Reversal of granulocyte adherence to nylon fibers using local anesthetic agents: possible application to filtration leukapheresis

Abstract The effects of the cationic anesthetic agents tetracaine and lidocaine on granulocyte function, morphology, and adherence to nylon fibers were studied in an attempt to improve current methods of granulocyte collection by filtration leukapheresis (FL). When dissolved in acid- citrate-dextrose (ACD) plasma, these drugs significantly increased granulocyte elution from the fibers in a dose-related fashion. Granulocytes exposed to tetracaine and lidocaine remained more than 95% viable, retained normal bactericidal capacity after the drugs were washed from the cells, and had preserved membrane integrity, as evidenced by the normal ultrastructural appearance of tetracaine- exposed cells and an absence of leakage of lysozyme or lactic dehydrogenase. Granulocytes eluted with the anesthetic agents were rounded in shape with a reduction in the number of filopodial cytoplasmic projections and a relative absence of cytoplasmic vacuolization when compared to granulocytes eluted with ACD plasma alone. Dose-related inhibition of phagocytosis and adherence, which was largely reversible after washing the granulocytes, was noted. Greater than 95% of the lidocaine could be removed from the eluate with a single centrifugation and resuspension, indicating that granulocytes prepared by FL with anesthetic-enhanced elution could be potentially transfusable.

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Reversal of granulocyte adherence to nylon fibers using local anesthetic agents: possible application to filtration leukapheresis

Blood ◽

10.1182/blood.v50.2.213.bloodjournal502213 ◽

1977 ◽

Vol 50 (2) ◽

pp. 213-225

Author(s):

CA Schiffer ◽

FT Sanel ◽

VB Young ◽

J Aisner

Keyword(s):

Local Anesthetic ◽

Membrane Integrity ◽

Lactic Dehydrogenase ◽

Anesthetic Agents ◽

Cytoplasmic Vacuolization ◽

Bactericidal Capacity ◽

Ultrastructural Appearance ◽

Acid Citrate Dextrose ◽

Citrate Dextrose ◽

Granulocyte Function

The effects of the cationic anesthetic agents tetracaine and lidocaine on granulocyte function, morphology, and adherence to nylon fibers were studied in an attempt to improve current methods of granulocyte collection by filtration leukapheresis (FL). When dissolved in acid- citrate-dextrose (ACD) plasma, these drugs significantly increased granulocyte elution from the fibers in a dose-related fashion. Granulocytes exposed to tetracaine and lidocaine remained more than 95% viable, retained normal bactericidal capacity after the drugs were washed from the cells, and had preserved membrane integrity, as evidenced by the normal ultrastructural appearance of tetracaine- exposed cells and an absence of leakage of lysozyme or lactic dehydrogenase. Granulocytes eluted with the anesthetic agents were rounded in shape with a reduction in the number of filopodial cytoplasmic projections and a relative absence of cytoplasmic vacuolization when compared to granulocytes eluted with ACD plasma alone. Dose-related inhibition of phagocytosis and adherence, which was largely reversible after washing the granulocytes, was noted. Greater than 95% of the lidocaine could be removed from the eluate with a single centrifugation and resuspension, indicating that granulocytes prepared by FL with anesthetic-enhanced elution could be potentially transfusable.

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Variations in Levels of Transaminases and Lactic Dehydrogenase in Bank Blood

Clinical Chemistry ◽

10.1093/clinchem/8.6.626 ◽

1962 ◽

Vol 8 (6) ◽

pp. 626-629 ◽

Cited By ~ 1

Author(s):

Leonard V Crowley

Keyword(s):

Blood Donors ◽

Lactic Dehydrogenase ◽

Screening Procedure ◽

Dextrose Solution ◽

Acid Citrate Dextrose ◽

Citrate Dextrose ◽

Bank Blood

Abstract Results of the assay of transaminases and lactic dehydrogenase in blood, proposed as a screening procedure for subclinical hepatitis in blood donors, show that the method of sampling influences the analytic results. Determinations of GOT and GPT may prove useful if made on specimens separated from cells soon after collection. Plasma in contact with cells from blood preserved in acid citrate dextrose solution appears to yield good results up to 21 days. Lactic-dehydrogenase levels are of limited value for this screening.

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Effects of Anticoagulants and Storage on Granulocyte Function in Bank Blood

Blood ◽

10.1182/blood.v43.2.207.207 ◽

1974 ◽

Vol 43 (2) ◽

pp. 207-217 ◽

Cited By ~ 28

Author(s):

Jeffrey McCullough ◽

Sandra J. Carter ◽

Paul G. Quie

Keyword(s):

Whole Blood ◽

Normal Function ◽

Bactericidal Assay ◽

Acid Citrate Dextrose ◽

Citrate Dextrose ◽

Granulocyte Function ◽

And Storage ◽

Citrate Phosphate ◽

Bank Blood

Abstract Storage of granulocytes for transfusion has not been practical because it has been considered that granulocyte function in bank blood is retained for only a few hours after collection. In the present study, granulocyte function was evaluated using the bactericidal assay and the quantitative nitroblue tetrazolium (NBT) method. Granulocytes from whole blood collected into acid citrate dextrose (ACD), citrate phosphate dextrose (CPD), heparin, ion exchange, and sodium citrate anticoagulants showed no functional impairment after 24 hr of storage at 4°C. With further storage, all granulocytes showed a loss of NBT activity. However, after 48 and 72 hr, granulocytes from whole blood stored in ACD and CPD killed the expected number of bacteria in the bactericidal assay. Thus, when tested in vitro, granulocytes maintain normal function, at least during the first 24 hr after collection when stored in certain anticoagulants under standard blood bank conditions.

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Conditions Affecting the Responses of Human Platelets to Epinephrine

Thrombosis and Haemostasis ◽

10.1055/s-0038-1647031 ◽

1988 ◽

Vol 60 (02) ◽

pp. 209-216 ◽

Cited By ~ 23

Author(s):

Chantal Lalau Keraly ◽

Raelene L Kinlough-Rathbone ◽

Marian A Packham ◽

Hidenori Suzuki ◽

J Fraser Mustard

Keyword(s):

Platelet Rich Plasma ◽

Shape Change ◽

Human Platelets ◽

Dense Granule ◽

Primary Phase ◽

Lag Phase ◽

Acid Citrate Dextrose ◽

Two Phases ◽

Citrate Dextrose ◽

Thromboxane Formation

SummaryConditions affecting the responses of human platelets to epinephrine were examined. In platelet-rich plasma prepared from blood anticoagulated with hirudin or PPACK (D-pheny- lalanyl-L-prolyl-L-arginine chloromethyl ketone), epinephrine did not cause shape change or aggregation. In a Tyrode-albumin- apyrase solution containing a concentration of Ca2+ in the physiological range, and fibrinogen, epinephrine in concentrations as high as 40 μM did not induce platelet shape change, caused either no primary aggregation or very slight primary aggregation, and did not induce thromboxane formation, release of dense granule contents, or secondary aggregation. In contrast, in citrated platelet-rich plasma, epinephrine induced two phases of aggregation. This is not attributable to the generation of traces of thrombin since the same effects were evident when blood was taken into a combined citrate-hirudin anticoagulant or a combined citrate-PPACK anticoagulant. In a modified Tyrode-albu- min-apyrase solution containing approximately 20 μM Ca2+, 1 mM Mg2+, and fibrinogen, epinephrine induced extensive aggregation after a lag phase, but no primary phase was evident; thromboxane formation and release of dense granule contents accompanied the aggregation response. These responses were also observed when PPACK was included with the acid-citrate- dextrose anticoagulant, and in the washing and resuspending fluids. In the presence of aspirin or the thromboxane receptor blocker BM 13.177 a few small aggregates were detected by particle counting and by scanning electron microscopy; with the latter inhibitor, the platelets in the aggregates retained their disc shape; secondary aggregation and the responses associated with it did not occur. Thus thromboxane A2 formation is not necessary for the formation of these small aggregates, but is required for extensive aggregation and release. As with other weak agonists, the close platelet-to-platelet contact in the low Ca2+ medium appears to be necessary for full secondary aggregation. Omission of fibrinogen from the low Ca2+ medium prevented both primary and secondary aggregation in response to epinephrine. An antibody (10E5) to the glycoprotein Ilb/IIIa complex was completely inhibitory in the presence of fibrinogen. Thus the response of human platelets to epinephrine is influenced by the concentration of Ca2+ and the presence of fibrinogen in the medium in which they are suspended.

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ChemInform Abstract: SEARCH FOR NEW LOCAL ANESTHETIC AGENTS, III. SYNTHESIS OF 2-ALKYLARYLAMINOACETYLAMINO-4-PHENYLTHIAZOLES

Chemischer Informationsdienst ◽

10.1002/chin.197806182 ◽

1978 ◽

Vol 9 (6) ◽

Author(s):

P. K. SRIVASTAVA ◽

R. D. SHARMA ◽

S. K. RAI

Keyword(s):

Local Anesthetic ◽

Anesthetic Agents

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RISTOMYCIN (AGGRISTIN) PRECIPITATION TEST: A NEW, RAPID AND RELIABLE METHOD FOR THE DETECTION OF INTRAVASCULAR CLOTTING

10.1055/s-0038-1643133 ◽

1987 ◽

Author(s):

G Pfliegler ◽

J Arnout ◽

J Vermylen

Keyword(s):

Reliable Method ◽

Degradation Products ◽

Laboratory Diagnosis ◽

Blood Collection ◽

Vein Thrombosis ◽

Fibrin Degradation Products ◽

Acid Citrate Dextrose ◽

Citrate Dextrose ◽

Precipitation Test

The rapid and specific detection of fibrin monomers (fm) and fibrin degradation products (fdp) is of major importance in the laboratory diagnosis of disseminated intravascular coagulation, deep vein thrombosis or pulmonary embolism. Most methods in use are either time-consuming, needing special techniques, or insensitive and poorly specific. Some time ago, Watanabe and Tullis described a simple and rapid, semiquantitative test to detect fm and fdp in plasma, based on the finding that ristocetin in low concentrations (1.0-1.5 mg/ml) can specifically precipitate fm and fdp. To 0.4 ml ACD plasma, 0.1 ml ristocetin (2.5 mg/ml) is added and vortexed. The mixture is then incubated for 30 min at 20°C and centrifuged at 50xg for 5 min. The test is considered to be positive when fibrin-like strands or small or large pellets are observed on the bottom of the tube. More recently, Pfliegler et al. reported that ristomycin (AGGRISTIN), a structural analogue of ristocetin, can replace ristocetin in this test.Here we report on further results with the ristomycin (AGGRISTIN) precipitation test in 138 patients with various intravascular thrombotic events. The results of this test, performed on ACD plasma, were compared to the serum fdp values detected by immunoelectrophoresis (IEF) and by the haemagglutina-tion inhibition test (HIT). In all 30 cases with serum fdp above 30 ug/ml (HIT) or 28 pg/ml (IEF), the precipitation test was positive; at lower fdp concentrations, as detected by HIT or IEF, the test still was positive in 70 per cent of these thrombosis patients, suggesting a superior sensitivity. In 16 patients with elevated fibrinogen levels (but no evidence of thrombosis), the test was positive in only 3. No false positive results were detected in 16 healthy controls. Preliminary results show that the minor disadvantage of the test (blood collection on acid citrate dextrose) may readily be overcome by the in vitro adjustment of the pH of citrate plasma, commonly used for other haemostatic tests, to between 7.0 and 7.4.On the basis of our results we suggest that the AGGRISTIN (ristomycin) precipitation test is a simple, rapid and reliable method for the laboratory diagnosis of intravascular clotting.

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Evaluation of Premedicants as Protective Agents Against Convulsive (LD50) Doses of Local Anesthetic Agents in Rats

Anesthesia & Analgesia ◽

10.1213/00000539-197101000-00025 ◽

1971 ◽

Vol 50 (1) ◽

pp. 127???130 ◽

Cited By ~ 5

Author(s):

J. ANTONIO ALDRETE ◽

WALTER DANIEL

Keyword(s):

Local Anesthetic ◽

Protective Agents ◽

Anesthetic Agents

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Implementation of a Simplified Regional Citrate Anticoagulation Protocol for Post-Dilution Continuous Hemofiltration Using a Bicarbonate Buffered, Calcium Containing Replacement Solution

Blood Purification ◽

10.1159/000452755 ◽

2016 ◽

Vol 42 (4) ◽

pp. 349-355 ◽

Cited By ~ 6

Author(s):

Christopher J. Kirwan ◽

Ross Hutchison ◽

Sherif Ghabina ◽

Stephanie Schwarze ◽

Abigail Beane ◽

...

Keyword(s):

Metabolic Alkalosis ◽

Calcium Supplementation ◽

Regional Citrate Anticoagulation ◽

Prospective Audit ◽

Bleeding Events ◽

Continuous Hemofiltration ◽

Citrate Anticoagulation ◽

Replacement Solution ◽

Acid Citrate Dextrose ◽

Citrate Dextrose

Background/Aims: Recent updates to the Nikkiso Aquarius continuous renal replacement therapy (CRRT) platform allowed us to develop a post-dilution protocol for regional citrate anticoagulation (RCA) using standard bicarbonate buffered, calcium containing replacement solution with acid citrate dextrose formula-A as a citrate source. Our objective was to demonstrate that the protocol was safe and effective. Methods: Prospective audit of consecutive patients receiving RCA for CRRT within intensive care unit, who were either contraindicated to heparin or had poor filter lifespan (<12 h for 2 consecutive filters) on heparin. Results: We present the first 29 patients who used 98 filters. After excluding ‘non-clot' filter loss, 50% had a duration of >27 h. Calcium supplementation was required for 30 (30%) filter circuits, in 17 of 29 (58%) patients. One patient discontinued the treatment due to metabolic alkalosis, but there were no adverse bleeding events. Conclusion: Post-dilution RCA system is effective and simple to use on the Aquarius platform and results in comparable filter life for patients relatively contraindicated to heparin.

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