Effect of storage conditions on function of granulocytes.
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Biomedical subjects
Publications and source records attributed to J McCullough.
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A 6-month-old girl developed severe, but transient neutropenia due to an antibody against a neutrophil-specific antigen, NA1. The mean absolute neutrophil count was 380/microliter for 2 months and spontaneously returned to normal as the anti-NA1 antibody disappeared. After recovery, the NA1 antigen was demonstrated on the patient's neutrophils. No etiology was found. There was no therapy directed at suppressing the antibody or increasing the neutrophil count. Life-threatening infection did not occur during the neutropenia.
We report the results of a new test, indium oxine in 111 scanning, in the diagnosis of postoperative infection. Indium 111 was used to label autologous polymorphonuclear leukocytes, which when reinjected migrate to sites of infection, inflammation, or both. Standard scintigraphy localizes the labeled inflammatory cells at these sites. Sixty-six scans were performed in 43 surgical patients. Thirty-seven scans were categorized as true-positive; 19 scans were categorized as true-negative. Therefore, the accuracy rate was 85%. Two scans (3%) in one patient represented false-positive results. Two scans (3%) were positive for inflammation but there was no infection present; this group was denoted as equivocal. Six scans (9%) were false-negative; false-negative scans are more likely in old lesions with poor blood supply and in areas that overlap regions of normal uptake. The noninvasive nature of the test, high accuracy rate, and ease of administration make it a potentially useful tool in the diagnosis of postoperative infection.
Specific binding of [3H] PGE2 and [3H] PGF2 alpha to smooth endoplasmic reticulum (SER) membranes prepared from rat skin at various stages of the development of essential fatty acid (EFA)-deficiency syndrome was demonstrated in these studies. The normal SER-binding capacity for [3H] PGE2 per mg of membrane protein was approximately two-fold that for [3H] PGF2 alpha in control rats fed a diet rich in essential fatty acids (EFA). Two weeks following the feeding of the animals with the EFA-deficient diet: histological evaluation revealed hyperplasia and acanthosis of the epidermal layer; a two-fold increase in labeling index (a marker for DNA synthesis) and no alteration in the SER-binding capacity for [3H] PGE2 or [3H] PGF2 alpha. By the 12th week however, labeling index had increased six-fold, accompanied by a five-fold increase in SER-binding capacity for [3H] PGF2 alpha. SER-binding for [3H] PGE2 was relatively unaltered. These results have demonstrated a relationship between marked alteration of skin SER-binding capacity for PGF2 alpha and the increase in epidermal hyperplasia and proliferation characteristic of the skin of the EFA-deficient rat. Refeeding of the animals with EFA-rich diet restored the epidermal proliferation and the SER-binding capacity to PGF2 alpha to normal conditions.
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Saccharomyces kluyveri is a heterothallic yeast with two allelic mating types denoted as a-k and alpha-k by analogy with Saccharomyces cerevisiae and from the work described here. S. kluyveri produces mating pheromones analogous to those of S. cerevisiae, but which appear to have different specificity. S. kluyveri thus differs from S. cerevisiae, Hansenula wingei, and Schizosaccharomyces pombe in that it exhibits both strong constitutive agglutination and mating pheromones. alpha-k cells produce a pheromone ("alpha-k-factor") which causes a-k cells to arrest in the G1 phase of the cell cycle and to undergo a morphological change. After a period of time dependent on the concentration of alpha-k-factor, cells exposed to the factor resume cell division. alpha-k-factor has no effect on a-k/alpha-k diploids or on alpha-k cells, but at high concentration does induce G1 arrest of S. cerevisiaea cells (a-c). a-k cells produce a pheromone ("a-k-factor") which causes alpha-k cells to exhibit a morphological change. In addition, a-k cells exhibit the Bar phenotype with respect to alpha-k-factor. Partially purified preparations of S. cerevisiae alpha-factor are more active in inducing G1 arrest of a-k cells than of a-c cells. A more purified preparation of alpha-c-factor is less active against a-k cells than a-c cells, suggesting that an additional factor (KRE, kluyveri response enhancer) may be lost during purification. Attempts to mate S. kluyveri and S. cerevisiae cells by prototroph selection and by cell-to-cell mating have been unsuccessful with all combinations of mating types. Thus, S. cerevisiae and S. kluyveri are incompatible for mating even though their pheromones exhibit some physiological cross-reaction.
Granulocyte transfusion is becoming widely used in the treatment of infections in granulocytopenic patients. Several techniques are available for granulocyte collection. Some involve centrifugation of the whole blood and one removes granulocytes from whole blood by reversible adhesion to nylon fibers. The risks to the donor from leukapheresis do not appear to be greater than from whole blood donation. Granulocytes collected by centrifuge techniques function normally in vitro and have normal intravascular recovery and disappearance following transfusion. Granulocytes collected by filtration leukapheresis function almost normally in vitro but have a reduced intravascular recovery and abnormal kinetics as they leave the circulation. The role of leukocyte typing and compatibility testing for granulocyte transfusion is controversial. When the recipient has circulating antibody against donor leukocytes, transfused leukocytes do not circulate or migrate to sites of infection but are sequestered in the liver and spleen. Clinical studies have not defined whether patients benefit equally well clinically from transfusion of compatible or incompatible granulocytes. Initial reports of clinical trials of granulocyte transfusion were promising. However, similar patients who did not receive granulocytes were not studied. Most subsequent controlled trials showed a clear benefit from granulocyte transfusion while others did not. Differences in antibiotic therapy, chemotherapy, use of laminar flow rooms, and grouping of patients make it difficult to compare these clinical trials. Some, but not all, infected granulocytopenic patients benefit from transfusion. Granulocyte transfusions improve survival of granulocytopenic patients with gram negative sepsis and prolonged bone marrow aplasia. Studies are now attempting to identify other patients who should receive granulocytes, the optimum dose and schedule of transfusions, the optimum time to begin transfusion, and the value, if any, of prophylactic transfusions.
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Studies of the in vivo kinetics of granulocytes labeled in vitro with indium-111 were carried out in 10 normal subjects. The granulocyte suspension was prepared with a Ficoll-Hypaque density gradient and labeled with indium-111-oxine. No elution or reutilization of the radioisotope was demonstrable in vitro. The average intravascular recovery of indium-labeled granulocytes was 30% +/- 6, and the t 1/2 was 5.0 +/- 1.6 hr. Normal in vitro function of these cells was demonstrated by bacterial killing and chemotaxis assays. Because indium-111 decays by gamma emission, the fate of in vivo labeled granulocytes can be followed with scintigraphic techniques. Images obtained indicated normal uptake of activity in the liver and spleen. Effective in vivo function of indium-labeled granulocytes was demonstrated in four patients by the localization of radioactivity at sites of inflammation or abscess. Although the intravascular recovery with this method is lower than that reported for some other radioisotope methods, the comparison of abnormal findings with normal values by this technique is probably valid. Indium-labeled granulocytes should prove useful in the study of granulocyte collection, transfusion, histocompatibility, and storage.
Zoster immune plasma (ZIP) was evaluated for treatment of cutaneous disseminated zoster in immunocompromised hosts. Twenty patients were studied: 13 were enrolled in a double-blind protocol, five received ZIP under an open protocol, and two were observed without receiving a transfusion. In the double-blind study, eight patients actually received ZIP; five were given plasma lacking varicella-zoster virus antibodies (control plasma). The clinical course of zoster in the group given ZIP was the same as that of patients given control plasma or no transfusions. Because ZIP did not alter the clinical course of zoster and because zoster patients produced high-antibody titers without ZIP, we concluded that ZIP is not useful for treatment of cutaneous disseminated zoster and should be reserved for prevention or modification of varicella in exposed, susceptible immunocompromised patients.
First heart sound (S1) energy spectra in isovolumic systole, hemodynamics, and angiographic left ventricular wall motion (LVWM) at rest and with atrial pacing were compared in 27 patients who underwent diagnostic cardiac catheterization and angiography because of chest pain. Eighteen patients were found to have coronary artery disease (CAD) and nine patients, normal coronary arteries. Eleven of the 18 CAD patients (61%) had a mean reduction in the spectral energy of S1 of 6.5 +/- 1.4 (SEM) dB below control (-52%), during interruption of ischemic stress of rapid atrial pacing, compared to only one of nine patients without CAD (P less than 0.05). Only five CAD patients (28%) had an abnormal rise (greater than or equal to 5 mm) in left ventricular end-diastolic pressure (LVEDP) either during or upon interruption of pacing, and six (33%) had ischemic ST-segment depression greater than or equal to mv in the ECG. Similarly two patients free of CAD (22%) had an abnormal increase in LVEDP, and none had ECG evidence of ischemia. Seventeen CAD patients (94%) had segmental LVWM abnormalities at rest or with interruption of pacing, while three patients with normal coronary arteries (33%) had abnormal angiographic LVWM (P less than 0.01). Thus, reduction is S1 spectral energy is a common accompaniment of myocardial ischemia. In the present study, it was more frequently observed than abnormalities in either the ECG or LVEDP, but was not was consistently seen as segmental left ventricular wall motion abnormalities.
Granulocytes for transfusion can now be obtained from normal donors by one of four techniques that involve either centrifugation or reversible adhesion of granulocytes to nylon fibers. The leukapheresis process appears to be safe for donors and standards for the selection and care of donors are being formulated. It appears desirable to transfuse granulocytes that are compatible in a leukoagglutination crossmatching, however, better methods for histocompatibility testing must be developed. Granulocyte transfusions clearly are of benefit to patients with Gram-negative sepsis and granulocyte counts of less than 500/cu mm for at least ten days. They may be valuable for granulocytopenic patients with other severe infections; however, there is no indication that granulocyte transfusions are indicated prophylactically or for febrile granulocytopenic patients without evidence of infection.
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With the increasing use of granulocyte transfusion it is becoming important to determine if granulocytes can be preserved for a few days. If so, the optimum storage conditions must be identified. We studied the function in vitro of granulocytes collected as they would be for transfusion by continuous-flow centrifuge leukapheresis (CFCL) and filtration leukapheresis (FL). Granulocytes collected by CFCL maintained normal ability to phagocytose and kill bacteria after 48 hr and normal chemotaxis after 24 hr of storage at 20 degrees--24 degrees C. Neither 1 degrees--6 degrees C nor 37 degrees C were as effective in preserving chemotactic response. Agitation of the granulocyte suspension during storage caused reduced bacterial killing and chemotaxis. Granulocytes collected by FL functioned very poorly after 24 hr storage at all temperatures studied. These studies suggest that it may be possible to store CFCL granulocytes at 20 degrees--24 degrees C for 24 hr. FL granulocytes should not be stored at all.
Zoster immune plasma (ZIP) was given to 31 susceptible immunocompromised children one to seven days (median, two days) following household, playmate, or hospital exposures to varicella. The average amount of ZIP transfused was 7 ml/kg. Twenty-one children did not develop varicella or persistent antibodies to varicella-zoster virus (VZV). Eight (26%) of the 31 contracted clinical varicella. Seven cases were mild, but in one child, who was given ZIP seven days after exposure, visceral disease developed and the child died. Two children had subclinical varicella that was documented by persistence of VZV antibodies for at least ten months after passive immunization. Because none of the 30 children given ZIP one to six days following exposure had severe varicella, we conclude that ZIP is effective in preventing or modifying varicella in immunocompromised patients if given shortly after exposure.
Transfusion of massive amounts of citrate anticoagulated blood during plateletpheresis with the intermittent flow centrifuge can produce symptoms and electrocardiographic changes suggestive of hypocalcemia. Following 15 procedures the ionized calcium decreased by an average of 32.4%, the average postpheresis serum citrate was 26.7 mg/dl and the QT interval was prolonged by 0.08 sec. Twelve plateletphereses performed with half-strength ACD solution caused an average decrease in ionized calcium of 16%, serum citrate levels of 12.5 mg/dl and QT prolongation of 0.04 sec. No donors experienced significant clinical symptoms with citrate infusion rates of less than 65 mg/kg/h. Solutions with citrate concentrations lower than ACD-A should be developed for use in plateletpheresis procedures involving citrate infusion rates greater than this.