Biomedical subjects

J A Bertrand

Publications and source records attributed to J A Bertrand.

At least 19 recordsLinked to original sources

PubMed2006

The effect of carbohydrate source on nitrogen capture in dairy cows on pasture.

The objective of this study was to determine if feeding carbohydrate supplements with faster degradation rates than corn to dairy cows grazing ryegrass would improve nitrogen capture, milk production, and components. Treatments were grain supplements based on: 1) corn (CORN), 2) barley and molasses (BM), or 3) citrus pulp and molasses (CM). For BM and CM, the diet composition was the same as that of CORN except that a portion of the corn was replaced with barley and molasses or citrus pulp and molasses, respectively, on a dry matter basis. Cows grazed ryegrass (Lolium multiflorum Lam.) pasture. Yield of milk, 3.5% fat-corrected milk, energy-corrected milk, and milk fat, as well as milk fat percentage, were not different among treatments. True milk protein percentage was higher for CORN (2.81%) compared with CM (2.70%), but was not different for BM (2.77%). However, true milk protein yield was not different among treatments. Milk urea N was higher for BM (11.43 mg/dL) compared with both CORN and CM (average: 9.95 mg/dL). There were no differences among CORN, BM, and CM treatments for overall BUN (average: 10.60 mg/dL). At 0400 h, however, cows on CORN had higher BUN than cows on CM (11.43 vs. 9.96 mg/dL), but there were no differences between CORN and BM (average: 11.21 mg/dL) or BM and CM (average: 10.48 mg/dL), and there were no differences among treatments at other time points. The CM diet might have shown more advantage if the pasture crude protein content was higher. Partial replacement of corn with citrus pulp for grazing cows should be further studied using pasture with higher crude protein content. Although cows receiving CM and BM did not produce more milk than cows on CORN, if barley or citrus pulp is less expensive than corn, they may be viable replacements for a portion of the corn supplement for grazing cows.

Animal Nutritional Physiological Phenomena↗

PubMed2005

Nutrient content of whole cottonseed.

The objective of this study was to determine if the nutrient and gossypol contents and in vitro digestibility of 3 types of genetically modified whole cottonseed differed from traditional whole cottonseed. Samples of seed from traditional (no genetic modifications) and genetically modified varieties of cotton grown in 1999 and 2000 were analyzed. Genetic modifications included the insertion of genes to protect cotton from insect pests (Bt), and damage from glyphosate herbicides (RR), and from both (Bt/RR). Year effects were significant for in vitro dry matter (DM) digestibility, gossypol, DM, crude protein (CP), fat, neutral detergent fiber (NDF), acid detergent fiber (ADF), and ash. Higher rainfall resulted in higher CP, fat, and ash and lower NDF and gossypol. There were no differences among seed types for ground or whole seed digestibility, DM, CP, fat, NDF, ADF, ash, lignin, net energy for lactation, amino acids, total fatty acids, or seed index. Overall, the nutrient content and digestibility of varieties of genetically modified seed were similar to that of varieties of traditional whole cottonseed.

Animal Feed↗

PubMed2004

Ruminal biohydrogenation in Holstein cows fed soybean fatty acids as amides or calcium salts.

Fatty amides of high oleate fats and calcium salts of palm oil were reported to resist biohydrogenation by ruminal microorganisms. This study was conducted to determine whether converting polyunsaturated fat sources to amides and calcium salts had equal ability to resist biohydrogenation. A total mixed ration consisting of forage and concentrate contained (dry basis): 1) 2.45% soybean oil (SBO), 2) 2.75% calcium salt of SBO, 3) 2.75% amide of SBO, or 4) 2.75% of a mixture of the calcium salt and amide (80:20, wt/wt) of SBO. The 4 diets were fed ad libitum to 4 multiparous lactating Holstein cows fitted with ruminal cannulas in a 4 x 4 Latin square with 21-d periods. Omasal samples were taken to measure postruminal fatty acid content and determine the extent of ruminal biohydrogenation. Adding SBO to the diets as either calcium salts or amides increased omasal flow of C18:2 (n-6) from 25 to 39 g/d. Omasal flow of C18:1 increased from 36 to 49 g/d when SBO was fed to cows as calcium salts, but increased to 86 g/d when SBO was fed as amides. Adding the soybean amide to the diet more than doubled the delivery of C18:1 (n-9) to the omasum of lactating cows, but it also increased trans fatty acid production in the rumen accompanied by milk fat depression. In this study, calcium salts and amide derivatives of fatty acids were both effective in enhancing omasal flow of unsaturataed fatty acids in lactating dairy cows. Amides were more effective than calcium salts for increasing the postruminal flow of oleic acid.

Amides↗

PubMed2003

Structural characterization of the GSK-3beta active site using selective and non-selective ATP-mimetic inhibitors.

GSK-3beta is a regulatory serine/threonine kinase with a plethora of cellular targets. Consequently, selective small molecule inhibitors of GSK-3beta may have a variety of therapeutic uses including the treatment of neurodegenerative diseases, type II diabetes and cancer. In order to characterize the active site of GSK-3beta, we determined crystal structures of unphosphorylated GSK-3beta in complex with selective and non-selective ATP-mimetic inhibitors. Analysis of the inhibitors' interactions with GSK-3beta in the structures reveals how the enzyme can accommodate a number of diverse molecular scaffolds. In addition, a conserved water molecule near Thr138 is identified that can serve a functional role in inhibitor binding. Finally, a comparison of the interactions made by selective and non-selective inhibitors highlights residues on the edge of the ATP binding-site that can be used to obtain inhibitor selectivity. Information gained from these structures provides a promising route for the design of second-generation GSK-3beta inhibitors.

Adenosine Triphosphate↗

PubMed2002

Effects of feeding time on nitrogen capture by lactating dairy cows grazing rye pasture.

The objective of this experiment was to determine whether varying times at which a partial mixed ration was fed, either before or after grazing, affected N utilization from rye pasture and thus affected milk yield and components. Sixteen Holstein cows were fed a partial mixed ration (PMR) either at 0700, 0830, or 1100 h. Cows were milked at 0900 h and turned out to graze at 0930 h. Treatments represented feeding times 2.5 h and 1 h before grazing and immediately after grazing. The study was conducted as a 3 x 3 Latin square with three 17-d periods. There were no significant differences among treatments for pasture intake or yield of milk or milk components. Milk yield, fat %, and protein % were 29.4, 29.6, and 29.3 kg, 3.5, 3.5, and 3.4%, and 3.4, 3.5, and 3.4% for treatments, respectively. The milk urea levels were 15.6, 15.1, and 15.5 mg/dl, and were not different among treatments. Blood samples were collected on the last day of each period at 0645, 0845, 1045, 1200, and 1400 h. Blood urea nitrogen (BUN) was measured as an indicator of ruminal N capture. Concentrations were not significantly different among diets before grazing; however, they were significantly different among all treatments approximately 1 h after cows were removed from pasture. Cows fed at 0700 h, 2 h before grazing, maintained lower BUN levels across the 7 h during which the blood samples were collected. Cows that ate the PMR immediately after grazing maintained the highest BUN. Feeding a PMR to cows that graze at different times before and after grazing affected the capture of ruminal N, as indicated by differences in the levels of BUN, but there was no effect on yield of milk or milk components.

Animal Nutritional Physiological Phenomena↗

PubMed2001

Comparison of fatty acid content of milk from Jersey and Holstein cows consuming pasture or a total mixed ration.

Holstein (n = 19) and Jersey (n = 18) cows were used to study effects of two feeding systems on fatty acid composition of milk. Confinement cows were fed a total mixed ration with corn silage and alfalfa silage and pastured cows grazed a crabgrass (90%) and clover (10%) pasture and were allowed 5.5 kg of grain per head daily. Two milk samples were collected from each cow at morning and afternoon milkings 1 d each week for four consecutive weeks in June and July 1998. One set of milk samples was analyzed to determine fatty acid composition, and the second set was used for crude protein and total fat analyses. Data were analyzed by the general linear models procedure of SAS, using a split-plot model with breed, treatment, and breed x treatment as main effects and time of sampling and week as subplot effects along with appropriate interactions. Milk from pastured cows was higher than milk from confinement cows for the cis-9, trans-11 octadecadienoic acid isomer of conjugated linoleic acid (CLA). Also, milk from Holsteins was higher than milk from Jerseys for C16:1, C18:1, and CLA and lower than Jerseys for C6:0, C8:0, C10:0, C12:0, and C14:0. Several treatment x week interactions existed, but main effects were still important; for example, proportions of CLA in milk of grazed cows were relatively constant across weeks (0.66, 0.64, 0.64, and 0.69% +/- 0.02%, respectively), but the CLA in milk of confinement cows increased in wk 4 (0.35, 0.31, 0.31, and 0.48% +/- 0.02% for wk 1 to 4, respectively). There are potentially important differences in fatty acid composition of milk from cows consuming a warm season pasture species compared with milk from cows consuming a total mixed ration, as well as differences between Holstein and Jersey breeds.

Animal Feed↗

PubMed2000

"Open" structures of MurD: domain movements and structural similarities with folylpolyglutamate synthetase.

UDP-N-acetylmuramoyl-l-alanine:d-glutamate (MurD) ligase catalyses the addition of d-glutamate to the nucleotide precursor UDP-N-acetylmuramoyl-l-alanine (UMA). The crystal structures of Escherichia coli in the substrate-free form and MurD complexed with UMA have been determined at 2.4 A and 1.88 A resolution, respectively. The MurD structure comprises three domains each of a topology reminiscent of nucleotide-binding folds. In the two structures the C-terminal domain undergoes a large rigid-body rotation away from the N-terminal and central domains. These two "open" structures were compared with the four published "closed" structures of MurD. In addition the comparison reveals which regions are affected by the binding of UMA, ATP and d-Glu. Also we compare and discuss two structurally characterized enzymes which belong to the same ligase superfamily: MurD and folylpolyglutamate synthetase (FGS). The analysis allows the identification of key residues involved in the reaction mechanism of FGS. The determination of the two "open" conformation structures represents a new step towards the complete elucidation of the enzymatic mechanism of the MurD ligase.

Adenosine Triphosphate↗

PubMed2000

Mechanism of calcite crystal growth inhibition by the N-terminal undecapeptide of lithostathine.

Pancreatic juice is supersaturated with calcium carbonate. Calcite crystals therefore may occur, obstruct pancreatic ducts, and finally cause a lithiasis. Human lithostathine, a protein synthesized by the pancreas, inhibits the growth of calcite crystals by inducing a habit modification: the rhombohedral (10 14) usual habit is transformed into a needle-like habit through the (11 0) crystal form. A similar observation was made with the N-terminal undecapeptide (pE(1)R(11)) of lithostathine. We therefore aimed at discovering how peptides inhibit calcium salt crystal growth. We solved the complete x-ray structure of lithostathine, including the flexible N-terminal domain, at 1.3 A. Docking studies of pE(1)R(11) with the (10 14) and (11 0) faces through molecular dynamics simulation resulted in three successive steps. First, the undecapeptide progressively unfolded as it approached the calcite surface. Second, mobile lateral chains of amino acids made hydrogen bonds with the calcite surface. Last, electrostatic bonds between calcium ions and peptide bonds stabilized and anchored pE(1)R(11) on the crystal surface. pE(1)R(11)-calcite interaction was stronger with the (11 0) face than with the (10 14) face, confirming earlier experimental observations. Energy contributions showed that the peptide backbone governed the binding more than did the lateral chains. The ability of peptides to inhibit crystal growth is therefore essentially based on backbone flexibility.

Amino Acid Sequence↗

PubMed2000

In situ disappearance of malate from alfalfa and bermudagrass hay.

The objectives of this study were to determine the rate of malate and dry matter disappearance from different forages in the rumen. Four nonlactating, ruminally cannulated Holstein cows were fed a hay-based diet. Samples of early and late harvested alfalfa, Coastal bermudagrass, and Tifton 85 bermudagrass hays were ground, placed in nylon in situ bags, and ruminally incubated for 0, 0.5, 1, 2, 4, 8, 12, 24, and 48 h. After incubation, samples were rinsed, freeze-dried, extracted, and analyzed for malate content by HPLC with an organic acid column. When forages were incubated in the rumen, malate concentrations were less than 0.55 mg/g of dry matter at 0.5 h and remained low for the 48-h incubation period. These results suggest that malate was solublized and utilized within 30 min after reaching the rumen. Dry matter digestibility of both forages increased with time and was different across forages. Both alfalfa samples were digested to a greater extent between 0.5 and 24 h than either type of bermudagrass, but after 48 h the early maturity Tifton 85 digestibility was similar to alfalfa. Even though it is more common to feed unground forages to ruminants, these in situ results suggest that once malate is available in the rumen it will disappear quickly.

Animal Feed↗

PubMed1999

Role of the ortholog and paralog amino acid invariants in the active site of the UDP-MurNAc-L-alanine:D-glutamate ligase (MurD).

To evaluate their role in the active site of the UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase (MurD) from Escherichia coli, 12 residues conserved either in the Mur superfamily [Eveland, S. S., Pompliano, D. L., and Anderson, M. S. (1997) Biochemistry 36, 6223-6229; Bouhss, A., Mengin-Lecreulx, D., Blanot, D., van Heijenoort, J., and Parquet, C. (1997) Biochemistry 36, 11556-11563] or in the sequences of 26 MurD orthologs were submitted to site-directed mutagenesis. All these residues lay within the cleft of the active site of MurD as defined by its 3D structure [Bertrand, J. A., Auger, D., Fanchon, E., Martin, L., Blanot, D., van Heijenoort, J., and Dideberg, O. (1997) EMBO J. 16, 3416-3425]. Fourteen mutant proteins (D35A, K115A, E157A/K, H183A, Y194F, K198A/F, N268A, N271A, H301A, R302A, D317A, and R425A) containing a C-terminal (His)(6) extension were prepared and their steady-state kinetic parameters determined. All had a reduced enzymatic activity, which in many cases was very low, but no mutation led to a total loss of activity. Examination of the specificity constants k(cat)/K(m) for the three MurD substrates indicated that most mutations affected both the binding of one substrate and the catalytic process. These kinetic results correlated with the assigned function of the residues based on the X-ray structures.

Amino Acid Sequence↗

PubMed1999

Determination of the MurD mechanism through crystallographic analysis of enzyme complexes.

UDP -N- acetylmuramoyl- L -alanine: D -glutamate (MurD) ligase catalyses the addition of d -glutamate to the nucleotide precursor UDP -N- acetylmuramoyl- L -alanine (UMA). The crystal structures of three complexes of Escherichia coli MurD with a variety of substrates and products have been determined to high resolution. These include (1) the quaternary complex of MurD, the substrate UMA, the product ADP, and Mg2+, (2) the quaternary complex of MurD, the substrate UMA, the product ADP, and Mn2+, and (3) the binary complex of MurD with the product UDP - N- acetylmuramoyl- L -alanine- D -glutamate (UMAG). The reaction mechanism supported by these structures proceeds by the phosphorylation of the C-terminal carboxylate group of UMA by the gamma-phosphate group of ATP to form an acyl-phosphate intermediate, followed by the nucleophilic attack by the amino group of D-glutamate to produce UMAG. A key feature in the reaction intermediate is the presence of two magnesium ions bridging negatively charged groups.

Adenosine Diphosphate↗

PubMed1999

Interaction of tallow and hay particle size on ruminal parameters.

Four nonlactating ruminally cannulated Holstein cows were used in a 4 x 4 Latin square experiment with 4 21-d periods to determine if the effects of dietary fat would be affected by hay particle length. Treatments consisted of two levels of tallow (0 and 5%) and two hay particle lengths (short-cut and long-cut) in a 2 x 2 factorial. Diets contained alfalfa hay, corn silage, and concentrate [1:1:2, dry matter (DM) basis] fed as a total mixed ration (TMR) once per day. Samples of the 0 and 5% tallow TMR were ground and incubated in situ in polyester bags for 24 and 48 h. Ruminal samples were taken on day 21 at 0800 h and at 2-h intervals until 1600 h. The total tract digestibilities of acid detergent fiber (ADF) and neutral detergent fiber (NDF) were not affected by tallow or by hay by tallow interactions. There was a trend for tallow to improve total tract digestibility of crude protein (CP) (70.2 vs. 74.7%). After 48 h of ruminal incubation, tallow significantly decreased the digestibilities of DM, ADF, and NDF. No hay length by tallow interactions for DM, NDF, ADF or CP digestibilities occurred after 24 or 48 h. Tallow increased concentrations of propionate and decreased concentrations of acetate and valerate and the acetate-to-propionate ratio. Total volatile fatty acids increased when tallow was added to diets with short-cut hay, which suggests that when unprotected fat is added to diets with a high level of hay, a short-cut hay length may be advantageous. This result may be due to shorter rumen retention time of feed particles, which reduces the time for fatty acids to exert antimicrobial effects. Or, it may because the increased surface area of the hay particle provides more area for microbial attachment and increased fermentation.

Adipose Tissue↗

PubMed1998

Oxyanion-mediated inhibition of serine proteases.

Novel aryl derivatives of benzamidine were synthesized and tested for their inhibitory potency against bovine trypsin, rat skin tryptase, human recombinant granzyme A, human thrombin, and human plasma kallikrein. All compounds show competitive inhibition against these proteases with Ki values in the micromolar range. X-ray structures were determined to 1.8 A resolution for trypsin complexed with two of the para-substituted benzamidine derivatives, 1-(4-amidinophenyl)-3-(4-chlorophenyl)urea (ACPU) and 1-(4-amidinophenyl)-3-(4-phenoxyphenyl)urea (APPU). Although the inhibitors do not engage in direct and specific interactions outside the S1 pocket, they do form intimate indirect contacts with the active site of trypsin. The inhibitors are linked to the enzyme by a sulfate ion that forms an intricate network of three-centered hydrogen bonds. Comparison of these structures with other serine protease structures with noncovalently bound oxyanions reveals a pair of highly conserved oxyanion-binding sites in the active site. The positions of noncovalently bound oxyanions, such as the oxygen atoms of sulfate, are distinct from the positions of covalent oxyanions of tetrahedral intermediates. Noncovalent oxyanion positions are outside the "oxyanion hole." Kinetics data suggest that protonation stabilizes the ternary inhibitor/oxyanion/protease complex. In sum, both cations and anions can mediate Ki. Cation mediation of potency of competitive inhibitors of serine proteases was previously reported by Stroud and co-workers [Katz, B. A., Clark, J. M., Finer-Moore, J. S., Jenkins, T. E., Johnson, C. R., Ross, M. J., Luong, C., Moore, W. R., and Stroud, R. M. (1998) Nature 391, 608-612].

Amino Acid Sequence↗

PubMed1998

Interactions of tallow and hay particle size on yield and composition of milk from lactating Holstein cows.

An 18-wk lactation study was conducted to determine whether the effects of tallow on the lactation performance of dairy cows were influenced by particle size of hay in the ration. A total mixed ration containing 50% concentrate, 25% corn silage, and 25% alfalfa hay (dry matter basis) was fed to Holstein cows. Four total mixed rations were developed based on differences in the percentage of tallow in the concentrate and particle size of alfalfa hay: 1) 0% tallow, long-cut hay; 2) 0% tallow, short-cut hay; 3) 5% tallow, long-cut hay; and 4) 5% tallow, short-cut hay. Ration had no effect on dry matter intake, body weight gain or change in body condition score. Tallow increased milk and milk protein yields but reduced milk protein concentration. However, the effects of tallow on milk and milk protein yields were the same, regardless of hay length in the ration. A tendency for an interaction of tallow and hay particle size was detected for fat-corrected milk (FCM) because tallow increased FCM more when hay was short. Ration had no effect on volatile fatty acids in ruminal samples collected via a stomach tube. In this study, the effects of tallow on milk yield and composition from Holstein cows were the same, regardless of hay particle size in the ration. The tendency for tallow to increase FCM more when hay was short suggests at least a limited role of forage particle size in the determination of how fat supplements in dairy rations affect lactation performance.

Animal Feed↗

PubMed1998

Effect of ruminally protected amino acids on milk yield and composition of Jersey cows fed whole cottonseed.

The objectives of this experiment were to determine whether ruminally protected amino acids (AA) increased milk protein when this content was depressed by the addition of whole cottonseeds in the diets of early lactation Jersey cows. Treatments were 1) a control diet, 2) a diet containing whole cottonseed, and 3) a diet containing whole cottonseed and ruminally protected lysine and methionine. Cows were assigned to treatments at a mean of 7 d postpartum and remained on the experiment for 18 wk. Dry matter intake and yields of milk, milk fat, fat-corrected milk, and energy-corrected milk were not affected by treatment. Milk fat content tended to decrease for cows fed diets containing whole cottonseed. However, the percentages of milk protein, total N, and casein N were depressed by the addition of whole cottonseed and were increased by the addition of ruminally protected AA. Plasma concentrations of methionine, but not lysine, were increased when ruminally protected AA were fed, suggesting that lysine was the most limiting.

Amino Acids↗

PubMed1997

Crystal structure of UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli.

UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase (MurD) is a cytoplasmic enzyme involved in the biosynthesis of peptidoglycan which catalyzes the addition of D-glutamate to the nucleotide precursor UDP-N-acetylmuramoyl-L-alanine (UMA). The crystal structure of MurD in the presence of its substrate UMA has been solved to 1.9 A resolution. Phase information was obtained from multiple anomalous dispersion using the K-shell edge of selenium in combination with multiple isomorphous replacement. The structure comprises three domains of topology each reminiscent of nucleotide-binding folds: the N- and C-terminal domains are consistent with the dinucleotide-binding fold called the Rossmann fold, and the central domain with the mononucleotide-binding fold also observed in the GTPase family. The structure reveals the binding site of the substrate UMA, and comparison with known NTP complexes allows the identification of residues interacting with ATP. The study describes the first structure of the UDP-N-acetylmuramoyl-peptide ligase family.

Adenosine Triphosphate↗

PubMed1997

Influence of tallow and Aspergillus oryzae fermentation extract in dairy cattle rations.

Objectives were to determine the effects of adding 3 g/d of Aspergillus oryzae fermentation extract to diets with or without 5.6% added tallow. Twenty-eight Holstein cows (mean = 98 d of lactation) were assigned to a randomized block experiment in a 2 x 2 factorial arrangement of treatments. Treatments were the basal diet 1) without tallow or extract, 2) with extract but no tallow, 3) with tallow but no extract, and 4) with tallow and extract. Milk production, dry matter intake, 3.5% fat corrected milk, digestibility of neutral detergent fiber in the total tract were depressed for cows fed tallow. Addition of fermentation extract did not stimulate fiber digestion or milk production of cows fed diets with or without fat. Addition of extract did not overcome depression of fiber digestibility by cows fed tallow.

Analysis of Variance↗

PubMed1996

Crystal structure of human lithostathine, the pancreatic inhibitor of stone formation.

Human lithostathine (HLIT) is a pancreatic glycoprotein which inhibits the growth and nucleation of calcium carbonate crystals. The crystal structure of the monomeric 17 kDa HLIT, determined to a resolution of 1.55 angstroms, was refined to a crystallographic R-factor of 18.6%. Structural comparison with the carbohydrate-recognition domains of rat mannose-binding protein and E-selectin indicates that the C-terminal domain of HLIT shares a common architecture with the C-type lectins. Nevertheless, HLIT does not bind carbohydrate nor does it contain the characteristic calcium-binding sites of the C-type lectins. In consequence, HLIT represents the first structurally characterized member of this superfamily which is not a lectin. Analysis of the charge distribution and calculation of its dipole moment reveal that HLIT is a strongly polarized molecule. Eight acidic residues which are separated by regular 6 angstrom spacings form a unique and continuous patch on the molecular surface. This arrangement coincides with the distribution of calcium ions on certain planes of the calcium carbonate crystal; the dipole moment of HLIT may play a role in orienting the protein on the crystal surface prior to the more specific interactions of the acidic residues.

Amino Acid Sequence↗