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Biomedical subjects

C R Valeri

Publications and source records attributed to C R Valeri.

At least 199 records · Page 11Linked to original sources
PubMed1985

Thromboxane A2 moderates permeability after limb ischemia.

Reperfusion after limb ischemia results in muscle edema as well as excess secretion of thromboxane A2 (TxA2), an agent associated with permeability increase in other settings. This study tests whether TxA2 moderates the permeability following limb ischemia. A tourniquet inflated to 300 mmHg was applied for 2 hours around the hind limb of four groups of dogs. In untreated animals (N = 25), 2 hours following tourniquet release, plasma TxB2 values rose from 320 pg/ml to 2416 pg/ml (p less than 0.001), and popliteal lymph values rose from 378 pg/ml to 1046 pg/ml (p less than 0.001). Platelet TxB2 was unaltered and plasma 6-keto-PGF1 alpha levels did not vary. Following ischemia, lymph flow (QL) increased from 0.07 to 0.37 ml/h (p less than 0.05), while the lymph/plasma (L/P) protein ratio was unchanged at 0.41. These measurements indicate increased permeability since increase in hydrostatic pressure in a second group by tourniquet inflation to 50 mmHg (N = 7) led to a rise in QL from 0.07 to 0.22 ml/h, but a fall in the L/P ratio to 0.32, a value lower than the ischemic group (p less than 0.05). Pretreatment with the imidazole derivative ketoconazole (N = 11) reduced platelet Tx synthesis from 42 ng to 2 ng/10(9) platelets, but lymph TxB2 levels rose to 1703 pg/ml after ischemia, indicating an extravascular or vessel wall site of synthesis not inhibited by ketoconazole. Pretreatment with a lower molecular weight imidazole derivative OKY 046 (N = 9) inhibited all Tx synthesis after ischemia. Prior to tourniquet inflation, both OKY 046 and ketoconazole lowered plasma TxB2 levels as well as the L/P ratio (p less than 0.05). After ischemia, OKY 046, but not ketoconazole, maintained the L/P ratio at 0.33, a value below that of untreated animals (p less than 0.05). These results indicate that nonplatelet-derived TxA2 modulates both baseline and ischemia-induced increases in microvascular permeability in the dog hind limb.

6-Ketoprostaglandin F1 alpha↗
PubMed1985

Relationship between the third component of human complement (C3) bound to stored preserved erythrocytes and their viability in vivo.

In this study we evaluated whether the 4 degrees C storage-induced coating of red blood cells (RBC) with molecules of the third component of human complement, C3, affects the viability of the preserved RBC. To this end, we determined whether the amount of C3 bound to preserved RBC correlated with the 24-hour survival value. The % anti-C3c-induced agglutination of stored RBC provided an estimate of the amount of RBC-bound C3. In some cases, the number of RBC-bound C3c-containing molecules was also quantitated. The 24-hour survival of autologous RBC was measured in 114 cases. All units were initially stored at 4 degrees C as RBC concentrates followed in 21 cases by frozen storage and in 75 cases by biochemical rejuvenation and frozen storage. The data showed a significant correlation between % anti-C3c-induced agglutination of the preserved RBC and the length of 4 degrees C storage of the RBC concentrates. Neither freezing nor rejuvenation cleaved the C3c fragment from stored RBC. The 24-hour survival was significantly and negatively correlated with both the storage length of RBC concentrates at 4 degrees C and with the amount of RBC-bound C3 but not with RBC ATP level. These data suggest that the RBC-bound C3 either contributes to or is a marker for the extent of the preservation injury of RBC.

Adenosine Triphosphate↗
PubMed1985

Quantitative differential agglutination method using the Coulter Counter to measure survival of compatible but identifiable red blood cells.

The method described here using a centrifuge and Coulter Cell Counter for the quantitative differential agglutination of human red cells uses commercial anti-A and anti-B antisera for the ABO system, and for the Rh system a commercial anti-D serum, a low ionic strength solution and an anti-human IgG antiserum. We compared this Coulter Counter method with the Technicon AutoAnalyzer method which utilizes bromelin and polyvinyl pyrrolidone, and anti-A, anti-B and anti-CD antisera, and found this new method to be the simpler of the two. The nonagglutinable count with the Coulter Counter was 1.07% for A1 red cells, 2.26% for A2 red cells, 1.06% for B red cells, and 1.78% for Rh-positive red cells, results similar to those seen with the Technicon AutoAnalyzer. Results with the Coulter Counter method were consistently accurate whether the ACD red cells were studied on the day of collection, after 10 days of 4 degrees C storage, or after 4 degrees C storage for up to 6 days followed by cryopreservation with 40% (w/v) glycerol at-80 degrees C, thawing and washing. In this study, red cell samples obtained from recipients who had received compatible but identifiable donor red cells were frozen with 40% glycerol and stored at-80 degrees C for 10 months, thawed and washed. Survival measurements on these washed previously frozen red cells were similar to the values in liquid-stored red cells.

Agglutination Tests↗
PubMed1985

Cryopreservation of human platelets using 6% dimethyl sulfoxide and storage at -80 degrees C. Effects of 2 years of frozen storage at -80 degrees C and transportation in dry ice.

Platelet studies were done in healthy male volunteers and in thrombocytopenic patients. Some of the platelets used in the study were isolated by mechanical apheresis using either the Haemonetics blood processor 30, the IBM blood processor 2997 or the Fenwal CS-3000 blood processor before freezing. Other platelets were isolated from individual units of whole blood and pooled before freezing. The platelets were frozen with a 6% cryoprotectant (DMSO) in a polyvinylchloride (PVC) plastic bag or a polyolefin plastic bag at -80 degrees C in a mechanical freezer and stored for as long as 3 years. Some of the frozen platelets were transported in dry ice in polystyrene foam containers to determine whether they would be adversely affected by such treatment. Platelet recovery after freezing, thawing and washing was about 75%. In the healthy male volunteers, in vivo recovery of autologous platelets 1-2 h after transfusion was about 33%, and the life span was about 8 days. In the thrombocytopenic patients, in vivo recovery values were 50% of those from fresh platelets. The transfusion of previously frozen washed platelets reduced clinical bleeding in the thrombocytopenic patients with bleeding. There was no evidence of quality deterioration in platelets after storage at -80 degrees C for at least 2 years, as determined from in vivo recovery and in vivo survival values, nor was there any adverse effect as a result of shipment of the frozen platelets in dry ice in polystyrene foam containers from one facility to another.

Blood Platelets↗
PubMed1985

Isolation of mononuclear leukocytes in a plastic bag system using Ficoll-Hypaque.

Mononuclear cells, present in bone marrow and peripheral blood, have been isolated from red cells and granulocytes using a ficoll-hypaque density centrifugation process. Cells isolated by this process which uses centrifuge tubes may become contaminated. In 19 studies in our laboratory we used Ficoll-Hypaque treatment to isolate mononuclear cells from cellular residues obtained during plateletpheresis using a modified 600-ml polyvinyl-chloride (PL-146) plastic bag with the Haemonetics blood processor V-50 or the Fenwal CS-3000 blood processor. The 600-ml PVC plastic bag was modified by sealing its vertical edges using radio frequency to form a narrow bag with a volume of approximately 200 ml. A 125-volume of diluted apheresis cellular residue was collected, and the mononuclear cells were isolated as follows: the diluted cellular residue was layered onto 75 ml of Ficoll-Hypaque with a specific gravity of 1.077 and was centrifuged at 260 g for 30 min at 22 degrees C. The supernatant plasma was removed. The mononuclear cell layer was transferred to a sterile 600-ml transfer bag, and the cells were washed with saline. Of the 4.24 +/- 0.9 X 10(9) mononuclear cells applied to the gradient, approximately 3.73 +/- 0.8 X 10(9) cells were recovered. The recovered cells consisted of 77.3 +/- 8% lymphocytes, 19.0 +/- 7% monocytes, and 3.6 +/- 3% granulocytes. There was no significant difference in tissue culture growth in the CFU-GEMM assay of mononuclear cells whether the plastic tube or the plastic bag system was used. Aerobic bacteriologic cultures were negative. The PL-146 plastic bag system used in this study proved to be a significant aid in isolating mononuclear cells from plateletpheresis residue.

Blood Component Removal↗
PubMed1985

Cryopreservation of human platelets and bone marrow and peripheral blood totipotential mononuclear stem cells.

Human platelets in sufficient numbers for a therapeutic transfusion can be collected for preservation either by pooling ABO- and Rh-compatible platelets or by apheresis procedures using mechanical cell-separating machines. Human platelets have been frozen successfully with 5 or 6% dimethyl sulfoxide (DMSO) and stored at -150 or -180 degrees C, respectively. Platelets frozen with 5% DMSO have been stored at -150 degrees C for at least 3 years, and platelets frozen with 6% DMSO have been stored at -80 degrees C for at least 2 years. Approximately 95% of the DMSO usually is removed by washing the platelets after thawing, and the residual DMSO produces no untoward effects. Washed platelets resuspended in plasma can be stored at room temperature for 6 to 8 hr before transfusion. Platelets thus frozen have freeze-thaw-wash recovery values of about 80%. In vivo survival values are only about 50% those seen with fresh platelets, and it is necessary to transfuse twice as many to achieve comparable results. Studies have shown that these platelets have satisfactory circulation, reduce clinical bleeding, and shorten the prolonged bleeding times associated with thrombocytopenia. Studies are now being made on human bone marrow and peripheral blood, from which totipotential cells devoid of immunocompetent cells can be isolated and frozen.

Blood Platelets↗
PubMed1985

Effect of erythrocyte storage and oxyhemoglobin affinity changes on cardiac function.

Storage of blood can depress erythrocyte 2,3-diphosphoglycerate (DPG) levels and thereby increase oxyhemoglobin affinity and potentially decrease capillary-to-tissue oxygen transport. We measured myocardial function and metabolism in isolated rabbit hearts with fixed coronary flow under basal conditions and during isoproterenol stress at 37 and 30 degrees C, comparing high and low oxyhemoglobin affinity (OHA) erythrocytes. The high OHA state resulted from standard storage conditions, which caused depressed values of DPG and P50 (the oxygen tension at which hemoglobin is 50% saturated). The low OHA erythrocytes were initially stored and then underwent biochemical treatment to restore the DPG and P50 values to normal. The low OHA cells released more oxygen, and myocardial oxygen consumption and contractile function were increased relative to the high OHA cells during both the basal and stress states at both 37 and 30 degrees C. These observations may be relevant for patients with limited coronary flow when such patients receive large transfusions of stored blood.

Animals↗
PubMed1985

Serum antielastase and neutrophil elastase levels in PiM phenotype cigarette smokers with airflow obstruction.

In order to assess blood factors which might explain why some cigarette smokers develop airflow obstruction while others do not, we compared two groups of PiM phenotype volunteers matched for age, sex and total pack-years of cigarette smoking; one group had airflow obstruction and the other did not. Functional levels of alpha-2-macroglobulin (alpha-2-M) and alpha-1-protease inhibitor (alpha-1-PI) were separately assessed by a protease binding procedure. Neutrophils were isolated from blood by counterflow centrifugation, and their elastase content was assayed with 3H-elastin-SDS (sodium dodecyl sulfate). The obstructed and nonobstructed groups were not different with respect to functional or immunoreactive levels of alpha-1-PI and alpha-2-M or elastase levels in their neutrophils. We do not find imbalances of circulating elastase or antielastase levels in PiM phenotype smokers with airflow obstruction.

Adult↗
PubMed1985

Subtle alternating electrocardiographic morphology as an indicator of decreased cardiac electrical stability.

Observations from finite-element computer models, together with analytic developments based on percolation theory have suggested that subtle fluctuations of ECG morphology might serve as an indicator diminished cardiac electrical stability. With fixed-rate atrial pacing in canines, we have previously observed a pattern of alternation in T wave energy which correlated with cardiac electrical stability. We report here on a series of 20 canine experiments in which cardiac electrical stability (measured via Ventricular Fibrillation Threshold determination) was compared to a non-degenerate, multidimensional measurement of the degree of alternating activity present in the ECG complex morphology. The decrease in cardiac electrical stability brought on by both coronary artery occlusion and systemic hypothermia was consistently accompanied by subtle alternation in ECG morphology, with the absolute degree of alternating activity being significantly (negatively) correlated with cardiac electrical stability.

Animals↗
PubMed1985

Influence of platelet volume on the ability of prostacyclin to inhibit platelet aggregation and the release reaction.

The influence of mean platelet volume (MPV) and platelet count on in vitro platelet sensitivity to prostacyclin (PGI2) was studied with human size-dependent platelet subpopulations prepared by counterflow centrifugation. The original unfractionated platelet suspension and each of five size-dependent platelet fractions were suspended in buffer at a platelet count of 2 X 10(8)/ml. The percent decrease in the extent of platelet aggregation and adenosine triphosphate (ATP) release in response to 10 micrograms/ml collagen was determined over a range of PGI2 concentrations in a Lumi-Aggregometer. A significant positive correlation between MPV and the concentration required to give 50% inhibition for both platelet aggregation and ATP release (r = 0.99, p less than 0.001 and r = 0.99, p less than 0.001, respectively) was observed. In separate experiments, the effect of platelet count on the ability of a given dose of PGI2 to inhibit platelet aggregation and ATP release was determined, and a significant inverse correlation was noted (r = 0.99, p less than 0.01 and r = 0.98, p less than 0.01, respectively). Our data indicate that the sensitivity of human platelets to the inhibitory effects of PGI2 is dependent on both the platelet volume and the platelet count. Thus, the presence of a greater platelet mass, resulting from either an increased MPV or an increased platelet count, decreases the inhibitory effectiveness of PGI2 on both platelet aggregation and the release reaction.

Adenosine Triphosphate↗
PubMed1985

Prostacyclin and thromboxane A2 moderate postischemic renal failure.

Since prostacyclin (PGI2) is known to regulate renal cortical blood flow and since ischemia stimulates thromboxane (Tx) A2 synthesis, the role of these prostanoids in moderating the response to renal ischemia was studied in the rat. At baseline, plasma TxB2 concentration in untreated animals (n = 13) was 357 pg/ml. The left renal pedicle was clamped for 45 minutes after a right nephrectomy (n = 16), which led after 5 minutes of reperfusion to a rise in TxB2 to 2825 pg/ml (p less than 0.001), but there was no change in 6-keto-PGF1 alpha. After 24 hours creatinine levels rose from 0.4 to 3.0 mg/dl (p less than 0.001), and left renal weight rose from 94% to 117% (p less than 0.001) relative to the weight of the right kidney. In nephrectomized but nonischemic sham control rats (n = 7), creatinine level was 0.9 mg/dl and kidney weight 91% after 24 hours. Pretreatment with OKY 046 (n = 13) (2 mg/kg administered intravenously) blocked ischemia-induced TxB2 synthesis, while 6-keto-PGF1 alpha levels rose from 96 to 302 pg/ml (p less than 0.001). There was no increase in creatinine levels or kidney weight relative to the sham group. Pretreatment with ibuprofen (n = 10) (12 mg/kg) or OKY 046 and ibuprofen (n = 9) inhibited TxB2 and 6-keto-PGF1 alpha synthesis, but creatinine levels and renal weight rose (p less than 0.001). Renal histology in OKY 046-pretreated animals was equal to that in nephrectomized controls, while all other ischemic groups showed tubular necrosis. Results indicate that a high PGI2/TxA2 ratio protects against renal ischemia.

6-Ketoprostaglandin F1 alpha↗
PubMed1985

The viability of young and old baboon red cells stored in the liquid state at 4 C.

Baboons were given intravenous injections of 50-70 microCi of 59Fe citrate to label young and old red cells. Blood subsequently collected from these animals was stored at 4 C in ACD, CPD, or CPDA-1 for up to 36 days. The 59Fe-labelled young and old red cells from the stored blood samples were labelled in vitro with 51Cr and 99mTc and then separated by counterflow centrifugation-elutriation into 7 fractions which differed in red cell volume. 59Fe-labelled young red cells were found in the fractions with the larger red cell volumes; 59Fe-labelled old red cells were found in the fractions with the smaller red cell volumes. 51Cr and 99mTc labelled young and old red cells in the 7 fractions in a similar manner. When salivary antigens were used to identify ABO compatible baboons, rapid removal of the donor red cells from the circulation was seen in 7 of the 15 homologous transfusions. This prompted us to assess compatibility from the survival of 51Cr-labelled donor red cells. In transfusions in which compatibility was identified from 51Cr T50 values of 8 days or greater, 24-hour posttransfusion survival values were similar for the 59Fe young and old red cells. Our data show that in vivo aging of red cells is a different process from that occurring with in vitro deterioration of red cells during liquid storage at 4 C.

Animals↗
PubMed1984

Monoclonal antibody testing of lymphocytes after overnight storage.

Often monoclonal antibody testing of lymphocytes is not performed until the day after blood collection for reasons of convenience or due to the need to transport the blood to other facilities. In order to determine whether accurate results can be obtained on the day after collection, we compared results obtained after storage overnight at 4 degrees C or 22 degrees C with results obtained with fresh lymphocytes. Lymphocytes from 24 normal individuals were evaluated with 10 monoclonal antibodies using an immunofluorescence technique with analysis by flow cytofluorometry. There were markedly altered results obtained with lymphocytes separated on the day after collection from whole blood stored at 4 degrees C. Lymphocytes separated from whole blood stored at 22 degrees C showed moderate changes in reactivity with some monoclonal antibodies. Lymphocytes that were separated from fresh blood and then stored at 4 degrees C or 22 degrees C showed results similar to fresh lymphocytes. These results underscore the importance of proper processing of blood samples to avoid misinterpretation of results.

Antibodies, Monoclonal↗
PubMed1984

A method for the separation of erythrocytes on the basis of size using counterflow centrifugation.

A method has been developed for separation of erythrocytes on the basis of size using counterflow centrifugation. Human red blood cells with an original mean corpuscular volume (MCV) of 89.2 +/- 4.1 fl were isolated, free of plasma proteins and other cell contaminants, into seven fractions ranging in size from 77.0 +/- 2.7 fl to 98.5 +/- 4.8 fl. The ratio of the age-related enzyme, erythrocyte glutamic oxaloacetic transferase (EGOT), to hemoglobin (Hb) increased progressively through the fractions, suggesting a correlation between erythrocyte volume and age. Reticulocytes, though present in all fractions, were selectively enriched in the larger subpopulations. To verify the biochemical evidence that erythrocytes decrease in volume with aging, in vivo cohort labeling of red blood cells with 59Fe was performed in baboons. A similar relationship of EGOT to Hb was observed to that in the human subpopulations. The peak activity of 59Fe/RBC appeared initially in the red blood cells with the highest MCV and progressed from the erythrocytes with the largest MCV to the erythrocytes with the smallest MCV over the next 10-12 weeks, confirming the hypothesis that red blood cells decrease in volume as they age. The technique of counterflow centrifugation appears to provide a simple, rapid, and reproducible method for the separation of erythrocytes on the basis of size.

Adult↗
PubMed1984

Aspirin decreases platelet uptake on Dacron vascular grafts in baboons.

The influence of a single dose of aspirin (5.4-7.4 mg/kg) on platelet uptake on 4-mm Dacron interposition grafts was studied in a baboon model using gamma camera scanning for 111-Indium labeled platelets. In vitro assessment of platelet function after aspirin administration revealed that in the baboon, as in the human, aspirin abolished arachidonic acid-induced platelet aggregation, prolonged the lag time between exposure to collagen and aggregation, and decreased plasma thromboxane B2 levels. Aspirin also prolonged the template bleeding time. Scans for 111-Indium labeled platelets revealed that pretreatment with a single dose of aspirin decreased platelet uptake on 4-mm Dacron carotid interposition grafts. This decrease in platelet uptake was associated with a significant improvement in 2-hour graft patency and with a trend toward improved 2-week patency.

Animals↗
PubMed1984

Inhibition of ischemia-induced thromboxane synthesis in man.

The ability of the imidazole derivative, ketoconazole, to inhibit thromboxane (Tx)A2 synthesis in response to ischemia was tested in ten volunteers. Two hours after taking placebo or ketoconazole 400 mg by mouth, plasma levels of the stable degradation product of TxA2, TxB2, were 300 +/- 129 pg/ml (mean +/- SEM) and 297 +/- 80 pg/ml, respectively. Arm ischemia for 10 min induced by inflation of a cuff to 220 mm Hg led to a rise in TxB2 levels to 657 +/- 157 pg/ml after placebo (p less than 0.05) and 337 +/- 81 pg/ml after ketoconazole. One hour after cuff deflation, TxB2 returned to pre-ischemia levels in both groups. Platelet TxB2 concentrations were 27 +/- 6 ng in the placebo and 35 +/- 6 ng/10(9) platelets in the ketoconazole group, and were unchanged by cuff inflation. The fact that plasma and platelet TxB2 values were not lower 2 hr after ketoconazole treatment was explored in another group of four nonstressed volunteers who received 400 mg of drug. After 2 hr, TxB2 values had fallen from 170 +/- 30 pg to 120 +/- 10 pg; at 4 hr, 6 hr, and 8 hr they were 30 +/- 20 pg, 5 +/- 5 pg, and 5 +/- 5 pg/ml, respectively. These results indicate that tourniquet ischemia provokes TxA2 synthesis, and that the source of this prostanoid is likely to be ischemic tissue and not platelets. Finally, ketoconazole can profoundly inhibit both background and stimulated TxA2 synthesis.

6-Ketoprostaglandin F1 alpha↗
PubMed1984

Arachidonic acid metabolites mediate early burn edema.

Standard burns were sequentially produced on the backs of Sprague-Dawley rats at 0, 1, 2, and 2 1/2 hr, followed by the IV injection of Evans blue dye. All animals were killed at 3 hr, and burns evaluated by wet/dry weight ratios, and Evans blue extravasation scored 1-4 by two observers. Five groups of rats were compared to controls. Rats made neutropenic by exposure to 137cesium showed no significant difference in wet/dry weight ratio or Evans blue extravasation compared to controls. At 1 1/2 hr four other groups were treated with various inhibitors of arachidonic acid metabolism including ibuprofen, a cyclo-oxygenase inhibitor; FPL 55712, a leukotriene (LT) receptor antagonist; ketoconazole, an inhibitor of thromboxane (Tx) synthetase; and lodoxamide, a calcium channel inhibitor. All treated groups showed significant reduction of Evans blue dye extravasation. Wet/dry weight ratios were significantly reduced in rats treated with FPL 55712 and ketoconazole before or after burning. These data support the postulate that oxygenation products of arachidonic acid, particularly Tx and LT, are important mediators in early burn edema.

Animals↗