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PubMed2000

The isolated blood and perfusion fluid perfused heart.

The isolated heart is deservedly one of the most popular experimental models in cardiovascular research, both in terms of cost and the quality and quantity of data it provides. However, it is a deceptively simple model, capable of throwing many problems in the path of the inexperienced or unwary perfuser. The following article discusses the advantages and disadvantages of the various types of isolated heart perfusion (Langendorff and working; blood and buffer perfused). We attempt to give an insight into the many factors which must be taken into consideration when first establishing these preparations, we identify the range of indices that can be measured and the potential pitfalls which, with a little care, can be readily avoided.

Animals↗
PubMed1975

Ultrastructural demonstration of phosphatases by perfusion fixation followed by perfusion incubation of rat liver.

An alternative to previous methods (tissue chopper, frozen sections) for the ultrastructural demonstration of phosphatases is described. The present approach is based on a short vascular perfusion of rat liver with glutaraldehyde through the inferior caval vein, followed by vascular perfusion incubation with a medium containing the enzyme substrates. The effect of glutaraldehyde on three different types of phosphatases was investigated, namely a lysosomal enzyme (acid phosphatase) a tightly bound microsomal enzyme (G6Pase) and a loosely bound microsomal enzyme (IDPase). It is demonstrated that by perfusion with glutaraldehyde for three minutes good cellular morphology is obtained and that 50-60% of the initial activity of glucose-6-phosphatase, inosine-diphosphatase and acid phosphatase remains. The localization and deposition of G6Pase activity were distinct and observed throughout the endoplasmic reticulum and the nuclear envelope. For acid phosphatase, the reaction product was confined to various types of lysosomes including presumed autophagic vacuoles. No signs of enzyme diffusion were noted. The present approach seems to offer some advantages: it is simple and requires no extra equipment, penetration of the fixative and incubation enzyme medium is good, and finally freeze artifacts are avoided.

Acid Phosphatase↗
PubMed1992

Maximisation of perfusion systems and process comparison with batch-type cultures. Maximisation of perfusion cultures.

A comparison of cell yields and monoclonal antibody productivity from the same hybridoma has been made in a wide range of cell bioreactors including both batch and continuous perfusion cultures. The most productive systems were based on porous microcarriers in fixed and fluidised beds which can be operated with a high degree of efficiency and reliability from the physico-chemical engineering point of view. Further improvements should be possible by improving the physiological environment in dense cell cultures, as indicated by the preliminary studies that are described. These include experimental data showing the relationship between monoclonal antibody production rates with glucose, glutamine, ammonia, and oxygen levels in microporous beads. The results strongly indicate that perfusion processes that are scaleable in both volume and cell density can significantly reduce production costs. Manufacturers of biologicals from animal cells now have a choice between the proven batch-type processes and reliable perfusion systems based on microporous beads.

Animals↗
PubMed1986

Thallium myocardial perfusion scintigraphy: influence of perfusion, scatter, and photon energy on the detection of lesions.

Experimental studies have been made of lesion detectability in myocardial perfusion studies using thallium-201. A series of images (AP view) was generated mathematically. Images were simulated with 100k, 200k, and 300k counts for the complete image and either non- or half-perfused lesions (lesion volume 1-6 1/2% of total myocardial volume). In addition thallium images were generated excluding scatter; images were also generated with a radiopharmaceutical using 140, 184 and 296 keV, assuming the biological distribution of thallium. All images were interpreted by seven observers. Count density did not affect the true-positive fraction of the interpretations. However, the false-positive fraction decreased significantly when count density increased. Lesions located far from the camera were detected worst. The intraobserved variability was largest for interpretation of these regions. Small lesions were poorly detected. Perfusion of lesions significantly affected their detection, independent of the count density, lesion volume, and lesion location. Scatter did not affect the detection of lesions. No significant difference was found when the performance of the observers was compared for 80 keV and 140 keV. The use of 184-keV and 296-keV photons resulted in a lower detection rate probably because of the use of a high energy collimator.

Coronary Circulation↗
PubMed1995

Retrograde cerebral perfusion does not perfuse the brain in nonhuman primates.

BACKGROUND: Recently retrograde cerebral perfusion (RCP) has been advocated as an alternative to complete circulatory arrest during aortic arch surgery. METHODS: In 19 baboons, we compared brain protection using hypothermic circulatory arrest or RCP. Animals were placed on cardiopulmonary bypass, cooled to 18 degrees C, underwent 1 hour of circulatory arrest or RCP, and were reperfused for 3 hours. Biochemical variables, cerebral blood flow (colored microsphere technique), and brain histology were assessed. RESULTS: Release of the brain-specific ischemic marker CK-BB was similar in both groups (peak values, 123 +/- 97 U/L in the circulatory arrest group and 164 +/- 88 U/L in the RCP group; p > 0.05), as were the arteriovenous differences in glucose uptake and lactate production (p > 0.05). During RCP, significant brain flow could not be detected (0.5 +/- 0.5 mL.min-1 x 100 g-1). About 90% of the blood was shunted to the inferior caval vein, and an equilibrium in circulating microspheres was found between RCP inflow and caval vein outflow. Less than 1% of the RCP inflow returned to the aortic arch. Histologic signs of brain damage were minimal in both groups, although slightly more glial edema was found in the RCP group. CONCLUSIONS: These data suggest that in nonhuman primates, retrograde cerebral perfusion does not perfuse the brain because of venovenous shunting.

Animals↗
PubMed1978

[Pulmonary gas exchange model: influence of the heterogeneity of distribution on the ventilation-perfusion and diffusion-perfusion ratios of oxygen transfer].

The purpose of this pulmonary gas exchange model is to study the effect produced by an inhomogeneous distribution of the ventilation-perfusion (V A/Q) and diffusion-perfusion (D/Q) ratios on the oxygen transfer. We calculate partial pressures of oxygen and carbon dioxide in venous blood, in capillary blood and alveolar gas of each element as the unique solution of a non-linear system, the parameters of which are the local values of ventilation, perfusion and diffusion. We show that an inhomogeneous distribution of any ratio leads to a decrease of the mixed arterial concentration of oxygen and that the greater the inhomogeneity, the greater the decrease. We show by numerical stimulation that if two inhomogeneities (V A/Q) and (D/Q) are associated, the oxygen arterial concentration decrease is rather less important if the diffusion-ventilation ratio has a distribution almost homogeneous, i.e. if the V A/Q and D/Q inhomogeneities are almost identical.

Carbon Dioxide↗
PubMed1991

Conversion of p-tyrosine to p-tyramine in the isolated perfused rat kidney: modulation by perfusate concentrations of p-tyrosine.

We used the isolated perfused rat kidney to evaluate the role of renal decarboxylation of p-tyrosine as the source of urinary p-tyramine. Kidneys were perfused with concentrations of p-tyrosine ranging from 0.02 mM to 2.0 mM. p-Tyramine was measured by a sensitive and specific radioenzymatic assay. An increase in the perfusate concentration of p-tyrosine resulted in a significant increase in p-tyramine production that was blocked by the addition of NSD-1015, an inhibitor of aromatic-1-amino decarboxylase (AADC). We conclude p-tyrosine is the precursor for the renal production of p-tyramine, renal AADC catalyzes the formation of urinary p-tyramine, synthesized p-tyramine is predominantly excreted in the urine, and p-tyramine synthesis is modulated by the arterial delivery of p-tyrosine to the kidney.

Animals↗
PubMed1983

Neuropeptide Y (NPY) reduces myocardial perfusion and inhibits the force of contraction of the isolated perfused rabbit heart.

Isolated rabbit hearts, perfused under constant pressure (Langendorff technique) were used to study the effect of neuropeptide Y (NPY) on heart rate, force of heart contraction and rate of myocardial perfusion. No significant net change in heart rate was noted. A dose-dependent negative inotropic effect was consistently demonstrated which was characterised by slow onset and was often preceded by a transient positive inotropic response. Addition of small doses of NPY resulted in a prompt reduction in flow of the perfusate through the coronary vasculature. Since NPY is present locally in cardiac nerves, these effects may have physiological importance.

Acetylcholine↗
PubMed1982

Dosimetry in isolation perfusion of the limbs by assessment of perfused tissue volume and grading of toxic tissue reactions.

The optimal single dosage of melphalan in isolation perfusion of the limbs for malignant melanoma was assessed. For this purpose a method to determine the volume of the isolated region in the individual patient and a grading system for the reaction of the normal tissues were introduced. A strictly standardized pharmacosurgical routine was developed that permitted an analysis of the correlation between dosage and the grade of toxic reaction in 90 perfusions. The optimal dosage of a cytostatic drug was considered to be the highest amount tolerated at an acceptable risk. Melphalan at 10 mg/l perfused tissue was determined as the likely optimum. This dose provoked remarkably little variation in toxicity, all reactions falling within a safe range. No exception to the applicability of this dosage was encountered.

Arm↗
PubMed1994

Effect of perfusate pH on reduction of quinidine capillary permeability by albumin in isolated perfused rat heart.

It has been suggested that albumin reduces quinidine capillary permeability (PS) in the single-pass perfused heart preparation by reducing paracellular transport of quinidine ions. Using this preparation, we examined the effect of albumin (0.1 per cent) on quinidine PS at perfusate pH's of 7.1 and 7.9 during uptake of quinidine (19 microM) and also during washout of the drug using a randomized design. Quinidine PS was approximately 16 ml/min/g heart at pH 7.9 and was not altered by the presence of albumin in perfusate. At pH 7.1, in the absence of albumin, quinidine PS was also 16 ml/min/g, but in the presence of albumin (0.1 per cent) PS was reduced significantly to approximately 5 ml/min/g (P < 0.001). In the absence of albumin PS was the same at pH 7.1 and 7.9 in spite of a greater degree of ionisation of quinidine at pH 7.1. This suggests that there is significant uptake of ionised quinidine at pH 7.1. The greater effect of albumin on PS at pH 7.1 supports the hypothesis that albumin reduces paracellular transport of quinidine ions.

Animals↗
PubMed1975

[Selective coronary perfusion scintigraphy. II. Abnormal coronary perfusion pattern (author's transl)].

Integrated assessment of selective coronary angiorgrams and perfusion scintigrams of 86 patients with coronary-artery insufficiency revealed anatomical and topographical details which could not be achieved with one of these methods alone. Pathological perfusion patterns were quite different from normal ones. The functional effects of single, centrally located stenoses, of more peripheral and multiple narrowings, as well as of anastomoses and collaterals were adequately demonstrated by selective coronary perfusion scintigraphy and prevented diagnostic errors. It is, therefore, an additional aid in the diagnosis of coronary-artery disease.

Collateral Circulation↗
PubMed1994

Matched ventilation-perfusion becomes mismatched ventilation-perfusion lung imaging after resolution of carcinoma of the bronchus.

Airway occlusion results in regional alveolar hypoxia followed by reflex hypoxic pulmonary arterial constriction. Ventilation-perfusion imaging demonstrates matched defects. In the two patients reported, occlusion of the main bronchus by bronchogenic carcinoma was resolved--in one case by radiation therapy, and in the other by removal of the tumor mass in the bronchus by bronchial slitted biopsy. The initially matched ventilation-perfusion findings became mismatched. While ventilation was presumed to be normal, pulmonary perfusion was not completely reversed.

Aged↗
PubMed1999

Identification of kidneys subjected to preretrieval warm ischemic injury by simultaneous monitoring of glomerular filtration and perfusate flow during hypothermic perfusion preservation.

BACKGROUND: Historically, ex vivo physiological evaluation of cadaveric renal allografts has been limited to assessing perfusate flow (PF) during hypothermic perfusion preservation (HPP). Using a small animal model, we have previously described a method for continuous monitoring of glomerular filtration rate (GFR) during HPP. Our study was undertaken to determine if monitoring GFR and PF during HPP distinguished kidneys subjected to preretrieval warm ischemic (WI) injury more reliably than PF alone. METHODS: In situ WI was induced in Lewis rats (n=10) by extrinsic occlusion of the suprarenal aorta for 30 min. After in situ cold perfusion and retrieval, the left kidney underwent 16 hr of HPP. Nonischemic (NI) control kidneys (n=10) were retrieved in the absence of suprarenal aortic occlusion. Longitudinal changes in PF, GFR, and filtration fraction (FF) during HPP were compared in WI versus NI kidneys (FF=GFR/PF x 100%). RESULTS: PF remained the same in both cohorts throughout HPP. GFR, however, increased to a significantly greater degree in WI versus NI kidneys during the first 4 hr of HPP (713+/-401 vs. 26+/-23%, respectively) (P<0.05). The increase in FF at 4 hr was 1203+/-696% in the WI kidneys versus 83+/-46% in the NI controls (P<0.05). CONCLUSIONS: In contrast to PF alone, measurement of both PF and GFR distinguished kidneys subjected to pre-WI from NI controls. The data provide a means to determine if monitoring of both GFR and PF during HPP will predict short- and long-term renal allograft function more reliably than PF alone.

Animals↗
PubMed1987

Characterization of alpha-adrenoreceptor mediated reduction of flow in a saline-perfused constant pressure model of rat perfused hindquarters.

An analysis of vasopressor responses elicited by alpha-adrenoreceptor agonists and KCl in a constant pressure saline perfused model of rat perfused hindquarters is presented. Using methoxamine (alpha 1), cirazoline (alpha 1), B-HT 920 (alpha 2) and St 587 (alpha 1) as alpha-adrenoreceptor agonists, and rauwolscine and prazosin as selective alpha 2- and alpha 1-adrenoreceptor antagonists, respectively, it is concluded that alpha-adrenoreceptor mediated vasoconstriction in this model is mediated by the alpha 1-subtype. Pretreatment of the animals with reserpine (i.p., at various doses and dose-regimens) or addition of angiotensin II (10(-9)-10(-6) M), PGF2 alpha(10(-8) & 5 X 10(-7) M) or KCl (2 & 4 X 10(-3) M), or changing [Mg2+] did not significantly affect the potency or intrinsic activity of the alpha-adrenoreceptor agonists studied. Experiments with the calcium entry blocking drugs gallopamil and nifedipine, or deletion of CaCl2 from the perfusion solution, did not reveal a contribution from the influx of extracellular calcium towards alpha 1-adrenoreceptor-mediated reduction of flow in RPH. KCl-induced vasoconstriction in RPH was found to be sensitive to the blockade of calcium entry by calcium entry blocking drugs, but not to alpha-adrenoreceptor-blocking drugs and calmodulin antagonist. It is concluded that the model described here is of potential value in the characterization of alpha-adrenoreceptor antagonists and calcium entry blocking drugs under in vitro conditions in resistance vessels of the rat.

Acid-Base Equilibrium↗
PubMed1986

The deuterium isotope effect on ethanol metabolism in perfused rat liver: effect of reversed perfusion on ethanol and oxygen uptake.

Livers from rats fasted for 24 hr were subjected to nonrecirculating perfusion with Krebs-Ringer bicarbonate solution containing 10 mM ethanol. The deuterium isotope effect was measured using (1-R)-[1-14C,1-2H]ethanol. A value of 2.57 +/- 0.09 (SD) was obtained independent of the direction of perfusion. Oxygen uptake and ethanol metabolism in contrast were significantly increased when reverse perfusion (i.e., from vena cava to vena portae) was used. The magnitude of the isotope effect indicates that contribution from microsomal ethanol-oxidizing system if this is the only supplementary system is 9.8% under the experimental conditions. At high ethanol concentrations, the contribution would approach 18%. Equal activities of microsomal ethanol-oxidizing system and catalase under the experimental conditions would mean that both contribute 7.3% of the total ethanol metabolism. At high ethanol concentrations (80 mM), however, catalase will be 6.8% and microsomal ethanol-oxidizing system is calculated to 13.3%. Preliminary experiments with rats pretreated with phenobarbital showed no change in the isotope effect or in the rate of ethanol metabolism, but a 40-50% increase in oxygen consumption. The acetaldehyde concentration in the effluent medium was below 1 microM.

Alcohol Dehydrogenase↗
PubMed1989

The use of a cell-free perfusate in the perfused rat hindquarter: methodological concerns.

The viability of using a cell-free perfusate in a rat hindlimb preparation to assess skeletal muscle glycogenesis was investigated. A perfusate containing 10 mM glucose and 10 microCi (1 Ci = 37 GBq) of D-[5-3H]glucose was recycled for a 60-min period. In agreement with other studies using more complex media, oxygen uptake of the preparation indicated adequate tissue oxygenation (8 mumol.min-1.g-1). Skeletal muscle fiber type heterogeneity in basal glycogen synthesis from glucose was shown (slow oxidative greater than fast oxidative glycolytic greater than fast glycolytic fibres). Insulin (4.2 mU/mL) markedly stimulated glycogenesis from D-[5-3H]glucose in the soleus (slow oxidative fiber), red gastrocnemius (fast oxidative glycolytic fiber), and white gastrocnemius muscles (p less than 0.05). A recent report indicates that tissue edema in this preparation did not affect insulin responsiveness of the tissue. In contrast, our observations indicate that glucos uptake was enhanced by insulin when edema was absent (p less than 0.05), but not when edema was present (p less than 0.05). In addition, the presence of tissue edema negated insulin-mediated glycogenesis in slow oxidative and fast oxidative glycolytic muscle (p less than 0.05 compared with control) but not in fast glycolytic muscle (p less than 0.05). These data warrant caution when using a cell-free media in the perfused rat hindquarter; however, in the absence of edema, normal responses of glucose metabolism are observed.

Animals↗
PubMed2003

Whole-brain CT perfusion measurement of perfused cerebral blood volume in acute ischemic stroke: probability curve for regional infarction.

PURPOSE: To determine the probability curve for regional cerebral infarction as a function of percentage normalized perfused cerebral blood volume (pCBV) in patients with acute ischemic stroke. MATERIALS AND METHODS: The authors retrospectively analyzed whole-brain computed tomographic (CT) perfusion scans from 28 patients with acute stroke (<6 hours) due to major arterial occlusion, without intracranial hemorrhage. Each patient had a positive follow-up CT scan 1-4 days later, without interval thrombolysis. Normalized pCBV, expressed as a percentage of contralateral normal brain pCBV, was determined in the core infarction and in regions just inside and outside the boundary between infarcted and noninfarcted brain. These regions were dichotomized into infarcted (core and inner band) and noninfarcted (outer band) categories. Logistic regression analysis was then used to create a reference curve of probability of infarction as a function of percentage normalized pCBV. RESULTS: Normalized pCBV values in the core, inner band, and outer band were 24.5% +/- 2.3, 36.3% +/- 2.4, and 72.1% +/- 2.4, with corresponding probabilities of infarction of .99, .96, and .11. The normalized pCBV at which the probability of survival reached .5 was 58.0% +/- 0.5. Sensitivity, specificity, and accuracy of the reference probability curve were 90.5% (209 of 231), 89.5% (212 of 237), and 90.0% (421 of 468), respectively. Negative and positive predictive values were 90.6% (212 of 234) and 89.3% (209 of 234), respectively. R2 was 0.73, and differences in perfusion between core and inner and outer bands were highly significant (P <.0001). CONCLUSION: A probability of infarction curve can help predict the likelihood of infarction as a function of percentage normalized pCBV.

Aged↗