The reaction of methylglyoxal with human and bovine lens proteins.

Methylglyoxal is an endogenous metabolite that increases in diabetes and has been implicated in some of its long-term complications such as retinopathy, neuropathy and cataract. We investigated the reaction of methylglyoxal with isolated human and bovine lens crystallins (alpha, beta H, beta L and gamma). After 7 days incubation at 37 degrees C and pH 6.9, the reaction of methylglyoxal with lens proteins yielded stable adducts that exhibited fluorescent properties. SDS-polyacrylamide gel electrophoresis was used to monitor aggregation and crosslinking of the modified protein and autoradiography showed that [14C]methylglyoxal was incorporated into all the protein bands. Bovine gamma-crystallin was the most reactive towards methylglyoxal. Reaction of methylglyoxal with bovine gamma II-crystallin, which is found mainly in the lens nucleus, could alter the change surface network of the molecule, resulting in aggregation, increased light scattering and hence cataract. Modification of gamma II-crystallin by methylglyoxal produced an overall loss of positive charge and an increase in molecular weight and non-disulfide covalent crosslinking. Amino acid analysis of the modified gamma II-crystallin showed a loss of 47% of arginine residues.

The reaction of malondialdehyde with lens proteins and the protective effect of aspirin.

Malondialdehyde, a product of lipid peroxidation and a by-product of thromboxane synthesis increases in human cataract. Malondialdehyde bound to soluble lens proteins over 4 h of incubation. Pre-incubation of lens proteins with aspirin offered protection against reaction with MDA. Gel chromatography was used to monitor aggregation of the modified protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the reaction with malondialdehyde led to non-disulphide covalent cross-linking of gamma-crystallin, which was decreased by incubation with aspirin. Malondialdehyde has two carbonyl groups which could react with primary amino groups, forming Schiff-base conjugates and covalently cross-link proteins. The modification and cross-linking could initiate the cataractogenic process.

Failure of dietary restriction to influence natural killer activity in old rats.

Natural killer activity of Sprague-Dawley rats maintained on an ad libitum versus restricted diet was compared using an 18 hour 51Cr-release assay, against the K562 erythroleukemic line, Yac-1 lymphoma cells and SV40-3T3 cells. The results indicated that no enhancement of natural killer function was induced by dietary restriction of 10.5-month-old rats from weaning. Prolongation of the restricted diet into late life (24 months) similarly did not enhance basal natural killer activity over levels observed in the ad libitum controls. This suggests that the improved resistance to some tumours seen after prolonged dietary restriction depends on another defensive mechanism, reduced metabolic activity and/or a reduction of available nutrients at cancerous foci.

Selection for lambing rate and clean fleece weight in sheep.

This study was designed to compare the effectiveness of selection for lambing rate (LB) and clean fleece weight (CFW) separately or by a combination of both traits at once. Two flocks of approximately 200 western white faced ewes per flock were maintained at two locations. Each flock was divided into four equal selection groups of similar initial birth type and age. The basis of selection in each flock was as follows: Line I, single trait selection for LB; Line II, single trait selection for CFW; Line III, multiple trait selection for LB and CFW; and Line IV; visual selection based on conformation and general fleece quality. Repeatability and heritability estimates were .18 and .16 +/- .04, respectively, for LB and .66 and .60 +/- .09, respectively, for CFW. The genetic correlation between LB and CFW was .26 +/- .16. Linear regression coefficients (number born/year) of annual response for LB in Lines I to IV were .018 +/- .008, .009 +/- .006, .011 +/- .005, and .009 +/- .005 (P > .05). Linear regression coefficients (kg/year) of annual observed response for CFW in lines I to IV were -.001 +/- .006, .027 +/- .009, .019 +/- .006, and .008 +/- .006. Response in Lines II and III was significant for CFW (P < .05) compared with Lines I and IV. In this study, CFW was increased by selection in white-faced sheep with little effect on lambing rate.

Palmitate esterification and glycerophosphate acyltransferase activity in adipose tissue of growing lambs.

The purposes of this study were to determine the effects of BW in growing lambs on rates of palmitate esterification (PE) and activity of glycerophosphate acyltransferase (GPAT) in subcutaneous and intermuscular adipose tissue in vitro and to determine the relationship between these in vitro measurements and carcass lipid accretion. At 37, 49, 60, and 71 kg of live weight, nine, nine, eight, and seven Columbia-Rambouillet-cross ewe lambs, respectively, were slaughtered and adipose tissue was dissected for glycerolipid biosynthesis assays. One-half of each carcass was boned, and carcass soft tissue lipid was measured. In both adipose tissues, PE and GPAT activity were highest at 37 kg (P = .0001); the greatest accumulation of carcass fat occurred between 37 and 49 kg (P = .0001). Differences in PE and GPAT were less well marked between 49-, 60-, and 71-kg lambs, but both were generally decreased with increased BW. Significant R2 from regressions of mass and percentage of carcass fat on either PE, GPAT, or steady-state glycerolipid synthesis (GLS) were from .39 to .44 (PE), .40 to .58 (GPAT), .50 to .75 (GLS), and .60 to .77 (PE, GPAT, and GLS combined). Changes in PE, GPAT, and GLS with BW were consistent with changes in rates of carcass fat accretion, which indicated a close relationship between glycerolipid biosynthesis in vitro and lipid deposition in vivo in growing-finishing lambs.

Growth and slaughter characteristics of ram and wether lambs implanted with zeranol.

Forty-nine Columbia ram and wether lambs born in April 1990 and 46 born in April 1991 were studied to determine the effects of zeranol implants on growth, difficulty of pelt removal, and carcass characteristics. Implanting ram and wether lambs once (1990) or twice (1991) with 12 mg of zeranol did not change live weight or ADG but gain/feed decreased (P < .05) in ram lambs slaughtered at approximately 50 kg. Testes weight was reduced approximately 50% by implanting. Two implants reduced (P < .05) the force needed to pull the pelt from the hind legs of ram lambs, but implanting tended to increase the force required to pull the pelt from wether lambs. Data for pelt weight, force required to pull the pelt, percentage of the carcass in the shoulder or splenius muscle, and Warner-Bratzler shear values showed that zeranol implants resulted in ram lambs becoming more like wethers and wether lambs becoming more like rams. Implanting with zeranol did not affect closure of the metacarpal growth plate in ram or in wether lambs. Difficulty of pelt removal can be reduced by implanting ram lambs with 12 mg of zeranol at approximately 114 d of age and reimplanting zeranol 28 d later.

Effects of maternal undernutrition during early pregnancy on growth, muscle cellularity, fiber type and carcass composition in lambs.

Seventy-two Rambouillet ewes were fed one of two different levels of energy and protein during gestation to determine the effects of maternal undernutrition on growth and development in their offspring. Levels of energy and protein for the two groups of ewes were: 70% of National Research Council (NRC) requirements 30 d prior to breeding and the first 100 d of gestation and then alfalfa hay fed at 70% of the average consumed by group 2, and 100% of NRC requirements 30 d prior to breeding and throughout gestation. Dissection data from the leg and muscle characteristics were recorded for each lamb. Neither dissected muscle and fat percentages nor proximate analyses of dissected muscles was influenced by ewe undernutrition during early pregnancy. However, the reduced conception rate, increased embryonic/fetal mortality, decreased birth weights and increased lamb mortality resulted in a severe reduction in kilograms of lamb per ewe bred. Feeding ewes 70% of NRC requirements during gestation resulted in lambs with heavier semitendinosus muscle weights, larger muscle fiber diameters and shorter sarcomere lengths than in lambs from adequately fed ewes. No influence of ewe diet, birth type or sex was observed for proportions of muscle fiber type (beta R, alpha R or alpha W). Therefore, low level feeding of ewes during early gestation had no detrimental effects on the carcass or muscle fiber characteristics of their lambs at slaughter.

Factors influencing difficulty of removing pelts from lamb carcasses.

Forty-eight Suffolk x white-faced ram and wether lambs approximately 5, 7, or 9 mo of age were slaughtered to evaluate the effects of age and gender on difficulty of pelt removal, pelt damage, and leg damage. A commercial belt-type pelt puller and a scale that recorded force required to remove the pelt from the thickest part of the legs was used as lambs hung suspended from their front legs. Rams required more force (P less than .05) to remove the pelt than wethers, and the difference between genders became larger as age increased. Neither pelt damage due to grain crack nor leg damage judged by amount of fell and fat removed by the pelt puller changed with age. Rams possessed thicker pelts (P less than .05) than wethers; this plus a greater amount of collagen crosslinking in ram skins could be responsible for the slightly smaller amount of grain crack observed in ram pelts. Factors involved in difficulty of pelt removal in ram lambs included age, splenius weight, and overall maturity. Difficulty of pelt removal in wether lambs was best predicted by including age and splenius weight in the model. These data tend to support packers' common practice of discounting rams over 5 mo of age because rams develop masculine characteristics and become harder to dress with increasing age.

Bone ossification and carcass characteristics of wethers given silastic implants containing estradiol.

Administration of growth promotants with estrogenic activity via hastening closure of the growth plate could have an economic impact on lambs because closure results in ovine carcasses being classified as yearlings. Twelve wether lambs approximately 12 mo old were given silastic implants filled with estradiol-17 beta and allotted randomly to be slaughtered 30, 60, 90, or 220 d after implantation to determine time of growth plate closure in relation to date of implanting. Seven comparable wethers not implanted served as controls. Four wethers implanted at 2 mo of age and slaughtered 220 d later also were included. Implanted wethers had serum concentrations of estradiol averaging 13.9 pg/ml over the 220-d implant period; controls averaged 2.7 pg/ml. Estradiol implants increased carcass maturity scores but fat deposition was not altered. Growth plate widths decreased (P less than .05) as wethers grew older and implants were in place for longer periods of time. Metacarpal growth plates in 12-mo-old lambs were completely ossified 220 d after implanting, but control wethers and wethers implanted at 2 mo maintained growth plate widths. Neither metacarpal nor metatarsal bone lengths differed (P less than .05) between control and implanted wethers. Growth plate ossification was not complete until 570 d of age in implanted lambs, even though bone length had stopped increasing by 408 d. Therefore, even though estradiol and other growth stimulants with estrogenic activity increase rate of ossification of the metacarpal growth plate, bone length or mature size is not limited.(ABSTRACT TRUNCATED AT 250 WORDS)

Gonadal function, sexual behavior, feedlot performance, and carcass traits of ram lambs actively immunized against GnRH.

The effect of active immunization against GnRH on production, carcass, and behavioral traits was examined in ram lambs fed to a uniform slaughter weight. Lambs (initial BW = 32.6+/-1 kg) were stratified by BW and assigned at random to one of four treatment groups (n = 12 lambs/group). Lambs were untreated, castrated, or actively immunized against GnRH using a GnRH-keyhole limpet hemocyanin conjugate (1 mg) emulsified with either Freund's complete adjuvant (FCA) or another oil-based adjuvant (ISA). Animals were housed individually and slaughtered at 58 kg BW. Immunoneutralization of GnRH reduced (P < .05) testes weight and the concentration of testosterone in serum at slaughter. Suppression of testicular size and function was most clearly evident in animals immunized using FCA. Final anti-GnRH titer was also highest in lambs immunized using FCA. Several measures of sexual behavior (frequency of mounts and ejaculations) were also reduced (P < .05) in animals immunized using FCA. The duration of the feeding period was greater (P < .05) for castrated lambs than for untreated lambs, and intermediate feeding periods were required for FCA and ISA lambs. Average daily gain was greater (P < .05) in untreated than in castrated, FCA, or ISA lambs. Similarly, feed efficiency for untreated lambs was greater (P < .05) than for castrated, FCA, or ISA lambs, but feed efficiency did not differ among castrated, FCA, or ISA lambs. Longissimus muscle area, lean and bone maturity, overall quality, muscling score, flank streaking, and color of fat did not differ among treatments. Intact, FCA, and ISA lambs had more (P < .05) desirable yield grades, less (P < .05) backfat, and less (P < .05) marbling than castrated lambs. In summary, immunization against GnRH decreased testicular weight and reduced (P < .05) feedlot performance and sexual behavior to levels comparable to those of castrated males. Partitioning of nutrients for growth and deposition of fat, however, seems to differ among immunologically castrated and physically castrated lambs. This difference in nutrient partitioning may be due to residual testicular activity in immunized lambs.

Relationship of body composition of mature ewes with condition score and body weight.

Fourteen mature, ovariectomized, western-range ewes with an initial mean BW of 72 +/- 4.5 kg and mean condition score (CS) of 7.5 +/- .3 were used to evaluate the relationship between CS and body composition. Diets of chopped straw and alfalfa hay were formulated to provide either maintenance energy or less than maintenance energy (100 or 60% of ME) to induce changes in BW and CS. After 180 d, ewes were weighted, scored for body condition, and slaughtered. All carcass components, viscera, and organs were analyzed for lipid, DM, and ash, and protein was determined by difference. Body weight and CS values were related by regression analysis to percentage of composition and weights of carcass components, carcass, and empty body. Body weight and CS were highly correlated (r = .89) and analysis indicated that each increase in CS resulted in an increase of 5.1 kg of BW. Condition score accounted for more variation of percentage of lipid in the empty body (R2 = .95) and carcass (R2 = .90) than did BW (R2 = .84 and .80, respectively). In contrast, BW accounted for more of the variation in carcass weight (R2 = .97) and empty BW (R2 = .99). Inclusion of both BW and CS in regression models did not increase the variation accounted for with the single best predictor. With mature western-range ewes in this study, CS was highly related to carcass lipids and could be used to describe energy reserves available to ewes.

Effects of age, castration, and season on difficulty of pelt removal in lambs.

Sixty-four white-faced rams and wethers were dressed with the aid of a commercial pelt puller. The effects of age, castration, and season on difficulty of pelt removal and pelt damage were evaluated. Lambs were divided into two age groups (5 and 12 mo) within gender (ram and whether) and season (spring and fall). A greater force (P less than .05) was required to remove pelts from rams than from wethers in both 5- and 12-mo-old groups. Older lambs slaughtered in the fall required more force (P less than .05) to remove their pelts than did those slaughtered in the spring, but differences by season did not exist for 5-mo-old lambs. The difference between rams and wethers in percentage of live weight that was closely shorn pelt weight was not significant (P greater than .05). The area of grain crack in the flank expressed as a percentage of total area of the skin was lower (P less than .05) for skins from 5-mo-old lambs and ram lambs than it was for skins from 12-mo-old lambs and wether lambs, respectively. Factors involved in difficulty of pelt removal in ram lambs included crosscut shoulder weight, fat firmness, and carcass weight. Difficulty of pelt removal in wether lambs was best predicted by including crosscut shoulder weight and bodywall thickness in multiple regression equations.

Influence of age and testosterone levels on masculine development in rams.

Fifty wethers and 51 spring-born rams were divided into five groups and slaughtered at different seasons of the year at average ages of 271, 361, 459, 557 or 652 d to determine the age and season at which differences in secondary sex characteristics could be detected. Serum testosterone concentrations and testes weights were low in January when the rams were 271 d of age and again in April at 361 d of age. By July, at 459 d of age, testosterone concentrations and testes weights had peaked and then decreased the following November at 557 d and February at 652 d. In contrast with plasma testosterone concentrations and testes weights, buckiness scores, splenius to semimembranosus or semitendinosus muscle ratios, splenius muscle weights and neck and shoulder percentages were not seasonal. All of these measures increased significantly up to July and continued to increase slowly, but not significantly, thereafter. Muscle color and texture scores and rib eye color scores tended to increase in a linear manner for both rams and wethers as age increased. Subcutaneous fat from rams was yellower and softer than that from wethers over all age groups. Ram fat firmness did not change (P greater than .05) with age, and the only significant change in ram fat color was between the groups at 271 and 361 d of age. Overall, season of year coupled with higher levels of serum testosterone was related to initial development of secondary sex characteristics in ram lambs.

Influence of gender, breed and age on maturity characteristics of sheep.

Fifty-one ewes, 50 wethers and 51 rams of Finn x whiteface or Suffolk x whiteface breeding were slaughtered at an average age of 271, 361, 459, 557 or 652 d to study changes in maturity characteristics with changes in age. When metacarpal epiphyseal cartilage (growth plates) ossifies with age, break joints on the distal end of the metacarpals fuse and the end of the bone then appears as a spool joint rather than as a break joint. This fusion occurs earlier chronologically in ewes than in rams or wethers. Most Finn x whiteface-cross ewes, which normally reach reproductive maturity earlier than Suffolk x whiteface-cross ewes, exhibited ossified metacarpal growth plates by 459 d of age; break joints of Suffolk x whiteface crosses were not fused until 557 d of age. Both breed groups of rams exhibited fused joints by 557 d of age, but only 3 of 10 wethers exhibited spool joints at 652 d of age. Microscopic examination showed that even in these three wethers the epiphyseal growth plate could still be detected. Yearling 2-yr-old classifications of sheep from teeth were not gender-dependent within the discrete age classes of this study. About half the sheep in each gender group were classified as yearlings at 459 d of age and about half in each group were classified as 2-yr-olds by 652 d of age. Stepwise multiple regression equations to estimate age of sheep in days were developed. Overall maturity, lean color and texture and rib bone maturity were the best carcass measures of maturity. Maturity scores for characteristics of the break joint did not enter any of the equations studied. When eye lens weight was included in equations with the best measures of carcass maturity, accuracy of estimates for age of sheep was improved.

Effects of extruded corn or grain sorghum on intake, digestibility, weight gain, and carcasses of finishing steers.

We conducted two trials to evaluate the effects of extruding vs dry-rolling either corn or grain sorghum on intake, digestibility, and performance of finishing steers. In Trial 1, 92 crossbred steers (average BW 413 kg) were used in a 2 x 2 factorial design. Diets contained either dry-rolled corn (RC), extruded corn (EC), dry-rolled grain sorghum (RGS), or extruded grain sorghum (EGS). Diets were fed for 110 d and contained 78.6% of the respective grain, 9% alfalfa pellets, 8.2% molasses, and 4.2% protein-mineral supplement. Daily gain was highest (P < .049) for steers fed RC, and the ADG of steers fed RGS was higher than that of steers fed extruded diets; there was no difference in ADG between steers fed EC and those fed EGS. Steers fed dry-rolled diets consumed more DM (P = .001) than steers fed extruded diets. Feed efficiency was not affected (P = .18) by processing method, but steers fed corn utilized the diets more efficiently (P = .006) than steers fed grain sorghum. Except for carcass weight, carcass data were not affected by grain type (P > .20). Dressing percentage, quality grade, and longissimus muscle area were lower (P < .09) in steers that received extruded grain than in those that received dry-rolled grain. In Trial 2, five ruminally cannulated crossbred steers (average BW 518 kg) were used in a 4 x 4 + 1 Latin square design to evaluate the ruminal and total tract digestion characteristics of the diets used in Trial 1. Type of grain had no effect (P > .16) on intake, total tract digestibility, or ruminal pH. Extruding corn or grain sorghum decreased intake (P < .001) but increased (P < .074) DM and starch digestibility compared with dry rolling; steers fed extruded diets had lower (P < .032) ADF and NDF digestibilities. Ruminal in situ DM and starch disappearance were higher (P < .03) and ruminal pH was lower (P < .052) in steers fed extruded grains than in those fed dry-rolled grains. Data from this study indicate that extruded corn and extruded grain sorghum are highly degradable feeds; however, decreased DM intake and lower ruminal pH levels resulted in lower performance.

Effect of plane of nutrition and age on carcass maturity of sheep.

One hundred twenty ewe and wether lambs were fed high (76% TDN, 12.3% crude protein) or low (70.6% TDN, 16.1% crude protein) energy diets and slaughtered at average ages of 261, 356 or 469 d. High planes of nutrition for lambs at 356 or 469 d of age resulted in heavier carcasses, thicker fat depth and higher flank streaking scores compared to lambs fed low planes of nutrition. Rib bone, break joint and overall maturity scores were not influenced (P greater than .05) by diet. Metacarpal and metatarsal bones from lambs fed high planes of nutrition were slightly longer and heavier and required more force to break (P less than .05) compared with bones from lambs fed low planes of nutrition. The majority of the ewes fed high or low planes of nutrition had fused growth plates and possessed spool joints by 469 d; none of the wethers had spool joints. An additional 27 wethers were slaughtered at an average of 480, 578 or 662 d. Rib bone, break joint and overall maturity scores were not different (P greater than .05) for animals fed the high or low energy diets, but bone weight and shaft break force values were higher for those fed high energy diets. At 662 d of age, only 20% of the wethers on each dietary regimen possessed fused growth plates. Overall, plane of nutrition in market-weight lambs and yearlings had very little influence on the presence or absence of a break joint or on other bone maturity scores.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of delayed bleeding on sensory characteristics of beef.

Sensory characteristics of bovine longissimus dorsi and biceps femoris muscles from 30 heifers exsanguinated 0, 3 or 6 min after stunning and 36 steers exsanguinated 0, 3 or 30 min after stunning were evaluated to determine if delayed bleeding influenced palatability. More blood was retained in carcasses of animals where exsanguination was delayed after stunning than in control animals, but no consistent differences existed for total pigment or per cent hemoglobin in muscle. Tenderness, juiciness, flavor, shear values and cooking loss were not affected by treatment.