Spectroscopic studies of myo-inositol monophosphatase with a novel fluorescent substrate.

myo-Inositol monophosphatase catalyzes dephosphorylation of the synthetic substrate anthraniloyl-2'-AMP. Binding of this fluorescent substrate to Tb(III)-monophosphatase was monitored by luminescence spectroscopy. The anthraniloyl chromophore excited at 330 nm sensitizes the long lived luminescence of enzyme bound Tb(III) at 490, 545, 585 and 620 nm. Assuming a mechanism of radiationless energy transfer, the actual distance of separation between the donor anthraniloyl moiety and the acceptor Tb(III) was calculated to be R = 10 angstroms. The binding studies support the earlier observation of Bone et al. (Proc. Natl. Acad. Sci. USA 89 (1992) 10031-10035) that the substrate and the lanthanide Gd(III) interact with a common binding domain of the protein. The catalytic activity of the monophosphatase is completely dependent upon Mg(II) ions which elicit changes in the secondary structure of the protein as revealed by circular dichroism measurements. Binding of Mg(II) ions tend to stabilize the secondary structure of the phosphatase against guanidinium-HCl denaturation.

Aedes aegypti (Diptera: Culicidae) vision: response to stimuli from the optical environment.

Although Aedes aegypti (L.) are capable of some wavelength discrimination, behavioral experiments demonstrated that color was not a stimulus to which they responded. Optical stimuli that did affect Ae. aegypti behavior were luminous reflectance, vertical contrast, and movement. Stationary objects of low reflectance and solid color were the most attractive to males and females.

Aedes aegypti (Diptera: Culicidae) vision: spectral sensitivity and other perceptual parameters of the female eye.

From measurements of the mean ommatidial diameter (17.2 microns) and mean interommatidial angle (6.2 degrees) of the eye of female Ae. aegypti (L.), a minimum resolvable angle of 12.3 degrees and an eye parameter of 1.6 microns were calculated. The eye had relatively poor acuity but high overall sensitivity to light. Electroretinographs revealed spectral sensitivity ranging from ultraviolet (323 nm) to orange-red (621 nm), with sensitivity peaks in the ultraviolet (lambda max323-345 nm) and green (lambda max523 nm) wavelengths. The retina is capable of some wavelength discrimination.

Wheat fiber and laxation: dose response and equilibration time.

We studied the dose response to soft white winter wheat fiber on fecal output in a group of healthy volunteers whose breakfasts consisted of wheat fiber cereals in amounts that provided 0.3 g, 5.6 g, 9.5 g, 11.2 g, 19.0 g, and 28.4 g dietary fiber per day for 14 days; no other aspects of their diet were altered. A linear dose response was observed between the six levels of fiber intake (r = 0.983, p less than 0.01) with a 1-g increase in wheat fiber, producing a mean 2.7-g increase in fecal weight. This increase was independent of the initial daily fecal weight of the volunteer (mean 117 +/- 64 g/day, range 5-297 g/day, n = 73). The maximum increase in fecal output due to cereal fiber was reached after the first week on the supplement. These data support the use of graded amounts of cereal fiber in the management of constipation.

The effect of starch-protein interaction in wheat on the glycemic response and rate of in vitro digestion.

To determine the effect on blood glucose of removal of protein from wheat products, healthy volunteers took test meals of white bread made from either regular or gluten-free flour. After bread made from gluten-free flour, the blood-glucose rise was significantly greater. This corresponded with a significantly more rapid rate of digestion in vitro and reduced starch malabsorption in vivo as judged by breath-H2 measurements. Addition of gluten to the gluten-free bread mix did not reverse these effects. Factors associated with unprocessed wheat flour, such as the natural starch-protein interaction, may therefore be important in wheat products in reducing both their rate of absorption and glycemic response. They may have implications in the dietary management both of diabetes and of diseases where small intestinal absorptive capacity is impaired.

Effect of modifying the rate of digestion of a food on the blood glucose, amino acid, and endocrine responses in patients with cirrhosis.

To see whether foods with slower rates of digestion may benefit the metabolic abnormalities seen in cirrhosis, the same food, processed in two different ways, was fed to seven patients with cirrhosis. The breakfast of lentils processed by prolonged heating, to produce more rapid in vitro digestion, resulted in a significantly higher incremental rise in large neutral amino acid levels at 60 min (p less than 0.02) and a tendency for a more rapid rise in total amino acid concentrations by comparison with conventionally cooked lentils with slower in vitro digestion rates. After more rapidly digested lentils, incremental levels of branched-chain amino acids were also higher at 60 min (67 +/- 9, p less than 0.001) despite a greater overall insulin response. Comparable incremental amino acid areas after both meals suggested that the total amount of amino acids absorbed was not influenced by processing. Greater blood glucose, insulin, and gastric inhibitory polypeptide responses were seen after the more processed meal with no significant differences in pancreatic glucagon, entroglucagon, or neurotensin levels. Processing a food to alter the rate of digestion may therefore be used to manipulate amino acid, glucose, and endocrine responses in cirrhosis.

Ileal loss of available carbohydrate in man: comparison of a breath hydrogen method with direct measurement using a human ileostomy model.

The breath hydrogen technique has suggested that a considerable amount of available plus unavailable carbohydrate enters the large intestine after the consumption of starchy foods (white bread 11%, wholemeal bread 8%, and red lentils 18%). Direct measurement of the available carbohydrate in ileal effluent after the consumption of test meals by three individuals with ileostomies gave values similar to those determined by the breath hydrogen technique (white bread 10%, wholemeal bread 8%, and red lentils 22%). These studies confirm that considerable amounts of "available carbohydrate" may be lost to the small intestine and physiologically must be considered as dietary fiber. The implications of this with respect to metabolism and colonic disease remain to be investigated.

The relationship between glycemic response, digestibility, and factors influencing the dietary habits of diabetics.

A significant relationship was found between the rate of release of the sugars; glucose, maltose, and maltotriose from amylitic digestion of 10 foods tested in vitro (expressed as the digestibility index) and the blood glucose response to 50-g carbohydrate portions of the same foods eaten by diabetics (expressed as the glycemic index), (r = 0.815, n = 10, p greater than 0.01). The glycemic index related to both the palatability of the foods (r = 0.731, p less than 0.05) and their frequency of use (r = 0.698, p less than 0.05). However, in this group of motivated diabetics food use was not related directly to palatability, but rather to health belief (r = 0.689, p less than 0.05). The results suggest that carbohydrate foods of potential use to the diabetic may be identified by their in vitro digestion characteristics but to a large extent their acceptance will depend on health belief and possibly ease of preparation.

Slowly digested carbohydrate food improves impaired carbohydrate tolerance in patients with cirrhosis.

To test whether impaired carbohydrate tolerance in cirrhosis could be modified by dietary means ten cirrhotic patients, five of them taking insulin, took as breakfast either lentils or wholemeal bread and cottage cheese containing the same amount of carbohydrate and protein. Lentils resulted in significantly diminished blood glucose, insulin (in those not on insulin) and gastric inhibitory peptide responses. Enteroglucagon and neutrotensin levels were high with lentils, suggesting that absorption of lentil carbohydrate continued into the ileum with perhaps some malabsorption, so confirming the results of earlier studies in vitro. However, breath hydrogen studies on a separate group of eight healthy volunteers indicated that the difference in carbohydrate malabsorption between lentil, and wholemeal bread was insignificant. It is suggested that slowly digested carbohydrate foods, such as leguminous seeds, may minimize carbohydrate intolerance in patients with cirrhosis.

Factors affecting starch digestibility and the glycemic response with special reference to legumes.

Large differences exist in the degree to which different starch containing foods affect the blood glucose levels of both normal volunteers and diabetics. These differences appear to relate to the digestibility of the starch and the factors determining this, including: the interaction of starch with fiber, antinutrients (eg, phytate) and protein in the food, together with the nature of the starch itself and its physical form (eg, raw or cooked, ground or whole). In this respect legumes exemplify a class of foods, high in fiber, protein and antinutrients, with a starch which is digested slowly in vitro. They also produce relatively small blood glucose rises after consumption by both normals and diabetics and in the longterm result in improved diabetic control. Identification of more such foods and further understanding of factors determining starch digestibility will allow greater therapeutic use of diet in the management of diabetics and disorders of carbohydrate metabolism.

The glycaemic index of foods tested in diabetic patients: a new basis for carbohydrate exchange favouring the use of legumes.

Recently diabetic patients have been encouraged to increase their carbohydrate intake, but exact details of which foods to use are lacking. To determine whether sufficiently large differences existed to justify more specific dietary advice, we compared the glycaemic responses to 50 g carbohydrate portions of different foods, taken as breakfast test meals by groups of five to seven diabetic patients. Two- to threefold differences were seen amongst the 15 foods tested. The glycaemic responses for spaghetti, 'All-bran', rice and beans were significantly below those for bread, while 'Cornflakes' were above. Factors predicted to influence this were without effect, including: substituting wholemeal for white bread, increasing substantially the simple sugars (using 'All-bran' or bananas instead of wholemeal bread) and doubling meal protein by adding cottage cheese to bread. Paired comparisons of the glycaemic response to the five legumes with those of the seven other starchy foods (breads, spaghetti, rice, Cornflakes, oatmeal porridge and potatoes) showed that the mean peak rise in blood glucose concentration and mean area under the glucose curve after beans were 23 and 28% lower, respectively, than the mean for the other foods (p less than 0.001). Such results suggest a potentially valuable role for dried leguminous seeds in carbohydrate exchanges for individuals with impaired carbohydrate tolerance. These large differences in blood glucose response to different food cannot at present be predicted directly from tables of chemical composition. Nevertheless, physiological testing may both aid in understanding the factors responsible and help selection of the appropriate carbohydrate foods for the diabetic diet.

Effect of processing on digestibility and the blood glucose response: a study of lentils.

To test the effect of processing on digestibility and the glycemic response to a leguminous seed, a group of eight healthy volunteers took a series of breakfast test meals containing either lentils which had been processed in four different ways or the same amount of carbohydrate as white bread. Lentils, boiled for 20 min, resulted in a flattened blood glucose response by comparison with bread. This was unaltered by blending the lentils to a paste or boiling them for an additional 40 minutes. However the blood glucose response was significantly enhanced by drying the boiled blended lentils for 12 h at 250 degrees F. In vitro digestion with human saliva showed the rate of sugars released from the food related positively to the blood glucose rise. Breath hydrogen studies indicated that carbohydrate malabsorption was too small to account for differences in the blood glucose response. These results emphasize the importance of processing in determining digestibility and hence the glycemic response to a food.