Composition Of Intestinal Fluids Research Articles

Characterization of neonatal and infant enterostomy fluids - part II: Drug solubility

Previous research revealed marked differences in the composition of intestinal fluids between infants and adults. To explore the impact on the solubilization of orally administered drugs, the present study assessed the solubility of five poorly water-soluble, lipophilic drugs in intestinal fluid pools from 19 infant enterostomy patients (infant HIF). For some but not all drugs, the average solubilizing capacity of infant HIF was similar to that of HIF obtained from adults (adult HIF) in fed conditions. Commonly used fed state simulated intestinal fluids (FeSSIF(-V2)) predicted fairly well drug solubility in the aqueous fraction of infant HIF, but did not account for the substantial solubilization by the lipid phase of infant HIF. Despite similarities in the average solubilities of some drugs in infant HIF and adult HIF or SIF, the underlying solubilization mechanisms likely differ, considering important compositional differences (e.g., low bile salt levels). Finally, the huge variability in composition of infant HIF pools resulted in a highly variable solubilizing capacity, potentially causing variations in drug bioavailability. The current study warrants future research focusing on (i) understanding the mechanisms underlying drug solubilization in infant HIF and (ii) evaluating the sensitivity of oral drug products to interpatient variations in drug solubilization.

Characterization of Aspirated Duodenal Fluids from Parkinson's Disease Patients.

Parkinson's disease, one of the most common neurodegenerative diseases, may not only affect the motor system, but also the physiology of the gastrointestinal tract. Delayed gastric emptying, impaired motility and altered intestinal bacteria are well-established consequences of the disease, which can have a pronounced effect on the absorption of orally administered drugs. In contrast, no studies have been performed into the composition of intestinal fluids. It is not unlikely that Parkinson's disease also affects the composition of intestinal fluids, a critical factor in the in vitro and in silico simulation of drug dissolution, solubilization and absorption. In the current study, duodenal fluids were aspirated from Parkinson's disease (PD) patients and age-matched healthy controls (healthy controls, HC) consecutively in fasted and fed conditions. The fluids were then characterized for pH, buffer capacity, osmolality, total protein, phospholipids, bile salts, cholesterol and lipids. In a fasted state, the intestinal fluid composition was highly similar in PD patients and healthy controls. In general, the same was true for fed-state fluids, apart from a slightly slower and less pronounced initial change in factors directly affected by the meal (i.e., buffer capacity, osmolality, total protein and lipids) in PD patients. The absence of a fast initial increase for these factors immediately after meal intake, as was observed in healthy controls, might result from slower gastric emptying in PD patients. Irrespective of the prandial state, a higher relative amount of secondary bile salts was observed in PD patients, potentially indicating altered intestinal bacterial metabolism. Overall, the data from this study indicate that only minor disease-specific adjustments in small intestinal fluid composition should be considered when simulating intestinal drug absorption in PD patients.

Characterization of neonatal and infant enterostomy fluids

The composition of gastrointestinal (GI) fluids is crucial for the dissolution, solubilization, and absorption of orally administered drugs. Disease- or age-related changes in GI fluid composition could significantly affect the pharmacokinetics of oral drugs. However, limited studies have been conducted on the characteristics of GI fluids in neonates and infants due to practical and ethical challenges. The current study collected enterostomy fluids from 21 neonate and infant patients over an extended period of time and from different regions of the small intestine and colon. The fluids were characterized for pH, buffer capacity, osmolality, total protein, bile salts, phospholipids, cholesterol, and lipid digestion products. The study found a large variability in the fluid characteristics among the different patients, in line with the highly heterogeneous study population. Compared to adult intestinal fluids, the enterostomy fluids from neonates and infants had low bile salt concentrations, with an increasing trend as a function of age; no secondary bile salts were detected. In contrast, total protein and lipid concentrations were relatively high, even in the distal small intestine. These findings suggest marked differences in intestinal fluid composition between neonates and infants versus adults, which may affect the absorption of certain drugs.

Photoperiod and dietary treatment in freshwater modulate the short-term intestinal response to seawater in Atlantic salmon (Salmo salar)

Stimulation and timing of smoltification are essential for successful Atlantic salmon (Salmo salar) aquaculture. This study investigated intestinal responses during dietary and photoperiod manipulation in freshwater (FW) and after a subsequent seven days residence in seawater (SW). “Small” and “large” Atlantic salmon parr (∼40 g and ∼ 130 g respectively) were treated in FW for 12 weeks and thereafter transferred to SW for seven days. During the FW phase, fish underwent two different light conditions, 24 L:0D – 24 L (“LL-LL” groups) and 7 L:17D – 24 L (“SP-LL” groups) or fed with either regular feed (“LL-LL C" and “SP-LL C" groups) or feed enriched with a salt mix plus free tryptophan (“LL-LL + diet” and “SP-LL + diet” groups). We analyzed Na+/K+-ATPase (NKA) activity, tissue bioelectrical properties in Ussing chambers, and intestinal fluid composition. The NKA activity showed minor variations in relation to fish size, treatments, or intestinal region (anterior or posterior). Photoperiod modulated epithelial bioelectrical properties (Isc and Rt) of the anterior and posterior intestine, particularly transepithelial resistance (Rt). Pharmacological experiments, targeting apical Na+/K+/2Cl− (NKCC2) and Na+/Cl− (NCC) co-transporters revealed intestinal region- and water salinity-dependent effects. In addition, stimulation of the intracellular cAMP with forskolin and IBMX showed intestinal region-, water salinity, and treatment-dependence responses with clear functional specialization of the anterior and posterior intestine. The intestinal fluid composition reflected the ability to process ingested SW and showed little variation in large fish. In summary, our data suggest a better pre-adaptation of the intestine during light-stimulated smoltification (SP-LL groups), and the combination of light and diet might give, in an industrial aquaculture setting, an advantage to smaller, but not larger smolts. Intestinal fluid composition in small fish can be used as an index of intestinal function and may act as a long-term performance proxy in SW Atlantic salmon.

Fasted intestinal solubility limits and distributions applied to the biopharmaceutics and developability classification systems

After oral administration, a drug’s solubility in intestinal fluid is an important parameter influencing bioavailability and if the value is known it can be applied to estimate multiple biopharmaceutical parameters including the solubility limited absorbable dose. Current in vitro measurements may utilise fasted human intestinal fluid (HIF) or simulated intestinal fluid (SIF) to provide an intestinal solubility value. This single point value is limited since its position in relation to the fasted intestinal solubility envelope is unknown. In this study we have applied a nine point fasted equilibrium solubility determination in SIF, based on a multi-dimensional analysis of fasted human intestinal fluid composition, to seven drugs that were previously utilised to investigate the developability classification system (ibuprofen, mefenamic acid, furosemide, dipyridamole, griseofulvin, paracetamol and acyclovir). The resulting fasted equilibrium solubility envelope encompasses literature solubility values in both HIF and SIF indicating that it measures the same solubility space as current approaches with solubility behaviour consistent with previous SIF design of experiment studies. In addition, it identifies that three drugs (griseofulvin, paracetamol and acyclovir) have a very narrow solubility range, a feature that single point solubility approaches would miss. The measured mid-point solubility value is statistically equivalent to the value determined with the original fasted simulated intestinal fluid recipe, further indicating similarity and that existing literature results could be utilised as a direct comparison. Since the multi-dimensional approach covered greater than ninety percent of the variability in fasted intestinal fluid composition, the measured maximum and minimum equilibrium solubility values should represent the extremes of fasted intestinal solubility and provide a range. The seven drugs all display different solubility ranges and behaviours, a result also consistent with previous studies. The dose/solubility ratio for each measurement point can be plotted using the developability classification system to highlight individual drug behaviours. The lowest solubility represents a worst-case scenario which may be useful in risk-based quality by design biopharmaceutical calculations than the mid-point value. The method also permits a dose/solubility ratio frequency distribution determination for the solubility envelope which permits further risk-based refinement, especially where the drug crosses a classification boundary. This novel approach therefore provides greater in vitro detail with respect to possible biopharmaceutical performance in vivo and an improved ability to apply risk-based analysis to biopharmaceutical performance. Further studies will be required to expand the number of drugs measured and link the in vitro measurements to in vivo results.

Characterisation of fasted state gastric and intestinal fluids collected from children

Fundamental knowledge about the composition of intestinal fluids in paediatric populations is currently unavailable. This study aimed to characterise gastric and intestinal fluid from paediatric populations.Gastric and intestinal fluid samples were obtained during routine clinical endoscopy from paediatric patients at a large teaching hospital. These fluids were characterised to measure the pH; buffer capacity; osmolality; bile acid concentration and composition.A total of 55 children were recruited to the study aged from 11 months to 15 years of age where 53 gastric fluid samples and 40 intestinal fluid samples were obtained. pH values recorded ranged from pH 0.57 to 11.05 (median: 2.50) in gastric fluids and from 0.89 to 8.97 (median: 3.27) in intestinal fluids. The buffer capacity did not change significantly between gastric and intestinal fluids with median values of 12 mM/L/ΔpH for both fluids. Gastric fluid osmolality values ranged from 1 to 615 mOsm/kg, while intestinal fluid values ranged from 35 to 631 mOsm/kg.Gastric fluid bile acid concentrations ranged from 0.002 to 2.3 mM with a median value of 0.017 mM whilst intestinal fluid bile acid concentrations ranged from 0.0008 to 3.3 mM with a median value of 0.178 mM. Glycocholate; taurocholic acid; glycochenodeoxycholate and taurochenodeoxycholate were the most commonly identified bile acids within paediatric intestinal fluids.All compositional components were associated with large inter-individual variability. Further work is required to develop simulated paediatric media and to explore the impact of these media on drug solubility and dissolution.

Effect of composition of simulated intestinal media on the solubility of poorly soluble compounds investigated by design of experiments

The composition of the human intestinal fluids varies both intra- and inter-individually. This will influence the solubility of orally administered drug compounds, and hence, the absorption and efficacy of compounds displaying solubility limited absorption.The purpose of this study was to assess the influence of simulated intestinal fluid (SIF) composition on the solubility of poorly soluble compounds.Using a Design of Experiments (DoE) approach, a set of 24 SIF was defined within the known compositions of human fasted state intestinal fluid. The SIF were composed of phospholipid, bile salt, and different pH, buffer capacities and osmolarities. On a small scale semi-robotic system, the solubility of 6 compounds (aprepitant, carvedilol, felodipine, fenofibrate, probucol, and zafirlukast) was determined in the 24 SIF.Compound specific models, describing key factors influencing the solubility of each compound, were identified. Although all models were different, the level of phospholipid and bile salt, the pH, and the interactions between these, had the biggest influences on solubility overall. Thus, a reduction of the DoE from five to three factors was possible (11–13 media), making DoE solubility studies feasible compared to single SIF solubility studies. Applying this DoE approach will lead to a better understanding of the impact of intestinal fluid composition on the solubility of a given drug compound.

Displacement of itraconazole from cyclodextrin complexes in biorelevant media: In vitro evaluation of supersaturation and precipitation behavior

Intestinal fluids contain several constituents with affinity for cyclodextrins that have the potential of displacing drugs from the cyclodextrin cavity by competition. In this study, the solubilizing capacity of 2-hydroxypropyl-β-cyclodextrin (HP-β-CD) for itraconazole was studied in presence of selected bile salts and phosphatidylcholine. Despite the fact that these competing agents significantly lowered the solubility of itraconazole in presence of cyclodextrins, the addition of concentrated solutions of these bile constituents to a solution containing itraconazole solubilized by HP-β-CD did not result in precipitation, even at bile salt and phospholipid concentrations where itraconazole precipitation would be anticipated based on solubility studies. This phenomenon was further investigated in more dynamic conditions via in vitro transfer studies, mimicking the gastrointestinal transfer of HP-β-CD solutions saturated with itraconazole. Intestinal supersaturation upon transfer was observed for all conditions tested and a concentration dependent impact of bile salts and phospholipids on the precipitation behavior of itraconazole was demonstrated: high concentrations of bile salts and phospholipids generated the highest degrees of supersaturation shortly after the transfer step but also resulted in stronger itraconazole precipitation at later time points. These findings demonstrate the possible impact of the variable intestinal fluid composition on the behavior of cyclodextrin containing formulations.

Statistical investigation of simulated intestinal fluid composition on the equilibrium solubility of biopharmaceutics classification system class II drugs

A drug’s solubility and dissolution behaviour within the gastrointestinal tract is a key property for successful administration by the oral route and one of the key factors in the biopharmaceutics classification system. This property can be determined by investigating drug solubility in human intestinal fluid (HIF) but this is difficult to obtain and highly variable, which has led to the development of multiple simulated intestinal fluid (SIF) recipes. Using a statistical design of experiment (DoE) technique this paper has investigated the effects and interactions on equilibrium drug solubility of seven typical SIF components (sodium taurocholate, lecithin, sodium phosphate, sodium chloride, pH, pancreatin and sodium oleate) within concentration ranges relevant to human intestinal fluid values. A range of poorly soluble drugs with acidic (naproxen, indomethacin, phenytoin, and piroxicam), basic (aprepitant, carvedilol, zafirlukast, tadalafil) or neutral (fenofibrate, griseofulvin, felodipine and probucol) properties have been investigated. The equilibrium solubility results determined are comparable with literature studies of the drugs in either HIF or SIF indicating that the DoE is operating in the correct space. With the exception of pancreatin, all of the factors individually had a statistically significant influence on equilibrium solubility with variations in magnitude of effect between the acidic and basic or neutral compounds and drug specific interactions were evident. Interestingly for the neutral compounds pH was the factor with the second largest solubility effect. Around one third of all the possible factor combinations showed a significant influence on equilibrium solubility with variations in interaction significance and magnitude of effect between the acidic and basic or neutral compounds. The least number of significant media component interactions were noted for the acidic compounds with three and the greatest for the neutral compounds at seven, with again drug specific effects evident. This indicates that a drug’s equilibrium solubility in SIF is influenced depending upon drug type by between eight to fourteen individual or combinations of media components with some of these drug specific. This illustrates the complex nature of these fluids and provides for individual drugs a visualisation of the possible solubility envelope within the gastrointestinal tract, which may be of importance for modelling in vivo behaviour. In addition the results indicate that the design of experiment approach can be employed to provide greater detail of drug solubility behaviour, possible drug specific interactions and influence of variations in gastrointestinal media components due to disease. The approach is also feasible and amenable to adaptation for high throughput screening of drug candidates.

Comparison of the Solubility and Dissolution of Drugs in FastedState Biorelevant Media (FaSSIF and FaSSIF-V2)

The equilibrium solubility values and dissolution profiles in each of the media (FaSSIF or FaSSIF-V2) produced using methylene chloride and the corresponding SIF Powder are equivalent for the drugs tested representing acid, basic, and neutral compounds. Therefore, it is practical to switch from the more labor-intensive solvent-evaporation method to the use of standardized instant powders for preparing biorelevant media without affecting the solubility and dissolution results. Testing in the two versions of FaSSIF media with different bile salt/lecithin ratios will give an indication on how the drug and formulation could be affected in vivo by the variations in bile salt/lecithin ratio within the range found in human small intestinal fluids. INTRODUCTION Biorelevant media are artificial in vitro media designed to share physicochemical properties with their corresponding fluids found in vivo in terms of osmolarity, pH, buffer capacity, and surfactant content, for example. Specifically, intestinal biorelevant media are widely used to mimic the properties of gastrointestinal fluids for in vitro solubility and dissolution studies. These media can also be used to simulate gastrointestinal stability and to perform permeability studies (1). Biorelevant media simulate the gastrointestinal fluids by containing the physiological surfactants, bile salt and lecithin. These media are particularly important for the development of poorly soluble drugs, because these surfactants significantly increase solubility. Biorelevant intestinal media were first proposed by Dressman et al. (2) at the University of Frankfurt in collaboration with Reppas from the University of Athens who identified the deficiencies of standard pharmacopeial media. These two groups also demonstrated the clear advantage of biorelevant media over standard conventional media for studying the solubility and dissolution of poorly soluble compounds. Biorelevant intestinal media simulate intestinal fluids secreted under fasted(FaSSIF) or fed-state conditions (FeSSIF). Food intake alters the physiochemical properties and composition of intestinal fluids for digestion. That is why separate intestinal media are defined for fastedand fed-state conditions. The two states of media enable formulation scientists to obtain solubility values and dissolution profiles of drugs and their formulations in vitro under both fasted and fed states. This can help detect food effects and optimize formulation performance using standard pharmacopeial dissolution test apparatus (e.g., USP Apparatus 2 and 4). There are biorelevant media representing fluids found along all regions in the gastrointestinal tract. However, simulated upper small intestinal media are the most commonly used, particularly for immediate-release drugs, because they simulate the fluids in the intestinal region from which most drugs are absorbed. FaSSIF and FaSSIF-V2 are the two most frequently used compositions for biorelevant intestinal media reflecting fasted-state conditions. The main difference between FaSSIF and FaSSIF-V2 is the reduced amount of lecithin in FaSSIF-V2 (Table 1) as compared with FaSSIF. The level of lecithin in both versions falls within the range typically found in aspirates sampled from healthy human volunteers. The purpose of this study was to evaluate the equilibrium solubility of four poorly water soluble drug substances (danazol, dipyridamole, ketoconazole, miconazole) and the drug dissolution from five drug products containing drugs with various pKa values under fasted-state conditions (i.e., without food interference) using the FaSSIF and FaSSIF-V2 media. The drug products contained acidic (mefenamic acid), basic (ketoconozole), or neutral (danazol, metoprolol, paracetamol) compounds. In addition, the solubility and drug dissolution behaviors were compared in media made from commercially available instant powders (SIF Powder Original and SIF Powder FaSSIF-V2 from biorelevant.com, UK) and prepared from methylene chloride as described by Marques (3). MATERIALS AND METHODS Materials Drugs Used for Solubility Testing Danazol was purchased from Jai Radhe (Ahmedabad, India). Dipyridamole was from Sigma Aldrich Chemie GmbH *Corresponding author. Contact: www.biorelevant.com/contact dx.doi.org/10.14227/DT200313P44

Quantitation of major protein constituents of murine intestinal fluid

The gastrointestinal tract is a hostile biological environment, yet not all ingested materials are destroyed. The minute differences that determine whether a substance persists or is digested, liberated, adsorbed, excreted, or taken up are still poorly understood. Most attempts to investigate the events occurring during an orogastrointestinal passage rely on simplified in vitro systems where an analyte is exposed to artificial intestinal fluids. To closely mimic the events in the gastrointestinal tract, the exact intestinal fluid composition and the in vivo concentration of its constituents must be known. The widely used lavage procedures, however, dilute the intestinal fluids to an extent that precludes recalculation to the original concentrations. Thus, we developed procedures with which undiluted murine intestinal fluid can be harvested; determined the in vivo concentrations of the digestive enzymes trypsin, chymotrypsin, and elastase and the adsorbents mucin and immunoglobulin A in small intestinal fluid of fasted and unfasted female Balb/c mice; and identified chymotrypsin and immunoglobulin A as valid endogenous dilution markers for the recalculation of aqueous lavages. With these technologies and information at hand, more reliable investigations on the fate of allergens, pathogens, food, and anthropogenic xenobiotics in the gastrointestinal tract will be possible.

Determination of intraluminal theophylline concentrations after oral intake of an immediate-and a slow-release dosage form

The purpose of this study was to evaluate a protocol which enables determining luminal drug concentrations after oral drug administration in man. Human intestinal fluids were aspirated from two sampling sites (duodenum and jejunum) at different time points after oral intake of theophylline; an immediate- and a slow-release dosage form were used to demonstrate the feasibility of discriminating between different formulations. Osmolarity and pH of the aspirates were measured and theophylline concentrations were determined by HPLC-UV. After intake of the immediaterelease formulation of theophylline, duodenal maximum concentrations up to 3 mM were reached within 30 min. Theophylline appeared to be almost completely absorbed before it reached the second sampling site in the jejunum, as observed jejunal concentrations were lower than 10% of the maximal duodenal concentrations. These results are in agreement with fast dissolution and fast absorption through the intestinal mucosa, which could be expected as theophylline belongs to class I of the Biopharmaceutical Classification System. In contrast to the immediate-release formulation, administering the slow-release dosage form resulted in a gradual appearance of theophylline, reaching maximal intestinal concentrations below 300 muM. The proposed methodology can be used to assess luminal drug concentrations and to monitor the time- and site-dependent composition of intestinal fluids after intake of an oral dosage form. This approach may contribute to a better understanding of the behaviour of oral drug formulations in the gastrointestinal tract and may be exploited to further unravel the complexity of the gastrointestinal absorption process. In addition, knowledge of luminal drug concentrations may assist in the selection of drug concentrations applied in in-vitro permeability assays.

Evolutionary aspects of intestinal bicarbonate secretion in fish

Experiments compared intestinal HCO 3 − secretion in the intestine of marine teleost Gulf toadfish, Opsanus beta, to representatives of early chondrostean and chondrichthyan fishes, the Siberian sturgeon, Acipenser baerii, and white-spotted bamboo shark, Chiloscyllium plagiosum, respectively. As seen in marine teleosts, luminal HCO 3 − concentrations were 10-fold plasma levels in all species when exposed to hyperosmotic conditions. While intestinal water absorption left Mg 2+ and SO 4 2− concentrated in intestinal fluids up to four-fold ambient seawater concentrations, HCO 3 − was concentrated up to 50 times ambient levels as a result of intestinal HCO 3 − secretion. Reduced luminal Cl − concentrations in the intestine of all species suggest that HCO 3 − secretion also occurs via Cl −/HCO 3 − exchange in chondrostean and chondrichthyan fishes. Sturgeon began precipitating carbonates from the gut after only 3 days at 14‰, a mechanism utilized by marine teleosts to reduce intestinal fluid osmolality and maintain calcium homeostasis. Analysis of published intestinal fluid composition in the cyclostome Lampetra fluviatilis reveals that this species likely also utilize intestinal HCO 3 − secretion for osmoregulation. Analysis of existing cyclostome data and our results indicate that intestinal Cl −/HCO 3 − exchange plays an integral role in maintaining hydromineral balance not only in teleosts, but in all fish (and perhaps other animals) with a need to drink seawater.

Predicting Pharmacokinetic Food Effects Using Biorelevant Solubility Media and Physiologically Based Modelling

Food-induced changes in gastric emptying time, gastric pH and/or intestinal fluid composition may have an impact on the pharmacokinetics of drugs. The aim of this work was to use mathematical models describing physiology in fed and fasted states together with biorelevant solubility and degradation data to simulate food effects for six compounds from recent Roche projects. The solubility of each compound was measured in different biorelevant media: simulated human gastric fluid for the fasted and fed state, simulated human intestinal fluid for the fasted, fed and high-fat state, and simulated human colonic fluid for the upper and the lower colon. A physiologically based absorption model was developed in GastroPlustrade mark for each compound using permeability, solubility, metabolism and distribution data. By incorporating the appropriate physiological parameters and solubility data into the model, the oral pharmacokinetics of each drug was simulated under fasted, fed and/or high-fat conditions. Predicted and observed plasma concentration-time profiles and food effects were compared for a range of doses to assess the accuracy of the simulations. The models were able to distinguish between minor and significant food effects. The simulation captured well the magnitude of the food effects and for the six compounds correctly predicted the observed plasma exposure in fasted, fed and high-fat conditions. Biorelevant solubility tests can be used together with physiologically based absorption models to predict clinical food effects caused by solubility and/or dissolution rate limitations.

Effects of prolonged copper exposure in the marine gulf toadfish ( Opsanus beta) : I. Hydromineral balance and plasma nitrogenous waste products

Acute (96 h) and prolonged (30 days) copper exposure induced osmoregulatory disturbance and impaired nitrogenous waste excretion in the marine teleost, the gulf toadfish ( Opsanus beta), which was found to be extremely tolerant to acute copper exposure with a 96 h LC50 exceeding 340 μM but exhibited disturbed mineral balance in response to both acute and prolonged exposure to ∼12 μM copper. The main cause of copper toxicity was found to be Na + and Cl − regulatory failure leading to elevated plasma [Na +] and [Cl −] and osmolality which in turn led to fluid loss from muscle tissue. Analysis of intestinal fluid composition revealed a complicated pattern of effects of copper exposure. Intestinal transport physiology was directly influenced by copper exposure with Cl − absorption being the most sensitive parameter. Evidence for increased Na + and fluid absorption when the fish exhibited elevated plasma osmolality indicates that the intestine may also exhibit a compensatory response to impairment of branchial transport processes, suggesting at least two target organs (gill and intestine) for copper toxicity in marine fish. Plasma Mg 2+ was elevated from approximately 1.5 mM to as much as 4.0 mM, likely as a result of increased branchial permeability. While plasma [ammonia] clearly responded to copper exposure, plasma [urea] exhibited a much more sensitive and pronounced response to both acute and prolonged copper exposure, resulting in as much as a three-fold increase in circulating urea levels. This response is most likely the result of the unique ability of this teleost to convert ammonia to urea.

Arachidonic acid intestinal absorption: mechanism of transport and influence of luminal factors of absorption in vitro.

The mechanism and characteristics of intestinal absorption of arachidonic acid were studied in vitro using everted intestinal sacs of the rat. Arachidonic acid absorption was studied at concentrations of 5 micron to 8.36 mM. The plot of absorption rate vs. concentration fitted best to a rectangular hyperbola at low micron concentrations and to a straight linear relationship in the mM range of concentrations. Metabolic inhibitors and uncouplers did not change absorption in either range of concentrations. The absorption of arachidonic acid increased with thinning of the unstirred water-layer, decrease in the pH, or the substitution of sodium taurocholate by Pluronic F 68 OR Tween 80. Absorption decreased following the equimolar additions of oleic, linoleic, and linolenic acids. Absorption rate did not change when the taurocholate concentration was varied from 5-15 mM or following the additions of butyric or glutamic acids, leucine, lysine, or dextrose. It was concluded that arachidonic acid is absorbed by a concentration-dependent dual mechanism of transport which is not energy dependent. At the low micron range of concentrations, facilitated diffusion is predominant, while at mM concentrations, simple diffusion is the dominant mechanism of absorption. Changes in the intestinal fluid composition, flow rate, and pH can modify the rate of absorption of arachidonic acid.

WATER AND ELECTROLYTE LOSSES DUE TO CHOLERA IN INFANTS AND SMALL CHILDREN: A RECOVERY BALANCE STUDY

Complete recovery balance studies on six infants with severe cholera are reported. Supporting data from a larger group of children with cholera, with particular reference to stool and small intestinal fluid composition, are also reported. The mean and range (in parentheses) for net retention at the end of the balance studies were: water, 96 gm (47—119); sodium, 10.5 mM (5.3—16.7); chloride, 8.9 mM (5.8—12.5); and potassium, 6.5 mM (4.3—9.7); all values are per kilogram of final body weight. Mean values for average evaporative water losses over day and night ranged from 31 gm to 63 gm/kg/24 hours. Stool output requiring parenteral replacement continued for 24 to 60 hours in five of six patients, and 48-hour stool output equaled 1:3 to 22% of final body weight. Stool composition in pediatric cholera is compared to that in infantile diarrhea and adult cholera. Significant differences from each of the latter two are described in this paper.