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Matrix vesicles and calcification.

Matrix vesicles (MVs) are extracellular, 100 nM in diameter, membrane-invested particles selectively located at sites of initial calcification in cartilage, bone, and predentin. The first crystals of apatitic bone mineral are formed within MVs close to the inner surfaces of their investing membranes. Matrix vesicle biogenesis occurs by polarized budding and pinching-off of vesicles from specific regions of the outer plasma membranes of differentiating growth plate chondrocytes, osteoblasts, and odontoblasts. Polarized release of MVs into selected areas of developing matrix determines the nonrandom distribution of calcification. Initiation of the first mineral crystals, within MVs (phase 1), is augmented by the activity of MV phosphatases (eg, alkaline phosphatase, adenosine triphosphatase and pyrophosphatase) plus calcium-binding molecules (eg, annexin I and phosphatidyl serine), all of which are concentrated in or near the MV membrane. Phase 2 of biologic mineralization begins with crystal release through the MV membrane, exposing preformed hydroxyapatite crystals to the extracellular fluid. The extracellular fluid normally contains sufficient Ca2+ and PO4(3-) to support continuous crystal proliferation, with preformed crystals serving as nuclei (templates) for the formation of new crystals by a process of homologous nucleation. In diseases such as osteoarthritis, crystal deposition arthritis, and atherosclerosis, MVs initiate pathologic calcification, which, in turn, augments disease progression.

Bone Matrix↗

Endogenous digitalis-like factors. An historical overview.

The sodium pump is a ubiquitous cell surface enzyme, a Na/K-ATPase, that maintains ion gradients between cells and the extracellular fluid. The extracellular domain of this enzyme contains a highly conserved receptor for a plant-derived family of compounds, the digitalis glycosides, used in the treatment of congestive heart failure, and certain cardiac arrhythmias. The concept that an endogenous modulator of this enzyme, analogous to the cardiac glycosides, emerged from work on two separate areas: the regulation of extracellular fluid (ECF) volume by a natriuretic hormone (NH), and the regulation of peripheral vascular resistance by a circulating inhibitor of vascular Na/K-ATPase. These two areas merged with the hypothesis that natriuretic hormone and the vascular Na/K-ATPase inhibitor were the same factor, and furthermore, that this factor played a causative role in the pathophysiology of certain types of hypertension. In this communication, the development of this field from its beginnings is traced; evidence for the existence of and efforts to identify the structure of this factor are briefly reviewed, and suggestions for future development of the field are put forward.

Animals↗

HPLC analysis of 6-mercaptopurine and metabolites in extracellular body fluids.

A convenient HPLC assay, which allows for the simultaneous measurement in extracellular fluids of 6-mercaptopurine and four of its metabolites, 6-thioguanine, 6-mercaptopurine riboside, 6-thioxanthine and 6-thiouric acid is described. Solid phase extraction allows for the clean isolation of analytes from plasma, urine or cerebrospinal fluid. The simultaneous determination of 6-mercaptopurine and some of its major metabolites in extracellular fluids may contribute to the monitoring of patient compliance, bioavailability, and individual variation in metabolism and absorption.

Adolescent↗

[Serum concentration of uric acid, a diagnostic 'must' in patients with hyponatremia].

The renal handling of urate involves complete glomerular filtration followed by reabsorption and secretion mainly in the proximal renal tubule. The amount of urate excreted depends on the extracellular fluid volume. Expansion of extracellular fluid volume increases and contraction of extracellular fluid volume decreases the clearance of urate. In the syndrome of inappropriate secretion of antidiuretic hormone, there is volume expansion associated with low uric acid. A comparable situation is observed whenever exaggerated amounts of free water are consumed. Similarly, hypouricaemia and a tendency to low serum sodium concentrations are observed in normal pregnancy, a state with volume expansion, a physiological situation reversed when eclampsia occurs. On the other hand, when hyponatraemia is associated with contraction of extracellular fluid volume as it is often the case when thiazides or loop-diuretics are administered, hyponatraemia is paralleled by increased serum uric acid concentrations.

Blood Volume↗

Plasma clearance of polyfructosan and extracellular body fluid distribution in idiopathic dilated cardiomyopathy and after heart transplantation.

The total extracellular fluid volume and distribution in plasma and interstitial spaces, and the microvascular permeability properties were studied in 16 nonedematous patients with congestive heart failure (CHF) due to idiopathic dilated cardiomyopathy and 17 such patients who underwent heart transplantation (HT) by analyzing the 3-hour plasma disappearance curve of polyfructosan. Eighteen healthy subjects served as controls. Polyfructosan (3.5 kD) is an extracellular marker and inulin analog transported almost solely by diffusion. The initial capillary membrane plasma clearance (i.e., the permeability-surface area product), the interstitial plasma clearance determined at 10 minutes (clearance[10), and the extracellular volume were determined from the polyfructosan curves. I-131-albumin was used as a plasma volume reference. Permeability-surface area product was elevated in both patient groups (6.6 +/- 1.9 ml/ kg/min in the CHF group and 6.7 +/- 2.0 ml/kg/min in the HT group vs 5.1 +/- 1.3 ml/kg/min in controls, p <0.01 for both), whereas clearance(10) was normalized in the HT group (4.5 +/- 0.9 ml/kg/min in the HT group, 4.4 +/- 0.7 ml/kg/min in controls vs 5.0 +/- 0.9 ml/kg/ min in the CHF group, p <0.1 and p <0.05, respectively). The normalization of interstitial plasma clearance of polyfructosan was associated with time since HT (r = 0.49, p <0.05). Plasma volumes were similar in all 3 groups (41 +/- 8 ml/kg in controls, 44 +/- 13 in the CHF group and 39 +/- 8 in the HT group). In contrast, total extracellular volume was elevated in both patients groups (177 +/- 27 ml/kg in the CHF group and 173 +/-27 in the HT group vs 152 +/- 12 in controls, p <0.01). The results strongly suggest a microvascular permeability defect in both patient groups that perhaps plays a role in the extravascular distribution of the excess extracellular fluid volume.

Adult↗

Arterial hypertension--a disease of the juxtaglomerular apparatus?

From a special strain of genetically hypertensive rats, the Milan hypertensive strain (MHS), arterial hypertension can be transplanted with the kidney to the Milan normotensive strain (MNS). During development of hypertension in MHS rats there was an activation of the tubuloglomerular feedback control that reduced glomerular filtration rate, leading to retention of electrolytes and fluid. This increased extracellular fluid volume reduces feedback sensitivity, but in a fashion that gives rise to chronic extracellular fluid expansion and can thereby raise the blood pressure. In a limited sense, arterial hypertension in these animals exists to prevent the kidney from retaining more extracellular fluid volume. The altered function in the juxtaglomerular apparatus of the MHS rats thus may explain the rise in arterial blood pressure.

Animals↗

Basolateral Na(+)-H+ antiporter. Mechanisms of electroneutral and conductive ion transport.

The basolateral Na-H antiporter of the turtle colon exhibits both conductive and electroneutral Na+ transport (Post and Dawson. 1992. American Journal of Physiology. 262:C1089-C1094). To explore the mechanism of antiporter-mediated current flow, we compared the conditions necessary to evoke conduction and exchange, and determined the kinetics of activation for both processes. Outward (cell to extracellular fluid) but not inward (extracellular fluid to cell) Na+ or Li+ gradients promoted antiporter-mediated Na+ or Li+ currents, whereas an outwardly directed proton gradient drove inward Na+ or Li+ currents. Proton gradient-driven, "counterflow" current is strong evidence for an exchange stoichiometry of > 1 Na+ or Li+ per proton. Consistent with this notion, outward Na+ and Li+ currents generated by outward Na+ or Li+ gradients displayed sigmoidal activation kinetics. Antiporter-mediated proton currents were never observed, suggesting that only a single proton was transported per turnover of the antiporter. In contrast to Na+ conduction, Na+ exchange was driven by either outwardly or inwardly directed Na+, Li+, or H+ gradients, and the activation of Na+/Na+ exchange was consistent with Michaelis-Menten kinetics (K1/2 = 5 mM). Raising the extracellular fluid Na+ or Li+ concentration, but not extracellular fluid proton concentration, inhibited antiporter-mediated conduction and activated Na+ exchange. These results are consistent with a model for the Na-H antiporter in which the binding of Na+ or Li+ to a high-affinity site gives rise to one-for-one cation exchange, but the binding of Na+ or Li+ ions to other, lower-affinity sites can give rise to a nonunity, cation exchange stoichiometry and, hence, the net translocation of charge. The relative proportion of conductive and nonconductive events is determined by the magnitude and orientation of the substrate gradient and by the serosal concentration of Na+ or Li+.

Animals↗

Incomplete coverage of mammalian bone matrix by lining cells.

A functional "membrane" is thought to exist that separates the general extracellular fluid from the bone extracellular fluid. Many investigators have concluded that bone lining cells form this barrier. It is posited that alterations in this barrier may lead to the recruitment of osteoclasts and contribute to the control of extracellular calcium homeostasis. The present ultrastructural studies, however, demonstrated that significant portions of the resting bone are in contact with the general extracellular fluid. In certain regions of the cochlea, up to 88% of the bone surface is in contact with extracellular fluid. These anatomic observations require a critical reevaluation of current theories of the physiologic roles of bone lining cells in osteoclast recruitment and calcium homeostasis.

Animals↗

Extracellular matrix metalloproteinase inducer is increased in smokers' bronchoalveolar lavage fluid.

Extracellular matrix metalloproteinase inducer (EMMPRIN), also called basigin, is present in the lung during development, but its expression in normal adult lung is minimal. Increases of EMMPRIN have been found in various forms of experimental lung injury. To determine whether EMMPRIN might be involved in alveolar injury/repair associated with smoking, we developed an ELISA for EMMPRIN and applied it to bronchoalveolar lavage fluids from never-smokers (n = 7), former smokers (n = 16), and current smokers (n = 58). The smoker groups included subjects with emphysema, as determined by high-resolution chest computed tomography. EMMPRIN levels were significantly elevated in current and former smokers (315 +/- 20 and 175 +/- 15 pg/ml SEM, respectively, compared with 31 +/- 7 pg/ml in never-smokers), but the EMMPRIN levels of smokers with emphysema were not different from smokers without emphysema. Immunohistochemistry of smokers' lung tissue showed EMMPRIN in bronchiolar epithelium and alveolar macrophages, but EMMPRIN mRNA in alveolar macrophages was not different between current and never-smokers. Matrix metalloproteinase-1 was also detectable in the bronchoalveolar lavage fluid from some smokers but not in never-smokers. These findings indicate that smoking is associated with increased intrapulmonary EMMPRIN. Whether EMMPRIN is involved in smoking-induced lung pathology remains to be determined.

Adult↗

Distribution of fluid between extracellular and intracellular compartments in the heart, lungs, liver and spleen of neonatal rats.

The volume of fluid in various tissue compartments of thoracic and abdominal viscera in neonatal rats has been calculated from the steady state distribution of radioindicators. During the first 3 weeks after birth, the volume of extracellular fluid (3H-inulin space) decreases at a similar rate (ca. 0.04 ml/g tissue/week) in heart, lungs, liver and spleen. Over the same postnatal period, the 51Cr tagged erythrocyte space (an estimate of vascularity) remains relatively constant in liver, increases slightly in the heart and spleen, and rises substantially (3-fold increase) in the lungs. The volume of parenchymal cell water, calculated from data for tissue water content and radioindicator spaces, tends to increase with age. However, the distribution of fluid between extracellular and intracellular compartments in 3-week-old animals is similar to that in adults.

Animals↗

Volume studies in men with mild to moderate hypertension.

The importance of salt and water in the pathophysiology of the hypertensive state is well recognized. The current study is the first to report simultaneous measurements of red blood cell mass, plasma volume, extracellular fluid and total body water levels. Studies were performed in 82 white men, 14 with normal blood pressure and 16 with low renin and 52 with normal renin hypertension. The results indicate that subjects with normal renin hypertension compared with age-matched controls are characterized by an absolute increase (1.5 liter/m2) in intracellular fluid (total body water minus extracellular fluid). Furthermore, the ratio of extracellular fluid to total body water is decreased (0.43 to 0.38). No volume differences were found between subjects with low renin hypertension and age-matched subjects with normal renin hypertension. We conclude that subjects with normal renin hypertension compared with age-matched peers are characterized by an expanded intracellular fluid and that subjects with low renin hypertension do not exhibit a unique volume disorder.

Adult↗

Four-compartment cellular level body composition model: comparison of two approaches.

OBJECTIVE: The aim of this study was to develop and compare two DXA-based four-compartment [body weight=body cell mass (BCM)+extracellular fluid (ECF)+extracellular solids (ECS)+fat] cellular level models. RESEARCH METHODS AND PROCEDURES: Total body potassium (TBK) model: BCM from TBK by whole-body counting-ECF(TBK)=LST-[BCM(TBK)+0.73 x osseous mineral (Mo)]. Bromide model: ECF from sodium bromide dilution-BCM(BROMIDE)=LST-(ECF(BROMIDE)+0.73xMo); Mo and LST measurements came from DXA. The two approaches were evaluated in 99 healthy men and 118 women. RESULTS: BCM estimates were highly correlated (r=0.97, p<0.001), as were ECF estimates (r=0.87, p<0.001); a small statistically significant mean difference was present (mean+/-SD; BCM(TBK) model, 30.4+/-8.9 kg; BCM(BROMIDE), 31.4+/-9.3 kg; Delta=1.0+/-2.8 kg; p<0.001; ECF(TBK), 18.5+/-4.2 kg; ECF(BROMIDE), 17.5+/-3.6 kg; Delta=1.0+/-2.8 kg; p<0.001). A high correlation (r=0.97, p<0.001) and good agreement (38.9+/-9.5 vs. 38.9+/-9.5 kg; Delta=0.0+/-2.4 kg; p=0.39) were present between TBW, derived as the sum of intracellular water from TBK and ECW from bromide, and measured TBW by 2H2O dilution. DISCUSSION: Two developed four-compartment cellular level DXA models, one of which is appropriate for use in most clinical and research settings, provide comparable results and are applicable for BCM and ECF estimation of subject groups with hydration disturbances.

Absorptiometry, Photon↗

Some aspects of sodium movement across porcine aortic walls in vitro.

1. Sodium movement across the porcine aortic arch in vitro is greater from the intimal to the medial surface than in the reverse direction. 2. A short-circuit current can be measured across the aortic arch and the calculated energy is greater than the energy required to support the net outward sodium movement, suggesting the possibility of active transport of other ions. 3. Net sodium movement is not significantly altered by an adverse gradient on the outside (medial side) but is reduced by a gradient in the opposite direction. 4. Sodium retention within the tissue is increased by a gradient on the medial side. 5. The movement of [14C]inulin is greater from the medial to the intimal surface, suggesting greater porosity on the medial side. A slight rise in both total tissue fluid and extracellular fluid occurred when the sodium concentration on the medial side of the preparation was increased. This did not occur with the sodium concentration increased on the intimal side. The changes were only significant statistically for total tissue fluid. 6. The short-circuit current can be enhanced by adrenaline and reduced by a variety of beta-adrenoreceptor-blocking agents. 7. It appears that aortic tissue can actively transport sodium and it is suggested that this mechanism could play a part in blood pressure control.

Alprenolol↗

The composition of extracellular interstitial fluid collected with a push-pull cannula from the testes of adult rats.

1. A push-pull cannula has been used to obtain extracellular interstitial fluid from the testes of anaesthetized adult rats. 2. Assuming or having demonstrated that appropriate radioactive markers had equilibrated between the vascular and extracellular fluid compartment of the testis, the dilution of the cannula infusate by extracellular interstitial fluid of the testis has been determined. These dilutions have then been used, with the measured concentrations of sodium, potassium, protein and testosterone in the perfusate, to calculate the concentrations of these substances in undiluted extracellular interstitial fluid of the testis. 3. Sodium, potassium and protein levels in testicular interstitial fluid calculated in this way were similar to blood plasma levels. Testosterone concentrations were certainly no greater than in testicular venous blood, and may have been even less. The results with testosterone and potassium contrast with earlier results obtained with less-physiological techniques.

Animals↗

Base deficit estimation in umbilical cord blood is influenced by gestational age, choice of fetal fluid compartment, and algorithm for calculation.

OBJECTIVE: The purpose of this study was to explore the influences of gestational age, the choice of fetal fluid compartment, and the algorithm for calculation on the estimation of the base deficit in umbilical cord arterial blood at birth. STUDY DESIGN: From 1995 to 2002, cord arterial blood gases and obstetric data were available for 43,551 newborn infants at 37+ weeks of gestation (cohort I). The mean base deficit in blood and the base deficit in extracellular fluid were estimated from pH and PCO2 values in 28,213 newborn infants with a 5-minute Apgar score of > or = 9 (cohort II) with the use of 3 different calculation algorithms (base deficit in blood, base deficit in extracellular fluid [A], and base deficit in extracellular fluid [B]). RESULTS: In cohort II, the base deficit in blood, the base deficit in extracellular fluid (A), and the base deficit in extracellular fluid (B) increased with advancing gestational age (linear regression; P < .0001). The curves run almost parallel, with the base deficit in blood being higher than the base deficit in extracellular fluid (A) and (B). With the use of receiver operating characteristic curves in cohort I, the area under curve to indicate a 5-minute Apgar score of < 7 and < 4 showed the area under curve-pH to be greater than the area under curve-base deficit in extracellular fluid (A) and (B), the area under curve-base deficit in blood to be greater than the area under curve-base deficit in extracellular fluid (A) and (B) for a 5-minute Apgar score of < 7, and the area under curve-base deficit in blood to be greater than the area under curve-base deficit in extracellular fluid (A) and (B) for an Apgar score of < 4. The cutoffs with highest sensitivity and lowest false-positive rate for a 5-minute Apgar score of < 7 and < 4 were, for both scores, a pH value of 7.15, a base deficit in blood of 10 mmol/L, a base deficit in extracellular fluid (A) of 8 mmol/L, and a base deficit in extracellular fluid (B) of 6 mmol/L. CONCLUSION: The calculated values of the base deficit in umbilical cord arterial blood are influenced decisively by gestational age, the choice of fetal fluid compartment, and the calculation algorithms that are used. The power of the base deficit to indicate neonatal distress depends on the choices of fluid compartment and the algorithm that is used to calculate the base deficit.

Acidosis↗

Fluid therapy for children: facts, fashions and questions.

Fluid therapy restores circulation by expanding extracellular fluid. However, a dispute has arisen regarding the nature of intravenous therapy for acutely ill children following the development of acute hyponatraemia from overuse of hypotonic saline. The foundation on which correct maintenance fluid therapy is built is examined and the difference between maintenance fluid therapy and restoration or replenishment fluid therapy for reduction in extracellular fluid volume is delineated. Changing practices and the basic physiology of extracellular fluid are discussed. Some propose changing the definition of "maintenance therapy" and recommend isotonic saline be used as maintenance and restoration therapy in undefined amounts leading to excess intravenous sodium chloride intake. Intravenous fluid therapy for children with volume depletion should first restore extracellular volume with measured infusions of isotonic saline followed by defined, appropriate maintenance therapy to replace physiological losses according to principles established 50 years ago.

Child↗