Exploring parental perspectives after commencement of flash glucose monitoring for type 1 diabetes in adolescents and young adults not meeting glycaemic targets: a qualitative study.

AIMS: To explore parental perspectives after flash glucose monitoring commencement in adolescents and young adults with type 1 diabetes who were not meeting glycaemic targets. METHODS: Twelve semi-structured interviews were conducted among parents of adolescents and young adults between the ages of 14 and 20 years (inclusive) with type 1 diabetes and not meeting glycaemic targets [HbA1c 81-130 mmol/mol (9.6-14.0%)] participating in a randomized controlled trial. Interviews were transcribed, then thematic analysis was performed to identify themes regarding parental experiences. RESULTS: Four key themes were found: flash glucose monitoring improved parental emotional well-being; flash glucose monitoring reduced diabetes-specific conflict within families; flash glucose monitoring facilitated the parental role in diabetes management; and sensor-related challenges, particularly sensors falling off, interfered with using flash glucose monitoring for diabetes management. The cost of self-funded sensors was the only barrier to continuing flash glucose monitoring that parents reported. CONCLUSIONS: This study provides new insights into the potential benefits and challenges of flash glucose monitoring use, drawn from the perspective of parents of adolescents and young adults not meeting glycaemic targets. As parents are often key partners in obtaining or purchasing this technology, these findings can be used to further inform parental expectations of this technology.

Defining ruminal and total-tract starch degradation for adult dairy cattle using in vivo data.

Literature was searched for studies performed in adult dairy cattle that simultaneously measured starch degradability in the rumen (RSDeg) and starch digestion in the total tract to compute postruminal starch digestion (PRSDig). Forty-one studies with 161 dietary treatments were used to form the data set. Of these diets, the major starch source was corn for 83 diets, small grain for 58 diets, and sorghum for 8 diets. Corn RSDeg was more variable than other sources. As measured in vivo across all starch sources, the percent RSDeg was influenced only by the amount of starch consumed, with the amount of degradation being approximately 75% at low starch intakes and decreasing to about 60% when 4 kg or more of starch were consumed. Small grain starch had greater RSDeg than corn or sorghum starch, which were approximately equal. The PRSDig of corn and small grain starches were approximately equal, but sorghum was about 15% less. Across all diets, models derived from the Cornell Net Carbohydrate Protein System predicted percentage of total-tract digestibility of starch very accurately, but overpredicted RSDeg and, as a result, underpredicted percent PRSDig. Calculation of RSDeg using a French model predicted the mean RSDeg with greater accuracy but less precisely. The relative differences in RSDeg percent among starch sources was correctly predicted by these models. A model using a revised rate of digestion as a way of combining effects of starch type and processing was developed, which predicted corn starch RSDeg and PRSDig with greater accuracy than nutrition models but only slightly better than using the mean observed degradation or the French calculation. Inaccuracies in prediction of RSDeg may be due mainly to processing effects and particle sizes, but these were not well reported in literature studies and were difficult to estimate. More accurate assessment of RSDeg and PRSDig will require better and more consistent reporting of grain processing. Based on this study, the French calculation is the most accurate of the models examined, although adjustments will be required to improve accuracy.

Intestinal digestibility of amino acids in rumen-undegraded protein estimated using a precision-fed cecectomized rooster bioassay: II. Distillers dried grains with solubles and fish meal.

The objectives of this experiment were to measure intestinal digestibility of AA in the rumen-undegraded protein fraction (RUP-AA) of distillers dried grains with solubles (DDGS) and fish meal (FM) samples and to determine whether these feeds contain a constant protein fraction that is undegradable in the rumen and indigestible in the small intestine, as assumed in the French Institut National de la Recherche Agronomique (Paris, France) and Scandinavian AAT-PBV (AAT = AA absorbed from small intestine; PBV = protein balance in the rumen) models. Five sources of DDGS and 5 sources of FM were obtained from Feed Analysis Consortium, Inc. (Champaign, IL). To obtain the rumen-undegradable protein fraction, samples were ruminally incubated in situ for 16 h in 4 lactating cows, and the collected rumen-undegraded residues (RUR) were pooled by sample. Subsamples of the intact feeds and RUR were crop-intubated to 4 cecectomized roosters, and total excreta were collected for 48 h. Intact feeds, RUR, and excreta were analyzed for AA. Basal endogenous AA loss estimates were obtained from fasted birds and were used to calculate standardized digestibility of RUP-AA and AA in the intact feeds. Indigestibility coefficients of the intact feeds were calculated as (100 - % standardized AA digestibility), and indigestibility of the RUR was calculated as [(100 - % ruminal degradation of AA) x (100 - % standardized RUP-AA digestibility)/100]. Results indicate that standardized digestibility of feed-AA differs from RUP-AA for DDGS samples but not for FM samples, and that standardized digestibility of individual AA differs within samples. For the DDGS samples, standardized feed-AA and RUP-AA digestibility values were most often lowest for His and Lys and highest for Met and Trp. For FM samples, standardized feed-AA and RUP-AA digestibility values were most often lowest for His and highest for Trp. Results also indicate that DDGS and most FM samples do not contain a constant protein fraction that is both undegradable in the rumen and indigestible in the small intestine. Indigestibility values of RUR were lower than in intact feeds, suggesting that the feed ingredients used in this experiment contain a protein fraction that is indigestible in the intestine but partly degradable in the rumen or digestible in the intestine after rumen incubation, or both.

Effects of providing two forms of supplemental methionine to periparturient Holstein dairy cows on feed intake and lactational performance.

Eighteen primiparous and 42 multiparous Holstein cows were blocked according to parity and expected calving date and assigned randomly to 1 of 3 dietary treatments: 1) a basal diet (negative control), 2) the basal diet plus 2-hydroxy-4-methylthio butanoic acid isopropyl ester (MetaSmart, Adisseo Inc., Antony, France), or 3) the basal diet plus rumen-protected Met (Smartamine M, Adisseo Inc., Alpharetta, GA). Treatments were initiated 21 d before expected calving and continued through 140 d postpartum. Diets were similar in ingredient and chemical composition, except for the content of Met in metabolizable protein. MetaSmart [0.35% prepartum and 0.54% postpartum in diet dry matter (DM)] and Smartamine M (0.06% prepartum and 0.10% postpartum in diet DM) were added to the basal diet in amounts needed to achieve a 3.0:1 ratio of Lys to Met in metabolizable protein. Prepartum DM intake (DMI; 13.5 kg/d), body weight (687 kg), body condition score (3.81), postpartum milk yield (42.0 kg/d), milk fat yield (1,549 g/d), milk fat content (3.66%), milk true protein yield (1,192 g/d), and milk urea N content (12.9 mg/dL) were not different among treatments. Postpartum DMI and body condition score were greater and the ratios of milk:DMI and milk N:feed N were less for cows fed the MetaSmart diet than for cows fed the control and Smartamine M diets. Milk protein content was greater for the Smartamine M (2.87%) and MetaSmart (2.81%) treatments than for the control treatment (2.72%). Concentrations of Met and Met + Cys in total plasma AA were different among treatments, with values for the Smartamine M treatment being the highest, followed by the MetaSmart and control treatments. The results indicated that both MetaSmart and Smartamine M are effective in providing metabolizable Met, but clarification of their relative contributions to metabolizable Met is still needed.

Intestinal digestibility of amino acids in rumen undegradable protein estimated using a precision-fed cecectomized rooster bioassay: I. Soybean meal and SoyPlus.

The objectives of this experiment were to measure intestinal digestibility of AA in rumen undegradable protein (RUP-AA) in soybean meal (SBM) and expeller SBM (SoyPlus, West Central, Ralston, IA; SP) and to determine if these feeds contain a constant protein fraction that is undegradable in the rumen and indigestible in the small intestine, as assumed in the French Institut National de la Recherche Agronomique (Paris, France) and Scandinavian AAT-PBV (AAT = AA absorbed from small intestine; PBV = protein balance in the rumen) models. Three samples of SBM and 3 samples of SP were obtained from the Feed Analysis Consortium Inc. (Savoy, IL). To obtain the RUP fraction, samples were ruminally incubated in situ for 16 h in 4 lactating cows, and the collected rumen undegraded residues (RUR) were pooled by sample. Subsamples of the intact feeds and RUR were crop intubated to 4 cecectomized roosters, and total excreta were collected for 48 h. Intact feeds, RUR, and excreta were analyzed for AA. Basal endogenous AA loss estimates were obtained from fasted birds and were used to calculate standardized digestibility of AA in the intact feeds and RUP-AA. Indigestibility coefficients of the intact feeds were calculated as (100 - % standardized AA digestibility), and indigestibility of the RUR was calculated as [(100 - % ruminal degradation of AA) x [(100 - % standardized RUP-AA digestibility)]/100]. Results indicated that standardized digestibility of feed-AA was similar to standardized digestibility of RUP-AA for SBM and SP samples and that standardized digestibility of individual AA differed within samples. Standardized feed-AA and RUP-AA digestibility values were lowest for Lys and Cys and highest for Trp and Met. Results also indicated that SBM and SP did not contain a constant protein fraction that was both undegradable in the rumen and indigestible in the small intestine. Indigestibility values of RUR were lower than in intact feeds, suggesting that SBM and SP contain a protein fraction that is indigestible in the intestine but partly degradable in the rumen, digestible in the intestine after ruminal incubation, or both.

In vitro digestibility of individual amino acids in rumen-undegraded protein: the modified three-step procedure and the immobilized digestive enzyme assay.

Three soybean meal, 3 SoyPlus (West Central Cooperative, Ralston, IA), 5 distillers dried grains with solubles, and 5 fish meal samples were used to evaluate the modified 3-step in vitro procedure (TSP) and the in vitro immobilized digestive enzyme assay (IDEA; Novus International Inc., St. Louis, MO) for estimating digestibility of AA in rumen-undegraded protein (RUP-AA). In a previous experiment, each sample was ruminally incubated in situ for 16 h, and in vivo digestibility of AA in the intact samples and in the rumen-undegraded residues (RUR) was obtained for all samples using the precision-fed cecectomized rooster assay. For the modified TSP, 5 g of RUR was weighed into polyester bags, which were then heat-sealed and placed into Daisy(II) incubator bottles. Samples were incubated in a pepsin/HCl solution followed by incubation in a pancreatin solution. After this incubation, residues remaining in the bags were analyzed for AA, and digestibility of RUP-AA was calculated based on disappearance from the bags. In vitro RUP-AA digestibility estimates obtained with this procedure were highly correlated to in vivo estimates. Corresponding intact feeds were also analyzed via the pepsin/pancreatin steps of the modified TSP. In vitro estimates of AA digestibility of the feeds were highly correlated to in vivo RUP-AA digestibility, which suggests that the feeds may not need to be ruminally incubated before determining RUP-AA digestibility in vitro. The RUR were also analyzed via the IDEA kits. The IDEA values of the RUR were good predictors of RUP-AA digestibility in soybean meal, SoyPlus, and distillers dried grains with solubles, but the IDEA values were not as good predictors of RUP-AA digestibility in fish meal. However, the IDEA values of intact feed samples were also determined and were highly correlated to in vivo RUP-AA digestibility for all feed types, suggesting that the IDEA value of intact feeds may be a better predictor of RUP-AA digestibility than the IDEA value of the RUR. In conclusion, the modified TSP and IDEA kits are good approaches for estimating RUP-AA digestibility in soybean meal products, distillers dried grains with solubles, and fish meal samples.

Evaluation of the furosine and homoarginine methods for determining reactive lysine in rumen-undegraded protein.

Three samples of soybean meal (SBM), 3 samples of expeller SBM (SoyPlus, West Central Cooperative, Ralston, IA), 5 samples of distillers dried grains with solubles (DDGS), and 5 samples of fish meal were used to evaluate the furosine and homoarginine procedures to estimate reactive Lys in the rumen-undegraded protein fraction (RUP-Lys). One sample each of SBM, expeller SBM, and DDGS were subjected to additional heat treatment in the lab to ensure there was a wide range in reactive RUP-Lys content among the samples. Furosine is a secondary product of the initial stages of the Maillard reaction and can be used to calculate blocked Lys. Homoarginine is formed via the reaction of reactive Lys with O-methylisourea and can be used to calculate the concentration of reactive Lys. In previous experiments, each sample was ruminally incubated in situ for 16 h, and standardized RUP-Lys digestibility of the samples was determined in cecectomized roosters. All rumen-undegraded residue (RUR) samples were analyzed for furosine and Lys; however, only 9 of the 16 samples contained furosine, and only the 4 unheated DDGS samples contained appreciable amounts of furosine. Blocked RUP-Lys was calculated from the furosine and Lys concentrations of the RUR. Both the intact feed and RUR samples were evaluated using the homoarginine method. All samples were incubated with an O-methylisourea/BaOH solution for 72 h and analyzed for Lys and homoarginine concentrations. Reactive Lys concentrations of the intact feeds and RUR were calculated. Results of the experiment indicate that blocked RUP-Lys determined via the furosine method was negatively correlated with standardized RUP-Lys digestibility, and reactive RUP-Lys determined via the guanidination method was positively correlated with standardized RUP-Lys digestibility. Reactive Lys concentrations of the intact samples were also highly correlated with RUP-Lys digestibility. In conclusion, the furosine assay is useful in predicting RUP-Lys digestibility of DDGS samples, and the guanidination procedure can be used to predict RUP-Lys digestibility of SBM, expeller SBM, DDGS, and fish meal samples.

Effect of incremental urea supplementation of a conventional corn silage-based diet on ruminal ammonia concentration and synthesis of microbial protein.

One primiparous and 3 multiparous lactating Holstein cows fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square design to determine the efficacy of adding urea to a corn silage-based diet on ruminal fermentation and microbial protein synthesis. Dietary treatments were 0, 0.3, 0.6, and 0.9% urea in diet dry matter (DM); urea was manually top dressed and incorporated into the ration. The basal diet contained (DM basis) 52% forage (with 61% of forage provided as corn silage) and 48% concentrate ingredients. The basal diet was formulated to meet National Research Council (NRC, 2001) requirements for energy and all nutrients except rumen-degradable protein (RDP) and metabolizable protein. Experimental periods lasted 14 d with the first 9 d for adaptation. The basal diet, without urea addition, contained 9.2% RDP in DM and had a predicted RDP balance of -167 g/d (NRC, 2001). There were no effects of dietary treatment on ruminal true digestibility of organic matter or ruminal apparent digestibility of neutral detergent fiber and acid detergent fiber. Total ruminal volatile fatty acid concentrations increased linearly with increasing urea level. Feeding increasing amounts of urea quadratically increased rumen ammonia N concentrations (9.0, 11.9, 12.8, and 17.4 mg/dL at 0, 0.3, 0.6, and 0.9% urea supplementation, respectively), passage of microbial N, and microbial N in duodenal digesta as a percentage of nonammonia N. The results of this study indicate that there were some positive effects of adding urea to the described lactating dairy cow diet, and that microbial protein synthesis was maximized at an average ruminal ammonia N concentration of 12.8 mg/dL when urea was added at 0.6% in diet DM.

Insulin-related growth factors stimulate proliferation of retinal progenitors in the goldfish.

The retina of the adult goldfish grows throughout the life of the animal, in part, by the continual addition of new neurons. Further, destruction of extant neurons in this tissue stimulates neuronal regeneration. In an attempt to identify growth factors that regulate both normal and injury-stimulated neurogenesis, we used organ culture techniques and tested nine peptide growth factors for their ability to modulate cell proliferation in both normal retinas and retinas with lesions. Of the growth factors tested, only the insulin-related peptides (insulin and insulin-like growth factors I and II) consistently stimulated proliferation, and this was restricted to the retinal progenitors within the circumferential germinal zone. None of the growth factors tested stimulated proliferation of rod precursors (cells in the mature retina whose progeny are exclusively rod photoreceptors) or the injury-stimulated retinal progenitors. Although the negative data are subject to multiple interpretations, these data suggest that in the retina of the adult goldfish, insulin-related peptides regulate proliferation of retinal progenitors within the circumferential germinal zone, but molecules that modulate the proliferation of the rod precursors or injury-induced retinal progenitors in the retina of the adult goldfish have yet to be identified.

Insulin-like growth factor-I binds in the inner plexiform layer and circumferential germinal zone in the retina of the goldfish.

Results of the previous study suggest that insulin-related peptides regulate proliferation of retinal progenitors in the adult goldfish. Because of their known roles in retinal neurogenesis, we have chosen to focus future studies on insulin-like growth factor I (IGF-I) and the IGF-I receptor. In the study described here, we characterized the spatial distribution and specificity of IGF-I binding sites in the retina of the adult goldfish by performing receptor-binding autoradiography with [125I]-IGF-I alone and with unlabeled IGF-I-related molecules (IGF-I, IGF-II, insulin, and des-[1-3]-IGF-I) as competitive inhibitors of [125I]-IGF-I binding. The results of these experiments show that IGF-I binds in two locations in the retina of the adult goldfish, within the inner plexiform layer of the differentiated retina and the circumferential germinal zone. The competition experiments suggest that [125I]-IGF-I binds at sites specific for IGF-I, and that both IGF-I receptors and IGF-I binding proteins are present in the retina.

Excystation of in vitro-derived Giardia lamblia cysts.

This is the first in-depth analysis of the excystation of Giardia lamblia cysts prepared in vitro. Its goals were both to achieve efficient excystation and to gain insights into this crucial but poorly understood process. To identify the critical elements of excystation, we tested the sequential low-pH induction and protease treatments which had been reported to be important for excystation of fecal cysts. The optimal pH for induction of excystation was 4.0. Emergence was greatly (approximately 10-fold) stimulated by subsequent exposure of in vitro-derived cysts to chymotrypsin, trypsin, or human pancreatic fluid. The stimulatory activity of each was abolished by soybean trypsin inhibitor, demonstrating that the activity of pancreatic fluid was due to these proteases. Excystation of in vitro-derived cysts was approximately 10 to 38%. Although the walls of in vitro-derived cysts were partially digested by protease treatment, trophozoites emerged only from one pole, as observed with fecal cysts. The conditions of encystation also determined the efficiency of excystation. Specifically, encystation in the presence of lactic acid, a major metabolite of colonic bacteria, stimulated excystation approximately fourfold, although it did not increase the total numbers of cysts. These experiments have shown that excystation of in vitro-derived cysts reflects that of cysts purified from human feces in that it is dependent upon conditions which simulate the passage of cysts through the human stomach (low pH) and into the small intestine (pancreatic proteases).

Giardia lamblia: the roles of bile, lactic acid, and pH in the completion of the life cycle in vitro.

Large numbers (10(4) to greater than 10(5)/ml) of Type I water-resistant Giardia lamblia cysts were produced in vitro under conditions that are characteristic of the human intestinal lumen. We define Type I cyst morphology as oval shaped, smooth, and refractile, with cyst wall, axostyle, and median body visible in relief by Normarski differential interference contrast optics. Human and porcine bile induced higher levels of encystation than bovine bile at the alkaline pH (7.8) which occurs in the human lower small intestine. High-pressure liquid chromatography analysis showed that the porcine bile had a preponderance of hyocholate, rather than cholate, while bovine bile had less chenodeoxycholate and more deoxycholate than human bile. Lactic acid, a major product of bacterial metabolism in the human colon, further stimulated encystation. Growth of the preencystation culture without bile also increased subsequent encystation. More than 90% of Type I cysts produced with porcine bile plus lactic acid were viable as indicated by the uptake and retention of fluorescein diacetate and exclusion of propidium iodide. Biological activity of in vitro-derived water-resistant cysts was demonstrated by the observation that 1 to 9.5% excysted in vitro. The percentage of excystation was greatly decreased following encystation at pH 7.0 or by omission of bile or lactic acid. This is the first quantitative in vitro demonstration of the complete life cycle of G. lamblia from humans.

Small-intestinal factors promote encystation of Giardia lamblia in vitro.

Bile salts and fatty acids stimulated differentiation of cultured Giardia lamblia trophozoites into water-resistant cysts at the slightly alkaline pH of the small intestinal lumen. Maximum encystation occurred at pH 7.8. Thus, specific small-intestinal factors may influence encystation in vivo as well as in vitro.