deviation]) but not when it was injected at 100 mg/kg (30.8 +/- 20.2 ng/ml) compared with that in the saline-treated rats (22.5 +/- 11.4 ng/ml). Although the subcutaneous administration of morphine (10 mg/kg) never increased the plasma histamine levels, the intravenous injection of vancomycin (100 mg/kg) 30 min after this morphine treatment markedly increased the plasma histamine levels (56.0 +/- 26.9 ng/ml). These findings provide experimental evidence that the combination of muscle relaxants or an opioid with vancomycin may increase the risk of anaphylactoid reactions by enhancing the release of histamine.     

Diagnostic value of some markers of infection in cardiosurgical patients in the early postoperative period

The diagnostic value of traditional markers of infection and procalcitonin test (PCT) in the early postoperative period was compared in 60 cardiosurgical patients with acquired cardiac diseases and at risk for postoperative infectious complications. The mean age of the patients was 51 +/- 11 years. Preoperatively, all the patients had no signs of infections. The patients were examined before and on days 1, 2, 3, and 6 after surgery. Along with the routine studies (thermometry, general blood analysis), the plasma concentration of PCT was determined by immunoluminometric technique (LUMI-test PCT, Brahms Aktiengesellschaft, Germany). The preoperative level of PCT did not exceed the normal values ( < 0.5 ng/ml). On postoperative days 2 to 17, 14 (23.3%) patients developed infectious complications (Group 2); the other patients were included into a group of comparison (Group 1). Just within the first postoperative days, the levels of PCT were significantly higher in Group 2 patients than in Group 1 (7.58 +/- 2.34 and 3.51 +/- 0.71 ng/ml, respectively; p < 0.05). A difference was found in the count of white blood cells between the groups only from day 3. There were no significant differences in body temperature between the groups. At the second stage of analysis of the data, in accordance with the level of PCT on the first day after surgery and its subsequent changes, all the patients were divided into 4 groups (A-D). The level of PCT on postoperative day was less than 0.5 ng/ml in Group A (n = 6), 0.5-2 ng/ml in Group B (n = 23) and more than 2 ng/ml in groups C (n = 26) and D (n = 5). Subsequently, it was in the normal range in Group A, decreased to the normal values in Groups B and C by day 6 following surgery. The persistence of the high level of PCT was observed in Group D where there were the bulk (60%) of infectious complications. As compared with the traditional clinical and laboratory criteria (fever, leukocytosis), PCT is the earliest and most specific marker of bacterial infection in cardiosurgical patients in the early postoperative period. The level of PCT > 3.5 ng/ml within the first 24 hours after surgery is shown to be a predictor of postoperative infectious complications.     

GM-CSF对脐血CD34+巨核祖细胞体外扩增及分化的影响

本实验旨在研究GM-CSF对脐血CD34^＋细胞诱导分化为巨核细胞的影响.采用免疫磁珠法分选CD34^＋细胞,在含有TPO＋IL-3＋SCF并添加了不同浓度（5、20、100ng/ml）的GM-CSF的无血清培养基中进行培养.培养6、10、14天后计数单个核细胞（MNC）,检测CD41^＋细胞比例和CFU-MK.结果表明,培养14天后3种不同浓度GM-CSF对MNC均有明显的扩增作用,其中以20和100ng/ml GM-CSF的扩增效果较好.3种不同浓度的GM-CSF均使CD41^＋细胞比例增加,20和100ng/ml与5 ng/ml GM-CSF相比更能提高CD41^＋细胞的比例.5和20 ng/ml的GM-CSF能促进CFU-MK的形成,但100ng/ml的GM-CSF却抑制CFU-MK的形成.结论：在TPO＋IL-3＋SCF细胞因子组合中添加GM-CSF有利于促进脐血CD34^＋细胞诱导分化为巨核细胞.     

Elimination of cytokine production in stored platelet concentrate aliquots by photochemical treatment with psoralen plus ultraviolet A light

BACKGROUND: Cytokines generated in platelet concentrates (PCs) during storage have been implicated as possible mediators of febrile nonhemolytic transfusion reactions. Two potential methods of white cell inactivation were compared for their ability to reduce cytokine synthesis in pooled random-donor PC aliquots: treatment with gamma-radiation and photochemical treatment (PCT) using psoralens and ultraviolet A light. STUDY DESIGN AND METHODS: ABO-matched PC aliquots were pooled and divided into separate aliquots. Aliquots (20 mL) were taken from each pool to serve as an untreated control and to undergo gamma-radiation. Aliquots were treated by using either gamma-radiation (2500 or 5000 cGy) or virucidal PCT. PCT with the psoralens 8-methoxypsoralen (8-MOP), aminomethyltrimethyl psoralen (AMT), and S-59 was investigated. PC aliquots were stored for 7 days and analyzed for levels of interleukin 8 by use of an enzyme-linked immunosorbent assay. Levels of DNA adduct formation were determined by using 3H-labeled psoralens. RESULTS: Levels of interleukin 8 in the untreated random-donor PC aliquots increased with increasing white cell counts, but they were not affected by pooling. The untreated control aliquots and the aliquots treated with gamma-radiation had significant increases in levels of interleukin 8 after 5 to 7 days of storage (p<0.05). PCT with S-59 resulted in a significant reduction in cytokine synthesis (p<0.05). Day 5 to 7 levels of interleukin 8 did not differ significantly from Day 0 levels. Inhibition of interleukin 8 production by PCT increased with increasing levels of DNA modification (S-59 > AMT > 8-MOP). CONCLUSION: PCT that utilizes S-59 has been developed to inactivate potential viral and bacterial pathogens in PC aliquots while maintaining in vitro platelet function. These data demonstrate that PCT of aliquots of pooled PC aliquots before storage also prevents white cell cytokine synthesis during storage. PCT may therefore offer the potential for reducing cytokine-associated febrile nonhemolytic transfusion reactions.     

Biologic activity of RANTES in apheresis PLT concentrates and its involvement in nonhemolytic transfusion reactions

BACKGROUND: RANTES, one of the PLT-derived biologic response modifiers, accumulates in PLT concentrates (PCs) during storage and may play a causative role in nonhemolytic transfusion reactions (NHTRs) after PC transfusion. STUDY DESIGN AND METHODS: To investigate the association of RANTES with NHTRs, the biologic activity of RANTES in the supernatant of stored PC at the intravascular concentration expected after PC transfusion was assessed by examining chemotaxis and histamine release in human basophils. In addition, the levels of RANTES in PCs involved in NHTRs were compared with those in PCs causing no transfusion reactions. RESULTS: The supernatant of PC diluted to contain 1 nM RANTES significantly increased the migration of and release of histamine from basophils. Neutralizing antibody to RANTES suppressed the PC-triggered migration, but not histamine release. The levels of RANTES in PCs involved in NHTRs after PC transfusion were comparable to those in PCs that did not cause any transfusion reactions. CONCLUSION: RANTES that accumulated in PCs during storage was biologically active in a basophil chemotaxis assay at the intravascular concentration expected after PC transfusion. However, the NHTRs after PC transfusion were not simply related to the RANTES level in PCs.     

The effects of chronic buprenorphine treatment on cocaine and food self-administration by rhesus monkeys.

The goal of this study was to determine if buprenorphine continues to reduce cocaine self-administration over long periods of treatment, or if tolerance develops to this effect. The effects of 30 to 120 days of buprenorphine treatment (0.32 mg/kg/day) on cocaine and food self-administration were examined in six rhesus monkeys. Saline control treatment was studied for 15 days before and after buprenorphine treatment. Intravenous cocaine (0.05 or 0.10 mg/kg) and food (1 g banana pellet) self-administration were maintained on a FR 4 (VR 16:S) schedule of reinforcement. Cocaine self-administration decreased significantly (P less than .0001) and remained 60 to 97% below saline treatment baseline levels (52 +/- 2 injections/day) throughout 120 days of buprenorphine treatment (P less than .01). After substitution of saline for buprenorphine, cocaine self-administration resumed and averaged between 21 (+/- 3.6) and 56 (+/- 6.5) injections per day over 20 days. Buprenorphine plasma levels averaged 18 (+/- 2.84) ng/ml (range 10.9-30 ng/ml) during buprenorphine treatment. Buprenorphine plasma levels usually decreased by 50% or more within 27 hr after the last buprenorphine dose. Low levels of buprenorphine (0.10-0.19 ng/ml) were measured for 30 to 74 days after abrupt termination of daily buprenorphine treatment. Food self-administration was initially reduced (P less than .01-.05), but tolerance to buprenorphine's suppression of food-maintained responding developed over 30 to 70 days of treatment. Food self-administration returned to and significantly exceeded (P less than .05-.01) saline treatment base-line levels, whereas cocaine self-administration remained significantly suppressed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Radioimmunometric determination of histamine in myeloproliferative syndromes

Quantitative analysis of histamine is increasingly used in clinical haematology. The present study demonstrates the properties of and potential indication for a novel histamine radioimmunoassay (RIA) in clinical haematology. The sensitivity of this test assay corresponds to a histamine level of 0.1 to 0.5 nM, the non-specific cross reaction with endogenous histamine metabolites appears to be less than 0.1%. The total histamine levels in the peripheral blood of healthy donors (n = 10) ranged from 10 to 100 ng/ml, the plasma histamine values from 0.02 to 0.6 ng/ml blood. Increased levels of total histamine were measured in myeloproliferative syndromes, i.e. in patients with chronic myeloid leukemia (CML) (8 of 9), myelofibrosis (OMS) (2 of 4), and polycythaemia vera (PCV) (1 of 2). An excessive increase in total histamine was observed in healthy rhesus monkeys (n = 10) treated with recombinant human interleukin-3 (rhIL-3). The total histamine value correlated with the absolute number of circulating blood basophils (correlation coefficient: 0.9). The calculated content of histamine per basophil was found to be 0.5 to 1.5 pg. Plasma histamine values in patients suffering from myeloproliferative syndromes were within the normal range. In contrast, a moderate to marked increase in plasma histamine values was observed in monkeys during IL-3 treatment. The radioimmunometric analysis of histamine clearly represents a useful new test system in clinical haematology, especially in the follow up of malignant as well as IL-3-induced myeloproliferation.     

Platelet-associated proteins in human breast cyst fluids

Thrombospondin, beta-thromboglobulin and platelet factor 4 were measured by radioimmunoassay in fluids from 50 human breast cysts. beta-Thromboglobulin concentrations were less than 4-330 ng/ml (median 13.5 ng/ml), while normal plasma contained 14-80 ng/ml. Cyst fluids contained 8-225 ng/ml platelet factor 4 with a median of 25 ng/ml (range in plasma 6.5-29 ng/ml). Thrombospondin levels in cyst fluids were 65-55 000 ng/ml (median 2 500 ng/ml) and 92% contained concentrations above the plasma range (58-215 ng/ml). When cyst fluids were classified by their electrolyte composition, group II cysts had similar levels of platelet factor 4 but significantly higher levels of beta-thromboglobulin and thrombospondin than group I. From the remarkably high concentrations of thrombospondin within cyst fluids and the differences between cyst types, it seems that either the protein is synthesised locally or selectively accumulated. Increased platelet activation is unlikely to be the source of the high thrombospondin levels in group II cysts.     

Assessment of complement activation during membrane-based plasmapheresis procedures

Previous studies have suggested that plasmapheresis procedures using a separation membrane may activate the complement system and release anaphylatoxins. This study determines the content in C3a/C3a(des Arg) and C5a/C5a(des Arg) in plasma donations obtained by the new Haemonetics Filter Core (FC) procedure and compares it to Baxter Autopheresis C (Auto-C). FC performs sequential blood centrifugation and plasma filtration on a microporous polyethersulfone membrane, while Auto-C removes blood cells by simultaneous gravitation and filtration on a rotating nylon membrane. One group of 34 donors donated on FC and two groups of 30 and 10 donors on Auto-C. Plasma aliquots were taken from the plasma units within 30 min of the end of the collection procedures, frozen at < -30 degrees C and assessed for C3a and C5a at various time points of storage. Mean C3a/C3a(des Arg) in FC plasma (N = 34) was 1,151 (range: 526-2,991), 1,092 (range: 349-3498), and 507 (range: 307-815) ng/ml at time of collection and after 6 and 12 months of storage, respectively. Respective C5a/C5a(des Arg) was 26.6 (range 4.9-74), 18.9 (9.5-42.6), and 30.9 (range: 10.7-62.3) ng/ml. Mean C3a/C3a(des Arg) was higher in Auto-C (P < 0.001): 4,724 ng/ml (N = 10; range: 2,400-7 ,360) and > 4,149 ng/ml (N = 30; 2,408- > 6,430) after 3 and 18 months storage, respectively. Mean C5a/C5a(des Arg) was 32.1 ng/ml (N = 30; range: 10.6-57.2) after 18 months of storage. Complement activation in FC plasmas appears limited compared to Auto-C, suggesting better biocompatibility of this collection device and/or a favourable impact of the sequential cell centrifugation/filtration technology used. Further studies are needed to explain differences in complement activation between apheresis procedures and to assess clinical impacts, if any.     

Interleukin-6, gamma interferon, and tumor necrosis factor receptors in typhoid fever related to outcome of antimicrobial therapy.

To study mechanisms of antibiotic effects in typhoid fever, levels of interleukin-6 (IL-6), gamma interferon (IFN-gamma), and cytokine receptors (tumor necrosis factor receptor [TNF-R] p55 and TNF-R p75) were measured in the plasma of 29 adult Nepalese with culture-positive typhoid fever before therapy and on days 4 and 15 after start of therapy with either ceftriaxone at 2 g/day for 3 days or chloramphenicol at 50 mg/kg of body weight per day for 14 days. Bacteriologic cure was defined as blood cultures testing negative on days 4 and 15 after start of therapy; clinical cure was defined as symptomatic improvement within 5 days after start of therapy and absence of relapse. Clinical and bacteriologic cures occurred in 24 patients. There were two clinical failures, two patients who failed to complete therapy because of leukopenia, and one relapse. Mean levels before therapy were elevated compared with those in healthy controls (IL-6, 11.4 pg/ml; IFN-gamma, 1.3 ng/ml; TNF-R p55, 3.8 ng/ml; and TNF-R p75, 6.1 ng/ml) and fell progressively during and after therapy. For six patients (three in each treatment group) who showed prolonged fever (> 5 days) or relapse, mean levels of IL-6 and TNF-R p55 before therapy (29.5 pg/ml and 6.1 ng/ml, respectively) and on day 4 (17.7 pg/ml and 4.0 ng/ml) were significantly greater than corresponding means for 23 patients who showed early defervescence (on admission, 6.7 pg/ml and 3.3 ng/ml, and on day 4, 1.8 pg/ml and 2.7 ng/ml, P < .05). These results indicate that the concentrations of plasma cytokines and their receptors are elevated in typhoid fever and that these concentrations can be useful in predicting outcome.     

Storage of pools of six and eight platelet concentrates

Platelet concentrates (PCs) prepared from units of whole blood are routinely stored singly at 20 to 24 degrees C and pooled prior to transfusion. Studies have been conducted to evaluate the in vitro properties of pools of six (n = 19) and eight (n = 17) ABO-identical PCs after storage, with comparative studies involving single units (n = 33). The pools were prepared using the sterile connecting device. One- day-old and 3-day-old PCs were pooled and stored for a total of 5 days in a container system consisting of two 1000-mL polyolefin containers. The pooled platelet suspension was divided approximately equally between the two containers. The platelet count was reduced by less than 5 percent during storage of the pools, which is similar to the reduction found with storage of control units of single PCs. The volume loss due to pooling was 9.6 +/− 1.9 percent (mean +/− 1 SD). The pH of the PC pools was approximately 7.0 after 5 days of storage, with no pool having a pH below 6.2. In vitro platelet properties, such as morphology score, extent of shape change induced by ADP, total ATP, aggregation response to ADP and collagen, response to hypotonic stress, lactate dehydrogenase discharge, and beta-thromboglobulin release, were similar for pools and control single PCs. In addition, comparable low levels of thymidine uptake were detected in the mononuclear leukocyte fraction of pooled and unpooled PCs that were stored for 5 days at 20 to 24 degrees C, which indicates that the mixing of lymphocytes in the pool did not stimulate in vitro immunologic reactions.(ABSTRACT TRUNCATED AT 250 WORDS)