Blood leukocyte responses to extracorporeal circulation. I. Short term extracorporeal circulation in dogs without and with extracorporeal irradiation

Short term (1 h) extracorporeal circulation without or with irradiation of blood was performed in two normal dogs in a series of experiments. The granulocyte count was constantly diminished, while the lymphocytes did not show any particular change in their concentration. In the majority of the experiments a decrease of the CFU-C content occurred to less than 70% of the initial level. There was no difference in the results of experiments with or without irradiation. In the "bag to bag" procedures, no significant change in the blood leukocyte counts including CFU-C, was established.     

Leukocyte isolation by sedimentation: the effect of rouleau-promoting agents on leukocyte differential count.

The rouleau-promoting agents dextran and polyvinylpyrrolidone (PVP) were used to accelerate erythrocyte sedimentation in order to harvest the leukocyte rich plasma (LRP). The objective of the work was to determine if agent concentration or blood: agent ratio had any effect on the leukocyte differential count and if so at what agent concentration and agent:blood ratio did the LRP leukocyte differential count most closely match the whole blood leukocyte differential count. With both sedimentation agents the data clearly indicate that both parameters effect LRP differential counts and that low concentrations of sedimentation agents are most important in obtaining LRP differential counts which most closely match the whole blood differential counts.     

Survival of transfused cryopreserved granulocytic progenitor cells (CFU-C) in recipient circulation

We have studied the pattern of CFU-C disappearance from the peripheral blood of normal and total-body-irradiated dogs given cryopreserved cell suspensions from bone marrow, foetal liver and peripheal blood containing known numbers of CFU-C under an autologous and allogeneic donor-recipient relationship. Only a small fraction of infused donor CFU-C could be detected in the circulation of recipients at the end of the infusion. There was an exponential fall in circulating CFU-C, indicating random loss of infused CFU-C. The CFU-C disappearance pattern in each experimental group was reproducible. The mean half life of autologous blood derived CFU-C in the circulating blood of normal recipients was 8.2 min and the mean blood CFU-C turnover was calculated to be 9.3 X 10(5) CFU-C/kg per day.     

Pineal control of immune status and hematological changes in blood and bone marrow of male squirrels (Funambulus pennanti) during their reproductively active phase

In addition to pineal control of reproduction in seasonal breeders, melatonin is also known to influence various immune parameters. In the present experiment, we assessed the effect of exogenous melatonin treatment on different hematological parameters of peripheral blood and bone marrow cells, together with histological observations of spleen and thymus blastogenic response and stimulation ratio, and hormonal assays (melatonin and testosterone) of Indian palm squirrel (Funambulus pennanti) during their reproductively active phase when endogenous melatonin levels are low. Daily subcutaneous injection of melatonin (25 microg/100 g body mass.) at 17.30-18.00 h to adult male squirrels for 60 consecutive days during May-June significantly increased the lymphocyte count of blood and bone marrow and the blastogenic response/percent stimulation ratio of spleen and thymus. Histological observation showed densely packed thymocytes and splenocytes. During this period, peripheral testosterone level was high and melatonin was low establishing an inverse relationship as noted earlier for this squirrel. In pinealectomized squirrels, decreased total leukocyte count and percent lymphocyte count in peripheral blood and bone marrow, along with a decreased cell density in spleen and thymus was observed histologically. Further, melatonin treatment of pinealectomized squirrels resulted in restoration of the immune parameters in line with a normal control level. We suggest that during the reproductively active period of male Indian palm squirrels the lymphoid organs were sensitive to melatonin; hence, the exogenous melatonin treatment had an immuno-enhancing effect.     

The r.b.e. of different-energy neutrons as determined by human bone-marrow cell-culture techniques.

The effect of X-rays and different-energy neutrons on human bone-marrow cells was studied using two different cell-culture techniques--diffusion chamber (DC) growth and colony formation in vitro (CFU-C). Based on the survival of proliferative granulocytes in DC on day 13, the D0 value was 80 rad with X-rays, and 117 rad as measured by the CFU-C assay. The D0 values for neutrons depended on the radiation source and the energy level. The r.b.e. values, which dropped with increasing energy levels of mono-energetic neutrons, were (i) 0.44 MeV; DC 3.7, CFU-C 4.1; (ii) 6 MeV; DC 1.8, CFU-C 2.0; (iii) 15 MeV; DC 1.6, CFU-C 1.6; (iv) fission neutrons; DC 2.6, CFU-C 2.4.     

Growth of in vitro colony-forming cells from normal human peripheral blood leukocytes cultured in diffusion chambers

The growth of granulopoietic progenitor cells (CFU-C) in diffusion chambers during culture of peripheral blood leukocytes from 10 normal subjects has been studied. At various times after initiation of diffusion chamber culture, cells harvested from the chambers were transferred to agar culture for measurement of CFU-C concentration. Under these conditions colonies could be grown successfully in agar culture provided pronase, necessary for the chamber harvesting procedure, was first removed by careful washing. A marked increase in the number of CFU-C, up to 25-fold the initial value, was observed in 8 out of 10 subjects. Here the growth pattern was similar, independent of the initial CFU-C values, with an immediate rise to a maximum between 6 and 13 days of culture followed by a decrease. In the other two subjects the growth of CFU-C throughout the diffusion chamber culture period was very poor. The growth of CFU-C from a given individual's blood was shown to be reproducible in repeated studies in 2 subjects, one of whom showed a proliferative and the other a non-proliferative pattern. Evidence suggests that the increase in CFU-C in diffusion chambers is the result of both self-renewal of these cells and influx from a more primitive compartment, although the present data do not allow an estimate of the relative magnitude of each.     

Hematological and clinico-biochemical characteristics of leukemia in Fischer 344 rats.

Seventy-one male and 52 female F 344 rats with leukemia used as controls in the 30-month inhalation studies were characterized by hematological and clinico-biochemical findings. Hematological findings revealed that the leukocyte count, mean corpuscular volume, and mean corpuscular hemoglobin increased in both sexes of leukemic rats showing profound anemia, while the platelet count, erythrocyte count, hematocrit, and hemoglobin concentration decreased. In these rats, the serum levels of low density lipoprotein, free cholesterol, total bilirubin, blood urea nitrogen, and triglyceride and the activities of glutamic oxalacetic transaminase, glutamic pyruvic transaminase, creatine phosphokinase, alkaline phosphatase, and lactate dehydrogenase increased markedly and the level of high density lipoprotein, the oxygen partial pressure, and the cholinesterase activity decreased. Clinical signs such as decrease in redness of the eyes, decrease in body weight, abdominal distension, staining of the public region, and debility were seen in most leukemic animals. These clinical signs and hematological and clinico-biochemical findings may be helpful in diagnosis of leukemia in long-term experiments.     

Alterations in the coronary circulation of man following ascent to 3,100 m altitude.

Alterations in coronary blood flow associated with adaptation to high altitude were examined. Three normal men native to low altitude were studied, first at sea level, and again after 10 days' sojourn at 3,100 m altitude. During rest at high altitude, a 32% decrease in coronary blood flow was largely offset by a 28% increase in coronary arterial O2 extraction to maintain myocardial O2 delivery. The increase in O2 extraction resulted mainly from a decrease in coronary sinus blood O2 content and saturation. However, coronary sinus O2 tension remained constant, implying a decrease in the affinity of hemoglobin for O2. These observations are consistent with the hypothesis that coronary blood flow is regulated to maintain constant myocardial tissue O2 tension (as reflected here by coronary sinus blood O2 tension). The absence of a decrease in coronary sinus O2 tension or a decrease in myocardial lactate extraction imply that myocardial hypoxia did not develop. Therefore, myocardial hypoxia is not the basis for the decrease in cardiac stroke volume at high altitude reported previously and also observed in the present study.     

Cerebrovascular circulation during blockade of angiotensin receptors

In experiments on anaesthetized rats, Losartan was found to cause an obvious decrease in the ABP in normotensive rats. The cerebral blood flow differed independence on cerebral vascular resistance and the ABP level. The autoregulation of the cerebral blood flow remained unaltered. In hypertensive rats Losartan caused a significant decrease in the ABP as compared with normotensive rats. A shift of lower limits of the cerebral blood flow autoregulation towards a lower ABP level, was observed.     

The effects of anticoagulants on hematological indices and blood cell morphology of common carp (Cyprinus carpio L.)

Hematological parameters (Ht, Hb, RBC, WBC, PLT), erythrocyte size, and osmotic fragility, differential leukocyte count, ROS production in common carp blood collected on three anticoagulants: heparin (10 IU/mL, Na2EDTA (0.1, 0.5, and 1 mg/mL), and sodium citrate (0.3 mg/mL) were compared. Na2EDTA caused partial blood hemolysis in Ht tubes which made Ht measurement impossible, and resulted in high variability of the results. Both, citrate and Na2EDTA increased sensitivity of red blood cells to hemolysis. Na2EDTA also induced erythrocyte anisocytosis and anisonucleosis. Na2EDTA significantly increased ROS production but no effect of anticoagulants on WBC, PLT or differential leukocyte count was observed. The obtained results show that Na2EDTA should not be used for evaluation of red blood cell parameters and erythrocyte morphology, and for ROS production measurement in common carp. Heparin proved to be the most appropriate anticoagulant to use for this species, although Na2EDTA and sodium citrate may be used for WBC and leukocyte differential count evaluations.     

Status of hemapoiesis in residents of the Techa riverside villages in the period of maximum radiation exposure. Report 1. Evaluation of the cellular composition of peripheral blood and the role of comorbidities in the oppression of hemapoiesis

Residents of the Techa riverside villages chronically exposed to ionizing radiation (the average dose rate in 1951-1956 was 0.047 Gy/year, the maximum reached 2.44 Gy/year) developed marked changes in the cellular composition of peripheral blood. The maximum reduction of peripheral blood counts occurred in the years 1951-1953, after which the beginning of the restoration of cellularity to the control level was observed. The dose rate at this point (1956) was about 0.02 Gy/year. The factors of radiation and non-radiation nature (gender, the age at the onset of exposure, health related disorders) in different combinations affect the number of peripheral blood cells. The influence of dose rate of a chronic radiation exposure on a platelet count takes precedence over other factors. The factors of gender and dose rate determine the number of erythrocytes in exposed persons. The changes accompanying the health status and dose rate significantly affect the number of neutrophils. The influence of comorbidity, age and dose rate on the number of monocytes was noted. A lymphocyte count was mainly determined by the age at the onset of exposure and concomitant diseases. A joint influence of chronic radiation exposure and concomitant diseases increases a mutual action on erythro- and thrombocytopoiesis. The decrease of the dose rate was followed by a gradual predominance of the somatic disease influence on leukocyte (neutrophils, lymphocytes and monocytes) counts.     

Paradoxical attenuation of leukocyte rolling in response to ischemia- reperfusion and extracorporeal blood circulation in inflamed tissue

In contrast to acute preparations such as the exteriorized mesentery or the cremaster muscle, chronically instrumented chamber models allow one to study the microcirculation under "physiological" conditions, i.e., in the absence of trauma-induced leukocyte rolling along the venular endothelium. To underscore the importance of studying the naive microcirculation, we implanted titanium dorsal skinfold chambers in hamsters and used intravital fluorescence microscopy to study venular leukocyte rolling in response to ischemia-reperfusion injury or extracorporeal blood circulation. The experiments were performed in chambers that fulfilled all well-established criteria for a physiological microcirculation as well as in chambers that showed various extents of leukocyte rolling due to trauma, hemorrhage, or inflammation. In ideal chambers with a physiological microcirculation (<30 rolling leukocytes/mm vessel circumference in 30 s), ischemia-reperfusion injury and extracorporeal blood circulation significantly stimulated leukocyte rolling along the venular endothelium and, subsequently, firm leukocyte adhesion. In contrast, both stimuli failed to elicit leukocyte rolling in borderline chambers (30-100 leukocytes/mm), and in blatantly inflamed chambers with yet higher numbers of rolling leukocytes at baseline (>100 leukocytes/mm), we observed a paradoxical reduction of leukocyte rolling after ischemia-reperfusion injury or extracorporeal blood circulation. A similar effect was observed when we superfused leukotriene B4 (LTB4) onto the chamber tissue. The initial increase in leukocyte rolling in response to an LTB4 challenge was reversed by a second superfusion 90 min later. These observations underscore 1) the benefit of studying leukocyte-endothelial cell interaction in chronically instrumented chamber models and 2) the necessity to strictly adhere to well-established criteria of a physiological microcirculation.     

Collection, storage and transfusion of blood stem cells for the treatment of hemopoietic failure.

Migration of hemopoietic stem cells via the blood to sites of stem cell need is a principle that becomes established during the embryonic development of hemopoiesis and can be observed in the adult whenever bone marrow transplantations are being performed. The regular presence of stem cells in the peripheral blood lends itself to the study of their collection, storage, and use for transfusion purposes in cases of bone marrow failure. Both in dog and in man, granulocyte-macrophage progenitor cells (CFU-C) can be collected by leukapheresis from the blood in large quantities, particularly if the yield is increased by the administration of mobilizing agents such as dextran sulfate, and appear to be an indicator for the presence of stem cells. For collection and storage, a closed plastic bag system has been developed that allows the safe handling of the cells. The loss of CFU-C from freezing and thawing with DMSO as a cryoprotective agent is only 10%-20%. If frozen and thawed mononuclear leukocytes are transfused into 1200 rad whole-body X-irradiated autologous or allogeneic recipient dogs, a hemopoietic take is observed when 0.2 X 10(5) CFU-C are present among the mononuclear leukocytes (MNC). Graft-versus-host disease can be avoided in the allogeneic situation when a purified CFU-C rich cell fraction is being transfused. In man collection and storage of MNC including CFU-C is feasible and may eventually become a therapeutic tool.     

General anesthesia and hepatic circulation

This article describes hepatic circulatory disturbances associated with anesthesia and surgical intervention. The material is presented in three parts: part 1 describes the effects of general anesthetics on the hepatic circulation; part 2 deals with different factors related to surgical procedures and anesthesia; and part 3 analyzes the role of hepatic circulatory disturbances and hepatic oxygen deprivation in anesthesia-induced hepatotoxicity. The analysis of available data suggests that general anesthesia affects the splanchnic and hepatic circulation in various directions and to different degrees. The majority of anesthetics decreases portal blood flow in association with a decrease in cardiac output. However, hepatic arterial blood flow can be preserved, decreased, or increased. The increase in hepatic arterial blood flow, when it occurs, is usually not enough to compensate for a decrease in portal blood flow and therefore total hepatic blood flow is usually decreased during anesthesia. This decrease in total hepatic blood flow has certain pharmacokinetic implications, namely a decrease in clearance of endogenous and exogenous substances with a high hepatic extraction ratio. On the other hand, a reduction in the hepatic oxygen supply might play a certain role in liver dysfunction occurring perioperatively. Surgical procedures-preparations combined with anesthesia have a very complex effect on the splanchnic and hepatic circulation. Within this complex, the surgical procedure-preparation plays the main role in developing circulatory disturbances, while anesthesia plays only a modifying role. Hepatic oxygen deprivation may play an important role in anesthesia-induced hepatotoxicity in different experimental models.     

Effect of intrarenal infusion of synthetic PAF-acether in dogs

The effect of intrarenal infusion of PAF-acether was studied in dogs. PAF-acether infusions caused a dose-dependent decrease in glomerular filtration rate, renal blood flow and urinary flow and electrolyte excretion, without significant changes in mean arterial pressures. In addition, the higher doses used caused also increases in packed cell volume, and decreases in plasma proteins and leukocyte and platelet count, whereas the lower doses did not elicit those changes. These data suggest that PAF-acether causes an impairment in renal function which is in part mediated by vasoactive substances released from platelets and leukocytes.     

Some selected haematological indices in Wistar rats in the vanadium-ethanol interaction

1. Wistar rats of both sexes daily received an ethanol solution of ammonium metavanadate (AMV) of 0.3 mg V/cm3 5% ethanol concentration as sole drinking liquid, for a period of 4 weeks. 2. The reference groups received for drinking aqueous AMV solution, or 5% ethanol, or water. 3. In animals drinking both water and ethanol AMV solution a decrease in the erythrocyte count and haemoglobin level was noted together with an increase of the percentage of reticulocytes and polychromatophilic erythrocytes in the peripheral blood. 4. A small rise of the percentage of polychromatophilic and orthochromatic erythroblasts was at the same time noted in the bone marrow of animals receiving ethanol AMV solution. 5. In the group of animals drinking 5% ethanol a fall of the erythrocyte count was observed and a rise of the leukocyte count, particularly of lymphocytes. 6. Substitution of water by 5% ethanol solution as solvent for AMV did not have any distinct influence on the toxicity of the tested compound.     

Hematological and enzymatic results of aluminum intoxication in rats

1. The study has been carried out on Wistar rats. The aim of the present study was to trace the effect of aluminum on enzyme activities and hematological parameters on erythrocytes.2. Aluminum decreased activities of acetylcholinesterase, glutathione reductase, glucose-6-phosphate dehydrogenase, and lactate dehydrogenase in the erythrocytes of the animals tested.3. In the peripheral blood, a significant decrease in the erythrocyte count, hemoglobin level and hematocrit index and increased percentage of reticulocytes and polychromatophilic erythrocytes were observed.4. The increase in the neutrophilic granulocyte and lymphocyte count was significant.5. An inhibitory effect of aluminum on the phagocytic activity of granulocytes was also observed.     

The in vivo effects of a culture medium. II. Influence of culture medium administered prior to irradiation on hemopoietic recovery of gamma-irradiated mice.

The culture medium administered to C57Bl/6 mice 18 h and 8 h before a single irradiation (9 Gy) had a radioprotective effect and clearly influenced postirradiation changes in haemopoiesis. Haemopoiesis recovery appeared to be faster in culture medium-pretreated animals than in those irradiated without such pretreatment. By 12-15 days after irradiation, the thymus cortex appeared to be repaired, on day 21 a multiple increase in extramedullary erythropoiesis, myelopoiesis and megakaryocytopoiesis in the red pulp was found and later, on day 28, the lymphopoiesis in the white pulp of spleen was restored. The rate of haemopoiesis proliferation of predominantly myeloid cells which reached a control level on day 28 following irradiation. Consequently, the regenerative processes in blood-forming organs were accompanied by considerable reticulocytosis and complete recovery of neutrophil and platelet counts in the peripheral blood as seen on day 21. Despite a slower rate complete recovery of the total leukocyte count was reached by day 180 after irradiation.     

Hematologic and serum biochemical values for Yucatan miniature swine

Hematologic and serum biochemical values were determined for healthy, mature Yucatan miniature swine, Sus scrofa. These values were similar to those reported for other breeds of swine. There was no effect on erythrocyte count, hematocrit, MCV, MCH, MCHC, RDW, platelet count, or leukocyte count attributable to sex (p greater than 0.05). Differential leukocyte counts generated on an automated multichannel blood cell counter, having a three part leukocyte differential capability, were compared to 100-cell manual leukocyte differentials. Determination of lymphocyte and non-lymphocyte fractions on this system were not significantly different from microscopic differentials (p less than 0.001). However, the mononuclear cell count did not correlate well with the percentage of monocytes determined manually (r = 0.084, p greater than 0.5). Leukocytes, erythrocytes, and platelets behaved properly with respect to counting thresholds as modified for counting cells of other common domestic species on this automated cell counter.     

Leucine aminopeptidase of neutrophils and lymphocytes in patients with cancer

The leucine aminopeptidase activity has been determined by using the cytochemical method of Burston and Folk in peripheral blood neutrophils and lymphocytes of 45 patients with various malignancies. Lung cancer, carcinoma of the stomach and cancer of the colon was diagnosed in 24, 16, and 5 patients, respectively. Patients with metastases showed a significantly higher activity of the enzyme if compared with that in the control group of healthy subjects and patients without metastases. The percentage of enzyme-positive lymphocytes was elevated significantly in patients with metastases whereas a total percentage of lymphocytes with regard to differential leukocyte count was diminished both in patients with and without metastases. The absolute count of neutrophils was elevated both in patients with and without metastases. The authors discuss the significance of their observation with regard to the antitumor cytotoxic effect of neutrophils and lymphocytes.