Michael Naim

Changes in IP3 and cytosolic Ca2+ in response to sugars and non‐sugar sweeteners in transduction of sweet taste in the rat.

1. The transduction pathways of sweet‐sensitive cells in rat circumvallate (CV) taste buds were investigated with assays for inositol 1,4,5‐trisphosphate (IP3) and with Ca2+ imaging. Stimulation with the non‐sugar sweeteners SC‐45647 and saccharin rapidly increased the cellular content of IP3 by 400 pmol (mg protein)‐1, while sucrose had a much smaller effect on IP3. As shown previously, sucrose, but not saccharin, increased the content of cyclic adenosine monophosphate (cAMP) of this preparation. 2. Stimulation of isolated CV taste buds with SC‐45647 increased the cytosolic Ca2+ concentration ([Ca2+]i) by 56.7 +/‐ 3.2 nM (n = 181). Due to the non‐confocality of the measuring system, these concentrations are underestimates. The increase in [Ca2+]i did not require the presence of extracellular Ca2+, suggesting that the Ca2+ release was from intracellular stores. 3. Individual cells responding to the non‐sugar sweeteners with Ca2+ release also responded to sucrose and to forskolin with an increase in [Ca2+]i. Such cells did not respond to the bitter tastant denatonium chloride. 4. Responses to sucrose were abolished by lowering the Ca2+ concentration of the stimulus solution, indicating Ca2+ uptake from the extracellular medium. 5. The responses of sweet‐sensitive cells to forskolin were also abolished when Ca2+ ions were omitted from the stimulus solution. They were partially inhibited by the presence of Co2+, Ni2+, D600 (methoxyverapamil) and amiloride, indicating multiple pathways of Ca2+ uptake activated by cAMP. 6. In conclusion, a sweet‐sensitive cell of the rat responds to sucrose with an increase in cAMP and Ca2+ uptake, but to non‐sugar sweeteners with an increase in IP3 and Ca2+ release. The increase in [Ca2+]i, common to both pathways, is presumably required for synaptic exocytosis and for signal termination.

A new isoflavone from soya beans

Abstract A new isoflavone was isolated from soya beans and shown to be 7,4′-dihydroxy, 6-methoxyisoflavone, for which the name glycitein is proposed.

Generation of Cyclic AMP in Taste Buds of the Rat Circumvallate Papilla in Response to Sucrose

Epithelial sheets rich in taste buds and free of muscle tissue and von Ebner’s washing glands were isolated as a U-shaped cleft which surrounds the circumvallate (CV) papilla of the rat tongue. The sheet of CV tissue from one tongue (total protein content: 8–14 µg) was cut in two approximately equal parts which were incubated with permeant phosphodiesterase inhibitor (IBMX; 0.3 mM) and 0 or 150–600 mM sucrose. After 6 min of incubation, the sheets were washed, the cells permeabilized and their cyclic AMP (cAMP) content determined by radioimmunoassay. Paired estimates with tissue from the same animal showed a significant sucrose-induced cAMP production (range 5–20 fmol/µg protein at 600 mM sucrose). This increase in intracellular cAMP was linearly dependent on the sucrose concentration and was suppressed by about 65% when 50 mM of a competitive antagonist of sucrose (methyl 4,6-dichloro-4,6-dideoxy-α-D-galactopyranoside) was added to the sucrose solution. Epithelial sheets free of taste buds did not respond to either sucrose or the inhibitor. These results are in line with previous suggestions that cAMP may be a second messenger in the transduction of sweet taste in the rat.

Interaction of MSG taste with nutrition: Perspectives in consummatory behavior and digestion

Studies in the taste system of mammals indicate that monosodium glutamate (MSG) produces a unique taste sensation termed umami. As a derivative of glutamic acid, MSG is a naturally occurring nutrient in many foods; its commercial use to improve food palatability for humans is well documented. Behavioral studies with experimental animals have revealed that preference for MSG in solutions and selection of MSG-flavored diets can be explained by sensory means with no appreciable effects on preference by postingestive consequences. However, preference for umami-flavored fluids is reduced by feeding rats low-protein diets or low quality protein. MSG-flavored diets, initially unpreferred, were subsequently highly selected. However, the adulteration of diets by MSG either did not or only slightly increased food intake. In light of the significant contribution of MSG to diet taste, apparently as a signal coupled to dietary proteins, physiological studies have been initiated to reveal its role as a stimulator of the cephalic phase of pancreatic exocrine secretion. Modified sham-feeding experiments with dogs have shown that oral stimulation by MSG produced significant stimulation of both pancreatic flow and protein output in conscious dogs.

Studies on the aroma of five fresh tomato cultivars and the precursors of cis- and trans-4,5-epoxy-(E)-2-decenals and methional.

Three tasty (BR-139, FA-624, and FA-612) and two less tasty (R-144 and R-175) fresh greenhouse tomato cultivars, which significantly differ in their flavor profiles, were screened for potent odorants using aroma extract dilution analysis (AEDA). On the basis of AEDA results, 19 volatiles were selected for quantification in those 5 cultivars using gas chromatography-mass spectrometry (GC-MS). Compounds such as 1-penten-3-one, ( E, E)- and ( E, Z)-2,4-decadienal, and 4-hydroxy-2,5-dimethyl-3(2 H)-furanone (Furaneol) had higher odor units in the more preferred cultivars, whereas methional, phenylacetaldehyde, 2-phenylethanol, or 2-isobutylthiazole had higher odor units in the less preferred cultivars. Simulation of the odor of the selected tomato cultivars by preparation of aroma models and comparison with the corresponding real samples confirmed that all important fresh tomato odorants were identified, that their concentrations were determined correctly in all five cultivars, and that differences in concentration, especially of the compounds mentioned above, make it possible to distinguish between them and are responsible for the differential preference. To help elucidate formation pathways of key odorants, labeled precursors were added to tomatoes. Biogenesis of cis- and trans-4,5-epoxy-( E)-2-decenals from linoleic acid and methional from methionine was confirmed.

Adenylate cyclase responses to sucrose stimulation in membranes of pig circumvallate taste papillae

1. Typical adenylate cyclase (AC) responses to guanine nucleotides were found in membranes of pig circumvallate (CV) taste papillae. 2. Sucrose stimulated AC activity in the CV membranes and this stimulation was GTP dependent and tissue specific. 3. The stimulatory effect of sucrose in the CV membranes was dependent on the concentration of membranes used in the AC assay. 4. This study provides the first biochemical data on cellular transduction of taste in the pig, compares positively to preliminary results in cattle and supports recent suggestions for a role of cAMP in sweet taste transduction.

The preference-aversion behavior of rats for nutritionally-controlled diets containing oil or fat

The preference-aversion behavior of Sprague-Dawley rats for semi-purified diets containing various levels of either corn oil, animal fat (lard) or hydrogenated vegetable oil (Crisco) was determined in brief and long-term, two-choice preference tests. After the first day of exposure, rats selected more calories from the control-unadulterated diet than from experimental diets containing either oil or fat. Only in the first hour of exposure (with diets containing either 5% Crisco, 15 or 25% lard) or in the first day (with diets containing 5% Crisco, 5 or 15% lard) was a preference for experimental diets observed. There were significant effects of oil and fat concentration and of exposure time upon preference for experimental diets, such that preference for the experimental diets was reduced as levels of oil or fat and as the time of experiment increased. Changes in diet selection had no major effect on total caloric intake measured as the sum of both experimental and control dietary choices, although total intake on a daily basis tended to decrease during the middle portion (2-4 days) of the study, then recover by the end of the study. The results suggest that postingestional factors related to the dietary oil and fat (rather than the sensory properties of the diets) directed the long-term preference behavior of rats for semi-purified diets adulterated with various levels of either corn oil, animal fat (lard) or hydrogenated vegetable oil (Crisco).

The effect of dietary raw and autoclaved soya-bean protein fractions on growth, pancreatic enlargement and pancreatic enzymes in rats

1. Raw soya-bean meal (RS) was fractionated into soya-bean lyophilized extract (SLE), soya-bean lyophilized residue (SLR), acid-precipitated proteins (APP) and whey proteins. 2. Trypsin (EC 3. 4. 21. 4)and chymotrypsin (EC 3. 4. 21.1) inhibitors (TI) were soluble at pH 8 and remained soluble after the extract was acidified to pH 4.4 Except for whey, heating abolished, almost totally, their inhibiting activity. 3. Feeding SLE diet (high TI content) and APP diet (low TI content) resulted in growth depression below the RS level. Feeding the SLR diet resulted in an optimal growth. Feeding diets containing heated fractions improved the growth rate though not to the level observed with heated RS (HS) diet. 4. RS, SLE, APP and whey diets produced similar pancreatic enlargement which could be totally (RS, whey) or partially (SLE, APP) abolished by heating. 5. Feeding the RS diet reduced pancreatic amylase content. The factor responsible for this effect cofractionated with SLE and whey proteins. 6. Two groups of factors in the various diets were probably responsible for the elevation in pancreatic proteases. The first group were the heat-labile factors present in RS, SLE and whey whereas the second group resisted the heat treatment and were found in APP and SLR. 7. The results suggest that for optimal growth rate of rats, heat treatment should be given to the unfractionated soya-bean proteins rather than to the isolated fractions. The results further indicated that TI are not the only factors that can lead to pancreatic enlargement and changes in pancreatic enzymes composition.

Preference of rats for food flavors and texture in nutritionally controlled semi-purified diets

Preference for nutritionally controlled, semi-purified diets modified by the addition of potent food flavors was determined for Sprague Dawley rats using two-choice diet preference tests. Intake of each food cup was monitored after 1 hr and for each 24 hr period thereafter up to 5 days. Preference was also determined for the flavored diets prepared in three forms differing in texture: powdered, and pellets of two sizes. Rats easily detected minor amounts of the food flavors, and the tests provided a catalog of 12 preferred flavors. Exposure time to the diets altered preference for a minority of flavors; diets initially avoided in the first hour test were likely to become less aversive upon continued exposure. Whether or not a specific flavored diet was preferred, total food intake was not affected during the 5 day period monitored. Rats displayed strong preference for diets of a pelleted texture compared to the same diets in a powdered form.

Taste responses to neohesperidin dihydrochalcone in rats and baboon monkeys

Preference-aversion behavior to solutions containing neohesperidin dihydrochalcone (NHDHC) was studied rats and baboon monkeys. Electrophysiological responses evoked by application of NHDHC solutions to taste receptors innervated by the chorda tympani and the glossopharyngeal nerves were also measured. As a group, rats were indifferent to solutions containing up to 1.2 x 10(-3) M NHDHC in short and long-term preference tests. A solution containing the very high concentration of 8.2 x 10(-3) M NHDHC was consumed less than water by all rats. The aversive behavior of rats to the 8.2 x 10(-3) M NHDHC solution appeared to be due to taste quality rather than olfaction. When percent preferences were calculated on an individual basis for the long-term preference tests, 59% of the rats were indifferent to solutions containing up to 1.2 x 10(-3) M NHDHC, 33% of the animals found this solution aversive and less than 8% showed preference. Behavioral responses to a solution of 3.4 x 10(-4) M aspartame also varied considerably among rats. The electrophysiological data were in line with the behavioral responses suggesting weak taste responses for NHDHC in rats. More pronounced responses observed in the glossopharyngeal nerve as compared to the chorda tympani. Baboon monkeys showed a strong preference for solutions containing 1.6 x 10(-5) M-1.6 x 10(-3) M NHDHC. A solution of 1.6 x 10(-2) M was consumed to a lesser extent than water. It is concluded that baboon monkeys present a better experimental model than rats for investigating the sweetness of NHDHC.