Recombinant factor IX (BeneFix®) by adjusted continuous infusion: a study of stability, sterility and clinical experience

The safety and efficacy of adjusted continuous infusion (CI) of recombinant factor IX (FIX; BeneFix) was assessed in vitro and in a clinical study. BeneFix was reconstituted at 100 IU mL-1 with or without unfractionated heparin (4 U mL-1) and stored at either 4 degrees C or room temperature. Reconstituted BeneFix retained at least 90% activity over 14 days if stored at 4 degrees C but stability was reduced at room temperature. BeneFix reconstituted in a sterile pharmacy was free of bacterial contamination. Six patients with haemophilia B received seven CIs of BeneFix to cover routine surgery and severe bleeding episodes. The CIs lasted between 3 and 10 days. In all cases, haemostasis was excellent and the desired therapeutic FIX level was easily maintained. No thrombotic episodes or inhibitor development occurred but two patients developed thrombophlebitis at the infusion site when heparin was not added to the infusion. BeneFix is not currently licensed for CI and we suggest that studies to enable licensing should be established as soon as possible.     

Loss of factor VIII activity during storage in PVC containers due to adsorption

Recombinant factor VIII concentrates are stable when administered in a reconstituted form according to the manufacturer's specifications, and undiluted via infusion with syringe mini-pumps. However many Haemophilia centres administer recombinant factor VIII further diluted in intravenous fluids for greater ease of administration. To investigate the stability of recombinant factor VIII during administration as a diluted infusion, reconstituted factor VIII was stored in polyvinylchloride (PVC) mini-bags undiluted (146 IU mL-1) and at factor VIII concentrations of 10 IU mL-1 and 2 IU mL-1. After 48 h of storage at room temperature in PVC mini-bags, the recoveries of factor VIII activity were 41.9% of the initial activity for the undiluted (146 IU mL-1) product and 43.7% of the initial activity for factor VIII diluted to 10 IU mL-1. For factor VIII diluted to 2 IU mL-1, the amount of factor VIII activity remaining at 48 h was only 1.8% of the initial activity. In contrast, 100% of factor VIII activity was recovered after 48 h when undiluted reconstituted product (146 IU mL-1) was stored in a syringe. To investigate the mechanism of factor VIII activity loss during storage, factor VIII samples collected after 0, 3 and 48 h of storage were analysed by immunoblotting with factor VIII antibodies. No evidence of factor VIII proteolytic degradation during storage was found, however, large amounts of factor VIII antigen were recovered from the empty PVC mini-bags following elution with denaturing detergent. We conclude that clinically significant losses of factor VIII activity occur during storage in PVC mini-bags and that the loss of activity is most likely due to protein adsorption onto the plastic surface. This loss of factor VIII activity during storage in PVC containers may substantially affect the safety and potential cost savings of administering recombinant factor VIII by continuous infusion.     

Porcine factor VIII (Hyate: C®): in vitro stability,microbiological safety and effect of heparin

The in vitro stability of porcine factor VIII (PF VIII) was evaluated when it was reconstituted with sterile water (PF VIII_SW) to ≈ 30 U PFVIII mL^?1, as per the manufacturer’s recommendations, and stored in plastic syringes at room temperature, with and without heparin and at four different dilutions. PF VIII was prepared antiseptically without laminar airflow and remained sterile at room temperature for 1 week. PF VIII_SW retained at least 88% of baseline activity for 48 h and 74–86% for 72 h. Addition of heparin 1 unit mL^?1 solution resulted in a decrease in the stability of PF VIII_SW to 72–74 % of baseline values by 24 h. Reconstituted PF VIII_SW, further diluted with normal saline to 10–24 U PF VIII mL^?1, retained 98% of baseline activity for 48 h and 84% of baseline for 72 h. PF VIII diluted to 6 U mL^?1, however, retained 100% baseline activity for only 24 h, and declined to 71% and 64 % of baseline by 48 and 72 h, respectively. PF VIII reconstituted with normal saline, instead of sterile water, retained 90% or more of baseline activity for a minimum of 4 days. Once reconstituted, PF VIII appeared to be more stable at room temperature than when stored in the refrigerator. These in vitro stability studies confirm that PF VIII (30 U mL^?1) can be given effectively by prolonged continuous infusion, since it retains 88% baseline activity at room temperature in a plastic syringe for a minimum of 48 h, remains sterile and will maintain baseline PF VIII levels when further diluted with saline.     

Rifampin in drug-incorporated diet: effect of duration and temperature of storage. Relevance to drug-susceptibility testing in mice inoculated with M. leprae

This paper is in two parts. Plasma concentrations of rifampin were assayed at 11 time points in 24 hr in mice fed one of three dosages of rifampin, either by gavage or by dietary incorporation. The drug-mixed diets had been stored for a maximum of 3 weeks at 4 degrees C or at room temperature (30 degrees C-35 degrees C). The peak concentration of rifampin produced by gavage was approximately 11/2 times higher than the maximum plasma concentration of the corresponding dosage in fresh diet. Plasma concentrations decreased with the increasing duration of storage of the drug-mixed diet, irrespective of whether the diet was stored at 4 degrees C or at room temperature. This decrease was less when the diet was stored at 4 degrees C than at room temperature. Drug levels were also assayed in another set of mice selected from ongoing drug-susceptibility experiments; these mice were fed a rifampin-incorporated diet stored at room temperature. The plasma concentrations in these mice, assayed at the time of foot pad harvest, were generally higher than in the 24-hr experiment. The harvest results from these mice were compared with the harvest results from a third set of mice, also from ongoing drug-susceptibility experiments, but fed a rifampin-mixed diet stored at 4 degrees C. Multiplication of Mycobacterium leprae in mouse foot pads was prevented by rifampin mixed in the diet at a dosage of greater than or equal to 0.003%, whether stored at room temperature or at 4 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

A matter of time: evaluating the storage of fecal samples for steroid analysis

The extraction and immunoassay of fecal steroids is an increasingly common technique, used in both captive and field studies to provide an approximation of an animal's circulating concentration of hormones through non-invasive methods. Storage of fecal samples is of critical concern because fecal bacteria metabolize fecal steroids within hours after deposit. Ethanol is often used as a preservative for fecal samples stored for several hours at room temperature. We examined the stability of fecal estrogen (fE) and glucocorticoid (fGC) metabolites from baboon (Papio cynocephalus) samples in a 95% ethanol solution at ambient temperature and at -20 degrees C over the course of six months, to determine the effect of storage on steroid concentrations. As measured by radioimmunoassay, fE metabolite concentrations increased by 122% at 90 days and fGC metabolite concentrations increased by 92% at 120 days. After peaking, both hormones declined to near initial concentrations by 180 days in ambient temperature samples. In samples stored at sub-zero temperatures, fGC metabolite concentrations showed a similar but dampened pattern, while fE metabolite concentrations exhibited small and variable changes with no consistent trend. We discuss explanations for the dynamic pattern of changing fecal metabolite concentrations and offer practical and analytical guidance to field workers for situations in which ideal conditions for stabilizing hormones are not available.     

Temperature behavior of infusion solutions

Since rheological qualities (density, viscosity) of solutions depend upon temperature and consecutively influence the conditions of current, i. e. among others the dropping speed of the solutions to be infused, the behaviour of the temperature of the solutions glucose 50, 200, alvesin "new", fructose 400, intralipid 10%, sorbitoli 100, electrolytic infusion 153 DAB 7 and as comparative fluid distilled water was investigated. All solutions showed qualitatively the same behaviour. In an infusion not running after 3 till 4 hours at all levels of the infusion bottle an assimilation of the temperature of refrigerator to room temperature is to be expected. In running infusion after about one hour the infusion solution in the canule reaches room temperature, whereas the temperature of the solution in the infusion bottle is about 10 degrees C lower. With increasing speed of infusion this difference in temperature decreases. A constancy of the rheological qualities of an infusion solution is, therefore, in non-running infusion to be reached after about three hours, in running infusion after about one hour, in case that before the solution had the temperature of the refrigerator (4 degrees C till 10 degrees C). The dropping speed is influenced in such a case only by the changing of the hydrostatic pressure.     

Phosphatidylethanol in human organs and blood: a study on autopsy material and influences by storage conditions

OBJECTIVE: Phosphatidylethanol (PEth) is an abnormal phospholipid that is formed and accumulated in mammalian cells that have been exposed to ethanol. PEth has been proposed as a marker of ethanol abuse. This study was conducted to investigate the concentration of PEth in blood and organs obtained during the autopsy of alcoholics. In addition, we performed experiments on rat tissues and human blood to evaluate the effect of various storage conditions on PEth concentrations. METHODS: Human tissues and blood from alcoholics and controls were obtained at autopsy and frozen at -20 degrees C until extraction. Blood from healthy donors was incubated with ethanol for 24 hr and thereafter either extracted directly or stored at room temperature, stored at 4 degrees C, frozen at -20 degrees C, or frozen in liquid nitrogen and stored at -80 degrees C before extraction. Rats were given intraperitoneal injections of ethanol and then killed, either while still intoxicated or when sober. Rat organs were homogenized and extracted directly, after a period of storage, and/or after freezing at -20 degrees C. PEth concentration was analyzed using HPLC and verified by mass spectrometry. RESULTS: In all rat organs studied, PEth was formed during freezing at -20 degrees C with ethanol present. PEth concentrations of 9 to 205 mumol/liter were observed in the blood obtained at autopsy. The highest value was found in the case with the highest blood alcohol concentration (114 mmol/liter) at the time of death. In the experiments on human blood stored with ethanol present, PEth concentrations were not affected after 72 hr at 4 degrees C or after freezing in liquid nitrogen and storage at -80 degrees C for up to 144 hr but were slightly elevated after 24 hr at room temperature and at -20 degrees C. PEth was found in all organs obtained from the cadavers of alcoholics. Storage of organs at 4 degrees C for 24 hr with ethanol present had no effect on the PEth concentration. The PEth concentration was unaffected when no ethanol was present at the time of freezing. CONCLUSIONS: The rat experiments indicated that the very high PEth concentrations found in the organs of the alcoholics were probably largely formed while the organs were frozen at -20 degrees C. Our data suggest that tissue material from bodies that were exposed to ethanol must be stored properly to obtain reliable results from subsequent analysis for PEth. Tissue should not be frozen at -20 degrees C but instead stored refrigerated until extraction, preferably within hours of autopsy, or frozen in liquid nitrogen and stored at -80 degrees C. Blood samples that contain ethanol can be stored refrigerated for up to 72 hr or frozen in liquid nitrogen and stored at -80 degrees C without affecting PEth levels.     

Effect of Plasma Storage on Erythrocyte Sodium Pump Measurements

The possibility that sodium handling in red cells may be an indicator of abnormalities associated with hypertension has encouraged many investigators to study erythrocyte sodium transport. The results have often been in conflict, perhaps because of the variety of laboratory techniques and procedures employed. Investigation into the effect of storage of red cells in plasma showed larger changes when cells were stored at 0°C than at room temperature (19°). At 0°C, sodium efflux via the pump increased 22.1% (p < 0.001) after 2 hours storage and 54.2% (p < 0.005) after 4 hrs of storage. At 19°C, sodium efflux values were more scattered after storage, but the mean value was not significantly different even after 4 hours. Intracellular sodium increased 17.3% in cells stored 4 hrs at 0°C; significant changes were not observed when cells were stored 2 hrs at 0°C or 2-4 hours at 19°C, The number of Na, K-ATPase sites per red blood cell did not change with either storage condition. For samples processed within one hour of drawing, or stored at room temperature, there were close correlations between the sodium efflux per site and the intracellular sodium concentration as well as between the intracellular sodium and the sites per red cell. These relationships were not evident for cells stored at 0°C. These data indicate that red cells to be used in pump measurements should be stored at room temperature and should be processed within 2 hours.     

The in vitro quality of washed, prestorage leucocyte-depleted red blood cell concentrates

BACKGROUND AND OBJECTIVES: No data are currently available on the quality of washed prestorage leucocyte-depleted red blood cell concentrates (RCCs). MATERIALS AND METHODS: Five groups of RCCs stored in additive solution (SAG-M) were washed. The groups differed in the age of RCCs (2-5 days or 11-15 days), the temperature during the washing procedure and a 6-h storage period (4 degrees C or room temperature) and the washing solution (saline, SAG-M or 5% albumin). We measured ATP, 2,3-diphosphoglycerate (2,3-DPG), haemolysis, blood cell count, Na(+), K(+), pH, pO(2), pCO(2) and lactate, before and after the washing procedure and hourly during the 6-h postwash storage period. RESULTS: The erythrocyte ATP content increased by 2-13%, relative to the baseline value, during the washing procedure. The 2,3-DPG level decreased by 15-35% in 2-6-day-old RCCs and by 30-40% in 11-15-day-old RCCs (relative to baseline values) during the washing procedure. In RCCs that were washed and stored at room temperature, and in 2-week-old RCCs, a further decrease in 2,3-DPG of up to 40%, relative to the baseline value, was observed during the 6-h postwash time-period. CONCLUSIONS: Washing of RCCs stored in SAG-M results in a considerable, significant loss of erythrocyte 2,3-DPG, especially in older RCCs. This loss increases in during a 6-h storage period postwash, even at 4 degrees C. This loss of erythrocyte quality might well outweigh the benefits of washed SAG-M RCCs during massive transfusion in neonates.     

Prolonged stability of endogenous cardiotrophin-1 in whole blood

Cardiotrophin-1 (CT-1) is a recently identified cytokine of the interleukin-6 (IL-6) family that signals through the gp130 signalling pathway. CT-1 may be of central importance to the pathogenesis of ventricular remodelling in patients with acute myocardial infarction (AMI) and therefore have clinical value in the identification of patients with impaired ventricular function. Central to the clinical use of CT-1 is in the in vitro stability of the peptide. Twelve subjects were recruited. A total of 25 mL of peripheral venous blood was collected into chilled polypropylene tubes containing EDTA and aprotinin and divided into 5 aliquots. One sample was spun in a prerefrigerated centrifuge (4 degrees C) at 3,000 rpm for 10 minutes and plasma separated and frozen at -70 degrees C immediately. Remaining samples were stored for 24 and 48 hours at room temperature or on ice. CT-1 in extracted plasma specimens was measured with a competitive chemiluminescent assay. The concentration of CT-1 in samples stored optimally was 43.1 +/- 6.05 fmol/mL. CT-1 levels for storage at room temperature compared with ice at the remaining time points were as follows: 24 hours, 41.5 +/- 5.76 v 37.5 +/- 8.66; and 48 hours, 42.6 +/- 6.28 v 41.0 +/- 5.42 fmol/mL. There were no significant changes in concentrations of CT-1 stored optimally or kept for up to 48 hours in aliquots of whole blood at room temperature or on ice. We conclude that CT-1 is stable in specimens of whole blood treated with EDTA and aprotinin and stored for up to 48 hours at room temperature or on ice, hence permitting its development in the routine clinical investigation of patients with heart failure.     

Survival of calcium pyrophosphate crystals in stored synovial fluids.

Eleven synovial fluids containing calcium pyrophosphate dihydrate (CPPD) were examined repeatedly over an eight week period to assess whether storage conditions and duration influenced the number of crystals present. Aliquots of each fluid were stored at room temperature, 4 degrees C, and -70 degrees C. At -70 degrees C there was no change in crystal count after eight weeks' storage. At room temperature and 4 degrees C crystal counts declined slowly over the eight week period, though CPPD crystals were still readily apparent after eight weeks in 10/11 (4 degrees C) and 8/11 (room temperature) fluids. No change in crystal morphology was detected and, apart from one fluid kept at room temperature in which fungal hyphae were noted at six weeks, no new crystals were seen. Calcium pyrophosphate dihydrate crystals in synovial fluid can be maintained for prolonged periods by freezing.     

The viability of Cysticercus cellulosae in Thai native food (NHAM)

The purpose of this study was to determine the viability of Cysticercus cellulosae in various forms of Thai native food (Nham). Four duplicated experiments were carried out and represent as in the average values. The results revealed that under the room temperature (27-30 degrees C), the intact C. cellulosae in Nham could survive as long as 12-18 hours after preparation while isolated C. cellulosae and C. cellulosae in a piece of 2 X 9 cm. of pork in 0.85% saline could survive as long as 60 and 66 hours, respectively. The Cysticercus in other ingredients of Nham and in saline solution (6%) could be viable for 48-96 and 12-18 hours, respectively. Under the temperature of 4 degrees C, Cysticercus in various recipes could survive for 96 hours while in the controls, it could survive as long as 20-30 days. However, C. cellulosae in various compositions of Nham could be viable 11-20 days which was longer than those in potassium nitrate or salt alone. The findings of this study might infer the suggest that optimum time for consumming Nham in raw condition with safety should be after preparation at least 2-3 days at room temperature.     

Chemical analysis of freshly prepared and stored capsaicin solutions: implications for tussigenic challenges

The purpose of this study was to assess the stability of stored capsaicin solutions and the actual concentrations of prepared solutions. Capsaicin solutions ranging in concentration from 0.5 to 128 microM were mixed and analyzed using high performance liquid chromatography. Samples of varying concentrations were then stored under 4 environmental conditions: 4 degrees C and protected from light, room temperature (RT) exposed to light, RT protected from light, and -20 degrees C and protected from light. The concentrations were measured every other month for 1 year. Actual concentrations of freshly prepared solutions were on average 88.3% of predicted. For solutions stored at 4 degrees C, there was a decrease only in the lower concentrations (0.5, 1, and 2 microM) after 2 months (P=0.003). Solutions stored at RT exposed to light decreased in concentration after 6 months (P=0.020), and solutions stored at RT protected from light decreased in concentration after 4 months (P=0.026). The group stored at -20 degrees C decreased in concentration after 1 year (P=0.033). We conclude that the actual concentration of capsaicin solution is less than predicted, and solutions of 4 microM or higher concentration are stable for 1 year if stored at 4 degrees C protected from light.     

Methodology of the H2 breath test. I. Collection and storage for gas measurement

The measurement of hydrogen and methane in expired air is widely used in the field of gastrointestinal diagnosis. Techniques as simple and as reliable as possible are therefore requested for the collection and storage of breath samples. As far as collection is concerned, we compared three systems of end-expiratory sampling: a modified Haldane-Priestley tube, a Y-piece device fitted to a plastic syringe and a commercially available two-bag system. There was a significant correlation between the results obtained with all three systems, suggesting that all are sufficiently reliable. However, the two-bag system does not require particular training on behalf of the operator or particular cooperation from the patients and also makes it possible to take samples from more than one patient at the same time. For the storage of breath samples plastic syringes are the most commonly used device. Nonetheless, at room temperature there is a leakage of hydrogen equal to 9% after 24 hours, increasing to 29% after 5 days of storage. Refrigeration of the syringes at -20 degrees C prevents any loss in the first 48 hours and limits it to 5% after 5 days. The stability of the methane was higher than that of the hydrogen: after 5 days the loss is 4% at room temperature and 2% at -20 degrees C. For both gases the losses increase significantly at a temperature of 37 degrees C and are not affected by the initial concentration of the stored gas.(ABSTRACT TRUNCATED AT 250 WORDS)     

Metabolic damage to human saphenous vein during preparation for coronary artery bypass grafting

Segments of saphenous vein from patients undergoing coronary artery by-pass graft surgery were frozen in liquid nitrogen immediately on dissection (control), after stripping of the adventitia and side branch ligation (manipulation), after distention with blood (distention), or at completion of the last proximal anastomosis (prepared vein). Vein was stored during the operation in patient's heparinised arterial blood at room temperature. Frozen vein was extracted with perchloric acid. ATP, ADP, and AMP, adenosine, inosine and hypoxanthine concentrations were measured by high pressure liquid chromatography. Prepared vein had ca 50% lower ATP concentrations and ATP/ADP ratio than control vein, higher concentrations of inosine and hypoxanthine and lower concentrations of AMP and adenosine. ATP concentration and ATP/ADP ratio did not correlate with the time elapsed between dissection and freezing of the prepared vein. The characteristic changes seen in prepared vein were not seen when control vein was simply stored in arterial blood at 23 degrees C, in normal saline at 23 degrees C or 4 degrees C, in Krebs-Ringer bicarbonate buffer at 37 degrees C or at St Thomas's Hospital cardioplegic solution at 4 degrees C. Distention with unlimited pressure did not distension at less than 300 mmHg gave rise to the same changes in ATP concentration and ATP/ADP ratio as in the prepared vein. These results show that vein suffered metabolic changes during preparation for bypass grafting and suggest that uncontrolled distention may contribute to these changes. Such biochemical measurements provide a quantitative estimate of tissue damage and allow objective comparison of different preparative techniques.     

Stability of ceftazidime in normal saline solution after exposure to light

Bactericidal activity of ceftazidime is determined by the time that concentrations in tissue and serum are above the MIC for the pathogens during the dosing interval. Thus, the most effective mode of administration of ceftazidime is continuous infusion. However, this agent is light sensitive which may result in instability when administered by this method without protection from light. Until now we have had no data to demonstrate the stability of this drug during continuous infusion. Therefore, the objective of this study was to provide such data. One gram of ceftazidime was mixed with 1,000 ml normal saline and exposed to two 36 watt fluorescence lights for 24 hours. The distance between ceftazidime solution and light source was 1 meter. Twenty samples (1 g-ceftazidime in normal saline) solution were evaluated. The mean ceftazidime concentrations in normal saline solution were decreased by only 1.69%, 4.44% and 7.19% after 6, 12 and 24 hours after exposure to light, respectively. Therefore, we conclude that the reduction of drug concentration was not considered to be significantly high, and this agent can be administered by continuous infusion.     

In vitro excystation of Haplorchis taichui (Trematoda: Heterophyidae)

The effects of trypsin, bile extract, temperature and acid-based condition for the in vitro excystation of Haplorchis taichui metacercariae were studied. At 37 degrees C, approximately half the number of metacercariae excysted when exposed to 1% trypsin for 15 minutes with no more excystation found beyond this time. Increasing trypsin concentration seemed to reduce the excystation rate while bile extract was, however, unlikely to be an absolute requirement. A temperature of 37 degrees-41 degrees C yielded a similar excystation result in combination with 1% trypsin; however, less excystation occurred at a lower temperature of 35 degrees C. The acid-based environment of pH 8 gave the best excystation result in association with 1% trypsin at a temperature of 39 degrees C. Higher and lower basicity produced a smaller excystation rate. An environmental condition of 1% trypsin at pH 8 and a temperature of between 37 degrees-41 degrees C was recommended for the in vitro excystation of H. taichui metacercariae. The relatively broad temperature and pH range condition for the excystation of H. taichui corresponded with various definitive hosts that were infected naturally by this fluke.     

Cold preservation of rat pancreatic islets just above the freezing point using University of Wisconsin solution

AIMS: To confirm whether rat islets stored at a temperature just above the freezing point using University of Wisconsin (UW) solution would remain viable for the short term. METHODOLOGY: Rat islets were stored for 24 hours in UW solution, either at 4 degrees C or at -0.6 degrees C (just above the specific freezing point of the UW solution). After cold storage, the islets were assessed for in vitro viability by static incubation and for in vivo viability by a transplantation study. One thousand islets preserved under different conditions were injected intraportally into a streptozotocin-induced diabetic rat as an isograft. Four weeks after the transplantation, an intravenous glucose tolerance test was performed. RESULTS: Islets stored at -0.6 degrees C showed higher insulin secretion rates than those stored at 4 degrees C on a static challenge. The interval from transplantation to the achievement of normoglycemia was also shorter in the -0.6 degrees C group than in the 4 degrees C group. After islet transplantation, the daily nonfasting plasma glucose concentration was higher in the 4 degrees C group than in the -0.6 degrees C group. When compared with the 4 degrees C group, the -0.6 degrees C group showed lower blood glucose values during all investigational periods on an intravenous glucose tolerance test. CONCLUSION: Islet preservation at -0.6 degrees C using UW solution is more advantageous for short term.     

Effect of cold-storage in the accumulation of bioreactive substances in platelet concentrates treated with second messenger effects.

BACKGROUND AND OBJECTIVES. The mandatory 5-day of shelf-life platelet concentrates (PCs) creates outdating and inventory control problems in blood banking. Moreover, storage of PCs at 22-24 degrees C has been associated with a time-dependent accumulation of pyrogenic cytokines, potentially harmful for recipients. Previous studies have shown that supplementation of PCs with ThromboSol, a mixture of second-messengers effectors, might allow storage of functionally active platelets at refrigerated temperature to be extended. This study further investigates this storage approach by comparing the accumulation of bioactive compounds in standard and refrigerated PCs. DESIGN AND METHODS. The PCs were supplemented with ThromboSol or a control solution and stored in parallel at 24 degrees C with continuous agitation or undisturbed at 4 degrees C. Samples were removed on days 1, 5, 9 of storage, and assayed for their content of interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-alpha, transforming growth factor (TGF)-beta1, and anaphylatoxins C3a and C4a. RESULTS. Throughout storage, refrigerated PCs, both ThromboSol-treated and untreated units, displayed a slightly lower level of IL-6 and significantly lower concentration of IL-8 than conventionally stored PCs. ThromboSol slightly reduced the level of these cytokines in PCs. Throughout storage at 22 degrees C, an accumulation of anaphylatoxins C3a and C4a was seen both in both control and ThromboSol-treated PCs. This accumulation was significantly reduced in control PCs stored at 4 degrees C, but not in refrigerated PCs supplemented with ThromboSol. Cold-storage, with or without ThromboSol, had a minor effect on the accumulation of TGF-beta1 in PCs. INTERPRETATION AND CONCLUSIONS. Our data confirm that release of bioactive compounds during in vitro storage of PCs is a temperature-sensitive process. The ThromboSol-refrigeration system could be a useful alternative for extending storage of PCs, without increasing the accumulation of cytokines (IL-6, IL-8), known to be involved in febrile reactions in recipients. Nevertheless, this storage system has no benefit on the level of other bioactive compounds (TGF-beta1, anaphylatoxins C3a and C4a) in PCs.     

Evaluation of self-taken samples for the presence of genital Chlamydia trachomatis infection in women using the ligase chain reaction assay

The aim of this study was to evaluate the sensitivity and acceptability of self-taken vulval-introital (VI) samples, first-catch urine (FCU) samples and clinician-obtained cervical samples for the presence of genital Chlamydia trachomatis infections in women using the ligase chain reaction (LCR) assay. One hundred and four patients were enrolled, of whom 54 patients had chlamydial DNA in at least one of the samples tested. The sensitivity of the cervical sample was 96.3%, vulval-introital sample in LCR buffer 92.6%, vulval-introital swab collected dry 88.9%, FCU stored at +2-8 degrees C 81.5%, FCU stored at room temperature 77.8% and FCU stored with 2% w/v boric acid at room temperature 87.0%. Self-taken vulval-introital LCR samples were shown to be an acceptable alternative to a clinician-obtained LCR sample. The addition of boric acid may overcome the need for a continuous cold chain for FCU samples.