Histamine levels in stored platelet concentrates. Relationship to white cell content

The histamine levels in samples from platelet concentrates (PC) were measured at various storage times by a radioenzymatic assay. Elevated histamine levels were detected in 5 of 14 PC after 3 days of storage (range, less than 1 to 13.3 ng/ml) and in 9 of 14 PC after 5 days (range, less than 1 to 22.2 ng/ml). A very good linear correlation (r = 0.913) was found between the initial white cell content of the PC and the histamine level at 5 days of storage. The rise in histamine content was not influenced by the type of plastic container. The results indicate a process of histamine release by the white cells during storage. Although histamine is metabolized rapidly in vivo, a critical histamine threshold could be reached in man by the rapid infusion of stored PC containing high levels of histamine. This could explain some unexpected transfusion reactions in patients receiving PC.     

Treatment of normal individuals with granulocyte-colony-stimulating factor: donor experiences and the effects on peripheral blood CD34+ cell counts and on the collection of peripheral blood stem cells

BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) has been used in patients to increase the level of circulating hematopoietic progenitors. Although G-CSF has been administered to some healthy individuals, the kinetics of mobilization of peripheral blood stem cells (PBSCs), the optimum dose schedule and the incidence and nature of adverse reactions in normal individuals are not completely defined. STUDY DESIGN AND METHODS: Normal individuals (n = 102) who received G- CSF for 5 or 10 days at doses of 2, 5, 7.5, or 10 micrograms per kg per day were studied. The subjects were observed for symptoms and physical changes, and blood samples were obtained for a variety of laboratory tests. After 5 or 10 days of G-CSF treatment, PBSCs were collected by apheresis and analyzed. RESULTS: Overall, 89 percent of the individuals completed the 5-day treatment protocol and 88 percent completed the 10- day protocol without modification of the dose of G-CSF administered. Ninety percent of donors experienced some side effect of G-CSF. The most frequent effects noted were bone pain (83%), headache (39%), body aches (23%), fatigue (14%), and nausea and/or vomiting (12%). The dose of G-CSF administered directly affected the proportion of people with bone pain (p = 0.025) or body aches (p = 0.045) or who were feeling hot or having night sweats (p = 0.02) or taking analgesics (p = 0.01). With the 5-day dose schedule, several changes in serum chemistries occurred, including increases in alkaline phosphatase (p = 0.001), alanine aminotransferase (p = 0.0013), lactate dehydrogenase (p = 0.0001), and sodium (p = 0.0001). Decreases occurred in glucose (p = 0.045), potassium (p = 0.0004), bilirubin (p = 0.001), and blood urea nitrogen (p = 0.0017). In donors who received G-CSF for 5 days, the absolute neutrophil count was increased after one G-CSF dose, and it reached a maximum on Day 6, as did the number of CD34+ cells (64.6 +/? 55.9 × 10(6) cells/L). In those same donors, the platelet count after apheresis on Day 6 was 32 +/? 13 percent lower than pretreatment values (250 +/? 42 × 10(9) cells/L). In donors receiving G-CSF for 10 days, the neutrophil count reached a maximum on Day 8, but the number of CD34+ cells peaked on Day 6 (58.3 +/? 52.1 × 10(5) cells/L) and then declined. The platelet count decreased from pretreatment values by 28 +/? 12 percent prior to apheresis on Day 11. When individuals were treated for 5 days with G-CSF, the quantity of CD34+ cells collected was directly related to the G-CSF dose. When 5 micrograms per kg per day was given, 2.80 +/? 1.81 × 10(8) cells were collected, compared with collection of 4.67 +/? 3.11 × 10(8) cells when 10 micrograms per kg per day was given (p = 0.04). More important, PBSCs collected after 10 days of G-CSF administration (5 micrograms/kg/day) had significantly fewer CD34+ cells (0.82 +/? 0.37 × 10(8) cells, p = 0.01) than did PBSCs collected after 5 days of G-CSF (5 micrograms/kg/day). CONCLUSION: Most normal donors receiving G-CSF experience side effects, but these are mild to moderate in degree. Some alterations in blood chemistries occur, but none were clinically serious. Because of the symptoms associated with G-CSF, these individuals must be monitored closely. The treatment of normal donors with G-CSF for more than 5 days significantly decreased the number of circulating CD34+ cells and the quantity collected by apheresis.     

Model for the physics and physiology of fluid administration

This article describes a model designed to provide an understanding of fluid flow in intravenous systems and human subjects. Experiments were developed which demonstrate that the model can represent common clinical situations. The model depicts physical devices as ideal resistors, pressure sources, and flow sources. The patient's venous system is depicted as a combination of ordinary and Starling resistors. For flows between 0 and 300 ml/hr, both physical devices and patients are adequately represented by a straight line representing the pressure-flow relationship (PFR): pressure = opening pressure + flow × resistance, where the slope is the resistance to fluid flow and the intercept is the opening pressure. The PFR for a normal vein is characterized by a flat slope (vein resistance =22±20 mm Hg/L/hr, mean ± SD) and a low intercept (opening pressure =15±8 mm Hg). The PFR for a partially obstructed vein has a resistance equal to that of an unobstructed vein and an opening pressure elevated approximately equal to the pressure obstructing the vein. For perivascular tissue, the PFR has a steep slope (tissue resistance =1,125±1,376 mm Hg/L/hr), while tissue opening pressure depends on the amount of fluid infused. At the onset of fluid extravasation (infiltration), tissue pressure usually is lower than venous pressure (8±8 versus 15±8 mm Hg), until fluid fills the distensible tissue compartment. In clinical practice, when infiltration or obstruction occurs, flow decreases and the clinician adjusts the roller clamp until correct flow resumes; no problem is obvious. The combined model for the intravenous tubing and venous systems explains the behavior of current clinical infusion devices.Presented in part at the Sixth Medical Monitoring Technology Conference, Vail, CO, March 1986; at the Annual Meeting of the American Society of Anesthesiologists, Las Vegas, NV, October 1986; at the Seventh Medical Monitoring Technology Conference, Vail, CO, March 1987; at a meeting on Computers in Critical Care and Pulmonary Medicine, San Diego, CA, June 1987; at the Annual Meeting of the American Society of Anesthesiologists, Atlanta, GA, October 1987; at the Regional Meeting of the Association for the Advancement of Medical Instrumentation, Cincinnati, OH, October 1987; at the Institute of Electrical and Electronics Engineers Ninth Annual Conference of the Engineering in Medicine and Biology Society, Boston, MA, November 1987; and at a meeting of the American Society of Hospital Pharmacists, Atlanta, GA, December 1987.Supported in part by grants from IVAC Corp.The author thanks the following individuals for important intellectual and/or technical assistance: Peter Basser, PhD, Avital Cnaan, PhD, Adriane Concus, MD, John Fox, MD, David Gissen, MD, David Joseph, MD, Anne Kamara, David Leith, MD, Leonard Lind, MD, Richard Morris, MB, BS, Barbara Orlowitz, MEE, Mary Anne Palleiko, RN, Beverly Philip, MD, Daniel Raemer, PhD, David Scott, MB, BS, John Stelling, MPH, and Marie vanRensberg, MB. At IVAC Corp: Walter Bochenko, BSEE, MBA, Robert Butterfield, BSEE, Douglas Christian, RPh, MBA, Alan Davison, BS, David Doan, PhD, Alan Somerville, BSEE, MS, Robin Wernick, BSEE, MS.     

Autoantibodies against bactericidal/permeability-increasing protein in patients with cystic fibrosis

Cystic fibrosis (CF), a genetic disorder, is characterized by chronic pulmonary infection/inflammation which leads to respiratory failure. The presence of anti-neutrophil cytoplasmic autoantibodies (ANCA) has previously been observed in the sera of patients with CF. In view of the known relationship of ANCA with primary vasculitis and of their putative pathogenetic role in these disorders, we studied the presence, specificity and isotype of ANCA and their clinical associations in 66 adult CF patients. None of the 66 CF samples had autoantibodies to the major ANCA antigens, proteinase 3 or myeloperoxidase. However, 60/66 (91%) CF samples contained IgG and 55/66 (83%) IgA, autoantibodies to bactericidal/permeability increasing protein (BPI), a recently characterized ANCA specificity. All the IgA anti-BPI-positive samples were also IgG anti-BPI-positive. The autoantibody specificity was confirmed by inhibition assay and immunoblotting of CF sera against a neutrophil granule preparation. Furthermore, in this cross-sectional study, anti-BPI levels were inversely correlated with the observed reductions in FEV1 and FVC (IgA anti-BPI and FEV1: r = 0.508, <it>p</it> &lt; 0.0001), and both IgG and IgA anti-BPI levels were higher in CF patients with secondary vasculitis (<it>n</it> = 6) than in those without (<it>p</it> &lt; 0.05). ANCA with specificity for BPI were present in the majority of CF sera in this study and autoimmune processes may be associated with the development of pulmonary injury in CF.      

Novel polymorphisms in mec genes and a new mec complex type in methicillin-resistant Staphylococcus aureus isolates obtained in rural Wisconsin

We determined allelic polymorphisms in the mec complexes of 524 methicillin-resistant Staphylococcus aureus isolates by partial or complete sequencing of three mec genes, mecA, mecI, and mecR1. The isolates had been collected over a 10-year period from patients living in rural Wisconsin, where the use of antibiotics was expected to be lower than in the bigger cities. Of the 18 mutation types identified, 16 had not been described previously. The five most common mutations were a mutation 7 nucleotides (nt) upstream from the start site (G-->T) in the mecA promoter (34.7%), an E246G change encoded by mecA (2.2%), a cytosine insertion at codon 257 in mecA (13.2%), an N121K change encoded by mecI (7.8%), and a major mecI-mecR1 deletion designated as a class B1 mec complex deletion type (25.4%). There was a high degree of conservation of the amino acid sequence of MecR1. Strains with the same mutations had variable resistance to oxacillin, and the median MIC for isolates that harbored the 7-nt-upstream mutation was lower than that for strains harboring other mutations. Our data suggest that the mecA promoter mutation plays a more important role in determining methicillin resistance than mutations in mecI and mecA do. Eighty-five percent of the tested isolates (n = 148) with the mecA promoter mutation and the class B1 mec complex deletion belonged to the same major clonal group, identified as MCG-2, and harbored the type IV staphylococcal cassette chromosome mec DNA. There was also a wide range of oxacillin MICs for strains with wild-type mecA, mecI, and mecR1 sequences, suggesting that the genetic backgrounds of clinical strains are significant in determining susceptibility to methicillin.     

A specific protocol of autologous bone marrow concentrate and platelet products versus exercise therapy for symptomatic knee osteoarthritis: a randomized controlled trial with 2 year follow-up

Background

Cell-based therapies have shown promise for the treatment of knee osteoarthritis (OA). The current study compared exercise therapy to autologous bone marrow concentrate (BMC) and platelet products for knee OA treatment.

Methods

Patients with symptomatic knee OA (N?=?48) were randomized into either an exercise therapy control group or treatment group with injection of autologous BMC and platelet products. Patients in the control group could crossover to BMC treatment after 3 months. Clinical outcomes were documented at baseline and at 6-weeks, 3, 6, 12 and 24 months, including the Knee Society Score (KSS), Pain Visual Analogue Scale, Short Form-12 Scales (SF-12), and Lower Extremity Activity Scale (LEAS).

Results

All patients in the exercise group crossed over to receive BMC treatment after 3 months (N?=?22 crossover). At 3 months, KSS-knee, SF-12 Physical, and LEAS improved significantly in the crossover group compared to exercise, similar to significant improvements on KSS-knee and LEAS for the treatment group (N?=?26) compared to exercise group at 3 months. After BMC treatment, patients’ clinical outcome scores (except SF-12 Mental Health), were significantly improved through the 2-year follow-up compared to baseline. No serious adverse events were reported.

Conclusion

The use of image-guided percutaneous BMC with platelet products yielded better results than exercise therapy as an effective alternative therapy for patients with symptomatic moderate to moderate-severe osteoarthritis of the knee.Trial registration NCT02034032. https://clinicaltrials.gov/ct2/show/NCT02034032. Registered 13 January 2014

     

Direct quantitation of IgG subclasses 1, 2, and 3 bound to red cells by Rh1 (D) antibodies

The authors developed quantitative radioimmunoassays to allow direct measurement of total human IgG and individual IgG subclasses among antibodies bound to cell surfaces. The assays use four mouse monoclonal radioiodinated antibodies, one that reacts equally well with all four human IgG subclasses and three that are specific for human IgG subclasses 1, 2, or 3. The assays were used to analyze IgG subclass composition in 21 high-titer anti-D samples from Rh-negative volunteers immunized for Rh immunoglobulin production. Anti-D activity was restricted primarily to the IgG1 and IgG3 subclasses. Eleven of 21 sera demonstrated red cell antibodies with a marked predominance of IgG1 (87 +/- 3.6% of total IgG antibody, +/- SEM) and low levels of IgG3 (1.4 +/- 0.73%). In the remaining 10 sera, IgG3 made up a greater proportion of total IgG antibody (32 +/- 3.8%), although IgG1 was still predominant (61 +/- 4.1%). This observed dichotomy in the IgG subclass profiles of different anti-D sera may be a consideration in the selection of anti-D sera for the production of the immunoglobulin used in the prophylaxis of Rh-incompatible pregnancies.     

The effect of cuff pressure deflation rate on accuracy in indirect measurement of blood pressure with the auscultatory method

The importance of cuff deflation rate in the auscultatory method of measuring blood pressure was investigated using a computer-based model. To determine the relationship between the cuff deflation rate and the measurement error, two cuff deflation protocols were used, one based on heart rate (mm Hg per heartbeat), the other on a constant rate (mm Hg per second). The different deflation protocols and rates were tested using a constant blood pressure of 120/80 mm Hg and heart rates ranging from 40 to 120 beats/min. It was confirmed that a cuff deflation rate that is time based will introduce larger errors at low heart rates. Using heart rate as a basis for cuff deflation rate yields a constant error that is independent of heart rate. The currently used standard of 3 mm Hg/s could result in a maximum error of 2.5 mm Hg in both systolic and diastolic pressures at a heart rate of 72 beats/min. The maximum systolic and diastolic errors increase to more than 4 mm Hg at 40 beats/min. A deflation rate of 2 mm Hg/beat, however, yields a maximum error of 2 mm Hg for both systolic and diastolic pressures, independent of heart rate. A cuff deflation rate based on heart rate is recommended to help minimize changes in measurement error when measuring blood pressure if a wide range of heart rates will be encountered.Supported by grants from IVAC, San Diego, CA, and Physio Control, Redmond, WA.     

Factors influencing capnography in the bain circuit

The Bain circuit provides continuous fresh gas flow near the airway. The potential mixing of this fresh gas with expired gas may prevent reliable analysis of expired gas. We therefore investigated the interaction of sampling site, fresh gas flow rate, expiratory flow rate, and sampling flow rate on expiratory capnography. Sampling near the fresh gas outlet yielded inaccurate results under several of these conditions. The magnitude of the error was related to the fresh gas and expiratory flow rates. A reliable sampling region near the endotracheal tube was identified.     

Transfusion-related acute lung injury caused by an NB2 granulocyte- specific antibody in a patient with thrombotic thrombocytopenic purpura

HLA and granulocyte-specific antibodies have been implicated in the production of transfusion-related acute lung injury (TRALI). Reported here is a case that suggests that the patient's preexisting condition may play an important role in determining whether TRALI develops upon transfusion of blood products containing anti-white cell (WBC) antibodies. A 29-year-old woman with thrombotic thrombocytopenic purpura (TTP) underwent an uneventful 1.5-volume plasma exchange, which was followed by the transfusion of 2 red cell (RBC) units. At the end of the second RBC transfusion, the patient developed clinical signs and symptoms of noncardiogenic pulmonary edema. Serologic studies demonstrated that the serum from the second RBC donor had no HLA antibodies but did have a granulocyte-specific antibody (anti-NB2) that caused the agglutination of the recipient's granulocytes, which were NB2 positive. Serum from the donor of the first RBC unit and serum from the donors of units used in the exchange had no HLA or granulocyte-specific antibodies that reacted with the recipient's WBCs. Because the donor implicated in this reaction had a history of 21 blood donations, none of which had been associated with a transfusion reaction, we suggest that the patient's preexisting condition played a significant role in this episode of TRALI, owing to the granulocyte-specific antibody.