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Basolateral transport pathways for K+ and Cl- in rabbit proximal tubule: effects on cell volume.

To characterize the nature of K+ and Cl- transport pathways across basolateral membrane of rabbit proximal convoluted tubule, we used quantitative video microscopy to measure cell volume changes induced by rapid basolateral K+ and Cl- concentration changes. Elevating basolateral K+ resulted in cell swelling, which was largely inhibited by replacement of basolateral Cl- with cyclamate (85%) or by addition of 2 mM Ba2+ (72%). Substitution of basolateral Cl- by NO3- enhanced cell swelling, whereas substitution of Cl- by I- did not affect the K(+)-induced volume changes. Removal of Cl- from the bath reversed the cell swelling induced by raising K+ in the bath. Steady-state cell volume was 28% greater in hypotonic medium (250 mosmol/kgH2O) than in hypertonic medium (350 mosmol/kgH2O), and the rate of increase in cell volume induced by raising K+ was three times higher in hypotonic than in hypertonic medium. Substitution of Cl- by NO3- did not alter the effect of medium osmolality on K(+)-induced cell swelling, whereas addition of 0.2 mM diphenylamine-2-carboxylate inhibited the response (63%). We conclude that K(+)-induced cell swelling results from entry of K+ and Cl- into the cell across the basolateral membrane; it is proposed that transport of KCl across the basolateral cell membrane proceeds largely through two separate conductive pathways for K+ and Cl-. Cell swelling activates KCl transport occurring via K+ and Cl- channels across the basolateral membrane.

Animals↗

Basolateral ionic permeabilities of macula densa cells.

It has recently been shown that membrane ionic transport pathways of macula densa cells can be measured using conventional microelectrodes. To determine if conductances could be identified at the basolateral membrane of macula densa cells, cortical thick ascending limbs (CTAL) with attached glomeruli were continuously perfused with a 25 mM NaCl bicarbonate-free Ringer solution. Individual basolateral Na+, Cl-, NaCl, and K+ concentrations were altered by isosmotic replacement with N-methyl-D-glucamine and/or cyclamate. Reduction in basolateral [Na+] from 150 to 25 mM hyperpolarized basolateral membrane potential (Vbl) by 9.9 +/- 1.3 mV (n = 10; all data are corrected for changes in liquid junction potential at bath electrode). A decrease in bath [Cl-] from 150 to 25 mM depolarized Vbl by 20 +/- 2.4 mV (n = 13), whereas decreases in bath [NaCl] from 150 to 25 mM depolarized Vbl by 29 +/- 6.8 mV (n = 5). In the presence of 150 mM NaCl bathing solution, a stepwise increase in [K+] from 5 to 15 mM (by replacement of 10 mM NaCl with 10 mM KCl) depolarized Vbl by 3.3 +/- 1.1 mV (n = 8). After correction for individual transepithelial diffusion potentials, Cl conductance averaged 59 +/- 19% of the total basolateral conductance, whereas K+ (23 +/- 8%) and Na+ (17 +/- 10%) contributed significantly less to the overall basolateral conductance. These results indicate that membrane potential of macula densa cells may be very sensitive to alterations in intracellular Cl- activity and suggest that apical transport of NaCl through a furosemide-sensitive Na(+)-K(+)-2Cl- transporter may affect membrane potential in macula densa cells via a change in intracellular Cl- activity.

Action Potentials↗

Vasopressin activates a chloride conductance in cultured cortical collecting duct cells.

In rabbit cortical collecting duct (CCD) cells, arginine vasopressin (AVP) causes a transient increase followed by a sustained depression of transepithelial potential difference (PDte). Mechanisms underlying the decrease in PDte are not well understood. In this study, we used primary cultures of rabbit CCD cells to study effects of AVP. Basolateral addition of AVP caused a dose-dependent increase in transepithelial conductance (Gte) and a corresponding decrease in PDte. A significant effect was observed at 1 pM AVP, and half-maximal response occurred at 30 pM AVP; 1 nM AVP increased Gte and decreased PDte. Replacement of apical Na+ with N-methyl-D-glucamine did not prevent the effect of AVP on Gte, suggesting that it is not mediated by an increase in apical Na+ conductance. Similarly, apical Ba2+ (1 mM) or 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS, 0.1 mM) failed to prevent the effect of AVP. On the other hand, 5-nitro-2(3-phenylpropylamino)benzoic acid (0.1 mM) caused partial inhibition, whereas substitution of apical Cl- with gluconate or cyclamate almost completely prevented the AVP-induced increase in Gte. In unidirectional ion-flux studies, 1 nM AVP caused only a modest increase in apical-to-basolateral (A-->BL) flux of 22Na and had no effect on transepithelial flux of 86Rb in either direction. On the other hand, AVP caused a pronounced increase in A-->BL flux and BL-->A flux of 36Cl, resulting in an increased net Cl- absorption. The effect of AVP on Gte could be mimicked by 8-bromo-adenosine 3',5'-cyclic monophosphate (8-bromo-cAMP) and isoproterenol, and effects of AVP and isoproterenol were not additive.(ABSTRACT TRUNCATED AT 250 WORDS)

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Cytosolic [Ca2+] signaling pathway in macula densa cells.

Previous micropuncture studies suggested that macula densa (MD) cells might detect variations in luminal sodium chloride concentration ([NaCl]l) through changes in cytosolic calcium ([Ca2+]c). To test this hypothesis, MD [Ca2+]c was measured with fluorescence microscopy using fura 2 in the isolated perfused thick ascending limb with attached glomerulus preparation dissected from rabbit kidney. Tubules were bathed and perfused with a Ringer solution, [NaCl]l was varied and isosmotically replaced with N-methyl-D-glucamine cyclamate. Control [Ca2+]c, during perfusion with 25 mM NaCl and 150 mM NaCl in the bath, averaged 101. 6 +/- 8.2 nM (n = 21). Increasing [NaCl]l to 150 mM elevated [Ca2+]c by 39.1 +/- 5.2 nM (n = 21, P < 0.01). This effect was concentration dependent between zero and 60 mM [NaCl]l. The presence of either luminal furosemide or basolateral nifedipine or 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB), a potent Cl- channel blocker, significantly reduced resting [Ca2+]c and abolished the increase in [Ca2+]c in response to increased [NaCl]l. Nifedipine failed to produce a similar inhibitory effect when added exclusively to the luminal perfusate. Also, 100 nM BAY K 8644, a voltage-gated Ca2+ channel agonist, added to the bathing solution increased [Ca2+]c by 33.2 +/- 8.1 nM (n = 5, P < 0.05). These observations suggest that MD cells may detect variations in [NaCl]l through a signaling pathway that includes Na+-2Cl--K+ cotransport, basolateral membrane depolarization via Cl- channels, and Ca2+ entry through voltage-gated Ca2+ channels.

Animals↗

Gustatory neural coding in the monkey cortex: the quality of sweetness.

1. We analyzed the activity of single neurons in gustatory cortex of alert cynomolgus monkeys in response to the four basic taste stimuli and to a range of chemicals, all of which are predominantly sweet to humans. 2. We recorded taste-evoked responses from a cortical area that measured 4.0 mm in its anteroposterior extent, 5.6 mm dorsoventrally and 2.2 mm mediolaterally. Taste-responsive neurons constituted 4.7% of the 3,066 neurons tested in the course of 66 recording tracks. Nongustatory cells included those responsive to mouth movement (34.1%), tongue touch (1.9%), stimulus approach (0.7%), and tongue extension (0.5%). The functions of 58.2% of the cells we isolated could not be determined. 3. The mean breadth of tuning of these cortical taste neurons was a moderate 0.59 (range 0.00-0.93). 4. There was no evidence that taste cells with similar functional attributes were clustered in the cortex, i.e., there was no apparent topographic organization of taste qualities. 5. A taste space was generated from the correlations among patterns of neural activity evoked by the stimulus array. Within the space, NaCl was most isolated from other stimuli; the profiles elicited by HCl, quinine HCl, and water were all moderately intercorrelated and were clearly distinct from the cluster of sweet stimuli. 6. The 19 sweet chemicals formed a coherent cluster centered on the simple carbohydrates (glucose, fructose, sucrose, maltose) and sorbitol. Nearest this core were calcium cyclamate, aspartame, and cran-raspberry juice. In the next concentric ring were acesulfame potassium, xylose, xylitol, sorbose, polycose, and myoinositol. Increasingly distant from the sugars were sodium saccharin, stevioside, neohesperidin DHC, L-tryptophan and monellin. 7. We compared these results with those of a human psychophysical study of sweet stimuli. Using the position of glucose as a reference, we measured the distances to all other stimuli that were common to the two studies (n = 15). The correlation between the human psychophysical data and those derived from evoked activity in the macaque cortex was +0.82. 8. The high correlation between human psychophysical and macaque electrophysiological data implies that the subtle distinctions among stimuli that are predominantly sweet are quite similar for these two species and reinforces the value of this neural model for human taste perception.

Action Potentials↗

Primate sense of taste: behavioral and single chorda tympani and glossopharyngeal nerve fiber recordings in the rhesus monkey, Macaca mulatta.

The responses of 51 chorda tympani proper (CT) and 33 glossopharyngeal (NG) neural taste units from the rhesus monkey (Macaca mulatta) were recorded during stimulation of either the anterior (CT) or posterior (NG) part of the tongue with 26 stimuli that taste salty, umami, sour, bitter, and sweet to humans. In the CT, hierarchical cluster analysis separated four major clusters. The N and S clusters were most populous, followed by the H cluster and a small Q cluster. NaCl, monosodium glutamate (MSG), and MSG with guanosine 5'-monophosphate were the best stimuli in the N cluster. Amiloride suppressed responses to NaCl. KCl did not stimulate fibers from this cluster. S cluster fibers were characterized by strong responses to all sweeteners. The H cluster responded best to acids but also to some of the sweeteners such as xylitol, fructose, and sucrose. Q fibers responded well to quinine hydrochloride (QHCl) and caffeine, but not to denatonium benzoate. In the NG, hierarchical cluster analysis separated three major clusters. Q fibers formed the largest cluster. QHCl, caffeine, and sucrose octa-acetate but not denatonium benzoate elicited very strong responses in these fibers. S fibers formed a second cluster. Although most of the sweeteners stimulated the S fibers, their responses were not so pronounced as in CT S fibers. The small M cluster was formed by fibers that responded best to MSG. They also responded to NaCl and acids. Two bottle preference tests showed a positive relationship between a sweetener's ability to stimulate the taste fibers and the animals' consumption. Thus the most-liked sweeteners stimulated the S cluster fibers of CT best, whereas less-liked sweeteners such as D-phenylalanine elicited a response in Q fibers and sodium cyclamate stimulated N fibers. The results show that both CT and NG taste fibers of M. mulatta group according to the human concepts of taste qualities.

Animals↗

Gustatory responses of the hamster Mesocricetus auratus to various compounds considered sweet by humans.

The taste of 30 compounds was studied in the golden hamster with three different methods: single-fiber recordings, two-bottle preference (TBP), and conditioned taste aversion (CTA) tests. On the whole, the results showed that the sense of taste in the hamster differs in many respects from that in humans because, of 26 tested compounds known as sweet to humans, 11 had no taste or tasted differently. The results also supported the notion that activity in S-fibers elicits liking and activity in Q- or H-fibers rejection. Specifically hierarchial cluster analysis of 36 single fibers from the chorda tympani proper nerve separated N-, H-, and S-clusters consisting of 11 sucrose-, 14 NaCl-, and 11 citric-best fibers. Ace-K, cyanosuosan, N-4-cyanophenyl-N'-cyanoguanidineacetate (CCGA), -tryptophan, N-3, 5-dichlorophenyl-N'-(S)-alpha-methylbenzylguanidineacetate (DMGA), saccharin, SC-45647, and suosan stimulated only the S-fibers, were significantly preferred in TBP tests, and generalized to sucrose in the CTA tests. Ethylene glycol stimulated the N-fibers in addition to the S-fibers. This explains its generalization to sucrose in CTA. Its toxicity may contribute to its rejection in TBP tests. Sodium cyclamate stimulated a few N- but no S-fibers, which may explain the nondiscriminatory TBP and CTA results. Glycine elicited its largest response in the S-fibers, although it also stimulated other fibers. The resulting mixed taste sensation may explain why it was not preferred in TBP, although it generalized to sucrose in the CTA. Alitame, aspartame, N-4-cyanophenylcarbamoyl--aspartyl-(R)-alpha-methylbenzylamine (CAM), N-4-cyanophenylcarbamoyl-(R, S)-3-amino-3-(3, 4-methylenedioxyphenyl) propionic acid (CAMPA), N-(S)-2-methylhexanoyl--glutamyl-5-amino-2-pyridinecarbonitrile (MAGAP), N-1-naphthoyl--glutamyl-5-amino-2-pyridinecarbonitrile (NAGAP), NHDHC, superaspartame, and thaumatin were among the compounds considered sweet by humans that gave no response, were not discriminated in the TBP test, and gave no generalization in the CTA tests.

Animals↗

Sense of taste in a new world monkey, the common marmoset: recordings from the chorda tympani and glossopharyngeal nerves.

Whole nerve, as well as single fiber, responses in the chorda tympani proper (CT) and glossopharyngeal (NG) nerves of common marmosets were recorded during taste stimulation with three salts, four acids, six bitter compounds and more than 30 sweeteners. We recorded responses of 49 CT and 41 NG taste fibers. The hierarchical cluster analysis distinguished three major clusters in both CT and NG: S, Q, and H. The S(CT) fibers, 38% of all CT fibers, responded only to sweeteners. The S(CT) fibers did not respond during stimulation with salts, acids, and bitter compounds but exhibited OFF responses after citric and ascorbic acids, quinine hydrochloride (QHCl), and salts (in 80% of S(CT) fibers). S(NG) fibers, 50% of all NG fibers, also responded to sweeteners but not to stimuli of other taste qualities (except for citric acid, which stimulated 70% of the S(NG) fibers). Some sweeteners, including natural (the sweet proteins brazzein, monellin) and artificial [cyclamate, neohesperidin dihydrochalcone (NHDHC), N-3,5-dichlorophenyl-N'-(S)-alpha-methylbenzylguanidineacetate (DMGA), N-4-cyanophenylcarbamoyl-(R,S)-3-amino-3-(3,4-methylenedioxyphenyl) propionic acid (CAMPA)] did not elicit responses in the S fibers. In general, the response profiles of the S(CT) and S(NG) clusters were very similar, the correlation coefficient between the responses to sweeteners in these clusters was 0.94. Both the Q(CT) and the Q(NG) fibers (40 and 46% of all fibers) were predominantly responsive to bitter compounds, although their responses to the same set of bitter compounds were quite different. Sweeteners with sweet/bitter taste for humans also stimulated the Q clusters. The H clusters (22 and 3% of all fibers) were predominantly responsive to acids and did not respond to stimuli of other taste qualities. However, bitter stimuli, mainly QHCl, inhibited activity in 70% of H(CT) fibers. Among a total of 90 fibers from both nerves there was only 1 NaCl-best fiber in CT. We found, however, that 35% of the CT fibers reacted to salts with inhibition of activity during stimulation, followed by an OFF response. This OFF response was diminished or eliminated by amiloride. These characteristics indicate that amiloride-sensitive sodium channels are involved in salt transduction in marmosets. In the two NG fibers responding to NaCl, we recorded neither suppression by amiloride nor OFF responses. Comparison of marmoset data with those of other nonhuman primates studied, rhesus and chimpanzee, demonstrates phylogenetic trends in the organization of taste system. This can help to uncover pathways of primate evolution.

Animals↗

Electrophysiological identification of cell types in cortical collecting duct monolayers.

Double-barrel microelectrodes were used to determine membrane voltages and the intracellular pH (pHi) in primary cultures of cortical collecting duct cells (CCD) grown in the absence of aldosterone. Electrophysiologically, two main cell types were identified. In cell type 1, the apical membrane voltage (Va) was -60 +/- 5 mV. The fractional resistance of the apical membrane (fRa) was 0.40 +/- 0.03, and pHi was 7.21 +/- 0.04. Exposure to 50 mM K+ on the apical side depolarized Va by 21 +/- 4 mV. When Cl- was replaced by cyclamate two types of responses were observed: (a) depolarization of Va by 26 +/- 3 mV while pHi remained unchanged, and (b) no change in Va. In cell type 2, Va was -36 +/- 5 mV, fRa was 0.91 +/- 0.03 and increasing apical [K+] from 5 to 50 mM did not change Va. Two subpopulations were distinguished by the response of pHi to lowering apical [Cl-]. In one of them pHi increased from 6.99 +/- 0.05 to 7.11 +/- 0.07. In the other, pHi was significantly decreased from 7.16 +/- 0.08 to 7.03 +/- 0.07. These results are compatible with the conclusion that about 50% of the impaled cells type 2 have a Cl-/HCO-3 exchanger at the apical membrane. In summary, two different cell types can be identified electrophysiologically in CCD monolayers. Cell type 1 has the electrical characteristics of principal cells. Cell type 2 resembles the intercalated cells. The cell alkalinization observed in approximately 50% of the cells type 2 in response to Cl- removal suggests the presence of an apical Cl-/HCO-3 exchanger. Thus, these cells should be the bicarbonate-secreting cells. The remaining cells should correspond to the acid-secreting cells.

Animals↗

Swelling-activated Cl- channels support Cl- secretion by bovine ciliary epithelium.

PURPOSE: To determine whether swelling-activated Cl- currents (ICl,swell) observed in isolated nonpigmented ciliary epithelial (NPE) cells contribute to Cl- secretion across the ciliary epithelium. METHODS: Ion transport across intact bovine ciliary epithelium was monitored electrically. Native isolated bovine NPE cells were harvested enzymatically. Cell volume changes were measured by calcein-fluorescence quenching. RESULTS: Bilateral reduction in osmolality transiently increased short-circuit current (Isc), averaging 60% to 70%. Bilateral pretreatment with 5-nitro-2-(phenylpropylamino)-benzoate (NPPB), a Cl- channel blocker, reduced Isc stimulation by approximately 60%, suggesting that transcellular ICl,swell largely mediates the increased current. The hypotonically-triggered Isc stimulation was also inhibited by phloretin, a blocker of swelling-activated Cl- channels and by flufenamic acid, a blocker of Cl- and nonselective cation channels. Cyclamate substitution for bath Cl- reduced the baseline Isc and the increase in hypotonically-triggered Isc. In that case, addition of either NPPB or flufenamic acid did not produce further inhibition. The transepithelial responses were correlated with regulatory volume responses of freshly harvested NPE cells. Hypotonicity elicited a regulatory volume decrease (RVD) over a period comparable to that of the hypotonicity-triggered increase in Isc. The RVD was also inhibited by Cl--channel blockers and by Cl- substitution. CONCLUSIONS: ICl,swell of NPE cells is functionally expressed in intact ciliary epithelium and is oriented to subserve aqueous humor formation. NPE cell volume can be measured with calcein-fluorescence quenching. ICl,swell may be stimulated by increased stromal fluid uptake and delivery to the NPE cells, facilitating Cl- secretion and increasing fluid release into the posterior chamber.

Animals↗

Characteristics of salt and water transport in superficial and juxtamedullary straight segments of proximal tubules.

The purpose of the present studies was to characterize the nature of salt and water transport out of the superficial (SF) and juxtamedullary (JM) straight segments of rabbit proximal tubules as examined by in vitro microperfusion techniques. When the perfusate consisted of a solution simulating ultrafiltrate of plasma, there were no differences between SF and JM straight tubules in either net reabsorption of fluid (SF=0.47 nl/mm per min; JM=0.56 nl/mm per min) or in transtubular potential difference (PD) (SF=-2.1 mV; JM=-1.8 mV). Removal of glucose and alanine from the perfusate had no effect on the magnitude of the PD in either straight segment. Ouabain decreased both the net reabsorptive rates and the PD. Isosmolal replacement of NaCL by Na-cyclamate (a presumed impermeant anion) in the perfusate and the bath caused an increase in luminal negativity in both segments wheras similar substitution of NaCL by choline-CL (nontransported cation) changed the PD TO NEAR ZERO. These studies, therefore, suggest that sodium is transported out of the proximal straight tubules by an active noncoupled process that generates a PD (electrogenic process). When the perfusate consisted of a solution with a high chloride concentration (resulting from greater HCO3 than CI reabsorption in the proximal convoluted tubule), different PDs in SF and JM tubules were generated: SF=+1.6 plus or minus 0.2 mV; JM=-1.3 plus or minus 0.3 mV. This difference in PD was attributed to relative differences in Na and CI permeabilities in these two segments. Electrophysiological and isotopic estimates of the chloride to sodium permeability revealed that the SF tubule is about twice as permeant to chloride than to sodium whereas the JM tubules are approximately twice as permeant to sodium than to chloride. It is concluded that the mechanism of active sodium transport in the straight segment of proximal tubule differs from that of the convoluted segment and that both the SF and JM straight segments differ from each other with respect os sodium and chloride permeability.

Animals↗

Mechanism of NaCl and water reabsorption in the proximal convoluted tubule of rat kidney.

The role of chloride concentration gradients in proximal NaCl and water reabsorption was examined in superficial proximal tubules of the rat by using perfusion and collection techniques. Reabsorptive rates (Jv), chloride concentrations, and transtubular potential difference were measured during perfusion with solutions (A) simulating an ultrafiltrate of plasma; (B) similar to (A) except that 20 meq/liter bicarbonate was replaced with acetate; (C) resembling late proximal fluid (glucose, amino acid, acetate-free, low bicarbonate, and high chloride); and (D) in which glucose and amino acids were replaced with raffinose and bicarbonate was partially replaced by poorly reabsorbable anions (cyclamate,sulfate, and methyl sulfate). In tubules perfused with solutions A and B, Jv were 2.17 and 2.7 nl mm-1 min-1 and chloride concentrations were 131.5 +/- 3.1 and 135 +/- 395 meq/liter, respectively, indicating that reabsorption is qualitatively similar to free-flow conditions and that acetate adequately replaces bicarbonate. With solution C, Jv was 2.10 nl mm-1 min-1 and potential difference was +1.5 +/- 0.2 mV, indicating that the combined presence of glucose, alanine, acetate, and bicarbonate per se is not an absolute requirement. Fluid reabsorption was virtually abolished when tubules were perfused with D solutions; Jv was not significantly different from zero despite sodium and chloride concentrations similar to plasma; chloride concentration was 110.8 +/- 0.2 meq/liter and potential difference was -0.98 mV indicating that chloride was close to electrochemical equilibrium. These results suggest the importance of the chloride gradient to proximal tubule reabsorption in regions where actively reabsorbable solutes (glucose, alanine, acetate, and bicarbonate) are lacking and provide further evidence for a passive model of NaCl and water transport.

Absorption↗

Characteristics of volume reabsorption in rabbit superficial and juxtamedullary proximal convoluted tubules.

Segments of superficial and juxtamedullary proximal convoluted tubules of the rabbit were perfused in vitro to examine the mechanisms responsible for net volume reabsorption. The very early postglomerular segments were not studied. Fluid reabsorptive rates and transepithelial potential differences were compared under various conditions: (a) with perfusate that simulated glomerular filtrate; (b) with perfusate that lacked glucose, amino acids, and acetate and that had HCO(3) and Cl concentrations of 5 and 140 mM, respectively; (c) with perfusate that lacked glucose, amino acids, and acetate but with 20 meq of NaHCO(3) replaced with 20 meq of Na cyclamate; (d) with the same perfusate as in b but in the presence of ouabain in the bath; (e) with ultrafiltrate of rabbit serum titrated with HCl to final HCO(3) and Cl concentrations of 2 and 134 mM, respectively. Tubules were perfused with this titrated ultrafiltrate at 37 degrees C, 21 degrees C, and in the presence of 0.1 mM ouabain in the bath. Bath fluid in all experiments was regular rabbit serum. Under conditions a and b superficial proximal convoluted tubule (SFPCT) and juxtamedullary proximal convoluted tubule (JMPCT) behaved similarly with the exception that SFPCT exhibited a lumen-positive and JMPCT a lumen-negative electrical potential under condition b. However, under condition c SFPCT failed to exhibit net volume reabsorption, whereas reabsorption in JMPCT continued unchanged. Ouabain did not affect volume reabsorption in SFPCT under condition d, whereas neither ouabain nor hypothermia affected SFPCT under condition e. In contrast, ouabain and hypothermia totally inhibited volume reabsorption in JMPCT under conditions d and e. These studies document heterogeneous mechanisms responsible for volume reabsorption in the major portions of SFPCT and JMPCT with passive forces predominating in SFPCT and active forces in JMPCT.

Absorption↗

Review of present and future use of nonnutritive sweeteners.

In response to growing consumer demand for better tasting, low-calorie, sugar-free food products, the number of food items containing nonnutritive sweeteners has grown markedly in recent years. In this paper, present sweetener consumption is reviewed; the history, properties, uses, advantages, and safety of approved sweeteners such as saccharin, aspartame, and acesulfame-K are presented, as well as those of sweeteners such as cyclamate, sucralose, and alitame that are awaiting FDA approval; the role of sweeteners in the dietary management of persons with diabetes is discussed; and counseling guidelines for safe consumption are given.

Diabetes Mellitus↗

Sulfadiazine-induced sialadenitis.

OBJECTIVE: To present a case of sialadenitis induced by sulfadiazine. CASE SUMMARY: A 50-year-old man with bronchitis developed swelling of the parotid glands and floor of the mouth as well as plugging in his ears 2 hours after ingestion of Bronco-aseptilex (sulfadiazine, cyclamate, and guaiacol). Skin-prick tests performed with serial dilutions of sulfadiazine and Bronco-aseptilex were all negative. We then performed oral challenge tests with Bronco-aseptilex, sulfadiazine, and other compounds of the commercial syrup. Enlargement of the salivary glands was observed after oral challenge tests with Bronco-aseptilex and sulfadiazine. CONCLUSIONS: We describe a patient who developed drug-induced sialadenitis. The oral challenge test confirmed that sulfadiazine was the etiologic agent.

Anti-Infective Agents↗

Two-generation saccharin bioassays.

The controversy regarding the safety of saccharin for human consumption started shortly after its discovery over 100 years ago and has yet to subside appreciably. The consumption of saccharin, particularly in North America, began to escalate when the U.S. Food and Drug Administration set new standards of identity which allowed foods containing artificial sweeteners to be promoted as "nonnutritive" or "noncaloric" sweeteners for use by the general public. In 1969, when cyclamates were banned, at least 10 single-generation feeding studies were undertaken with saccharin to more accurately assess the potential toxicological consequences resulting from the anticipated increase in its consumption. None of these studies resulted in any overt regulatory action. Subsequently, the introduction of the two-generation chronic toxicity/carcinogenicity bioassay added a new tool to the toxicologist's arsenal. Three two-generation studies using saccharin have since been conducted. The results from these studies clearly show that when rats were exposed to diets containing 5 or 7.5% sodium saccharin from the time of conception to death, an increased frequency of urinary bladder cancers was found, predominantly in the males. While some study results suggested that impurities in commercial saccharin or the presence of urinary tract calculi may have been responsible for the observed bladder tumors, it now appears that these possibilities are highly unlikely. The mechanism by which saccharin elicited the bladder tumors using the two-generation experiment has not been ascertained.

Animals↗

Effect of promoters on incidence of bladder cancer in experimental animal models.

Multistage models of carcinogenesis proposed to account for the observed patterns of tumor development in the skin, liver, lung, bladder and other organs involve initiation of neoplastic change in a few cells by a threshold dose of carcinogen followed by conversion of these latent tumor cells into an autonomous cancer by further doses of the same and/or other carcinogens, and/or noncarcinogenic promoting agents. In the rat urinary bladder, neoplastic change can be initiated by a few weeks treatment with low doses of N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) or N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) or by a single, low dose, intravesicular instillation of N-methyl-N-nitrosourea (MNU). Very few animals treated thus will develop bladder cancer unless exposed subsequently to some further regime which will promote tumor growth from the initiated cells. Many different factors will stimulate tumor growth in the initiated rat bladder, including further low doses of complete bladder carcinogens, dietary factors such as metabolites of tryptophan or deficiency of vitamin A, the food additives saccharin and cyclamate and some alkylating agents such as cyclophosphamide and methylmethane sulfonate. New and published evidence is reviewed which supports the belief that these and other factors are promoters or later stage carcinogens in the bladder. The difficulties of defining a promoter and of identifying markers of promotion, i.e., of distinguishing the second from the later stages of carcinogenesis in the urinary bladder, are discussed with reference to the action of promoters in the mouse skin initiation/promotion model. However, in terms of their effect on an initiated population on the risk of developing cancer, it is suggested that such a distinction is largely irrelevant. Since both second and later stage carcinogens accelerate tumor development in an initiated urothelium, they both have the potential to lower the age at which bladder cancer becomes symptomatic. They are thus as important as are initiating carcinogens in determining the patterns of age-related neoplastic disease in any population.

Adenocarcinoma↗

Promotion in urinary bladder carcinogenesis.

Aromatic amines, including 2-naphthylamine, 4-aminobiphenyl and benzidine, are known urinary bladder carcinogens in man and other species, but in rodents, aromatic amines and amides have usually induced liver tumors, occasionally also with tumors of the bladder and other tissues. Variations in organ specificity are related to differences in metabolism; for the production of bladder tumors, the rates of acetylation and deacetylation appear to be critical. Bladder specific carcinogens in rodents and other species have subsequently been identified, including N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) administered in the drinking water, N-[(4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) in the diet and N-methyl-N-nitrosourea (MNU) instilled intravesically. When low doses of several bladder carcinogens (BBN, FANFT, 2-acetylaminofluorene, and 3,3'-dichlorobenzidine) are administered to rats, either simultaneously or sequentially, a synergistic effect is observed with respect to bladder carcinogenesis. In addition, a multistage carcinogenesis process has been demonstrated for the rat bladder using MNU or FANFT as initiators, and dietary sodium saccharin, sodium cyclamate, or tryptophan as promoters. Calculi (or pellets) appear to enhance the promotion process but are not necessary for it to occur. Recent studies also indicate that urine has a role in the promoting process. The urothelium normally has a very low mitotic rate. If mucosal proliferation is increased, such as during fetal development or during regeneration and repair of an ulcer, the bladder appears to be considerably more sensitive to the effects of promoting substances. For example, if sodium saccharin is administered to rats after ulceration of the bladder, even without prior administration of an initiator, bladder carcinoma develops. Under these conditions, the substance appears as a carcinogen. Human populations with increased bladder epithelial proliferation, such as fetus, infants, patients with bacterial cystitis or men with partially obstructive prostatism, may have increased susceptibility to the action of carcinogenic or promoting stimuli.

Animals↗