Dialyzable Fraction Research Articles

Collection of nanoliter microdialysate fractions in plugs for off-line in vivo chemical monitoring with up to 2 s temporal resolution

An off-line in vivo neurochemical monitoring approach was developed based on collecting nanoliter microdialysate fractions as an array of “plugs” segmented by immiscible oil in a piece of Teflon tubing. The dialysis probe was integrated with the plug generator in a polydimethlysiloxane microfluidic device that could be mounted on the subject. The microfluidic device also allowed derivatization reagents to be added to the plugs for fluorescence detection of analytes. Using the device, 2nL fractions corresponding to 1–20ms sampling times depending upon dialysis flow rate, were collected. Because axial dispersion was prevented between them, each plug acted as a discrete sample collection vial and temporal resolution was not lost by mixing or diffusion during transport. In vitro tests of the system revealed that the temporal resolution of the system was as good as 2s and was limited by mass transport effects within the dialysis probe. After collection of dialysate fractions, they were pumped into a glass microfluidic chip that automatically analyzed the plugs by capillary electrophoresis with laser-induced fluorescence at 50s intervals. By using a relatively low flow rate during transfer to the chip, the temporal resolution of the samples could be preserved despite the relatively slow analysis time. The system was used to detect rapid dynamics in neuroactive amino acids evoked by microinjecting the glutamate uptake inhibitor l-trans-pyrrolidine-2,4-dicarboxylic acid (PDC) or K+ into the striatum of anesthetized rats. The resulted showed increases in neurotransmitter efflux that reached a peak in 20s for PDC and 13s for K+.

Involvement of NO in the inhibitory effect of Calotropis procera latex protein fractions on leukocyte rolling, adhesion and infiltration in rat peritonitis model

Aim of the study The latex of Calotropis procera has been used in the traditional medicinal system for the treatment of leprosy, ulcers, tumors, piles and diseases of liver, spleen, abdomen and toothache. It comprises of a non-dialyzable protein fraction (LP) that exhibits anti-inflammatory properties and a dialyzable fraction (DF) exhibiting pro-inflammatory properties. The present study was carried out to evaluate the effect of LP sub-fractions on neutrophil functions and nociception in rodent models and to elucidate the mediatory role of nitric oxide (NO). Material and methods The LP was subjected to ion exchange chromatography and the effect of its three sub-fractions (LP PI, LP PII and LP PIII) thus obtained was evaluated on leukocyte functions in the rat peritonitis model and on nociception in the mouse model. Results LP sub-fractions exhibit distinct protein profile and produce a significant decrease in the carrageenan and DF induced neutrophil influx and exhibit anti-nociceptive property. The LP and its sub-fractions produced a marked reduction in the number of rolling and adherent leukocytes in the mesenteric microvasculature as revealed by intravital microscopy. The anti-inflammatory effect of LP PI, the most potent anti-inflammatory fraction of LP, was accompanied by an increase in the serum levels of NO. Further, our study shows that NO is also involved in the inhibitory effect of LP PI on neutrophil influx. Conclusions Our study shows that LP fraction of Calotropis procera comprises of three distinct sets of proteins exhibiting anti-inflammatory and anti-nociceptive properties of which LP PI was most potent in inhibiting neutrophil functions and its effects are mediated through NO production.

In vivo-effect of intraadrenal nicotine and substance P application on rat adrenal medullary catecholamine secretion.

The present study was conducted to characterize in vivo the intraadrenal catecholamine (CA) secretion in rats. This was possible by using a microdialysis system (MDS) which mimics some properties of an artificial capillary. One end of this system was connected to a peristaltic pump, from the other end fractions were sampled at 5 min intervals. Concentrations of epinephrine (E) and norepinephrine (NE) in adrenal dialysate fractions were determined by HPLC electrochemical detection. Through this MDS nicotine was administered directly into the adrenal medulla of freely moving rats and the response of catecholamine release was determined. In the second part of the study the effect of exogenous substance P (SP) on spontaneous as well as on nicotine-stimulated CA release was investigated. Like nicotine, SP was administered directly into the adrenal medulla. At a flow rate of 25 microliter/min the transfer rates of CA and nicotine were approximately 1% whereas SP passed at a rate of 01.-0.2%. Under resting conditions CA release remained constant. In response to 2 x 10(-7) M nicotine (which resulted in local concentration of 2 x 10(-7) M), E and NE secretion increased 2.9 and 5.4-fold, respectively. However, due to an increased E response this difference attenuated with a later onset of the first stimulus. The higher concentrations of 10(-4) M resulted in 8.1 and 10.8-fold increases for E and NE. This latter response is clearly supraphysiologic and therefore the 2 x 10(-5) M concentration was used for further experimentation. CA secretion was stimulated with nicotine four times at 30 min intervals.(ABSTRACT TRUNCATED AT 250 WORDS)

Dialysability of Magnesium and Calcium from Hospital Duplicate Meals: Influence Exerted by Other Elements

Total and dialysable magnesium and calcium levels and corresponding dialysabilities were measured in duplicate meals (n = 108) during 36 consecutive days. The interaction exerted by other nutrients and energy on them was also performed. Total mean magnesium and calcium fractions of 113.9 +/- 98.3 and 337.2 +/- 278.9 mg/meal respectively, were found. The Mg and Ca levels supplied by meals are positively (p < 0.05) correlated with macronutrient contents (carbohydrates and proteins). The mean dialysable Mg and Ca fractions were 56.9 +/- 36.3 and 127.4 +/- 112.3 mg/meal (50.4 +/- 13.2 and 37.8 +/- 10.7% as dialysabilities, respectively). Total Mg and Ca levels are significantly correlated with corresponding element dialysabilities (p < 0.05). For both minerals, significant correlations between their total and dialysable fractions and between their dialysable level and dialysabilities were noted (p < 0.01). The mean Mg and Ca daily dietary intakes (DDI) were 341.7 +/- 68.0 and 1,011.6 +/- 424.4 mg/day, respectively. For Ca and Mg the existence of similarities in their behaviour in meals and absorptive processes has been found. Duplicate meals with raw vegetables are good sources of bioaccessible Mg. High Ca dialysability has been found in the analysed meals. The fish and products constitute a good source of bioaccessible Ca. Mg, Ca, zinc, and chromium levels enhanced significantly the Mg dialysability. The Ca dialysability rose significantly with dialysable Ca and chromium fractions (p < 0.05).

Speciation of aluminium, arsenic and molybdenum in excessively limed lakes

The possible existence of the potentially toxic oxyanions of Al (Al(OH) 4 −), As (HAsO 4 2−), and Mo (MoO 4 2−) was examined in excessively limed lakes. In-situ dialysis (MWCO 1 kDa) was performed in the surface and bottom waters of two excessively limed lakes (pH 7.1–7.7) and one acidic lake (pH ∼ 5.4). The dialysable metal concentrations were compared to the equilibrium distribution of species as calculated with the geochemical code Visual MINTEQ incorporating the CD-MUSIC and Stockholm Humic models for complexation onto colloidal ferrihydrite and dissolved organic matter. Arsenic and molybdenum in the excessively limed lakes were to a large extent present in the dialysable fraction (> 79% and > 92% respectively). They were calculated to exist as free or adsorbed oxyanions. Most of the Al was observed to reside in the colloidal fraction (51–82%). In agreement with this, model predictions indicated aluminium to be present mostly as colloids or bound to dissolved organic matter. Only a small fraction was modelled as Al(OH) 4 − ions. In most cases, modelled values were in agreement with the dialysis results. The free concentrations of the three oxyanions were mostly low compared to toxic levels.

Chromium and manganese levels in convenience and fast foods: In vitro study of the dialyzable fraction

Chromium and manganese presence was determined in a total of 170 samples of convenience and fast foods widely consumed in Spain. Electrothermal atomic absorption spectrometry was used as analytical technique. Reliability of the procedure was checked. Cr levels ranged from 0.01 to 1.10 μg/g (fresh weight of edible portion). The most elevated levels were found in beef and pork based-food, food with a greater content of spices and aromatic herbs, whole cereals, dry fruits and cheese. Mn levels ranged from 0.15 to 2.90 μg/g (fresh weight of edible portion). The most elevated levels of Mn were found in chicken and pork-based foods and sauces. Mean Cr and Mn dialyzable fraction estimated by in vitro assays ranged from 0.38% to 1.05% and from 7.75% to 15.60%, respectively. These findings revealed that certain convenience and fast foods contribute a considerable fraction of Cr and Mn dietary intake.

Use of DLE as treatment in murine endotoxic shock (41.54)

Abstract The LPS is a structural component of the external membrane of the Gram negative bacteria and is the most powerful of the inflammation inductors. High doses of LPS have as consequence a high production of inflammatory cytokines, which causes the endotoxic shock. The Dializable Leukocyte Extracts (DLE) is the dialyzable fraction with lower molecular weight to12 KDa which come from leukocytes lysates. In this work a model of endotoxic shock LPS was induced established in C57BL6 male 6 weeks of age, the dose for endotoxic shock induction was 130mg/kg mouse weight. The doses of 0.001ug and 0.0001ug mDLE showed protection against endotoxic shock death in 10% of the mice and the 0.01ug mDLE dose protected against of endotoxic shock death from 10 to 30% of the mice. Levels of TNF-alpha, IL-6, IL-12, MCP-1, IFN-gamma, IL-10 and TGF-beta were measured in serum of mice endotoxic shock induced and treated with mDLE, the groups in which LPS was administrated and then treated with 0.01ug, 0.001ug or 0.0001ug of mDLE showed smaller levels of inflammatory cytokines and more higher levels of antiinflammatory cytokines than the LPS treated control group. The splenic index was measured in the endotoxic shock induced and mDLE treated groups, the group LPS and the 0.001ug of mDLE treated showed the higher splenic index in comparison with the LPS control group. So the DLE can be use as a potential therapeutic agent as adjuvant in the treatment of sepsis.

Determination of daily dietary intake of chromium by duplicate diet sampling: In vitro availability study

Intake of chromium was estimated using a duplicate diet sampling method of 108 meals (36 breakfasts, 36 lunches and 36 dinners) from the restaurant of the Hospital of Motril (S.E. Spain), corresponding to 36 consecutive days. Total and dialyzable Cr levels were measured by a validated electro-thermal atomic absorption spectrometry (ETAAS) method. A mean Cr fraction of 26 ± 12 µg meal−1was found. The Cr uptake from meals was directly and significantly (p < 0.001) correlated with their macronutrient (carbohydrates, fibre and protein) content. Cereals and cereal by-products, legumes, dry fruits, meat, potatoes, dairy products and seafood are the primary sources of Cr. The mean Cr fraction dialyzed through dialysis tubing was 1.2 ± 1.1 µg meal–1(4.6 ± 3.8% as mean Cr dialysability). Cr intake for breakfasts was significantly lower (p < 0.001). A correlation between the logarithmic data of total and dialyzable fraction of Cr in meals (p = 0.020) was found and dialysis ratio enhancement and, therefore, bioavailability increased with total Cr. The dialysed element content present in meals was significantly correlated with fibre, protein, Fe, Na, I, F, sodium, ascorbic acid and vitamin A levels (p < 0.05). At Fe contents in meals higher than ≅7.5 mg meal−1, the net absorption of Cr decreased significantly. The mean Cr daily dietary intake (DDI) was 77 ± 17 µg day−1, which indicates that no adverse effects in relation to Cr nutrition (deficiency or toxicity) should occur in individuals from the area.

Antihypertensive Effect of Pleurotus nebrodensis in Spontaneously Hypertensive Rats

In this study, we examined the effects of Pleurotus nebrodensis on systolic blood pressure of spontaneously hypertensive rats. Single-dose and continuous-dose tests with sample diets made from the fruit body of the mushroom, P. nebrodensis were carried out on SHR and control rats. Sample diets included 6% dry powder of fruit body (6% dry powder), hot water extract, polysaccharide fraction, protein fraction, dialyzable fraction and non-dialyzable fraction. Polysaccharide and protein fractions were obtained by hot water extraction. The protein fraction was separated to the dialyzable fraction and non-dialyzable fraction by dialysis. In the single-dose test, protein fraction, hot water extract and polysaccharide fraction decreased systolic blood pressure. Blood pressure was lowered after administration for 2 h, and it returned to the pre-administration blood pressure after 48 h. In the continuous-dose test, spontaneously hypertensive rats were fed each of the diets for 16 weeks. The 6% dry powder group showed significantly inhibited elevation of blood pressure compared with the control group and there was no influence on total cholesterol and triglyceride levels. The non-dialyzable fraction showed suppression of increase in blood pressure from the start of the continuous oral administration. Effects on the rennin angiotensin system and renal function were also indicated. The antihypertensive action effect of P. nebrodensis can be expected to not only prevent but also to improve hypertension.

Total and dialyzable levels of manganese from duplicate meals and influence of other nutrients: Estimation of daily dietary intake

Both total and dialyzable Mn levels were determined in 108 duplicate meals during 36 consecutive days. Both mineral fractions were measured by a graphite furnace atomic absorption spectrometry (GFAAS) method previously optimized. A total mean Mn fraction of 1.03±0.49mg was found in the meals. The Mn supplied by the meals is directly and significantly (p<0.001) correlated with macronutrient content (carbohydrates, fibre and protein). The mean Mn fraction dialyzed through the dialysis membrane was 0.23±0.17mg (22.0±8.93% as bioaccessible fraction). The total and dialyzable Mn fractions found for breakfasts were significantly lower (p<0.001). Nevertheless, the Mn bioavailabilities expressed as the percentage of dialyzable element, were not significantly different among the three primary meals (breakfast, lunch and dinner). A significant correlation between the total and the dialyzable fraction of Mn in meals was found (p<0.001, r=0.78, r2=0.61). The dialyzed element fractions present in meals were significantly correlated mainly with carbohydrates, protein and several amino acid levels (p <0.01). Foods with higher carbohydrate and therefore energy contents, e.g. cereals, legumes, vegetables and fruits, would be primary sources of bioaccessible Mn in the diet. The bioaccessibility of Mn was only significant influenced by energy, carbohydrates and Se levels present in meals. The mean Mn daily dietary intake (DDI) was 3.05±0.61mgday−1.

Change in acetylcholine release from rat bladder with partial outlet obstruction

To investigate changes in acetylcholine release from the bladder of rats with partial bladder outlet obstruction (BOO), as partial BOO leads to hypertrophy and an alteration in the contractions of the detrusor smooth muscle, and acetylcholine plays an important role in urinary bladder contractions but there is little available information on acetylcholine release after BOO. Partial BOO was induced in adult female rats by ligating the proximal urethra over a 1 mm angiocatheter; sham-operated rats served as controls. The rats were killed 2 weeks, 3 and 6 months after induction of BOO. We investigated the contractions induced by carbachol, KCl (80 mm), ATP and electrical-field stimulation (EFS, 2.5-40 Hz), and collected the dialysate obtained from a microdialysis probe inserted into the muscle strips during EFS, and measured the amount of acetylcholine in the dialysate fraction by high-performance liquid chromatography with electro-chemical detection. S-100 immunohistochemical staining of the bladder preparations was used for histological examination in BOO and control rats. The bladder weight gradually increased after BOO. There were no significant changes in KCl-induced contractions throughout the experimental period in either group. There were no significant changes in carbachol-induced contractions until 3 months after BOO but there was a significant reduction at 6 months. ATP-induced contractions were significantly increased 2 weeks and 3 months after BOO. EFS-induced contractions were gradually reduced after BOO. Acetylcholine release from the bladder strips was not significantly different between the groups until 2 weeks after BOO. However, acetylcholine release in BOO rats was significantly decreased 3-6 months after BOO, being significantly lower than that of the control rats. In the histological study, the number of nerve fibres in the BOO rats was significantly lower than in the control rats. We suggest that the prolonged BOO caused a decrease in EFS-induced acetylcholine release and the number of nerves in the rat urinary bladder, which might contribute to bladder underactivity in BOO.

Estimate of the digestibility, assimilability and intestinal permeability of butyltins occurring in wine

An estimate of the digestibility and assimilability of butyltins occurring in contaminated wines (Port, red and white) was obtained by means of in vitro studies of gastrointestinal digestion. The influence of the wine matrix on the intestinal permeability was explored by studying the accumulation of butyltins in Caco-2 monolayers either when these species are dissolved in buffer only or in the dialysates of digested wines. Some important information about the fate of the butyltin compounds ingested from contaminated wines could be achieved. Only a very small fraction of the ingested DBT and TBT, the two most toxic forms, appear to be able to reach the epithelium as judged by the small dialyzable fraction found (<2%). This is probably independent from the food/drink matrix introducing these contaminants, since the influence of the involved enzymes appear to be dominant, especially for DBT and TBT. Additionally, the intestinal permeability of the three butyltins was also very low, the wine matrix possibly having a hindrance effect in a few cases.

Formation of dialyzable iron during in-vitro digestion and extraction of mycoprotein

Mycoprotein is the biomass of the fungus Fusarium venenatum. The ability of mycoprotein to enhance production of dialyzable iron following in-vitro digestion or extraction was studied, with the aim of identifying the components responsible. Digested mycoprotein, extracted mycoprotein and digested chicken breast muscle all produced about ten times as much dialyzable iron as the control. All of the dialyzable iron from mycoprotein was ferric. Essentially all the dialyzable iron passed through a 1 kDa molecular weight cut-off membrane. Iron binding components were extracted from mycoprotein using weak acid but not water. Extraction of mycoprotein produced both dialyzable and non-dialyzable components, which bound ferric iron. Analysis of the digests and the dialyzable fraction of acid extraction showed that they contained siderophores, which contributed to the formation of dialyzable iron.

Effect of l-DOPA on nitric oxide production in striatum of freely mobile mice

In Parkinson's disease, nitric oxide (NO) and other free radicals are thought to be involved in neuronal degeneration. Furthermore, l-DOPA is suggested to have a cytotoxic action on dopaminergic neurons. We studied 24-h NO production and the effect of l-DOPA on this in freely mobile mice using in vivo microdialysis. A microdialysis probe was implanted into the right striatum 12 h before the experiment. This dialysis probe was perfused with Ringer solution for 100 min, then with 20, 50, or 100 nM l-DOPA for 20 min, and finally with Ringer solution. Dialysate fractions were collected every 20 min for 4 h. Production of nitrite and total NO were significantly higher during daytime than during nighttime. Nitrate production was increased significantly by l-DOPA. NO production in the striatum appears to exhibit a diurnal rhythm and to increase with exposure to l-DOPA.

Management of the Cardinal Features of Andropause

There are several problems facing aging men, especially sexual dysfunction, hypogonadism, and psychologic changes. This constellation of changes is sometimes referred to as "manopause" or "andropause." Unlike the dramatic changes in the hormonal milieu occurring during menopause in women, the age-related changes in reproductive hormones of men are subtle and occur gradually throughout the years of mature life. It has been estimated that circulating testosterone (T) declines longitudinally from age 19 at an average rate of 1% per year. The free or dialyzable fraction of serum T and the bioavailable (the sum of free fraction and loosely bound to albumin fraction) T decline more rapidly with age. Although the essential role of androgens in reproductive tissue development and emergence of secondary sex characteristics is well known, their role in adult sexual function seems to be primarily facultative. The effect of T on the central nervous system extends beyond sexual behavior. T has been shown to alter mood, memory, ability to concentrate, and the overall sense of vigor and well being that may interact with a host of other psychologic changes associated with aging. Disordered erectile function is not generally an endocrine problem but rather vascular, neurologic, and psychogenic in origin. It also may be the first sign of systemic vascular disease. The clinical management of andropause requires an individualized approach. In some men, the main problem may be psychologic, whereas in others, hypogonadism may play an important role. Many with erectile failure, suffer silently regardless of its etiology. In this review, we suggest some practical guidelines for the management of these conditions.

Effects of Caffeine, Halothane, and 4-Chloro-m -cresol on Skeletal Muscle Lactate and Pyruvate in Malignant Hyperthermia–susceptible and Normal Swine as Assessed by Microdialysis

Skeletal muscle fibers from malignant hyperthermia (MH)-susceptible humans and swine are markedly more sensitive to ryanodine receptor (RyR1) agonists than those from normal individuals. Reproducible shifts in the dose-response of skeletal muscle to caffeine and halothane are the basis of the current in vitro diagnostic caffeine-halothane contracture test. In an attempt to develop a less invasive MH diagnostic test, the authors determined the effects of RyR1 agonists (caffeine, 4-chloro-m-cresol [4CmC], and halothane) on the adductor muscle with respect to the lactate-pyruvate (L/P) system that was percutaneously dialyzed using a microdialysis technique in homozygous MH-susceptible compared with normal swine. Animals were anesthetized (ketamine-propofol) and artificially ventilated. Sets of six CMA/20 microdialysis catheters were implanted; each catheter was perfused with different RyR1 agonist concentrations. After a 30-min equilibration after implantation, one of the catheters was perfused (2 microl/min) with vehicle (0.9% saline or lipid emulsion), and the other five were perfused with caffeine (1-64 mM), 4CmC (0.1-8 mM), or halothane (prepared in lipid emulsion; 10-500 mM). Outflow dialysate fractions collected at 10-min intervals and L/P parameters were measured enzymatically. Only in the MH-susceptible group did all RyR1 agonists increase dialysate L/P in a dose-dependent manner. The dose-effect relations were most prominent with 4CmC. With the halothane lipid emulsion, data scatter was high compared with that of the caffeine group and especially the 4CmC group. There were no signs of global muscle rigidity, systemic hypermetabolism, or a clinical MH episode during microdialysis RyR1 perfusion. The authors data demonstrate that the in vivo muscle microdialysis of the porcine L/P system reveals distinct differences between MH-susceptible and MH-normal muscle, especially in response to highly specific RyR1 agonists such as 4CmC. The microdialysis L/P technique seems to have an MH diagnostic potential in the clinical setting.

Identification of the dialysable serum inducer of germ-tube formation in Candida albicans.

Yeast cells of Candida albicans are induced by serum at 37 degrees C to produce germ tubes, the first step in a transition from yeast to hyphal growth. Previously, it has been shown that the active component is not serum albumin but is present in the dialysable fraction of serum. In this study, serum induction of germ-tube formation is shown to occur even in the presence of added exogenous nitrogen sources and is therefore not signalled by nitrogen derepression. The active component in serum was purified by ion-exchange, reverse-phase and size-exclusion chromatography from the dialysable fraction of serum and was identified by NMR to be d-glucose. Enzymic destruction of glucose, using glucose oxidase, demonstrated that d-glucose was the only active component in these fractions. Induction of germ-tube formation by d-glucose required a temperature of 37 degrees C and the pH optimum was between pH 7.0 and 8.0. d-Glucose induced germ-tube formation in a panel of clinical isolates of C. albicans. Although d-glucose is the major inducer in serum, a second non-dialysable, trichloroacetic acid precipitable inducer is also present. However, whereas either 1.4 % (v/v) serum or an equivalent concentration of d-glucose induced 50 % germ-tube formation, the non-dialysable component required a 10-fold higher concentration to induce 50 % germ-tube formation. Serum is, therefore, the most effective induction medium for germ-tube formation because it is buffered at about pH 8.5 and contains two distinct inducers (glucose and a non-dialysable component), both active at this pH.

Discovery and neurochemical screening of peptides in brain extracellular fluid by chemical analysis of in vivo microdialysis samples.

Endogenous peptides from brain extracellular fluid of live rats were analyzed using capillary liquid chromatography (LC)-tandem mass spectrometry (MS2). A 4-mm-long microdialysis probe perfused at 0.6 microL/min implanted into the striatum of anesthetized male rats was used to collect 3.6 microL dialysate fractions that were injected on-line into the capillary LC-MS2 system for analysis. A total of 3349 MS2 spectra were collected from 13 different animals under basal conditions and during localized depolarization evoked by infusion of a high-K+ solution through the microdialysis probe. Subtractive analysis revealed a total of 859 MS2 spectra that were observed only during depolarization. From these spectra, 29 peptide sequences (25 were peptides not previously observed) from 6 different protein precursors were identified using database searching software. Proteins identified include precursors to neuropeptides, synaptic proteins, blood proteins, and transporters. The identified peptides represent candidates for neurotransmitters, neuromodulators, and markers of synaptic activity or brain tissue damage. A screen for neuroactivity of novel proenkephalin fragments that were found was performed by infusing the peptides into the brain while monitoring amino acid neurotransmitters by microdialysis sampling combined with capillary electrophoresis. Three of the six tested peptides evoked significant increases in various neuroactive amino acids. These results demonstrate that this combination of methods can identify novel neurotransmitter candidates and screen for potential neuroactivity.

In vivo sampling for pharmacokinetic studies in small experimental animals: a combination of microdialysis, planar chromatography and digital autoradiography

Purpose We describe a method for measuring the pharmacokinetics of positron emission tomography (PET) tracers during rat studies using microdialysis (MD) in combination with planar chromatography and digital autoradiography with a phosphoimager plate (DAR). Procedures An MD probe was inserted into the jugular vein of the rat and perfused with physiological buffer solution (n≥3). An 18F-labeled radiopharmaceutical was injected intravenously, and dialysate fractions were collected, measured for radioactivity, and applied on a chromatography plate. This plate was exposed with an imaging plate for radioactivity imaging. Results Time activity curves for unbound, unchanged tracer and metabolites were determined. The amount of unchanged tracer in the last MD fraction was compared with the amount in the plasma sample collected after sacrifice of the animal. Conclusion MD continuously samples and monitors the amount of unbound tracer in the blood from one single experimental animal. MD combined with planar chromatography and DAR is a highly sensitive and linear method for simultaneous analysis of PET tracers and their metabolites in blood.

Hypoxia is not the sole cause of lactate production during shock.

Traditionally, elevated blood lactate after hemorrhage is interpreted as tissue hypoperfusion, hypoxia, and anaerobic glycolysis. The severity and duration of the increase in blood lactate correlate with death. Recent in vitro studies indicate that epinephrine stimulates lactate production in well-oxygenated skeletal muscle by increasing activity of the Na+-K+-adenosine triphosphatase (ATPase), which derives a significant amount of adenosine triphosphate from glycolysis. Using in vivo microdialysis, we tested whether inhibiting the Na+-K+ pump with ouabain could reduce muscle lactate production during local exposure, via the microdialysis probe, to epinephrine or during hemorrhage in rats. Microdialysis catheters were placed in the muscle of both thighs of pentobarbital-anesthetized male Sprague-Dawley rats (275-350 g) and perfused (1 microL/min) with Krebs-phosphate buffer (pH 7.4) containing ethanol (5 mmol/L) to permit assessment of changes in local blood flow. To inhibit the Na+-K+-ATPase, ouabain (2-3 mmol/L) was added to the perfusate of one leg. In one series of studies, epinephrine was added to the perfusate. In another series, rats were hemorrhaged to a mean arterial pressure of 45 mm Hg for 30 minutes, followed by resuscitation with shed blood and 0.9% sodium chloride. Dialysate fractions were analyzed for lactate and ethanol fluorometrically. Lactate rose during epinephrine exposure or during hemorrhage and resuscitation. Treatment with ouabain reduced dialysate lactate concentration significantly in both series of studies. Local blood flow was reduced by either epinephrine or hemorrhage, but returned toward baseline afterward. Ouabain had no apparent effect on local blood flow. Increased Na+-K+ATPase activity during epinephrine treatment or hemorrhage contributes to muscle lactate production. Hypoxia is not necessarily the sole cause of hyperlactatemia during and after hemorrhagic shock.