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New health-promoting grain can help combatting life-style related disease. Barley (<i>Hordeum vulgare</i>, <i>Hv</i>) has nutritional and health promoting effects due beta-glucan and other bioactive compounds (1). <i>Hordeum spontaneum (Hs) </i>is its wild ancestor and comparisons suggest that dietary fibers and other bioactives changed through crop domestication (2). <i>Hs</i> accessions differ from <i>Hv </i>by their higher levels of prebiotic carbohydrates. Predicted glycemic indices (pGI (3)) for grain from <i>H. spontaneum</i>, transgenic Amylose-Only (AO) barley (4) and normal barley using static and dynamic human gastro-intestinal <i>in vitro</i> models showed that <i>Hs</i> and AO were lower (17.5 and 15.8, respectively) than normal barley (35.0). The AO grain has low pGI due to resistant starch. For <i>Hs</i> lower pGI was hypothesized to be effects of 1. Inhibition of the salivary amylase by grain proteinous inhibitors and/or 2. mechanical inhibition by high viscosity. The viscosity of 10 % grain flour in heated in water, measured by Rapid Visco Analyzer (RVA) at 37 ºC. Higher viscosity was found for <i>Hs</i> than for <i>Hv</i> and AO. Addition of alpha-amylase resulted in a slower decrease for <i>Hv</i> than for <i>Hs</i> demonstrating the inhibition of amylolytic digestion. Protein extracts from flour decreased pancreatic amylolytic activity <i>in vitro</i>. Bread with 50% of AO or <i>Hs</i> flour and 50% wheat flour showed good alveolation. <i>In vitro </i>dynamic digestion of the breads revealed increased soluble and in-soluble fibers present in the digesta. Our work supports the potential to include health-promoting grain of both high/full amylose and ancient germplasm as health-promoting ingredients for a healthy future diets.