In vitro analysis of shed blood from patients undergoing total knee replacement surgery.

One method used to obtain autologous blood includes collection of wound drainage postoperatively. The decision to wash wound drainage before infusion is left to individual institutions. The composition of blood collected from joint spaces has not been adequately evaluated. Wound drainage from total knee replacement was collected into a cardiotomy reservoir, without anticoagulation, for 4 hours after surgery. Coagulation parameters were evaluated on the washed supernatant and unwashed supernatant. The most significant findings were the following: 1. Modified prothrombin time: washed supernatant and unwashed supernatant were substituted for tissue thromboplastin reagent. The unwashed supernatant initiated fibrin formation (mean, 108 seconds), whereas the washed supernatant did not (mean, > 150 seconds, P = .01). 2. Euglobulin lysis times: Mixtures containing 50% normal plasma and 50% washed supernatant or unwashed supernatant were used to determine plasmin activation (unwashed supernatant + normal plasma = 24 minutes; washed supernatant + normal plasma = 106 minutes; P = .03). Lower euglobulin lysis times indicates increased plasmin activity. 3. Fibrin degradation products: Concentrations were significantly elevated in unwashed supernatant (mean 10,240 micrograms/mL) versus washed supernatant (mean 5 micrograms/mL, P = .02). Fibrin degradation products are inhibitors of fibrin formation and platelets. The authors conclude the unwashed supernatant from wound drainage collected after total knee replacement contains activated components of the soluble coagulation and fibrinolytic systems, and these substances can be significantly reduced with washing.

Blood component therapy during the neonatal period: a national survey of red cell transfusion practice, 1985.

A questionnaire to determine patterns of neonatal red cell transfusion practice during 1985 was mailed to 2200 blood banks of American Association of Blood Banks (AABB) institutional members and children's hospitals. There were 915 responses (41.6%); 785 responses (86%) contained sufficient data for analysis. The majority (70.6%) of 785 responding hospitals were community/urban institutions. However, more highly specialized, pediatric hospitals were also represented by 92 university/tertiary-care hospitals (11.7% of respondents) and 29 children's hospitals (3.7% of respondents). Two-thirds of hospitals performed a major antiglobulin crossmatch (rather than an abbreviated one) before all neonatal red cell transfusions. The red cell preparation most frequently selected for small-volume transfusions was ABO and Rh group-specific red cell concentrates. When performing only large-volume exchange transfusions, 19.2 percent of hospitals used whole blood; all others prepared reconstituted units of red cells plus fresh-frozen plasma, a practice that frequently causes exposure to two donors per unit. Another practice likely leading to multiple donor exposure is the use of fresh-frozen plasma to adjust the hematocrit of red cell preparations to a predetermined value prior to a small-volume transfusion. Over one-half of hospitals adjusting hematocrits used plasma, presumably from one donor, to dilute packed red cells from another donor, a practice that has no apparent medical benefit. Most hospitals (63.4%) provided red cells with a reduced risk of transmitting cytomegalovirus; blood from seronegative donors was selected by 65 percent of hospitals. The majority of hospitals, including most of the community/urban hospitals, did not irradiate blood products before transfusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Reference intervals for plasma lipoperoxides: age-, sex-, and specimen-related variations.

We measured lipoperoxides, as malondialdehyde (MDA), by liquid chromatography in plasma from 230 male and 148 female adult blood donors, to establish reliable reference values and to compare possible sex-, age-, and specimen-related differences. Our studies show that mean have higher MDA concentrations in plasma than do women (P less than 0.05), older men have higher values than younger men (P less than 0.05), and older women have higher values than young women (P less than 0.001). These age-related results support earlier studies in experimental animals that lipid peroxidation increases with increasing age. In addition, plasma from liquid EDTA-anti-coagulated blood has significantly lower MDA concentrations than does serum or plasma from blood treated with lithium heparin, sodium citrate, or CPDA-1 (P less than 0.001).

Erythrocyte cation transport activities as a function of cell age.

Erythrocyte cation transport systems were evaluated on cell fractions from 17 subjects. Density centrifugation was used to separate washed red cells into fractions enriched with younger and older cells; the cell age differences in these fractions were verified by reticulocyte counts (means are 3.5% for younger cell fractions and 0.7% for older cell fractions). Red cell age has a pronounced effect on several cation transport activities. The older cell fractions had increases in lithium-potassium cotransport (p less than 0.001), the rate constant for the lithium-potassium cotransport (p less than 0.001) and cellular cation permeability. The older cells had decreases in the number of ouabain binding sites (p less than 0.001), the rate constant for sodium efflux via the sodium-potassium adenosine triphosphatase pumps (p less than 0.001) and the sodium-lithium countertransport (p less than 0.025). In subjects with markedly different cell ages, these effects should be considered when evaluating red cell cation transport activities.

Membrane cascade plasmapheresis: theoretical analysis and in vitro studies.

As the donor plasma required in plasmapheresis therapy is not only expensive but may also cause side effects and transfer of infections, attempt was made in the present work to generate continuously autologous purified plasma. A membrane cascade plasmapheresis (MCP) system was developed which: 1. Separates plasma from blood using a membrane plasma separator 2. Fractionates this plasma into two fractions using a secondary filter, one fraction rich in albumin and the other rich in immunoglobulins 3. Returns to the patient filtered blood combined with albumin-rich plasma fraction Ringer solution was used to compensate for the loss of plasma volume rejected as immunoglobulins. The main problem is to maximize the clearance of IgG and recovery of albumin. Theoretical analysis of the system shows that the clearance of IgG increases with the increase in Ringer solution flow rate. However, this increase in clearance of IgG is accompanied by a decrease in albumin recovery. Theoretical analysis presents a relationship between the clearance of protein as a function of their sieving coefficients of the secondary filter, plasma separation rate, and the priming volume. In vitro evaluation of the system using patient's plasma and a Kuraray 2A filter (secondary filter) show that the relative clearances of IgG and albumin lie in the range of 65-70% and 40-45%, respectively. Further improvement in the selectivity of the secondary filter is necessary to minimize the clearance of albumin and to increase the clearance of IgG.

Exchange transfusion as an alternative to granulocyte concentrate administration in neonates with bacterial sepsis and profound neutropenia.

A high rate of mortality is observed in the subset of neonates with early-onset bacterial sepsis who develop profound neutropenia and depletion of the marrow neutrophil reserve. Several studies suggest that transfusion of apheresis-collected adult neutrophils into such subjects can be beneficial, yet inherent delays in preparation of neutrophil concentrates can limit their clinical utility. In six neonates with sepsis and profound neutropenia, we performed double-volume exchange transfusions using freshly drawn donor blood and determined that a net gain of 0.3-0.7 X 10(9) neutrophils/kg body weight had occurred. The blood neutrophil count of the patients increased from 220 to 1,660/mm3 before the transfusion to 1,650-5,140/mm3 immediately afterward. We next attempted to further increase the number of neutrophils delivered by exchange transfusion by subjecting blood donors to 2 min of physical exercise in order to induce neutrophilia. After such exercise, 30% (range 18-55%) more neutrophils were harvested. The metabolic burst activity and in vitro migration of neutrophils after exercise did not differ from paired preexercise values.

Use of whole blood exchange transfusion to supply neutrophils to septic, neutropenic neonates.

When neutropenia due to exhaustion of the marrow neutrophil reserve, develops in a neonate with bacterial sepsis the likelihood of survival is very small. We report such a case who was treated with a double-volume exchange transfusion using fresh unstored whole blood. We were able to determine a net gain of 5 x 10(8) neutrophils per kg. Then, in neutropenic neonatal animals, neutrophil transfusion by double-volume exchange transfusion with unstored blood was investigated.

Granulocyte transfusions in neonates with bacterial infection, neutropenia, and depletion of mature marrow neutrophils.

During a three-year period, 26 neonates with bacterial infection and neutropenia were studied. In order to assess the marrow neutrophil reserves, bone marrow aspirates were obtained from each of these patients. The neutrophil storage pool (percent polymorphonuclear + band neutrophils + metamyelocytes in 1,000 nucleated marrow cells) was significantly greater in those who survived their infection (mean = 20.1%, range 3.2% to 60.8%) than in those who died (mean = 1.9%, range 0.4% to 5.2%, P less than .002). In an attempt to improve survival in this group, seven neutrophil-depleted patients with sepsis were given granulocyte transfusions and all survived. In contrast only one of nine nontransfused and all survived. In contrast only one of nine nontransfused, neutrophil-depleted infants with sepsis survived (P less than .01). The seven granulocyte recipients were examined for possible adverse effects of the transfusions and none were detected. It is suggested that infected, neutropenic neonates with depletion of mature marrow neutrophils are at high risk for death from sepsis, and that these infants may benefit from granulocyte transfusion therapy.

Donor-leukocyte imaging in granulocytopenic patients with suspected abscesses: concise communication.

Indium-111-labeled donor leukocytes were used for the detection of foci of suppuration in eight severely leukopenic patients with marrow suppression, either idiopathic or associated with chemotherapeutic regimens for leukemia. In three patients good correlation was found between the results of imaging and clinical signs or subsequent proof of inflammation. In the other five patients, in whom no evidence of localized suppuration occurred, no abnormal accumulations of radioactivity were demonstrable. Labeled donor leukocytes provide a method for locating suppurative foci in severely leukopenic patients in whom autologous leukocyte labeling is impractical.

Effect of plasma exchange and immunosuppression on Rho(D), viral, and bacterial antibody titers in Rh-immunized women.

Pharmacologic immunosuppression and plasmapheresis have both been advocated as adjunctive methods of treatment for Rh immunization, but a combined regimen of the two has not been attempted. We treated four nonpregnant, severely Rh-sensitized female volunteers with intensive plasma exchange and either promethazine 150 mg or prednisone 60 mg per day in an attempt to remove circulating anti-Rho(D) and prevent further synthesis of the antibody. Each patient received three or four exchanges. The Rho(D) antibody titer decreased by at least one dilution immediately following 11 of 14 plasma exchanges and was ultimately lowered at least two dilutions in all patients. In one case the titer was reduced from 1:512 to 1:16, and a low titer was maintained for the duration of treatment. However, this regimen could increase the risk of infection for a mother and/or infant, as evidenced by the concomitant lowering of viral and bacterial antibody titers in these women.

Review: deficiencies in the neutrophil system of newborn infants, and the use of leukocyte transfusions in the treatment of neonatal sepsis.

Bacterial sepsis is a major cause of morbidity and mortality in the neonatal period. Deficiencies in neutrophil chemotaxis, phagocytosis, and bacterial killing have all been proposed as factors possibly responsible for this problem. In addition, the neutrophil storage pool, or all segmented neutrophils, band neutrophils, and metamyelocytes/kg body weight stored within the marrow, appears to be very small in neonates compared to that in adults, and when an infection develops in a neonate, neutrophil production from stem cells may be limited. In this paper we review the evidence for these neonatal neutrophil deficiencies and examine the animal and clinical studies which have tested leukocyte transfusion as a possible adjunctive therapeutic modality for sepsis neonatorum.

Childhood chronic relapsing polyneuropathy: dramatic improvement following plasmapheresis.

We present clinical data on two boys with chronic relapsing polyneuropathy. Their recurrent episodes of weakness had produced marked disability which was unresponsive to continuous prednisone therapy. Plasmapheresis produced dramatic improvement in muscle strength and functional ability. The remissions induced have been sustained despite withdrawal of steroid therapy.

Isozymes of aldolase.

A new method of starch-gel electrophoresis using a technique of staining dependent on enzyme activity has been employed to demonstrate the isozymes of aldolase from a variety of human, rat, and frog tissues. Five of these isozymes were detected in man, seven in the rat, and at least four in the frog. The abilities of these isozymes to cleave fructose-1 ,6-diphosphate and fructose-1-phosphate were compared.