Recent advances in analytical construct resins.

The area of solid-phase synthesis has witnessed exponential growth in the last fifteen years, but difficulties associated with the monitoring and analysis of resin-bound reactions and products have been apparent due to a limited number of analytical methods available. With the substrate tethered to an insoluble support traditional chromatographic monitoring is only possible after cleavage. In order to address this 'analytical bottleneck' Geysen, in 1996 elaborated Merrifield's initial dual linker strategy by incorporating an encoding system between two in-line linkers. These analytical construct resins represented a new approach for both the quality control of solid-phase combinatorial libraries and for the development of new synthetic sequences on solid-support. This review will summarize the development and application of analytical construct resins focusing on recent applications of the technology.

Relationship between hepatic fatty acid oxidation and gluconeogenesis in the fasting neonatal pig.

Hepatocytes were isolated from sixteen fasting neonatal pigs and used in two experiments: (1) to determine the effect of various factors on the ability for hepatic oxidation of fatty acids and (2) to clarify the relationship between fatty acid oxidation and glucose synthesis. In Expt 1, newborn pigs were either fasted from birth for 24 h or allowed to suck ad lib. for 3 d followed by a 24 h fast. In the presence of pyruvate, oxidation of octanoate (2 mM) was about 30-fold greater than oleate (1 mM) regardless of age, but glucose synthesis was not enhanced beyond that observed for pyruvate alone. Inclusion of carnitine (1 mM), glucagon (100 nM) or dibutyryl cAMP (50 microM) in the incubation media did not stimulate either fatty acid oxidation (octanoate or oleate) or glucose synthesis. Extending the period of fasting to 48 h (Expt 2) failed to enhance the fatty acid oxidative capacity or glucose synthesis rate. Likewise, the redox potential of the gluconeogenic substrate (lactate v. pyruvate) did not influence glucose synthesis regardless of the oxidative capacity exhibited for fatty acids. These data indicate that fatty acid oxidative capacity is not the first limiting factor to full expression of gluconeogenesis in hepatocytes isolated from fasted newborn pigs.

Effect of colostrum intake on hepatic gluconeogenesis and fatty acid oxidation in the neonatal pig.

A total of 24 newborn pigs were used to determine 1) the relationship between the quantity of colostrum consumed and the capacity for gluconeogenesis and fatty acid oxidation and 2) whether fatty acid oxidation limits gluconeogenesis in isolated hepatocytes. Neonatal pigs were obtained prior to nursing and allotted to one of three treatment groups; fed (ad libitum), limit-fed (25% of fed group), or fasted. Hepatocytes were isolated when pigs were 24 h old. Colostrum intake altered the metabolic status of neonates such that the capacity for glucose synthesis and oxidation of octanoate increased with intake. Glucose synthesis with lactate as the substrate was greater (P less than .01) for fed pigs (10.79 mumol glucose.h-1.mg DNA-1) than for either limit-fed (6.56) or fasted counterparts (4.78), which were similar (P greater than .10). Colostrum intake failed to stimulate synthesis from alanine. The oxidation rate for octanoate was similar for fed and limit-fed pigs (.62 and .61 nmol CO2.h-1.mg DNA-1, respectively) but greater (P less than .05) than that observed for fasted counterparts (.36). Oxidation of octanoate (2 mM) was approximately 30-fold greater than for oleate (1 mM); oxidation of the latter was not affected by either colostrum intake or the addition of carnitine (1 mM). The increase in octanoate oxidation, however, did not elicit an increase in glucose synthesis by fasting pigs with either lactate or alanine as precursors. Thus, we conclude that gluconeogenesis is a function of colostrum intake and that reducing equivalents and(or) ATP may not be primary factors limiting glucose synthesis in pigs fasted from birth.

Proteins in whey: chemical, physical, and functional properties.

There is abundant information concerning the functional behavior of whey proteins in model systems. The data on functional properties reported by different researchers, however, reveal wide discrepancies in values. For example, in the case of comparable whey preparations, apparent solubilities may range from 10 to 100%; strength of gels from 0.3 to greater than 10 N, foam overruns from 250 to 1500%, and foam stabilities from 0.5 to 30 min. Many of the data are of limited value in assessing the true functional characteristics of different preparations, treatments, or processing effects. Reports to date are useful in indicating the relative behavior of different proteins; however, the data do not always predict the performance of such proteins in actual food systems. This reflects the fact that in foods, extensive interactions with other components may occur, resulting in modified behavior of the proteins. Harper, (1984) has advocated the testing of these various preparations in simulated food systems which should validly relate the behavior to performance in commercial systems. Emphasis on standardization of specific protocols, with regard to order of addition in ingredients, temperature, pH control, and amount of energy input during mixing, homogenization, emulsification, etc. deserves serious consideration. While this approach is justifiable in terms of providing valuable data to commercial users, it does not minimize the importance of examining these proteins in model systems where the physicochemical basis of each functional attribute can be described in molecular terms (Kinsella, 1987). Such information is necessary to expedite appropriate methods of processing in order to control compositional variability, extent of denatauration, and possible protein modification. In addition, rapid, reliable tests for routine quality assurance that can provide practical information concerning functional applications would be of great value. Whey protein preparations vary immensely in functional behavior and are presently relegated to limited use as functional ingredients in the food industry. This need not be the case since conventional and new technologies permit rigorous control of production protocols, e.g., careful control of heat treatments can result in the production of whey protein preparations with consistent, reliable functional properties (deWit, 1981, 1984; Harper, 1984; Morr, 1985). As the market for functional proteins continues to expand, the whey industry must seek the means to refine whey protein products; determine useful functional properties; develop standardized manufacturing protocols; demonstrate the effectiveness of whey as a functional ingredient; promote, and then market, whey on the basis of performance at competitive cost.

Effect of 1,3-butanediol and short chain acids in sow gestation diets on maternal plasma metabolites and fetal energy storage.

Gestating sows were fed diets in which 15% of the metabolizable energy was in the form of glucose monohydrate (control), 1,3-butanediol (BD) or an equimolar mixture of acetate and lactate (AL) in order to study the effects of ketogenic, glucogenic and lipogenic substrates on fetal energy storage. Diets were initiated on d 90 of gestation. Blood plasma was obtained from sows 2 and 8 h after feeding on d 102 of gestation. Sows receiving BD had a higher (P less than .001) concentration of beta-hydroxybutyrate (.54 vs .12, .14 mM) and a lower (P less than .05) concentration of glucose (72 vs 82, 86 mg/dl) after 8 h than sows in control or AL groups, respectively. Sows in the AL group had a higher (P less than .10) acetate concentration at 2 h, but no difference was observed by 8 h. Lactate concentration was lower (P less than .10) in AL sows when compared with those in the control group (69 vs 101 mg/dl). Two pigs/litter were killed at birth and two/litter were fasted for 36 h with blood samples obtained at 12-h intervals. Newborn pigs from AL and BS sows had more total liver glycogen than pigs derived from control sows (4.18, 4.07 vs 3.09 g, respectively); however, this difference was significant only for pigs from AL sows (P less than .10). Pigs in BD and AL groups had a higher (17 to 25%), though not significantly different, glucose concentration than controls after 24 and 36 h of fasting.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of exogenous glucagon and free fatty acids on gluconeogenesis in fasting neonatal pigs.

Thirty-two pigs (1 d old) were used to determine if exogenous glucagon and(or) free fatty acids (FFA oleic acid) would enhance gluconeogenesis and glucose homeostasis during fasting. Pigs were acquired at birth, fitted with an indwelling arterial cannula (via umbilicus) and fasted 24 h to deplete liver glycogen. A jugular cannula was inserted nonsurgically 8 to 10 h before initiation of a primed-continuous infusion consisting of control (excipient), glucagon (Glu), oleic acid (FFA), or both glucagon and oleic acid (Glu-FFA). Plasma Glu averaged 395 pg/ml preinfusion and was similar across treatments. The concentration increased fivefold (P less than .05) by 80 min for Glu and Glu-FFA pigs and remained constant thereafter (160 min: 2,379, 2,258 pg/ml; 240 min: 2,355, 2,274 pg/ml, respectively). Glucagon infusion did not alter plasma glucose after 240 min of infusion (control, 50 vs Glu, 51 mg/dl); however, Glu-FFA effected an increase (60 mg/dl, P less than .10). In contrast, pigs infused with FFA alone had a lower glucose concentration (40 mg/dl, P less than .10). Rate of glucose synthesis was determined using liver slices, acquired immediately postinfusion, with alanine and lactate as substrate (7.5 mM). The rate of synthesis was not altered by Glu or Glu-FFA infusion (2.91, 2.43 mumol glucose X g-1 X h-1 vs 2.91 for control). In contrast, exogenous FFA reduced synthesis to 1.85 mumol glucose X g-1 X h-1 (P less than .05) with lactate as substrate. It appears that Glu is not the primary factor limiting gluconeogenesis in fasting newborn pigs.(ABSTRACT TRUNCATED AT 250 WORDS)