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At least 559 records · Page 31Linked to original sources
PubMed2000

Closed circuit cardiopulmonary bypass with centrifugal pump for open-heart surgery: new trial for air removal.

The purpose of this study is to examine the efficiency of venous air removal with a new cardiopulmonary bypass (CPB) circuit design for conventional open-heart surgeries. A main concern with a closed circuit for open-heart surgeries is air entrainment into the venous line. A venous filter was placed proximal to the centrifugal pump. The circuit proximal to the centrifugal pump was divided into two lines; one line was attached to the venous reservoir outlet. By clamping the line to the reservoir, this circuit becomes closed. Negative pressure was applied to the purge line connected to the venous reservoir for venous air removal. Micro bubbles were measured at two locations, both distal to the venous and arterial filters. When the injection rate reached 100 ml/min, with the air-injection over 30 s, micro bubbles greater than 40 micro were observed at the outlet of venous filter. However, there was no micro bubble greater than 40 micro detected at the outlet of arterial filter. Although micro bubbles greater than 40 micro were not detected at the outlet of the arterial filter up to the injection rate of 300 ml/min, when the injection rate reached 400 ml/min, micro bubbles greater than 50 microm were detected distal to the arterial filter. From this examination, we determined that air entrained in the venous line up to approximately 300 ml/min is automatically removed by this method with the pressure-balanced condition. This pressure balance means that resistance of venous return, gravity siphon, negative pressure by centrifugal pump, and negative pressure applied to the air-purge line of the filter are balanced; that is, the venous return is sufficient, and the venous reservoir volume is kept stable. From this study we determined that this circuit design efficiently removes the entrained air in the venous line.

Air↗
PubMed2001

Estimation of left ventricular recovery level based on the motor current waveform analysis on circulatory support with centrifugal blood pump.

In a mock circulatory loop simulating the left heart bypass using a centrifugal blood pump, analysis of the motor current waveform of the centrifugal pump was performed to derive a useful parameter to evaluate the status of ventricular function. The relationship between the peak, amplitude, and the peak of the fundamental frequency of the power spectral density of the periodic motor current waveform (MCpsdP) that reflected the pulsatile ventricular pressure, and the peak of the left ventricular pressure (LVP) was examined. Although both peak and amplitude of the motor current waveform showed an excellent correlation with the peak LVP, they failed to predict the opening of the aortic valve. The MCpsdP that corresponds to the frequency of the heart rate showed an excellent correlation with the peak LVP throughout the LVP levels, but the slope between them changed with the opening of the aortic valve. Thus, it is possible to follow the change in the LVP and detect even the opening of the aortic valve, and, hence, the recovery of the left ventricle. However, the slope of the linear regression equation varied, depending on the pump speed. This result implies that the MCpsdP can be possibly used to follow the change of ventricular function during circulatory assistance with a centrifugal blood pump as well as to control the pump speed in response to varying ventricular function.

Centrifugation↗
PubMed2001

Platelet damage caused by the centrifugal pump: laser-light scattering analysis of aggregation patterns.

Platelets are damaged by shear force during centrifugal pumping; however, the mechanism of this damage has not been fully investigated. A new laser-light scattering (LS) assay method enables quantification of real-time changes in the number of platelet aggregates of different sizes. Using this method, we assessed the kinetics of platelet damage caused by the centrifugal pump from the standpoint of platelet aggregation capacity. Conventional optical density (OD) and LS methods were used to measure platelet aggregation with a Kowa AG-10 aggregometer. Platelet aggregation in fresh human blood was evaluated in a mock circuit for 3 h under a flow rate of 5.0 L/min and a pressure head of 100 mm Hg. Test samples were obtained before pumping for control, and at 1, 2, and 3 h after the start of pumping. The test series was begun 8 times. Aggregation after stimulation by 2.0 microg/ml collagen was determined; small (9 to 25 microm), medium (25 to 50 microm), and large (50 to 70 microm) aggregates were counted by the LS method. OD measurement at hourly intervals showed significant reduction in platelet aggregation. The LS method showed that generation of small and medium aggregates was not suppressed during pumping, but that generation of large aggregates was significantly reduced at 2 and 3 h of pumping. Platelet aggregation is significantly suppressed during centrifugal pumping, and the resulting platelet dysfunction is due mainly to inhibited development of small aggregates into larger aggregates.

Analysis of Variance↗
PubMed2002

Effects of scaling on centrifugal blood pumps.

Experimental studies on the effects of scaling on the performance of centrifugal blood pumps were conducted in a closed-loop test rig. For the prototype, eight different impellers of the same outer diameter of 25 mm were tested at 1,500, 2,000, and 2,500 revolutions per minute (rpm) using blood analog as fluid medium. This corresponds to Reynolds numbers (Re) of 25,900, 34,500, and 43,200, respectively. The results indicated that the nondimensional pump characteristic is a function of Re. This is understandable since the typical operating Re for centrifugal blood pumps is less than 100,000. Thus, the effects of scaling cannot be ignored for centrifugal blood pumps. Experiments on a 5x scaled-up model have also indicated that the scaled-up model is more efficient than the prototype model. Our results showed that in the range of Re tested, the nondimensional head versus flow curve is a function of Re to the power of approximately 0.25. It is observed that the nondimensional head versus flow is a function of diameter ratio to the power of 0.2.

Blood Flow Velocity↗
PubMedDate not supplied

In vitro function of granulocytes isolated from blood of normal volunteers using continuous-flow centrifugation in the IBM-aminco celltrifuge and adhesion-filtration leukapheresis using nylon fiber.

Granulocytes were harvested from each of five healthy male volunteers once by continuous flow centrifugation with the IBM-Aminco Celltrifuge, and once by adhesion filtration leukapheresis with nylon fiber. Granulocyte recovery and purity were significantly better with the filtration leukapheresis system than with continuous flow centrifugation. Measurements of trypan blue dye exclusion and muramidase activity were similar to those in control granulocytes regardless of the method of isolation. Granulocyte-stimulated oxygen consumption was diminished in granulocytes prepared by the adhesion filtration method, but normal in those prepared by continuous flow centrifugation with the IBM-Aminco Celltrifuge.

Blood Transfusion↗
PubMedDate not supplied

Is very fresh blood needed in preparing leukocyte-poor red blood cells by inverted centrifugation?

It is recommended that units selected for inverted centrifugal preparation of leukocyte-poor red blood cells be less than 24 hours old. The reason for this is unclear and the recommendation poses practical problems. We evaluated the effectiveness of inverted centrifugation in reducing the WBC:RBC ratio in 31 units stored up to six days at 4 C. Leukocyte removal was at least as effective with six-day stored units as with less than one day old, averaging 77 per cent reduction with 76 per cent recovery of red blood cells. No reduction in leukocytes occurred with six days storage alone. It appears to be unnecessary to restrict leukocyte removal by inverted centrifugation to units less than 24 hours old.

Blood Preservation↗
PubMedDate not supplied

Cryopreservation of human platelets isolated by discontinuous-flow centrifugation using the Haemonetics Model 30 Blood Processor.

Platelets were isolated from normal volunteers by discontinuous-flow centrifugation using the Haemonetics Model 30 Blood Processor. The numerical equivalent of about five single units of platelets collected at each pheresis were frozen together in a -80 C mechanical freezer with a 6% final concentration of dimethylsulfoxide (DMSO) as the cryoprotectant. Platelet freeze-thaw-wash recovery in vitro was about 80 per cent and the platelet recovery value depended upon the method used to enumerate the platelets. The 51Cr survival values in vivo were about 50 per cent less than those in fresh platelets. These values were not significantly different from those seen when platelets were isolated from single units of blood by differential serial centrifugation. Transfusion of two and one-half units of freeze-preserved platelets provided an increase in the recipient's circulating platelet count comparable with that from one unit of fresh platelets. The hemostatic effectiveness of freeze-preserved platelets isolated by discontinuous-flow centrifugation has not yet been studied.

Adult↗
PubMedDate not supplied

The effect of centrifugation time on cellular residues in fresh plasma.

Cellular residues were present in all fresh plasma prepared by a standard technique with centrifugation times of 5, 7, or 10 minutes. The amounts of cellular residues in the plasma were inversely proportional to the time of centrifugation. Platelet contamination was statistically different in the three times tested. In contrast, the residual amounts of erythrocytes and leukocytes were significantly different only when 5- and 10-minute centrifugation times were compared.

Blood Platelets↗
PubMedDate not supplied

Removal by centrifugation of leukocytes and red cells from apheresis platelets.

Platelets prepared by discontinuous flow centrifugation contain significant numbers of leukocytes and red cells. Platelet products often are centrifuged further to remove this cellular "contamination." A method is described by which platelet products obtained by cytapheresis were prepared using an additional slow centrifugation step (121 X g for 7.5 minutes). In a production environment, we removed an average of 97 percent of leukocytes and most of the contaminating red cells with this method, while retaining 89 percent of the platelets in the final product. This method is an improvement over methods described previously and was consistently effective in routine use.

Blood Platelets↗
PubMed2000

Isolation of monocytes from whole blood by density gradient centrifugation and counter-current elutriation followed by cryopreservation: six years' experience.

BACKGROUND: Monocyte purification by means of counter-current elutriation and subsequent cryopreservation for future use was initiated in 1986 and has been established as a routine since 1993. AIM: To sum up and evaluate our method for the isolation and preservation of monocytes. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from healthy donor blood by density gradient centrifugation, and monocytes were isolated from the PBMC by counter-current elutriation centrifugation using the Beckman J-6M/E centrifuge. The monocytes were then cryopreserved at 135 degrees C and thawed when required for experimental use. RESULTS: Results are given for the last 6 years, including 59 elutriations and the fractions containing monocytes. The mean purity of monocytes was 93% (range 64-98%); mean recovery was 51% (range 22-55%). Studies of CD14 expression and Annexin V indicate that there are no differences between elutriated fractions immediately upon purification or after freezing and thawing. The studies also indicate that interdonor variations are much larger than intradonor variations. DISCUSSION: Although it differs from other reports in certain respects, our procedure has nevertheless produced results in line with other findings. After extensive testing and use in different contexts we feel confident that we have established a method for producing a large number of purified and well-preserved monocytes. CONCLUSION: The goal of being able to perform a large number of experiments with monocytes of high purity and good functionality has been reached.

Annexin A5↗
PubMedDate not supplied

Comparison of centrifugation- and noncentrifugation-based techniques for recovery of motile human sperm in assisted reproduction.

To compare standard density gradient centrifugation sperm preparation with a novel non-centrifugation-based dual-chamber capillary dish in efficiency for motile human sperm separation, approximately 3 mL fresh ejaculate specimens was obtained from 21 men (median age = 32 years. range 26-42 years) undergoing infertility evaluation. For each specimen, half of the sample was processed with a standard 45%/90% density gradient preparation (PureSperm. Nidacon International, Gothenburg, Sweden) followed by semen analysis. The other half was incubated in the Zech glass capillary dish (Astromedtec, Salzburg, Austria) consisting of 2 concentric wells overlaid by a U-ring and coverglass. After approximately 3 h, a 1-mL sample was taken from the central chamber and analyzed. Percentage motile sperm recovery, absolute (motile) cell number, and path velocities were compared for spermatozoa obtained from both methods. Both techniques reduced overall sperm concentration while enriching specimens with more motile spermatozoa. A trend towards higher % recovery of motile spermatozoa (p = .264) was observed with the Zech device, but at a cost of fewer absolute numbers of higher velocity cells (p = .004). The Zech device, therefore, localized a very small population of motile sperm without exposure to centrifugation stress, which has been considered potentially harmful to spermatozoa. This technique could theoretically improve efficiency by reducing time required to identify motile cells in in vitro fertilization where intracytoplasmic sperm injection is planned. However, refinements in incubation interval and suspension volumes are needed before this technique can be considered comparable to the density gradient method in recovering sperm for use in intrauterine insemination.

Adult↗
PubMed1988

[The effect of different centrifugation conditions for the isolation of mixed rumen bacteria, on their nitrogen and diaminopimelic acid content].

Three cows were given two rations, a silage diet (3 animals) and a green forage diet (2 animals). Samples of rumen content were collected and aliquots of these were separated in a fraction of feed particles and protozoa (FP-fraction) and a fraction of mixed bacteria, varying the conditions of differential centrifugation. The low speed centrifugation was practised at 100 X g/5 min, 400 X g/10 min, 1000 X g/10 min, and 2000 X g/10 min. High speed conditions were 30,000 X g/30 min 4 degrees C. The lyophylisated sediments were used for determination of N and DAP. The content of N gave similar results for all fractions of mixed bacteria, the mean value being 7.43 +/- 0.48% (n = 20), while the N-content of the FP-fractions being 5.68 +/- 0.37% (n = 19). The N:DAP-ratio gave similar values for the cows fed the silage diet, the values were 29.45 +/- 1.56 (n = 12). The values for the cows receiving the green forage diet differed, the mean values were 23.08 +/- 0.88 and 42.01 +/- 5.81 (n = 5), respectively. In all five experiments highest ratios were found at 100 X g. Further investigations showed that storage at -20 degrees C rumen fluid after isolation of feed particles and protozoa decreased both the N- and DAP- content without affecting the N:DAP-ratio. Centrifugation at low speed with 100 X g resulted in a markedly decreased contamination with DAP in all the FP-fractions. Optimal conditions for separation of feed particles and protozoa from rumen fluid to get a fraction best reflecting the rumen bacterial populations are 100 X g/5 min.

Amino Acids, Diamino↗
PubMed1989

[The effect of different centrifugation conditions in the isolation of mixed rumen bacteria on their content of nitrogen and diaminopimelic acid: use of duodenal content as raw material].

The objective of the present study was, to investigate the effect of varying conditions of differential centrifugation of duodenal content on the isolation of bacteria (B-fraction) and feed particles + protozoa (FP-fraction). The treatments at low-speed centrifugation were as follows: 100 x g/5 min, 400 x g/10 min, 1000 x g/10 min and 2000 x g/10 min, high speed conditions were 30,000 x g/30 min/4 degrees C. The results of three experiments are given. Analytical examination gave similar results for N-contents for all treatments, the mean values being 7.90 +/- 0.27% (n = 12) for B-fractions and 6.53 +/- 0.73% (n = 12) for FP-fractions. Increasing the low-speed from 100 x g to 2000 x g lead to increasing DAP-contents and decreasing N:DAP-ratios of the bacterial isolates, the values being 2.43, 3.02, 3.22 and 3.39 mg DAP/g DM and 32.0, 27.4, 25.0 and 23.0 N:DAP-ratio. Decreased isolation of bacterial material in the B-fraction in conjunction with increased incorporation in the FP-fraction resulted in rising the speed of the low speed centrifugation. The rates of loss of DAP, measured by comparison with the total amount were 10, 32, 48 and 70% respectively. It was concluded to prefer the isolation of bacteria from rumen fluid.

Amino Acids, Diamino↗
PubMed1965

Isolation of nerve endings from the posterior pituitary gland. Electron microscopy of fractions obtained by centrifugation.

Bovine posterior pituitary glands were homogenized in 10 per cent sucrose and fractionated by differential centrifugation. The following centrifugation procedure resulted in the most satisfactory separation: 1000 g for 15 minutes-nuclei, connective tissue, basement membranes with associated endothelium, giant nerve endings, and whole pituicytes; 4200 g for 15 minutes-free nerve endings, including Herring bodies; 17,000 g for 15 minutes-mitochondria; 68,000 g for 15 minutes-neurosecretory granules. Electron microscopic examination was carried out on whole tissue and on the isolated fractions. Isolated nerve endings were examined also by negative staining techniques. Isolated nerve endings retain an apparently normal complement of mitochondria, neurosecretory granules, and microvesicles ("synaptic" vesicles). The free nerve endings closely resemble those observed in sections of intact posterior pituitary tissue. Free microvesicles were not observed in any of the fractions isolated and apparently sediment at centrifugal forces higher than those employed in this study.

Animals↗
PubMed1978

Isolation of rat liver lysosomes by isopycnic centrifugation in a metrizamide gradient.

A preparation, similar to the light mitochondrial fraction of rat liver (L fraction of de Duve et al, (1955, Biochem. J. 60: 604-617), was subfractionated by isopycnic centrifugation in a metrizamide gradient and the distribution of several marker enzymes was established. The granules were layered at the top or bottom of the gradient. In both cases, as ascertained by the enzyme distributions, the lysosomes are well separated from the peroxisomes. A good separation from mitochondria is obtained only when the L fraction if set down underneath the gradient. Taking into account the analytical centrifugation results, a procedure was devised to purify lysosomes from several grams of liver by centrifugation of an L fraction in a discontinuous metrizamide gradient. By this method, a fraction containing 10--12% of the whole liver lysosomes can be prepared. As inferred from the relative specific activity of marker enzymes, it can be estimated that lysosomes are purified between 66 and 80 times in this fraction. As ascertained by plasma membrane marker enzyme activity, the main contaminant could be the plasma membrane components. However, cytochemical tests for 5'AMPase and for acid phosphatase suggest that a large part of the plasma membrane marker enzyme activity present in the purified lysosome preparation could be associated with the lysosomal membrane. The procedure for the isolation of rat liver lysosomes described in this paper is compared with the already existing methods.

Acid Phosphatase↗
PubMed1988

Automation of routine coagulation testing using a random access centrifugal analyzer.

The results are reported of a clinical and laboratory evaluation of the use of a random-access centrifugal analyzer linked to a personal computer in the management of the routine workload of a hemostasis laboratory. Over a three-month period, prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin clotting time (TCT), and derived fibrinogen (Fib) were performed on a total of 929 samples. Included in the study were 448 samples from patients receiving anticoagulants (oral anticoagulants, 228; heparin, 166; heparin and warfarin, 130) and 351 samples from patients requiring coagulation screens (PT, APTT, TCT, Fib). Tests were done in parallel with tilt-tube manual techniques and the results correlated. The correlation coefficients were PT, 0.99; TCT, 0.72; APTT, 0.96; Fib, 0.97. Discrepancies were analyzed and were due to hypofibrinogenemia and hyperlipidemia. The poorer correlation coefficient of TCT was attributable both to lower reproducibility of the manual test and the effect of dysfibrinogenemia or FDPs in liver disease. In no case was an abnormality or diagnosis missed using the centrifugal analyzer. In several cases the increased sensitivity of the analyzer improved the detection of the lupus anticoagulant. The use of automation was accompanied by a major reduction in workload and reagent costs. The machine has been used to assay a wide range of coagulation tests by clot based and chromogenic substrate methods. In conclusion, a programmed centrifugal analyzer is a safe, efficient, and flexible way of automating routine coagulation tests. It widens the reportoire of tests performed in the Hemostasis laboratory by using a machine capable of being used in other areas of pathology.

Blood Coagulation Tests↗
PubMed1992

A rapid method to isolate platelets from human blood by density gradient centrifugation.

Platelets can be damaged easily or activated during isolation, making them unsuitable for functional studies. The most common technique for isolating platelets involves centrifugation. Although gentler methods have been devised to isolate platelets by density gradient centrifugation or electrophoresis, these techniques either result in a relatively dilute platelet preparation or are time-consuming. A simple, gentle technique for isolating concentrated platelet preparations for experimental or clinical use is reported. Freshly drawn whole blood was spun over a commercially available density gradient medium for 30 minutes. The mononuclear cell layer (which also contains most of the platelets) was collected and nucleated cells were pelleted by centrifugation. The recovery of platelets was about 60%. Contamination with leukocytes was less than 1%, and the platelet concentration was about 130% of blood concentration. Higher concentrations can be obtained if more whole blood is layered onto the Mono-Poly Resolving Medium (MPRM; Flow Laboratories, McLean, VA). About 10% of the platelets expressed the activation marker GMP-140 by flow cytometric analysis. They could be activated by thrombin so that 70% to 90% of the platelets expressed GMP-140. Thus, this technique can rapidly and easily yield a functionally intact platelet preparation. This preparation can be purified again if needed. No specialized skills or equipment are needed. A significant advantage of the method is that platelets can be obtained from thrombocytopenic patients in final concentrations that are high enough to use for platelet function testing.

Blood Platelets↗
PubMed1996

Centrifugation of bovine oocytes for nuclear micromanipulation and sperm microinjection.

The reproductive biotechnologies of intracytoplasmic sperm microinjection, nuclear transfer and DNA microinjection require the visualization of cytoplasmic components and nuclei of oocytes and early embryonic cells. Bovine oocytes were matured and fertilized in vitro, and then centrifuged at the germinal vesicle, metaphase II and pronuclear stages and at syngamy. These (n = 536) were examined using light and transmission electron microscopy. The organelles stratified in five distinct zones in a consistent pattern in both oocytes and zygotes, though relative fractions changed in organelle composition after fertilization. These comprised a centripetal lipid zone, below which was a vesicular zone, then a supra-equatorial band of smooth endoplasmic reticulum (SER), a clear zone and a centrifugal mitochondrial zone. Cortical granules were located peripherally, single or clumped together, in the clear and mitochondrial zones. The nuclei were usually found associated with the SER or Golgi membranes, and chromatin was clumped at one pole within the nucleus. The maturation spindles were often located beneath the oolemma in all zones, while the first mitotic spindle was usually located in the clear zone. Some of the oocytes were activated by centrifugation and completed maturation.

Animals↗