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Biomedical subjects

R Best

Publications and source records attributed to R Best.

At least 37 records · Page 2Linked to original sources

Additional value of measurement of urinary cortisone and unconjugated cortisol metabolites in assessing the activity of 11 beta-hydroxysteroid dehydrogenase in vivo.

OBJECTIVE: It has been shown recently that 11 beta-hydroxysteroid dehydrogenase (11 beta-HSD) is expressed as at least 2 isozymes. In the liver, 11 beta-HSD1 converts cortisone to cortisol; in the kidney, 11 beta-HSD2 converts cortisol to cortisone. Conventional assessment of 11 beta-HSD activity in vivo has relied on gas chromatographic measurement of the ratios of conjugated cortisol and cortisone metabolites. However, these do not permit distinction between the tissue-specific activities of the enzymes and do not reflect all forms of 11 beta-HSD deficiency. In this report, we have assessed the usefulness of measuring unconjugated cortisol metabolites and free cortisol and cortisone in urine as indices of renal 11 beta-HSD activity in man. DESIGN: Six healthy male subjects established in sodium balance were given either glycyrrhetinic acid (170 mg t.d.s., to inhibit 11 beta-HSD2), carbenoxolone (100 mg t.d.s., to inhibit both 11 beta-HSD1 and 11 beta- HSD2) or both inhibitors in combination. MEASUREMENTS: Urinary electrolytes were measured and the concentrations of total and unconjugated urinary cortisol and its metabolites were determined by gas chromatography mass spectrometry. RESULTS: Glycyrrhetinic acid and carbenoxolone inhibited renal 11 beta-HSD2 to a similar degree, as judged by similar sodium retention. As previously reported, conventional measurement of ratios of total cortisol to cortisone metabolites were influenced to a greater extent by glycyrrhetinic acid (100-200% increase in ratio from baseline) than by carbenoxolone (< 30% increase). However, the effect of carbenoxolone was readily detected by measurement of urinary unconjugated cortisol/cortisone (130-480% increase of ratio from baseline) and also by measurement of ratios of unconjugated cortisol metabolites (60-130% increase). CONCLUSIONS: Measurement of free cortisol and cortisone in urine provides the most sensitive index of renal 11 beta-HSD activity. Measurement of total and conjugated urinary steroids is insensitive in circumstances where 11 beta-HSD activity in liver or elsewhere may be abnormal.

11-beta-Hydroxysteroid Dehydrogenases↗

Seasonal variation in glucocorticoid activity in healthy men.

Many endocrine systems are subject to seasonal variation. However, studies of the hypothalamic-pituitary-adrenal axis in man have been limited to patients with psychiatric illness with conflicting results. We studied 105 healthy men, age 24-33 yr, during a 15-month period that included two winters. We measured cortisol and its metabolites by gas chromatography/mass spectrometry in plasma and urine and the intensity of dermal blanching after overnight topical application of beclomethasone dipropionate. There were no differences between subjects studied during the two winter periods, but marked differences between subjects studied in winter and summer. In winter, 0900-h plasma cortisol concentrations were higher (73 +/- 10 ng/mL, n = 41 vs. 35 +/- 4, n = 25 in summer; P < 0.01), total cortisol metabolite excretion was lower (678 +/- 67 micrograms/mmol creatinine vs. 900 +/- 98; P < 0.05), the ratio of metabolites of cortisol to those of cortisone was higher (3.0 +/- 0.2 vs. 2.1 +/- 0.1; P < 0.01), and dermal glucocorticoid sensitivity was higher (7.2 +/- 0.4 arbitrary units vs. 5.6 +/- 0.5; P < 0.02). Although blood pressure and fasting insulin/glucose relationships were not measurably different between seasons, these correlated with dermal vasoconstriction and cortisol metabolite excretion rate. We conclude that plasma cortisol and tissue sensitivity to glucocorticoids are higher in winter, but cortisol production rate is reduced. This could be explained by a reduction in cortisol clearance rate: urinary free cortisol/cortisone ratios were not different but A-ring-reduced metabolites of cortisol were higher in winter, suggesting that conversion of cortisone to cortisol by hepatic 11 beta-hydroxysteroid dehydrogenase 1 is enhanced. It is an intriguing possibility that increased glucocorticoid activity contributes to the increased prevalence of disease during the winter.

Adult↗

Dexamethasone and 11-dehydrodexamethasone as tools to investigate the isozymes of 11 beta-hydroxysteroid dehydrogenase in vitro and in vivo.

Dexamethasone is used in the clinic to test the sensitivity of the hypothalamic-pituitary-adrenal axis to negative feedback. It has also been proposed that metabolism of dexamethasone might differentiate between the activities of the two isozymes of 11 beta-hydroxysteroid dehydrogenase (11 beta HSD1 and 11 beta HSD2). We have developed a gas chromatographic mass spectrometric assay for dexamethasone and 11-dehydrodexamethasone and have confirmed in vitro that dexamethasone is a substrate for 11 beta-HSD2 but not 11 beta-HSD1 (conversion to 11-dehydrodexamethasone 0.6 +/- 0.3% in homogenates of rat liver with NADP+ for 11 beta-HSD1, and 29.4 +/- 10.3% and 40.0 +/- 2.0% in homogenates of rat and human kidney respectively with NAD+ for 11 beta-HSD2). However, we have also made the novel observation that 11-dehydrodexamethasone is a substrate for both isozymes (conversion to dexamethasone 65.0 +/- 20.4% for 11 beta HSD1 and 53.5 +/- 20.8% and 69.0 +/- 4.5% for 11 beta HSD2, rat and human respectively). In healthy humans, the concentrations of 11-dehydrodexamethasone in plasma after an intravenous bolus of dexamethasone were less than 10% of those of dexamethasone, and 11-dehydrodexamethasone was detected (at 0.8-65.0 nM) in plasma from only 11 of 20 subjects at 0900 h on the morning after oral dexamethasone (0.1-1 mg taken at 2400 h). Concentrations of 11-dehydrodexamethasone did not correlate with the degree of suppression of plasma cortisol. Thus dexamethasone is not useful in differentiating the activities of the isozymes of 11 beta-HSD in vivo and variations in 11 beta-HSD activity do not explain the interindividual variability in suppression of plasma cortisol by low doses of dexamethasone.

11-beta-Hydroxysteroid Dehydrogenases↗

Reciprocating gait orthosis powered with electrical muscle stimulation (RGO II). Part I: Performance evaluation of 70 paraplegic patients.

Seventy paraplegics were fitted with an improved Reciprocating Gait Orthosis powered with or without (low-level injury) electrical stimulation of the thigh muscles (RGO II) as a secondary rehabilitation phase after the acute period. The patients comprised a broad cross-section of the paraplegic population applying for medical services and varied in age from 16 to 55 years, time since injury ranging from less than 1 to 15 years, injury levels ranging from C-6/7 to T-11/12, and varying levels of spasticity, contractures, scoliosis and other related medical and physiologic problems. The success/failure ratio was dependent on the injury level, which was 1:1 for paraplegics with injury level at C-6/7; 1.67:1 for those with injury of T-1/3; and about 4:1 for paraplegics with injury level from T-3 to T-12. Lack of motivation and medical problems unrelated to the RGO II treatment were the primary reasons for failure. The duration of treatment (outpatient service three times per week) ranged from 2 to 48 weeks (mean: 16). Forty-one patients who completed the RGO II rehabilitation and were sent home with the orthosis for independent use (for at least 6 months and up to 3 years) were surveyed by a staff member for analysis of the meaning and impact of the RGO II on the patient's life and health, and potential problems. It was shown that 80.5% of the 41 patients were regular users and 19.5% were non-users. Thirty-eight of the 41 patients declined an offer to return the RGO II equipment for a full refund, while three patients were willing to return the orthosis. It was concluded that the RGO II is viable orthosis for restoring standing and limited walking in paraplegics while providing sufficient function, safety, and reliability. The most appropriate patients for the use of such an orthosis consist primarily of those with T-3 to T-12 injury level and good motivation, although highly selected patients with higher injury levels also can benefit from its use. Regular use of the RGO II, even for exercise only, had a general positive impact on the patients' health and outlook.

Adolescent↗

Reciprocating gait orthosis powered with electrical muscle stimulation (RGO II). Part II: Medical evaluation of 70 paraplegic patients.

Medical evaluation was performed on a group of paraplegics who were trained to walk with the Reciprocating Gait Orthosis powered with electrical muscle stimulation (RGO II). The evaluation included changes in spasticity, cholesterol level, bone metabolism, cardiac output and stroke volume, vital capacity, knee extensors torque, and heart rate at the end of a 30-meter walk. After an average of 14 weeks of training during which patients walked for 3 hours per week, significant reductions in spasticity, total cholesterol and low-density lipids, hydroxyproline/creatinine ratio, and increased knee extensor torque were evident. The data also showed that improvements occurred in the calcium/creatinine ratio, serum calcium and alkaline phosphatase levels, cardiac output and stroke volume, and vital capacity, yet these improvements were not statistically significant. The final heart rate at the end of a 30-meter walk showed that the RGO II required only a moderate level of exertion, which was found to be the lowest among the other mechanical or muscle stimulation orthoses available to paraplegics. It was concluded that the limited but reasonable level of functional regain provided by the RGO II is associated with a general improvement in the paraplegic's physiological condition if used for a minimum of 3 to 4 hours per week.

Bone Resorption↗

Accuracy of formula-derived creatinine clearance in paraplegic subjects.

The usefulness of formula-derived creatinine clearance (CC) is not well established among the paraplegic population. We prospectively collected 24-hour-urines from paraplegic subjects, obtaining 81 collections from 42 patients over 27 months. After validation, 57 collections were used to obtain measured CC. CCs were also estimated using several formulae. Measured CC and estimated CC were then compared. In the case of the Cockroft-Gault formula, the overall correlation was excellent, with an r value of 0.88 and p = 0.0001. In our study 40, 70, and 93% of the estimated CCs were within 10, 20, and 33% of the measured CCs. These numbers are similar to the ones reported by Cockroft and Gault. Also, the data on measured creatinine clearance in paraplegics compared favorably with parallel data obtained from nonparaplegics (91 24-hour-urine collections, 65 validated). We conclude that the Cockroft-Gault formula (and other formulae) estimate creatinine clearance as accurately in the paraplegic population as they do in the nonparaplegic population.

Adult↗

Synchronous 20 Hz rhythmic activity in hippocampal networks induced by activation of metabotropic glutamate receptors in vitro.

High frequency synchronized neuronal activity plays an important role in spatiotemporal processing of sensory inputs and possibly cognitive function. This activity, in the so called beta and gamma electroencephalographic frequency range is found in cortical areas and in the hippocampus. Here we show that activation of metabotropic glutamate receptors by the agonist 1-aminocyclopentane-(1S,3R)-dicarboxylate evokes synchronous 20 Hz rhythmic activity in rat hippocampal slices. Quisqualate was more potent than 1-aminocyclopentane-(1S,3R)-dicarboxylate which suggests involvement of either the metabotropic glutamate receptor subtypes I, V or both. The 20 Hz rhythmic activity was different from the theta-rhythmic effect induced by the cholinergic muscarinic receptor agonist carbachol which showed a dominant frequency of 9.0 +/- 0.5 Hz. The 20 Hz rhythmic activity is correlated with an increased firing rate of pyramidal neurons. The 20 Hz rhythmic activity was reduced by the GABA(A) receptor antagonist bicuculline, thus interneuron activity plays a major role in the 20 Hz oscillations. The 20 Hz rhythmic activity induced by activation of metabotropic glutamate receptors represents a new aspect of synchronized activity in the hippocampus and may be involved in the binding and representation of converging sensory inputs.

Animals↗

Increased vasoconstrictor sensitivity to glucocorticoids in essential hypertension.

Glucocorticoids raise blood pressure but were thought not to play a pathophysiological role in essential hypertension when it was demonstrated that cortisol secretion rates and circulating concentrations are normal in this disease. However, recent observations suggest that increased tissue sensitivity to cortisol, mediated by either abnormal glucocorticoid receptors or impaired inactivation of cortisol by 11 beta-dehydrogenase, may allow cortisol to raise blood pressure despite normal circulating concentrations. We studied 11 patients with essential hypertension and 11 matched normotensive control subjects. Dermal vasoconstriction after topical application of both cortisol (16 +/- 4 versus 32 +/- 5 U, control subjects versus hypertensive patients; P < .02) and beclomethasone dipropionate (75 +/- 10 versus 100 +/- 7 U; P < .05) was increased in the hypertensive patients. Hypothalamic-pituitary glucocorticoid receptor sensitivity was normal, as judged by basal cortisol secretion rates and suppression of plasma cortisol during sequential overnight dexamethasone suppression tests. 11 beta-Dehydrogenase activity was impaired in essential hypertension, as judged by prolonged half-lives of [11 alpha-3H]cortisol (44 +/- 4 versus 58 +/- 4 minutes, control subjects versus hypertensive patients; P < .02). However, this did not correlate with the dermal vasoconstrictor response. We conclude that vasoconstrictor sensitivity to glucocorticoids is increased in essential hypertension and that this may initiate and/or sustain the increased peripheral vascular resistance that characterizes this disease. The mechanism of increased sensitivity remains uncertain, but it will be important to establish whether it relates to genetic abnormalities of the glucocorticoid receptor that have been observed in animal models and young individuals who are predisposed to essential hypertension.

Administration, Oral↗

Force-velocity relations of nine load-moving skeletal muscles.

The relationship between maximal velocity and load was studied in nine muscles of the cat's hind limb using a technique in which the initial and final muscle lengths are determined by equilibrium of a suspended mass and the muscle's passive and active forces elicited by tetanic stimulation. The maximal velocities of shortening during contraction under each of various loads was used to fit a Hill model using the least-squares method. It was shown that different muscles varied significantly in their ability to generate maximal velocity over a range of loads. The tibialis anterior muscle generate the highest velocity (28.4 cms-1), whereas the tibialis posterior generated the lowest maximal velocity (4.2 cms-1). In general, muscles with predominantly fast twitch fibres and with the largest elongation/shortening range displaced the load at the highest velocities, as compared with muscles with predominantly slow twitch and short excursion range which respond with low velocities. The a/P0 ratio of Hill's equation, which defines the curvature of the force velocity, also varied widely, being most monotonic (0.927) for the soleus and the steepest (0.067) for the extensor digitorum longus, further suggesting that fibre composition is also highly influential on the force-velocity relations of the muscle.

Animals↗

Architecture-based force-velocity models of load-moving skeletal muscles.

A predictive model of muscle force-velocity relationships is presented based on functional architectural variables. The parameters of Hill's equation describing muscle force-velocity relationship of nine muscles were estimated by their relationships with variables extracted from the whole-muscle length-force relationship and the percentage of slow-twitch fibres. Specifically, the maximal unloaded velocity (Vo) was estimated through multiple linear regression, from each muscle's fibre composition and the shortening range through which each muscle could produce active force. The maximal isometric force (Po) was also extracted from each muscle's length-force relationship. The ratio of Hill's dynamic constanta to Po and b to Vo, which determines the degree of curvature of the relation, was determined solely by the percent of slow-twitch fibres. This model was verified by fitting it to experimental force-velocity curves of nine different muscles in the cat's hindlimb. It was found that reasonable fits of force-velocity curves would be obtained with correlation coefficient in the range of 0.61 to 0.92, with an average of 0.82. The model predicted that muscles with relatively long shortening ranges would achieve higher maximal velocity, and that muscles with higher percentage of slow-twitch fibres had less pronounced curvature and lower maximal velocity in their force-velocity relationships. RELEVANCE: The results have direct implications in the design of neuroprosthetic limb control systems, which use electrical stimulation to restore function to muscles paralysed from spinal cord injury. The designer is enabled to optimally calibrate the controller according to the predicted individual force-velocity curves of different muscles by using the length-tension curves and fibre composition data available in the literature.

Journal Article↗

Characterization of acute bone marrow graft rejection in T cell-depleted, partially mismatched related donor bone marrow transplantation.

The purpose of this study was to characterize the phenotype and clonality of the T cell population in patients who experience acute rejection (AR) following bone marrow transplantation (BMT) from a partially mismatched related donor (PMRD). Phenotypic analysis was performed using flow cytometry, assignment of donor/host lineage by cytogenetics or HLA-specific flow cytometry, and analysis of the T cell receptor (TCR) by reverse-transcriptase polymerase chain reaction (RT-PCR). We have previously reported the initial appearance in the blood of AR patients of host CD8+brightCD3low T cells that progressively express increasing amounts of CD3+ cells. We now report that this cell population can differentiate into either a cytotoxic T cell phenotype (CD3+CD8+HLA-DR+CD57-) usually associated with AR of grafts from matched unrelated donors or a suppressor T cell phenotype (CD3+CD8+CD57+HLA-DR-) usually associated with AR of grafts from matched sibling donors. Analysis of the TCR V beta subsets from two patients revealed sorted host CD3+CD8+ cells (purity 90-95%) from the first patient to express V beta 18 almost exclusively. In a second patient with late rejection (55 days post-BMT), the CD3+CD8+ cells were predominantly restricted to V beta 1, 5.1, 7, 9, and 18. Although CD3+CD8+ T cells are known to be associated with AR, cytotoxic and suppressor lineages in AR from the same type of BMT and clonal distribution of T cells in AR have not been reported. Preliminary results suggest that V beta expression in AR of PMRD grafts is restricted and host T cell phenotype may vary. Further studies will investigate whether specific mismatches correlate with specific V beta usage and/or host T cell phenotype.

Base Sequence↗

Isotonic length/force models of nine different skeletal muscles.

The isotonic length/force relationships of nine skeletal muscles in the cat's hindlimb were determined using electrical stimulation of the sciatic nerve branches. Large variability in the active, passive, total force patterns and elongation ranges was found. The lateral gastrocnemius (LG), medial gastrocnemius (MG), peroneus longus (PL), flexor digitorum longus (FDL), tibialis posterior (TP) and soleus (Sol) showed symmetric active force curves, whereas those of the extensor digitorum longus (EDL), tibialis anterior (TA) and peroneus brevis (PB) were asymmetric. The total force curves of the EDL, LG, MG, FDL, TP and Sol increased quasilinearly through the elongation range, whereas the PL and PB increased in a nonlinear fashion. The TA had an intermediate plateau. The ranges were generally asymmetric, with a longer shortening range than lengthening past the optimum length. A simple model of the active force was fitted to all except the MG, EDL and TA, which are complex, with at least two compartments. These were successfully fitted with a two-compartment model. The variabilities encountered in the various isotonic length/force curves confirm the need to represent muscles according to their architecture to account for the variety of properties exhibited, which reflect their adaptations to their functions.

Animals↗

Impact tolerances of the rigidly fixated maxillofacial skeleton.

A study was designed to determine how soon an athlete who undergoes rigid fixation of a facial fracture can return to full competition. The impact resistance of a rigidly fixated malar complex fracture was studied and compared with that of an intact malar complex. Twelve fresh human cadaver heads were used. A custom-designed impact device was used to deliver a blow of a specific energy to each intact malar complex. The subsequent fractures were rigidly fixated at three points using titanium miniplates and screws. A second impact of identical energy was delivered. The forces generated and the subsequent displacement of hard and soft tissues were recorded after each impact. It was concluded from this study that an impact to a rigidly fixated malar complex fracture produced less force and greater displacement of hard and soft tissues than an impact of identical energy to an intact malar complex. The potential for sustaining more severe maxillofacial injuries after an initial facial fracture should be seriously considered. The results suggest that sufficient time should be allowed for the bony healing of a facial fracture to occur, even after rigid fixation, before an athlete can resume full contact activities.

Age Factors↗

A computer program for motion analysis of single cardiac myocytes.

Single adult cardiac ventricular cells were prepared by collagenase perfusion of a rat heart. They were stimulated electrically in a perfusion chamber and their length changes were followed under a microscope. The motion was followed via a video camera and by a TV-line counting device and was recorded on-line by a personal computer. The program RECORD was used to calculate peak amplitude, base line drift and peak width at different peak heights allowing the determination of a number of variables of the cellular motion. The method was applied to drugs affecting the amplitude of contractions and the speed of relaxation. Results of beta-adrenergic stimulation, muscarinic inhibition and of the Ca(2+)-ATPase inhibitor cyclopiazonic acid (CPA) are shown. Besides its stimulatory effect on length, the beta-adrenergic agonist isoprenaline concentration-dependently shortened relaxation time. Carbachol reversed the increase in cellular shortening caused by isoprenaline in a concentration-dependent manner without fully reversing the shortened relaxation. CPA prolonged the return to diastole, presumably due to its inhibition of Ca(2+)-reuptake into the sarcoplasmatic reticulum.

Animals↗

The isometric length-force models of nine different skeletal muscles.

The length-force relations of nine different skeletal muscles in the hindlimb of the cat were determined experimentally, with electrical stimulation of the sciatic nerve as the activation mode. It was shown that the active-, passive-, and total-force patterns varied widely among the muscles. The tibialis posterior (TP), medial and lateral gastrocnemius (MG, LG) and flexor digitorum longus (FDL) had a symmetric active-force curve, whereas the tibialis anterior (TA), peroneus brevis (PB), peroneus longus (PL), extensor digitorum longus (EDL), and soleus (SOL) had an asymmetric curve which exhibits about 25% of the maximal isometric force at extreme lengths. The SOL, EDL, and LG had a low-level passive force which appeared at short muscle length, whereas all other muscles exhibited initial passive force just before the optimal length. The total force was rising quasi-linearly for the SOL, whereas the other muscles exhibited an intermediate plateau about the optimal length. The LG and FDL had a substantial but temporary intermediate dip in the total force as the muscle was elongated past the optimal length. The elongation range of the various muscles also varied, ranging from +/- 15 to +/- 30% of the optimal length. The elongation range was symmetric for the FDL, LG, MG, TP, SOL, and EDL, and asymmetric for the PL, PB, and TA, being -12 to + 17%, -12 to + 17%, and -35 to + 12%, respectively. Two different models which incorporate muscle architecture were successfully fitted to the experimental data of the muscles except for the MG and TA. The architecture of these two muscles is highly nonhomogeneous and contains compartments with two pennation patterns or two different optimal lengths. New models, which add spatially and temporally the individual characteristics of each compartment of the muscles, were constructed for these two muscles. The new models demonstrated high correlation to the experimental data obtained from the MG and TA. It was concluded that the length-force relation varies widely among various skeletal muscles and is probably dependent on the primary function of the muscle in the context of integrated movement; this is a manifestation of architectural factors such as fiber pennation pattern and angle, cross-sectional area, ratio of muscle to tendon length, distribution of the fiber length within the muscle and compartmental pennation.

Animals↗

The primary structure of the hemoglobin of the Brazilian manatee (Trichechus inunguis, Sirenia).

The hemoglobin of the Brazilian Manatee (Trichechus inunguis, Sirenia) consists of one component. We present the primary structures of the alpha- and beta-chains which have been separated by chromatography on carboxymethyl-cellulose CM-52. The sequences have been determined by automatic Edman degradation with the film technique, using the native chains, tryptic peptides and the C-terminal prolyl-peptide obtained by acid hydrolysis of the Asp-Pro bond of the alpha-chains. Compared to the corresponding human chains we found 27 substitutions in the alpha- as well as in the beta-chains. Three heme contacts and four alpha 1/beta 1 contacts between the subunits are affected by exchanges. The hemoglobin of Trichechus inunguis is compared with those of Elephas maximus, Loxodonta africana, and Procavia habessinica and the monophyletic origin of the superorder Paenungulata is discussed.

Amino Acid Sequence↗

Insulin and glucose levels during CPR in the canine model.

Cardiopulmonary arrest and resuscitation produces tremendous physiological stress with resultant biochemical derangements. We undertook this study to determine insulin and glucose levels during cardiopulmonary arrest in the canine model. Baseline insulin and glucose levels were obtained from an ascending aortic arch catheter in six mongrel dogs. Ventricular fibrillation was induced by an electrical stimulus and ventilation was terminated. After five minutes of fibrillation, cardiopulmonary resuscitation (CPR) was initiated using external, mechanical CPR and a continuous epinephrine infusion at 5 micrograms/kg/min. Serum insulin and glucose levels were repeated 15 minutes after beginning CPR. Mean blood glucose 15 minutes after initiation of resuscitation (379 +/- 114 mg/dL) was significantly increased from prearrest levels (124 +/- 29 mg/dL, P less than .01). Mean serum insulin 15 minutes after initiation of resuscitation (11.3 +/- 3.3 microU/mL) was significantly decreased compared to prearrest levels (16.2 +/- 6.0 microU/mL, P less than .05). During ischemia, the myocardium becomes dependent primarily on glucose as a source of energy. Inappropriately low insulin levels during CPR may adversely affect an already compromised myocardial glucose metabolism. Further investigation is needed to determine the utility of insulin infusion during CPR.

Animals↗