On the use of X-ray absorption spectroscopy to elucidate the structure of lutetium adenosine mono- and triphosphate complexes.

Although the physiological impact of the actinide elements as nuclear toxicants has been widely investigated for half a century, a description of their interactions with biological molecules remains limited. It is however of primary importance to better assess the determinants of actinide speciation in cells and more generally in living organisms to unravel the molecular processes underlying actinide transport and deposition in tissues. The biological pathways of this family of elements in case of accidental contamination or chronic natural exposure (in the case of uranium rich soils for instance) are therefore a crucial issue of public health and of societal impact. Because of the high chemical affinity of those actinide elements for phosphate groups and the ubiquity of such chemical functions in biochemistry, phosphate derivatives are considered as probable targets of these cations. Among them, nucleotides and in particular adenosine mono- (AMP) and triphosphate (ATP) nucleotides occur in more chemical reactions than any other compounds on the earth's surface, except water, and are therefore critical target molecules. In the present study, we are interested in trans-plutonium actinide elements, in particular americium and curium that are more rarely considered in environmental and bioaccumulation studies than early actinides like uranium, neptunium and plutonium. A first step in this strategy is to work with chemical analogues like lanthanides that are not radioactive and therefore allow extended physical chemical characterization to be conducted that are difficult to perform with radioactive materials. We describe herein the interaction of lutetium(III) with adenosine AMP and ATP. With AMP and ATP, insoluble amorphous compounds have been obtained with molar ratios of 1:2 and 1:1, respectively. With an excess of ATP, with 1:2 molar ratio, a soluble complex has been obtained. A combination of spectroscopic techniques (IR, NMR, ESI-MS, EXAFS) together with quantum chemical calculations has been implemented in order to assess the lutetium coordination arrangement for the two nucleotides. In all the complexes described in the article, the lutetium cation is coordinated by the phosphate groups of the nucleotide plus additional putative water molecules with various tridimensional arrangements. With AMP 1:2 and ATP 1:1 solid-state compounds, polynuclear complexes are assumed to be obtained. In contrast, with ATP 1:2 soluble compound, the Lu coordination sphere is saturated by two ATP ligands, and this favors the formation of a mononuclear complex. In order to further interpret the EXAFS data obtained at the Lu LIII edge, model structures have been calculated for the 1:1 and 1:2 ATP complexes. They are discussed and compared to the EXAFS best fit metrical parameters.

Localization and quantification of carbon-centered radicals on any amino acid of a protein.

A general strategy to localize and quantify carbon-centered radicals within proteins is described. The methodology was first exemplified on amino acids and then on a peptide. This method is applicable to any protein system regardless of size, and the site of hydrogen abstraction by *OH on all residues within proteins is easily and accurately detected.

Decreases in aggression in tryptophan-supplemented broiler breeder males are not due to increases in blood niacin levels.

In previous studies with feed-restricted broiler breeders, it was found that supplementation with dietary Trp decreases aggressive activity more in socially dominant males than in socially subordinate males. Although it is probable that this effect is mediated centrally by serotonin, an alternative possibility is that it is associated with other metabolic products of Trp such as niacin. The objectives of this study were to determine whether 1) supplemental dietary niacinamide decreases aggression in broiler breeder males, 2) elevated blood niacin levels are associated with a decrease in aggression, and 3) social status influences any of these effects on behavior. Using a randomized complete block design, a control (0.19% Trp, 22 mg niacin/kg), Trp (1.5% Trp, 22 mg niacin/kg), or niacinamide (0.19% Trp, supplemented with 140 mg niacinamide/kg) diet was assigned to each pen. There were 16 birds per pen and three pens per treatment. Birds were maintained on skip-a-day feed restriction throughout, and behavioral observations were conducted until the social hierarchy in each pen was determined. Treatment diets were then fed from 15 through 18 wk of age, and the number of aggressive pecks and threats per pen recorded in three 20-min observation periods/wk. Blood samples were collected from dominant and subordinate birds and assayed for niacin using Tetrahymena pyriformis. Supplementing the diet with Trp significantly (P < 0.05) decreased aggression. Niacinamide-treated birds did not differ from either Trp-treated or control-treated birds, and no decreases in aggression were seen in birds with elevated blood niacin levels. Blood levels of niacin were, however, higher in dominant niacin-treated birds than in dominant control birds (P < 0.05), although there were no differences among subordinate or Trp-treated birds. Thus, the modulatory effect of Trp on aggression does not appear to be mediated by increased niacin synthesis.

Effect of feeding raw and treated common vetch seed (Vicia sativa) on the performance and egg quality parameters of laying hens.

Two experiments were conducted to study the effect of feeding raw or treated common vetch seeds (V) on BW, feed intake (FI), egg production (EP), feed conversion (FC), egg weight (EW), shell thickness (ST), yolk color score (YC), and Haugh unit score (HU) of Single Comb White Leghorn hens for 56 d. In Experiment 1, diets contained 0, 7.5, 15, and 22.5% raw V. Compared with the control, the 22.5% V diet decreased (P < .05) BW, FI, and EP. In Experiment 2, intact or ground (G) V were either soaked (S) in water (1:5) for 24 h or autoclaved (A) at 103.5 x 10(3) Pa for 8 h, and then dried (D) at 55 C for 24 h. Eight diets were used, a corn soybean (control) and seven others, each containing 25% V, previously subjected to the forementioned treatments as follows: untreated V (UV), SDV, GSDV, ADV, GADV, SADV, and GSADV. Compared with the control, the UV diet decreased (P < .05) FI (75 vs 98 g) and EP (47.3 vs 88.7%), increased FC (1.99 vs 1.4 kg feed per dozen eggs), and induced BW loss (-63 vs 49 g). Soaking intact V improved (P < .05) FI (85 g) and EP (69%). Remaining treatments resulted in further improvement. Hens fed all V diets produced eggs with HU score 13 points better than that of the control (P < .05). In both experiments, EW, YC, and ST were not different among treatments. Results indicated that autoclaved V at 25% level was not detrimental to layers' performance.

Valine deficiency. 2. The effect of feeding a valine-deficient diet during the starter period on performance and leg abnormality of male broiler chicks.

Using force-feeding and pair-feeding techniques, two experiments were conducted to investigate the problem of leg abnormality associated with feeding a Val-deficient diet as compared with a diet deficient in all the branched chain amino acids (BCAA) or a Val-supplemented diet. The BCAA-deficient diet contained .96, .52, and .63% of Leu, Ile, and Val, respectively. The Val-deficient diet contained 1.37 Leu, .82 Ile, and .63% Val. The Val-supplemented diet contained 1.37 Leu, .82 Ile, and .83% Val. In both studies birds fed the Val-supplemented gained more weight than birds in the other two groups. Although birds on Val-deficient and BCAA-deficient diets had similar weight gain, the former were lethargic and showed feather and leg abnormalities. Bone ash and bone calcium for Val-deficient birds were the lowest among the three groups studied (P < .05). The levels of the BCAA in plasma reflected those of the treatment diets. Plasma hydroxyproline was lowest in birds fed the Val-deficient diet (P < .05), indicating a reduction in bone collagen breakdown. Kidney function measurements were the same for birds on the Val-supplemented and Val-deficient diets. Fractional excretion of calcium in Val-deficient birds (.13%), however, was three times higher than that of Val-supplemented birds (P < .05). The results indicated that Val deficiency per se increased calcium excretion in urine and induced leg abnormality in young chickens.

Valine deficiency. 1. The effect of feeding a valine-deficient diet during the starter period on performance and feather structure of male broiler chicks.

Experiments were designed to investigate the effect of feeding diets deficient in one or more of the three branched-chain amino acids (BCAA) on the performance of 3-wk-old male broilers. In the first experiment, levels of .96 and 1.46% Leu, .52 and .82% Ile, and .65 and .95% Val were used. Feeding the lowest combination of the three BCAA resulted in weight gain (WG) and feed conversion ratio (FC) of 344 g and 1.59 g:g, respectively. These parameters were not improved by adding dietary increments of the three BCAA individually. The greatest response, however, for both WG (435 g) and FC (1.41 g:g) was obtained by the addition of the three BCAA simultaneously. Chicks fed the low-Val diets in combinations with added levels of Ile and Leu exhibited feather and leg abnormalities. A second experiment was designed to investigate the effect of Val deficiency on feather protein, feather amino acids, and calcium content of the bone. Three treatments were used: a BCAA-deficient diet with .96% Leu, .52% Ile, and .63% Val; a Val-deficient diet, which contained 1.37, .82, and .63% of Leu, Ile, and Val, respectively; and a Val-supplemented diet, which was the same as the previous diets except that the Val content was .83%. Valine deficiency significantly decreased WG (243 g), FC (1.69 g:g), bone calcium (134 mg/g dry bone), and feather protein (82.7% of wet weight). Valine deficiency also decreased the level of Cys in feathers, but increased those of Asp, Glu, Met, Tyr, His, and Lys.(ABSTRACT TRUNCATED AT 250 WORDS)

Uniformly 13C-labeled algal protein used to determine amino acid essentiality in vivo.

The edible alga Spirulina platensis was uniformly labeled with 13C by growth in an atmosphere of pure 13CO2. The labeled biomass was then incorporated into the diet of a laying hen for 27 days. The isotopic enrichment of individual amino acids in egg white and yolk proteins, as well as in various tissues of the hen at the end of the feeding period, was analyzed by negative chemical ionization gas chromatography/mass spectrometry. The amino acids of successive eggs showed one of two exclusive enrichment patterns: complete preservation of the intact carbon skeleton or extensive degradation and resynthesis. The same observation was made in tissue proteins. These patterns were cleanly divided according to known nutritional amino acid essentiality/nonessentiality but revealed differences in labeling among the nonessential amino acids: most notable was that proline accretion was derived entirely from the diet. Feeding uniformly 13C-labeled algal protein and recovering and analyzing de novo-synthesized protein provides a useful method to examine amino acid metabolism and determine conditional amino acid essentially in vivo.

The effect of dietary tryptophan on aggressive behavior in developing and mature broiler breeder males.

Broiler breeders display high levels of aggressive activity as a result of feed restriction. The objective of this study was to determine if aggressive activity in developing and mature broiler breeder males is influenced by increased dietary L-tryptophan (Trp). Broiler breeder males were raised using a skip-a-day feed restriction regimen. In Experiment 1, three levels of dietary Trp, .19 (control), .75, and 1.5%, were fed from the start of feed restriction at 4 wk of age and throughout development. All occurrences of aggressive behavior were recorded during three 20-min observation sessions per week on mornings on which feed was not delivered through Week 20. The Trp significantly decreased (P less than .05) aggressive pecking in birds fed supplemental Trp as compared to controls. No differences in threatening behavior were found. In the second part of Experiment 1, the effect of continued Trp supplementation on mature flocks of male broiler breeders and time-of-day effects on aggression were investigated. A depressing effect of supplemental Trp on pecking and threatening was observed. Maximum aggressive activity in all treatment groups occurred on mornings when feed was delivered. In Experiment 2, four levels of Trp, .19 (control), .38, .75, and 1.5%, were fed to developing birds as in Experiment 1. Birds fed supplemental Trp pecked less (P less than .05) than controls and .38% Trp was as effective in decreasing pecking as was 1.5%. It was concluded that increased levels of dietary Trp decrease aggression in both developing and mature, socially stable flocks of broiler breeder males.

Dietary requirements of leucine, isoleucine, and valine in male broilers during the starter period.

Growth studies were conducted with 3-wk-old, male broiler chicks to determine their requirements for the branched-chain amino acids (BCAA) leucine, isoleucine, and valine during the starter period. The central-composite, rotatable design was applied. The data were analyzed using regression methodology. The results showed that the linear term of Leu and the cross-product terms of Leu with Ile or Val were nonsignificant (P greater than .05). Second-order equations described the relationship between the BCAA, weight gain (WG), and feed conversion (FC). The coefficient of determination values indicated that 80% or more of the variation in WG and FC was explained by the polynomial equations. The results suggested that the Leu requirement does not exceed 1.16% of a diet calculated to contain 3,200 kcal of metabolizable energy per kilogram. The optimum response, maximum in the case of WG (444 g) and minimum for FC (1.40), would be obtained by feeding a diet containing 1.16, .80, and .90% of Leu, Ile, and Val, respectively. The values of Ile and Val are the actual values obtained when the diets were hydrolyzed for 72 hours. For Leu, however, the hydrolysis time was 24 hours.

Comparison of the sensitivities of growth and digestibility studies using intact, cecectomized, and cannulated roosters.

The sensitivity of several bioassays was determined using soybean meal and two autoclaved soybean meals. Chick growth study results revealed that a four percent difference in weight gain was sufficient to show statistical significance (P less than .05) between the diets. Lysine quantities and digestibility values were compared using excreta obtained from intact (excreta collected on mats or in bags), cecectomized. and ileal-cannulated roosters. A 30% difference in lysine quantity in the excreta (collected on mats) was required to detect statistically significant effects of different diets fed to intact birds. Using intact birds (excreta collected in bags), cecectomized, and cannulated birds, differences in lysine quantity of 41, 42, and 67% were required to show statistically significant effects. Results showed that altering birds did not significantly change lysine values obtained nor reduce variability among birds (P greater than .05).

Feeding BAY Vi 7533 to hens.

BAY Vi7533 (2-chloro-N-[[[4-(trifluoromethoxy)phenyl] amino] carbonyl] benzamide) was fed to White Leghorn and Vedette Mini-broiler breeder hens at concentrations ranging from 0 to 25 ppm in the ration. Concentrations of 15 and 25 ppm resulted in greater than 95% mortality of house flies, Musca domestica L., seeded as larvae into the droppings. Concentration of 25 ppm reduced egg production in the White Leghorn but not in the Vedette Mini-broiler hens. At each concentration (except 5 ppm for Vedette Mini-broilers) residues of the compound were detected in the eggs. These residues, however, had no adverse effects on fertility or hatchability of the eggs.

Effect of coccidiosis on heart composition and function in young turkeys.

Heart composition and function were measured in turkey poults inoculated with Eimeria meleagrimitis or E. adenoeides oocysts. Heart weights of infected poults were significantly lower than those of uninfected controls. Lipid levels of the heart were significantly reduced on day 4 postinoculation (PI) with E. meleagrimitis and on day 6 PI with E. adenoeides. The reduced lipid levels were closely associated with decreases in body weight. The protein per gram of heart tissue increased as the lipid levels decreased. Heart glycogen, when expressed as microgram per milligram of protein, decreased. However, the concentration of glycogen per gram of tissue apparently changed very little. Moisture levels of hearts from infected and control poults did not differ significantly. Heart rates and mean blood pressures of poults infected with E. adenoeides were slightly (approximately 8%) lower than values in control poults. However, exposure to stress in the form of ice baths or intravenous injection of epinephrine caused more severe and prolonged fluctuations in heart rate and blood pressure of the infected poults than in those of the controls.

The amino acid excretion values in intact and cecectomized negative control roosters used for determining metabolic plus endogenous urinary losses.

Intact and cecectomized adult male, Single Comb White Leghorns were fasted for 24 hr prior to the initiation of the collection period. Excreta were quantitatively collected from each of the unfed control roosters for a 24 to 48 hr fasting period. The amino acids excretion values (mg/bird/24 hr) were not significantly different (P greater than .05) between the intact and cecectomized roosters for histidine or methionine. However, lysine and glutamic acid were significantly different (P less than .05) and all other reported amino acids were significantly different (P less than .01). The results of this preliminary experiment indicate the possible role of the ceca in altering the amino acid excretion values. Additional research is required in the elucidation of cecal involvement, as well as the selection of a proper control for this method of analysis for amino acid availability.

Effect of time on response to Eimeria adenoeides and Eimeria meleagrimitis infection in young turkeys.

Physiologic characteristics were measured on day 7 postinoculation (PI) in 2-week-old turkeys inoculated with 10(5) Eimeria adenoeides oocysts, in pair-fed controls, and in control turkeys fed ad libitum. Pathophysiologic responses were measured in turkeys inoculated with 7 x 10(4) E. adenoeides oocysts or 1.2 x 10(5) E. meleagrimitis oocysts and necropsied 2, 4, 6, 8, 10, 15, and 21 days PI. At day 7 PI, weight gains and heart weights (as a percentage of body weight) of E. adenoeides-infected turkeys were significantly lower and plasma glucose levels significantly higher than those of pair-fed counterparts. Plasma carotenoid and protein levels of the infected turkeys were significantly lower than those of the controls fed ad libitum and were consistently, but not always significantly, lower that those of the pair-fed controls. Significant responses were first observed in E. meleagrimitis-infected turkeys on day 4 PI and in E. adenoeides-infected birds on day 6 PI. Birds infected with both species commonly had reduced weight gains, heart weights, and plasma carotenoid levels and increased plasma aspartate aminotransferase levels. Plasma carotenoids in E. adenoeides-infected turkeys were significantly reduced on day 6 PI only, whereas plasma carotenoids in E. meleagrimitis-infected turkeys were sometimes reduced significantly as early as day 2 PI and remained significantly lower than control values through day 21 PI.

Growth depression of chicks fed a crude rye extract containing pectic substances.

A crude extract with a high proportion of pectic substances was prepared from rye. This was fed to chicks in two studies to determine if rye pectic substances are responsible for the growth depression of chicks fed rye. The growth of chicks was significantly (P greater than .01) depressed when fed a diet containing this extract. Addition of penicillin to the diet improved the growth of chicks. Increasing the concentration of the extract in the diet produced an incremental depression in weight gain. The intestines of chicks fed this extract contained large quantities of gas that were reduced by penicillin supplementation. These results suggest that rye pectic substances may be responsible for the growth depression of chicks fed rye.

Eimeria meleagrimitis in young turkeys: effects on weight, blood, and organ parameters.

Weight and biochemical studies were conducted on 2-week-old turkeys inoculated with 10(4) to 5 X 10(5) sporulated Eimeria meleagrimitis oocysts, on their pair-fed controls (equivalent food intake), and on control turkeys fed ad libitum. Food consumption and rate of weight gain of all inoculated and pair-fed turkeys fell sharply on day 4 postinoculation (PI), but deaths occurred primarily among the birds inoculated with 5 X 10(5) oocysts. Heart weights (expressed as percentage of body weight) were reduced in inoculated and pair-fed birds, but liver, spleen, and pancreas weights did not differ from those of either control group. Feed conversion (feed consumed/gain) was less efficient for inoculated turkeys than for ad libitum or pair-fed controls and was least efficient for turkeys inoculated with 5 X 10(5) oocysts. Plasma glutamic oxaloacetic transaminase (GOT, aspartate and aminotransferase) activity increased, and carotenoid and total protein levels decreased in inoculated turkeys but not in the pair-fed turkeys, indicating that these changes were caused by the infection and not by reduced food intake. Plasma glutamic pyruvic transaminase (GPT, alanine aminotransferase) remained stable in all groups. Plasma glucose levels of inoculated birds did not differ from those of the control groups, but liver glucose and glycogen levels decreased in both the inoculated and pair-fed birds.

The effect of diet on liver glycogen and body composition in the chick.

Experiments were conducted to study the effects of diet on liver glycogen and body composition in the three-week-old chick. When percent protein and percent lysine were varied, liver glycogen concentration declined significantly as protein and lysine increased, 32.6 vs. 19.4 mg/g for the 14 and 35% protein diets, respectively. Carcass fat decreased and carcass moisture increased in a pattern similar to glycogen as protein and lysine increased in the diets. When seven levels of lysine were fed in isocaloric, isonitrogenous diets, glycogen content was not significantly different in any of the treatments. Liver glycogen was highest at the lowest energy level when dietary energy was increased and protein held constant. Carcass fat varied with the energy level but was also influenced by the amino acid balance. The data suggest that percent carbohydrate to percent fat ratio in the diet (both calculated) is an important factor affecting glycogen, but more research is needed to determine the mechanisms controlling the glycogen content of the liver.

The available lysine requirements of 7-9 week old sexed broiler chicks.

The experiments were conducted to determine the available lysine requirement of broiler chicks during the 7-9 week growing period. The chicks were sexed at day old, and the sexes were reared in separate pens in the same facility. Commercial-type broiler rations were fed from day old to 7 weeks of age. At 7 weeks the sexing errors were removed and the pens equalized to 50 birds per pen. The eight dietary treatments were replicated three times per sex. Graded levels of L-lysine-HCL were added to the basal ration to give a stepwise increase of 0.07% lysine per level. The available lysine requirement for female during the 7-9 week period was 0.64% for both growth and feed conversion. When this was expressed as a function of energy, 0.187% available lysine per megacalorie of metabolizable energy (M.E) per kg. of diet was required. The available lysine requirement for males during this period was 0.69 and 0.66% for gain and feed conversion, respectively. Expressed as available lysine per megacalorie of M.E. per kg. of diet, the values would be 0.202 and 0.193%, respectively.

The arginine requirement of the 4-7 week old broiler.

Two floor pen studies were conducted to determine the minimum arginine level needed to support maximum weight gain and optimum feed conversion during the 4-7 week period of growth for both males and females. Floor feather counts were used as an additional criteria for determining the arginine requirement. The basal diets were calculated to contain 0.74 and 0.80% arginine based on the analyzed values for corn, corn gluten meal, skimmilk and soybean meal. Regression equations were calculated from the data obtained for the males only. Based on the regression equations, the calculated requirement for maximum weight gain, optimum feed conversion, and floor feathers is 1.09, 1.10, and 1.13% arginine, respectively. When expressed as a function of energy, the requirements would be 0.329, 0.332, and 0.341% arginine per megacalorie of metabolizable energy per kg. of diet, respectively. The differences in rate of gain data for the females were non-significant, indicating the requirement of the females to be less than the males. The female requirement, based on floor feathers, would appear to be 0.98% dietary arginine.