Downregulation of Squalene Synthase Broadly Impacts Isoprenoid Biosynthesis in Guayule.

Production of natural rubber by Parthenium argentaum (guayule) requires increased yield for economic sustainability. An RNAi gene silencing strategy was used to engineer isoprenoid biosynthesis by downregulation of squalene synthase (SQS), such that the pool of farnesyl diphosphate (FPP) substrate might instead be available to initiate natural rubber synthesis. Downregulation of SQS resulted in significantly reduced squalene and slightly increased rubber, but not in the same tissues nor to the same extent, partially due to an apparent negative feedback regulatory mechanism that downregulated mevalonate pathway isoprenoid production, presumably associated with excess geranyl pyrophosphate levels. A detailed metabolomics analysis of isoprenoid production in guayule revealed significant differences in metabolism in different tissues, including in active mevalonate and methylerythritol phosphate pathways in stem tissue, where rubber and squalene accumulate. New insights and strategies for engineering isoprenoid production in guayule were identified.

Low Acrylamide Flatbreads Prepared from Colored Rice Flours and Relationship to Asparagine and Proximate Content of Flours and Flatbreads.

Acrylamide is a potentially toxic compound present in many plant-based foods, such as coffee, breads, and potato fries, which is reported to have carcinogenic, neurotoxic, and antifertility properties in vivo, suggesting the need to keep the acrylamide content of widely consumed food as low as possible. As pigmented rice contains bioactive phenolic and flavonoid compounds, the objective of this study was to potentially enhance the beneficial properties of flatbreads by evaluating the acrylamide content and proximate composition of 12 novel flatbreads prepared from the following commercial pigmented rice seeds: Black Japonica, Chinese Black, French Camargue, Himalayan Red, Long Grain Brown, Purple Sticky, Short Grain Brown, Wehani, Wild, Indian Brown Basmati, Organic Brown Jasmine, and Organic Jade Pearl. Although acrylamide levels ranged from 4.9 µg/kg in Long Grain Brown to 50.8 µg/kg in Chinese Black, the absolute values were all low (though statistically significantly differences existed among varieties). Acrylamide content did not correlate with its precursor asparagine. The variations in protein, carbohydrate, fat, ash, dry matter, and water content determined by proximate analysis, and the reported health benefits of colored rice cultivars used to prepare the flatbreads, might also be useful for relating composition to nutritional qualities and health properties, facilitating their use as nutritional and health-promoting functional foods.

Low Acrylamide Flatbreads from Colored Corn and Other Flours.

Dietary acrylamide formed during baking and frying of plant-based foods such as bread and other cereal products, coffee, fried potatoes, and olives is reported to induce genotoxic, carcinogenic, neurotoxic, and antifertility properties in vivo, suggesting the need to keep the acrylamide content low with respect to widely consumed heat-processed food including flatbreads. Due to the fact that pigmented corn flours contain biologically active and health-promoting phenolic and anthocyanin compounds, the objective of this study was to potentially define beneficial properties of flatbread by evaluating the acrylamide content determined by high-performance liquid chromatography/mass spectrometry (HPLC/MS) with a detection limit of 1.8 µg/kg and proximate composition by standard methods of six experimental flatbreads made from two white, two blue, one red, and one yellow corn flours obtained by milling commercial seeds. Acrylamide content was also determined in experimental flatbreads made from combinations in quinoa flour, wheat flour, and peanut meal with added broccoli or beet vegetables and of commercial flatbreads including tortillas and wraps. Proximate analysis of flatbreads showed significant differences in protein and fat but not in carbohydrate, mineral, and water content. The acrylamide content of 16 evaluated flatbreads ranged from 0 to 49.1 µg/kg, suggesting that these flatbreads have the potential to serve as low-acrylamide functional foods. The dietary significance of the results is discussed.

Consumer sensory evaluation and quality of Sorghum-Peanut Meal-Okra snacks.

Healthful tasty high protein, vegetable, gluten-free snacks are needed for all as well as those sensitive to gluten for in between meals and for after school events. Peanut meal a low value farm by-product was used to increase protein content and to add value for growers. Bile acid binding okra with cholesterol lowering potential and jalapeno, turmeric and ginger with healthy phytonutrients were included to increase vegetable consumption. The objective was to have healthy tasty snacks with ≥24% protein content. Gluten-free, whole grain, high protein, Sorghum-Peanut meal-Okra (SPO) and SPO-Jalapeno, SPO-Turmeric root and SPO-Ginger root snacks were sensory evaluated by 73 volunteers. Physical testing of the snacks included water activity, true and bulk density, texture and proximate analyses. Taste and Odor of the SPO and SPO-Jalapeno snacks were similar and significantly (p ≤ 0.05) higher than SPO-Turmeric root and SPO-Ginger root. Acceptability of SPO and SPO-Jalapeno snacks were both 88%; this value is quite desirable. Acceptability of SPO-Turmeric and SPO-Ginger were only 56 and 51% respectively. Turmeric and ginger have been reported with many health benefits; however these snacks were not preferred by the tasters. Water activity (Aw) of the snacks tested was SPO (0.42) < SPO-Turmeric (0.52) < SPO-Jalapeno (0.54) < SPO-Ginger (0.62). Water activity indicates that all the snacks were crispy and had longer shelf life. Expansion of these snacks was SPO-Ginger root 84%, SPO-Turmeric root 76%, SPO-Jalapeno 42% and SPO only 14%. Data suggest snacks containing spices were fluffy and would give good presentation in packaging. The objective of attaining protein level was clearly attained, as values ranged 24-26%. These snacks are easy to make in house kitchens or by food companies. These healthy snacks offer a gluten-free, high protein, tasty choice for all, including vegetarians and individuals hypersensitive to gluten.

High-Protein Nutritious Flatbreads and an Option for Gluten-Sensitive Individuals.

Whole grain quinoa and wheat, high-protein vegetable flatbreads were evaluated by tasters and a physical analysis was conducted. The objective was to produce nutritious, tasty gluten-free (quinoa) as well as gluten-containing (wheat) flatbreads. Flatbreads were Quinoa Peanut Oilcake Broccoli (QPCBROC), Wheat Peanut Oilcake Broccoli (WPCBROC), Quinoa Peanut Oilcake Beets (QPCBEET) and Wheat Peanut Oilcake Beets (WPCBEET). Peanut Oilcake would increase protein and add value to this farm byproduct. Bile acid binding broccoli and beets with cholesterol-lowering potential were used. Tasters preferred QPCBROC flatbreads for all sensory parameters. Acceptance of flatbreads was QPCBROC (83%), WPCBROC (70%), QPCBEET (78%) and WPCBEET (69%); these values were statistically similar. The objective of ≥25% protein content was exceeded by 5-8% and that of ≥70% acceptance was adequately achieved. These flatbreads were low in fat (5-6%) and contained essential minerals (4%) with only ≤1% added salt. Porosity and expansion data suggest that these flatbreads would take up relatively little shelf space. These flatbreads require only three ingredients and can be made in a household kitchen or by commercial production. These flatbreads offer a nutritious, tasty choice for all, and quinoa flatbreads offer an option for gluten-sensitive individuals.

Acrylamide Content of Experimental Flatbreads Prepared from Potato, Quinoa, and Wheat Flours with Added Fruit and Vegetable Peels and Mushroom Powders.

Flatbreads are a major food consumed worldwide. To mitigate an undesirable safety aspect of flatbreads that might be associated with the potentially-toxic compound acrylamide, we recently developed recipes using a variety of grains that resulted in the production of low-acrylamide flatbreads. To further enhance the functionality of flatbreads, we have developed, in this work, new experimental flatbreads using potato, quinoa, and wheat flours supplemented with peel powders prepared from commercial nonorganic and organic fruits and vegetables (apples, cherry tomatoes, melons, oranges, pepino melons, sweet potato yams), potato peels, and mushroom powders (Lion's Mane, Hericium erinaceus; Reishi, Ganoderma lucidum; and Turkey Tail, Trametes versicolor). These additives have all been reported to contain beneficial compositional and health properties. The results of fortification of the baked flatbreads showed either no effect or increases in acrylamide content by unknown mechanisms. Since the additives did not increase the acrylamide content of the quinoa flour flatbreads for the most part, such supplemented quinoa flatbreads have the potential to serve as a nutritional, gluten-free, low-acrylamide, and health-promoting functional food. Mushroom powder-fortified wheat flatbreads with relatively low acrylamide content may also have health benefits.

Acrylamide Content of Experimental and Commercial Flatbreads.

Acrylamide, formed in baked and fried plant-based foods, is reported to induce numerous adverse effects in cells, animals, and humans. Examples from the literature show that processed potato- and cereal-based products are two major food types that seem to contribute the highest amounts of acrylamide to the diet worldwide. To meet both the demand for gluten-free products and the interest in alternative grains, we previously developed recipes for flatbreads using a variety of different grains. In this study, we determined the acrylamide content of 15 experimental flatbreads made from a variety of flours and 21 commercial flatbreads. The application of a validated, highly sensitive HPLC/MS method revealed that flatbreads made with the following flours baked at 195.5 °C for 2 min had very low (<10 µg/kg) levels of acrylamide: brown rice, buckwheat, cornmeal, millet, oat, and quinoa. The acrylamide levels of the following flatbreads were 14 to 59 µg/kg: rye, sorghum, soy, wheat, commercial pita, pita crackers, pizza, naan, and lavash. Wheat-based matzo breads, which are rapidly baked to a crisp texture at high heat (∼400 °C), contained 101 to 504 µg/kg acrylamide. Potato-based products were some of the highest of the products tested, ranging from 153 (potato pancakes) to 2,070 (potato-containing gluten-free matzos) µg/kg acrylamide. Except for the potato-containing products, the flatbreads made in this study were lower in acrylamide content (<3 to 21.3 µg/kg) than any of the commercial products tested. Of these experimental flatbreads, wheat- and sorghum-based products were the highest. Flatbreads from alternative grains can result in gluten-free products with high nutritional value and less acrylamide. PRACTICAL APPLICATION: Acrylamide formation is dependent on both the composition of the food product and the method of cooking. Flatbreads have the potential to be high in acrylamide due to cooking methods which lead to the development of desirable browning products. Flatbreads developed in this study using alternative and ancient grains were mostly lower in acrylamide content than their wheat counterpart, suggesting that they can serve as a low-acrylamide, gluten-free functional food.

Impact of pre-hospital electrocardiograms on time to treatment and one year outcome in a rural regional ST-segment elevation myocardial infarction network.

BACKGROUND: Pre-hospital electrocardiograms (ECGs) are believed to reduce time to reperfusion in ST Segment Elevation Myocardial Infarction (STEMI) patients. Little is known of their impact on clinical outcomes in a rural setting. Geisinger regional STEMI network provides percutaneous coronary intervention (PCI) care to over a 100-mile radius in rural central Pennsylvania. METHODS: A retrospective analysis identified 280 consecutive STEMI patients treated with PCI between 1/1/09 and 8/31/11. Comparison between two STEMI groups was performed: 205 patients who were taken by the emergency medical system (EMS) to the nearest hospital (a non-PCI center), underwent an ECG revealing a STEMI, and were transported immediately to Geisinger Medical Center (GMC) for PCI (transfer group) versus 75 patients in whom a pre-hospital ECG was obtained and who were transported by EMS directly to Geisinger for PCI, bypassing the nearest hospital that did not perform PCI (the pre-hospital ECG group). RESULTS: Analysis of baseline characteristics revealed that the pre-hospital ECG cohort was older (65 vs. 60 years); had a higher percentage of previous myocardial infarctions (MI) (28% vs. 15%), heart failure (11% vs. 4%), and prior PCI (23% vs. 13%; p < 0.05 all comparisons). Median time from EMS contact to pre-hospital ECG in the pre-hospital ECG group was 5 minutes; from pre-hospital ECG to the GMC ED was 34 minutes. Median time from first medical contact (EMS contact) to reperfusion (device activation) was 79 versus 157 minutes (P < 0.001), respectively in pre-hospital ECG vs. transfer groups. Mortality in the two groups at 1 year was 4.1% in the pre-hospital ECG group versus 8.3% in the transfer group (P-value = 0.34). After adjusting for the difference in age between the two groups, the 62% reduction in 1 year mortality associated with having obtained a pre-hospital ECG was still not statistically significant (P-value = 0.19). CONCLUSION: In a rural regional STEMI network, pre-hospital ECGs decreased time from first medical contact to reperfusion by 50% and were associated with an excellent clinical outcome at 1 year. © 2016 Wiley Periodicals, Inc.

Sensory evaluation of gluten-free quinoa whole grain snacks.

Sensory evaluation of quinoa gluten-free whole grain low fat and salt snacks was conducted. The snacks were Quinoa, Quinoa-Cayenne Pepper, Quinoa-Ginger and Quinoa-Turmeric. Cayenne pepper, ginger and turmeric are common spices that contain health promoting nutrients. Cayenne pepper has been associated with enhancing heat production. Ginger has been reported to improve blood flow and prevent joint pains. Turmeric has been observed to have wound healing potential. All the snacks contained 6% corn oil and 2% salt. Snack dough was prepared using 120 mL water for 100 g dry ingredients. About 20 g of snack dough was placed on center of preheated KrumKake Express Baker and cooked for 2 min. Seventy in-house volunteers judged Color/Appearance of Quinoa, Quinoa-Cayenne Pepper and Quinoa-Ginger snacks significantly (p ≤ 0.05) higher than Quinoa-Turmeric snacks. Odor/Aroma of Quinoa-Ginger snacks was significantly higher than other snacks tested. Texture/Mouth-feel of Quinoa-Cayenne Pepper, Quinoa-Ginger and Quinoa-Turmeric snacks was similar and significantly higher than Quinoa snacks. Taste/Flavor and Acceptance was similar in four kinds of snacks tested. Water activity of all the snacks tested ranged from 0.41-0.55 suggesting that these snacks were crispy with good antimicrobial stability. These snacks would be quite filling due to their expansion of 2.6-3.1 times due to high porosity. Acceptance of snacks tested was Quinoa 79%, Quinoa-Cayenne Pepper 77%, Quinoa-Ginger 73% and Quinoa-Turmeric 70%. These snacks contained only 3-4 ingredients and could be made in any house kitchen or commercial production. Acceptance of 70-79% is very desirable. These healthy nutritious gluten-free quinoa snacks offer choice for all including vegetarians and individuals hypersensitive to gluten.

Remodeling the isoprenoid pathway in tobacco by expressing the cytoplasmic mevalonate pathway in chloroplasts.

Metabolic engineering to enhance production of isoprenoid metabolites for industrial and medical purposes is an important goal. The substrate for isoprenoid synthesis in plants is produced by the mevalonate pathway (MEV) in the cytosol and by the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway in plastids. A multi-gene approach was employed to insert the entire cytosolic MEV pathway into the tobacco chloroplast genome. Molecular analysis confirmed the site-specific insertion of seven transgenes and homoplasmy. Functionality was demonstrated by unimpeded growth on fosmidomycin, which specifically inhibits the MEP pathway. Transplastomic plants containing the MEV pathway genes accumulated higher levels of mevalonate, carotenoids, squalene, sterols, and triacyglycerols than control plants. This is the first time an entire eukaryotic pathway with six enzymes has been transplastomically expressed in plants. Thus, we have developed an important tool to redirect metabolic fluxes in the isoprenoid biosynthesis pathway and a viable multigene strategy for engineering metabolism in plants.

Steam cooking significantly improves in vitro bile acid binding of collard greens, kale, mustard greens, broccoli, green bell pepper, and cabbage.

Bile acid binding capacity has been related to the cholesterol-lowering potential of foods and food fractions. Lowered recirculation of bile acids results in utilization of cholesterol to synthesize bile acid and reduced fat absorption. Secondary bile acids have been associated with increased risk of cancer. Bile acid binding potential has been related to lowering the risk of heart disease and that of cancer. Previously, we have reported bile acid binding by several uncooked vegetables. However, most vegetables are consumed after cooking. How cooking would influence in vitro bile acid binding of various vegetables was investigated using a mixture of bile acids secreted in human bile under physiological conditions. Eight replicate incubations were conducted for each treatment simulating gastric and intestinal digestion, which included a substrate only, a bile acid mixture only, and 6 with substrate and bile acid mixture. Cholestyramine (a cholesterol-lowering, bile acid binding drug) was the positive control treatment and cellulose was the negative control. Relative to cholestyramine, in vitro bile acid binding on dry matter basis was for the collard greens, kale, and mustard greens, 13%; broccoli, 10%; Brussels sprouts and spinach, 8%; green bell pepper, 7%; and cabbage, 5%. These results point to the significantly different (P < or = .05) health-promoting potential of collard greens = kale = mustard greens > broccoli > Brussels sprouts = spinach = green bell pepper > cabbage as indicated by their bile acid binding on dry matter basis. Steam cooking significantly improved the in vitro bile acid binding of collard greens, kale, mustard greens, broccoli, green bell pepper, and cabbage compared with previously observed bile acid binding values for these vegetables raw (uncooked). Inclusion of steam-cooked collard greens, kale, mustard greens, broccoli, green bell pepper, and cabbage in our daily diet as health-promoting vegetables should be emphasized. These green/leafy vegetables, when consumed regularly after steam cooking, would lower the risk of cardiovascular disease and cancer, advance human nutrition research, and improve public health.