Response of the renal inner medulla to hypoxia: possible defense mechanisms.

BACKGROUND/AIMS: Owing to the precarious blood supply to the renal medulla and the high metabolic requirement of the medullary thick ascending limb of Henle's loop, this nephron segment should be especially vulnerable when its supply of O(2) declines. METHODS: Rats were exposed to 8 or 21% O(2) at different time points up to 5 h, and samples were collected for urine flow rate, urine (U(osm)) and renal papillary (RP(osm)) osmolality, urinary excretion of Na(+), Cl(-), K(+) and Mg(2+), blood gases, erythropoietin and vasopressinase activity in plasma. Other groups of rats were pretreated with desmopressin acetate (dDAVP) or underwent bilateral nephrectomy (BNX) 1 h prior to the exposure. RESULTS: Hypoxic rats had water diuresis (WD) within 2.5 h, as evidenced by lower U(osm) (333 ± 42 mosm/l) and RP(osm) (869 ± 57 mosm/l), thus suggesting that hypoxia led to a failure to achieve osmotic equilibrium within the renal papilla. Circulating vasopressinase activity increased, which was partially renal in origin because it was lower after BNX. The renal concentrating ability during hypoxia was maintained with dDAVP pretreatment, suggesting that dDAVP may have improved O(2) delivery and the active reabsorption of Na(+) in the renal medullary region. CONCLUSIONS: WD or high vasopressinase activity may be valuable diagnostic tools to assess renal medullary hypoxia. Pretreatment with dDAVP may prevent these changes during hypoxia.

Non-natriuretic doses of furosemide: potential use for decreasing the workload of the renal outer medulla with minimal magnesium wasting in the rat.

BACKGROUND/AIMS: Since furosemide (FS) inhibits active Na(+) reabsorption by medullary thick ascending limb (mTAL) in the renal outer medulla, it may decrease its work during periods of low O2 supply to deep in the renal outer medulla. This study was designed to demonstrate that there may be a dose of FS would reduce its metabolic work while preventing the excessive loss of magnesium (Mg(2+)). Mg(2+) is important because the ATP needed to perform work must have bound Mg(2+) to it. METHODS: Rats were injected intraperitoneally with a range of doses of FS. The measured outcomes were urine flow rate and parameters of functions of the mTAL (i.e. urine and renal papillary osmolality and urinary excretion of Na(+), Cl(-), K(+) and Mg(2+), and concentrations of Mg(2+) in serum). RESULTS: The urine flow rate increased significantly starting at 2.4 mg FS/kg. The renal papillary osmolality decreased at ≥0.4 mg FS/kg, and the large detectable natriuresis started at 1.6 mg FS/kg. At this latter dose, the urinary excretion of Mg(2+) rose significantly. CONCLUSION: In rats, the non-natriuretic dose of FS may reduce the work of the mTAL. The earliest indicator of reduced work in the mTAL appears to be a decrease in urine osmolality rather than a rise in urine flow rate. Higher doses of FS should be avoided, as they induce high rates of Mg(2+) excretion, which can deplete the body of this essential electrolyte.

Importance of Residual Water Permeability on the Excretion of Water during Water Diuresis in Rats.

When the concentration of sodium (Na(+)) in arterial plasma (P(Na)) declines sufficiently to inhibit the release of vasopressin, water will be excreted promptly when the vast majority of aquaporin 2 water channels (AQP2) have been removed from luminal membranes of late distal nephron segments. In this setting, the volume of filtrate delivered distally sets the upper limit on the magnitude of the water diuresis. Since there is an unknown volume of water reabsorbed in the late distal nephron, our objective was to provide a quantitative assessment of this parameter. Accordingly, rats were given a large oral water load, while minimizing non-osmotic stimuli for the release of vasopressin. The composition of plasma and urine were measured. The renal papilla was excised during the water diuresis to assess the osmotic driving force for water reabsorption in the inner medullary collecting duct. During water diuresis, the concentration of creatinine in the urine was 13-fold higher than in plasma, which implies that ~8% of filtered water was excreted. The papillary interstitial osmolality was 600 mOsm/L > the urine osmolality. Since 17% of filtered water is delivered to the earliest distal convoluted tubule micropuncture site, we conclude that half of the water delivered to the late distal nephron is reabsorbed downstream during water diuresis. The enormous osmotic driving force for the reabsorption of water in the inner medullary collecting duct may play a role in this reabsorption of water. Possible clinical implications are illustrated in the discussion of a case example.

Is there escape from renal actions of vasopressin in rats with a hyponatremia for greater than 48 hours?

Escape from the renal actions of vasopressin is said to occur in rats with chronic hyponatremia. Our objective was to provide specific evidence to test this hypothesis. Hence the osmolality in the excised renal papilla and in simultaneously voided urine (U(Osm)) was measured in rats with and without hyponatremia. To induce hyponatremia, rats were fed low-electrolyte chow for 6 days. In the first 3 days, water was provided ad lib. On days 4 to 6, a long acting vasopressin preparation (dDAVP) was given every 8 hours to induce water retention. The hyponatremic rats drank 21 mL 5% sucrose on day 4 and 6 mL on day 5. On the morning of day 6, these rats were given 10 mL of 5% glucose in water (D5W) by the intraperitoneal route at 09:00 hour and at 11:00 hour. Analyses were performed in blood, urine, and the excised renal papilla at 13:00 hour on day 6. The concentration of Na(+) in plasma (P(Na)) in rats without intraperitoneal D5W was 140±1 mEq/L (n=7) whereas it was 112±3 mEq/L in the hyponatremic group (n=12). The hyponatremic rats had a higher osmolality in the excised papillary (1,915±117 mOsm/kg H(2)O) than the U(Osm) (1,528±176 mOsm/kg H(2)O, P<0.05). One explanation for this difference is that the rats escaped from the renal action of vasopressin. Nevertheless, based on a quantitative analysis, other possibilities will be considered.

Studies to identify the basis for an alkaline urine pH in patients with calcium hydrogen phosphate kidney stones.

BACKGROUND: Patients with CaHPO(4) kidney stones belong to a diagnostic category that has a high urine pH as its common feature. Our objective was to provide a new clinical approach to examine the basis for this high pH. METHODS: The study group consisted of 26 CaHPO(4) stone formers and 28 normal volunteers. Urine was collected q2h plus an overnight sample to identify patients with a urine pH > 6.5 for 12/24 h. Urine ammonium (U(NH4)), sulphate (U(SO4)) and citrate were measured and diet net alkali was calculated. RESULTS: Of the 26 patients, 13 had persistently alkaline urine. In 7/13, U(NH4) (68 +/- 13 mEq/day) and U(SO4) (57 +/- 7 mEq/day) were both high. In 6/13 patients, U(NH4) was the usual 31 +/- 3 mEq/day; in 4/6, U(NH4)/U(SO4) was 0.9 +/- 0.1; the cause of the alkaline urine pH seemed to be a dietary alkali load because the rise in urine pH was episodic and coincided with a high net diet alkali load and peak citrate excretion rates. The remaining two patients had a high U(NH4)/U(SO4) (2.2 and 1.6). Citrate excretion was very low in the male, but not in the female patient. CONCLUSIONS: There are heterogeneous causes for a persistently high urine pH. Two of the patients had a possible molecular basis: the lesion could be a low proximal convoluted tubule cell pH in the male and an increased entry of NH(3) into the late distal nephron in the female.

Control of potassium excretion: a Paleolithic perspective.

PURPOSE OF REVIEW: Regulation of potassium (K) excretion was examined in an experimental setting that reflects the dietary conditions for humans in Paleolithic times (high, episodic intake of K with organic anions; low intake of NaCl), because this is when major control mechanisms were likely to have developed. RECENT FINDINGS: The major control of K secretion in this setting is to regulate the number of luminal K channels in the cortical collecting duct. Following a KCl load, the K concentration in the medullary interstitial compartment rose; the likely source of this medullary K was its absorption by the H/K-ATPase in the inner medullary collecting duct. As a result of the higher medullary K concentration, the absorption of Na and Cl was inhibited in the loop of Henle, and this led to an increased distal delivery of a sufficient quantity of Na to raise K excretion markedly, while avoiding a large natriuresis. In addition, because K in the diet was accompanied by 'future' bicarbonate, a role for bicarbonate in the control of K secretion via 'selecting' whether aldosterone would be a NaCl-conserving or a kaliuretic hormone is discussed. SUMMARY: This way of examining the control of K excretion provides new insights into clinical disorders with an abnormal plasma K concentration secondary to altered K excretion, and also into the pathophysiology of calcium-containing kidney stones.

Requirements for a high rate of potassium excretion in rats consuming a low electrolyte diet.

Control mechanisms for potassium (K(+)) excretion in humans developed in Palaeolithic times when diets were sodium poor and episodically K(+) rich. Nevertheless, our understanding of the regulation of K(+) excretion comes from experiments in rats with large sodium and K(+) intakes. Our objective was to identify how K(+) excretion was regulated when rats consumed a low NaCl diet to reflect Palaeolithic conditions. Rats that were given mineralocorticoids plus either NaCl, mannitol, or NaHCO(3) had a small kaliuresis. In contrast, KCl load induced a large kaliuresis and a near-maximal luminal [K(+)] in the terminal cortical collecting duct ([K(+)](CCD)). The time course of events was important. The rise in the [K(+)](CCD) was prompt, but the initial kaliuresis was only modest. Over the next 4 h, kaliuresis increased markedly due solely to a higher calculated distal flow rate, which appeared to be due to diminished reabsorption of NaCl in the loop of Henle; of note, the measured papillary [K(+)] rose. In summary, the increase in the [K(+)](CCD) in rats given KCl is likely to be due to an increase in the number of luminal K(+) channels rather than to mechanisms that are known to induce a lumen-negative voltage in cortical distal nephron segments. The higher distal flow rate might be due to a higher interstitial [K(+)], which inhibited NaCl reabsorption in the loop of Henle. Thus, to understand which of the potential control mechanisms are operating, one must look very closely at the conditions imposed by the experimental setting.

Properties permitting the renal cortex to be the oxygen sensor for the release of erythropoietin: clinical implications.

The PO2 at this site where erythropoietin release is regulated should vary only when the hemoglobin concentration changes in capillary blood. The kidney cortex is an ideal location for this O2 sensor for four reasons. First, it extracts a small proportion of the oxygen that is delivered in each liter of blood; this makes the PO2 signal easier to recognize. Second, there is a constant ratio of the work performed (consumption of O2) to the renal blood flow rate (delivery of O2). Third, the high renal blood flow rate improves diffusion of O2 from capillaries to this O2 receptor. Fourth, a high renal cortical PCO2 prevents an additional shift of the O2:hemoglobin dissociation curve by other factors from being a confounding variable. This suggests that the GFR and the renal blood flow rate should be examined in patients with unexplained anemia or erythrocytosis.

Physiology of acid-base balance: links with kidney stone prevention.

Two processes permit the urine pH and the medullary interstitial pH to remain in an "ideal range" to minimize the risk of forming kidney stones. First, a medullary shunt for NH(3) maintains the urine pH near 6.0 to minimize uric acid precipitation when distal H(+) secretion is high. Second, excreting dietary alkali excreting alkali as a family of organic anions--including citrate--rather than as bicarbonate maintains the urine pH near 6.0 while urinary citrate chelates ionized calcium, which minimizes CaHPO(4) precipitation. In patients with idiopathic hypercalciuria and recurrent calcium oxalate stones, the initial nidus is a calcium phosphate precipitate on the basolateral membrane of the thin limb of the loop of Henle (Randall's plaque). Formation of this precipitate requires medullary alkalinization; K(+) -depletion and augmented medullary H(+)/K(+) -ATPase may be predisposing factors.

Defining conditions that lead to the retention of water: the importance of the arterial sodium concentration.

BACKGROUND: A water diuresis occurs when a large volume of water is ingested rapidly. Nevertheless, water conservation is required to provide a source of water for evaporative heat dissipation throughout the day. Therefore, the objective was to define conditions that permit the retention of ingested water. METHODS: Volunteers collected urine q2h plus an overnight specimen; water loading was conducted after overnight food and water restriction; paired arterialized and venous blood samples were analyzed. RESULTS: When 20 mL water/kg was consumed in <15 minutes, the peak urine flow rate was 11 +/- 0.6 mL/min. The volume of water retained after water intake stopped, and when the urine was hyperosmolar, correlated directly with the daily excretion of sodium plus potassium (r(2)= 0.63). The plasma sodium concentration (P(Na)) was 4.0 +/- 0.5 mmol/L lower in arterialized than paired venous blood 30 to 40 minutes after water ingestion began (P < 0.01). In preliminary studies, the smallest water load consumed in 15 minutes that would reproducibly cause a water diuresis was defined in each subject. This same acute water load was retained, however, if it contained 150-mmol/L fructose, but not glucose, or if it was consumed slowly (sipping). The arterialized P(Na) was not significantly lower than in paired venous samples when water was sipped. CONCLUSION: A large fall in arterialized and not venous P(Na) best reflected the signal to induce a water diuresis. Although a very large water load can induce a water diuresis, smaller water loads can be retained for future heat dissipation.

Dogmas and controversies in the handling of nitrogenous wastes: excretion of nitrogenous wastes in human subjects.

Two major nitrogenous waste products, urea and ammonium (NH(4)(+)), are produced in humans when proteins are oxidized, and in this manuscript their excretions are examined from two perspectives. First, the specific physiology of each nitrogenous waste is reviewed and the current dogmas summarized. Second, their excretions are considered in the context of integrative physiology, i.e. the need to ensure that the urine composition is appropriate to minimize the risk of kidney stone formation. After the latter analysis, weak links in our understanding of the overall physiology become apparent and a conundrum is defined. The conundrum for the excretion of urea focuses on the fact that urea is not an effective osmole in the medullary-collecting duct when vasopressin acts. As a result, it appears that urinary urea cannot prevent a large decline in the urine flow rate and thereby minimize the risk of forming kidney stones in electrolyte-poor urine. The conundrum for the excretion of NH(4)(+) is: high rates of NH(4)(+) excretion require a low urine pH, yet a pH approximately 6.0 must be maintained in order to reduce the risk of precipitating uric acid in the urine. Possible ways of resolving these conundrums require novel physiological interpretations.

Facilitating an understanding of integrative physiology: emphasis on the composition of body fluid compartments.

As a teaching exercise, we used deductive reasoning and a quantitative analysis to convert a number of facts into a series of concepts to facilitate an understanding of integrative physiology and shed light on the composition of the different body fluid compartments. The starting point was the central need to regenerate ATP to perform biologic work. Because a large quantity of O2 must be delivered to cells at a sufficiently high concentration to aid its diffusion into mitochondria, approximately one third of the O2 in inspired air was extracted; this led to a P(CO2) in arterial blood of 40 mmHg (1 mmHg = 133.322 Pa). Blood flow to individual organs must be adjusted precisely to avoid having too low or too high a P(O2) in mitochondria--the latter augments the formation of reactive O2 species. The extracellular fluid (ECF) bicarbonate concentration (E(HCO3)) must be high to minimize H+ buffering by proteins. This high E(HCO3) sets the ECF concentrations of ionized calcium (Ca2+) and inorganic phosphate (HPO4(2-)) because of solubility issues. Three features defined the intracellular fluid (ICF) volume and composition. First, expelling monovalent anions minimized its mass (volume). Second, controlling the tissue P(CO2) ensured a relatively constant net valence on intracellular proteins. Third, the range of ICF Ca2+ concentrations must both induce regulatory signals and avoid Ca3(PO4)2 formation. All the above were incorporated into the integrated response that optimized the capacity for vigorous exercise.

Effect of fasting for two days on the excretion of ammonium in dogs with chronic metabolic acidosis.

BACKGROUND: The source of glutamine for renal ammonium production is ultimately dietary protein in the fed state and body proteins in fasting. OBJECTIVE: Our objective was to determine if less NH(+)(4) would be excreted by fasted dogs with chronic metabolic acidosis resulting in conservation of lean body mass. METHODS: Acid-loaded fed and fasted dogs were given 10 mmol NH(4)Cl/kg for 5 days; the fasted group had food withheld on days 4 and 5. RESULTS: The renal production of NH(+)(4) was not significantly different in both acid-loaded groups, yet the rate of NH(+)(4) excretion was significantly lower in the fasted dogs (8 vs. 21 mmol NH(+)(4)/mmol creatinine). The urine pH was significantly higher (6.0 versus 5.5) while titratable acid and the urine flow rate were significantly lower in these fasted dogs. Despite nearly equal urine flow rates and Na(+) excretion rates after an infusion of saline, the fasted dogs failed to increase the rate of excretion of NH(+)(4) to rates seen in the fed group. CONCLUSIONS: The lower rate of excretion of NH(+)(4) in fasted, acidotic dogs appeared to be due to a lower distal H(+) secretion. This may help preserve lean body mass during fasting.

Influence of hypernatraemia and urea excretion on the ability to excrete a maximally hypertonic urine in the rat.

Rats normally excrete 20-25 mmol of sodium (Na+) + potassium (K+) per kilogram per day. To minimize the need for a large water intake, they must excrete urine with a very high electrolyte concentration (tonicity). Our objective was to evaluate two potential factors that could influence the maximum urine tonicity, hypernatraemia and the rate of urea excretion. Balance studies were carried out in vasopressin-treated rats fed a low-electrolyte diet. In the first series, the drinking solution contained an equivalent sodium chloride (NaCl) load at 150 or 600 mmol l(-1). In the second series, the maximum urine tonicity was evaluated in rats consuming 600 mmol l(-1) NaCl with an 8-fold range of urea excretion. Hypernatraemia (148 +/- 1 mmol l(-1)) developed in all rats that drank 600 mmol l(-1) saline. Although the rate of Na+ + K+ excretion was similar in both saline groups, the maximum urine total cation concentration was significantly higher in the hypernatraemic group (731 +/- 31 vs. 412 +/- 37 mmol l(-1)). Only when the rate of excretion of urea was very low, was there a further increase in the maximum urine total cation concentration (1099 +/- 118 mmol l(-1)). Thus hypernatraemia was the most important factor associated with a higher urine tonicity.

Hypercalcaemia and metabolic alkalosis with betel nut chewing: emphasis on its integrative pathophysiology.

BACKGROUND: Events in the gastrointestinal tract that might contribute to a high absorption of calcium were simulated in vitro to evaluate why only a small proportion of individuals who ingest alkaline calcium salts develop hypercalcaemia, hypokalaemia and metabolic alkalosis. METHODS: A patient who chewed and swallowed around 40 betel nuts daily developed hypercalcaemia, metabolic alkalosis, hypokalaemia with renal potassium wasting, and renal insufficiency. The quantities of calcium and alkali per betel nut preparation were measured. Factors that might increase intestinal absorption of calcium were evaluated. RESULTS: Hypercalcaemia in the index case was accompanied by a high daily calcium excretion (248 mg, 6.2 mmol). Circulating levels of 1,25-dihydroxyvitamin D(3) and parathyroid hormone were low. Hypokalaemia with a high transtubular K(+) concentration gradient, metabolic alkalosis, a low excretion of phosphate and a very low glomerular filtration rate were prominent features. CONCLUSIONS: Possible explanations for the pathophysiology of metabolic alkalosis and hypokalaemia are provided. We speculate that a relatively greater availability of ionized calcium than inorganic phosphate in the lumen of the intestinal tract could have enhanced dietary calcium absorption.

Mechanisms used to dispose of progressively increasing alkali load in rats.

Our objective was to describe the process of alkali disposal in rats. Balance studies were performed while incremental loads of alkali were given to rats fed a low-alkali diet or their usual alkaline ash diet. Control groups received equimolar NaCl or KCl. Virtually all of the alkali was eliminated within 24 h when the dose exceeded 750 micromol. The most sensitive response to alkali input was a decline in the excretion of NH(4)(+). The next level of response was to increase the excretion of unmeasured anions; this rise was quantitatively the most important process in eliminating alkali. The maximum excretion of citrate was approximately 70% of its filtered load. An even higher alkali load augmented the excretion of 2-oxoglutarate to >400% of its filtered load. Only with the largest alkali load did bicarbonaturia become quantitatively important. We conclude that renal mechanisms eliminate alkali while minimizing bicarbonaturia. This provides a way of limiting changes in urine pH without sacrificing acid-base balance, a process that might lessen the risk of kidney stone formation.

Studies on the pathophysiology of the low urine pH in patients with uric acid stones.

BACKGROUND: A very low urine pH is the major risk factor for uric acid stone formation. METHODS: A subgroup of patients with a history of uric acid stones and a persistently low urine pH (<5.5 for at least 12 h/day) were selected for detailed study. Based on their relative ammonium (NH(+)(4)) and sulfate (SO(2-)(4)) excretions, patients were divided into two groups. RESULTS: The first group (N = 2) excreted 173 and 139% more NH(+)(4) than SO(2-)(4). Their daily urinary unmeasured anion excretion was higher than their calculated net diet alkali input (38 and 61 vs. 24 and 49 mEq, respectively). In the second group (N = 12), NH(+)(4) excretion was 69 +/- 5% that of SO(2-)(4). In 2 of 12, decreased renal ammoniagenesis was suspected due to a plasma potassium of 5.3 mmol/L and/or a lower GFR (65 and 59 L/day); these patients had an extremely low citrate excretion (3 and 1 mEq/day). In contrast, citrate excretion was not low in the remaining 10 patients (10.4 +/- 1.3 mEq/day). CONCLUSIONS: Patients in group 1 needed a higher NH(+)(4) excretion possibly because of a H+ load from excessive renal excretion of organic anions. We speculate that an alkaline proximal tubular cell pH could be the basis for the low NH(+)(4) and high citrate excretions in 10 of 12 patients in group 2. Dietary factors and/or a molecular lesion may contribute to their pathophysiology.