Effects of alternate day feeding of dried distiller's grains plus solubles in forage-fed steers on intake, ruminal fermentation and passage rates, and serum nonesterified fatty acid.

Four ruminally and duodenally cannulated Holstein steers (BW = 449 ± 7.3 kg) were used to examine the effects of feeding either dried distiller's grains plus solubles (DG) or grass hay on alternate days (every other day) on intake, ruminal fermentation and passage rates, and serum NEFA in forage-fed steers. Steers were assigned to 1 of 4 dietary treatments in a 4 × 4 Latin square: 1) only hay (CON), 2) hay and 0.4% of BW as DG DM daily (DG7), 3) hay daily and 0.8% BW DG every other day (DG2), and 4) alternate day feeding of hay and 0.8% of BW as DG (DGA). Treatment periods consisted of 13 d of adaptation and 8 d of collecting digesta and blood. Over the entire collection period, DMI was decreased ( = 0.004) for DGA compared with other treatments (13.0 ± 0.8, 12.7 ± 0.8, 13.3 ± 0.8, and 10.9 ± 0.8 kg/d for CON, DG7, DG2, and DGA, respectively). Immediately after feeding on days supplement was fed to DG2 and DGA (supplemented days [SUP]), ruminal pH of DGA was less than other treatments but by the end of the day was greater than other treatments (treatment × time, < 0.001). At feeding time on nonsupplemented days (NSUP), ruminal pH of DGA steers was greater than other treatments but was similar (treatment × time, < 0.001) to DG2 and CON by 5 h after feeding. Total concentrations of VFA were similar ( = 0.09) among treatments on SUP; however, on NSUP, total VFA concentrations were least in DGA from feeding until 4 h after feeding (treatment × time, = 0.02). No differences ( ≥ 0.06) were observed among treatments for apparent ruminal, total intestinal, and total tract DM, OM, or CP digestibility. There were no differences ( = 0.36) in serum NEFA among treatments on SUP; however, on NSUP, steers fed DGA (209.5 ± 12.7 m) had greater ( < 0.01) NEFA compared with other treatments (84.4 ± 12.7, 88.0 ± 12.7, and 77.7 ± 12.7 m for CON, DG7, and DG2, respectively). The DGA feeding strategy influenced DMI and ruminal kinetics and circulating NEFA without impacting total tract digestibility.

Effects of alternate-day feeding of dried distiller's grain plus solubles to forage-fed beef cows in mid- to late gestation.

Forty-six nonlactating beef cows were used to examine effects of dried distiller's grains plus solubles (DG) supplementation strategies to cows fed grass hay during mid- to late gestation on BW, ultrasound body composition characteristics, concentrations of serum NEFA and urea, feeding behavior, and calf birth weight. Cows were assigned to dietary treatments in a completely randomized design: 1) control, where hay was fed each day of the week (CON), 2) both hay and DG fed daily during the week (DG7), 3) hay fed daily but DG fed 3 d of the week (DG3), and 4) hay fed 4 d of the week alternating with DG fed on the remaining 3 d (DGA). Hay was offered ad libitum on days it was fed. The DG were fed at 0.40% of BW when offered daily and 0.93% of BW when offered 3 d per week (Monday, Wednesday, and Friday). Feed intake was monitored continuously over the 84-d feeding period. Hay intake and total DMI were reduced (P < 0.05) in DGA compared with DG7 and DG3. Gain and G:F were decreased (P < 0.05) for CON compared with other treatments. No differences (P > 0.05) were observed among treatments for change in BCS, intramuscular fat, rib fat, or rump fat from d 1 to 84. On a day when DG7, DG3, and DGA all received DG (Friday), DGA had reduced (P < 0.05) concentrations of urea compared with DG3 and DG7. On a day when only DG7 received DG (Saturday), urea was greater (P < 0.01) for DG3 and DGA compared with DG7, and concentrations of NEFA were greater (P < 0.01) in CON and DGA compared with DG7. On the second consecutive day when only DG7 received DG (Sunday), concentrations of NEFA were less (P < 0.001) for DG7 compared with other treatments. On days when all cows received hay, DGA spent more time eating (P < 0.05) compared with DG7 and DG3. Cows fed DGA had greater (P < 0.05) hay intake per meal and time per meal compared with other treatments. On days when DG7, DG3, and DGA all received DG, cows in the DG3 and DGA treatments had greater (P < 0.05) number of DG meals, time spent eating, intake per meal, and time per meal but a slower (P < 0.05) rate of DG intake compared with DG7. No differences (P > 0.05) were observed in calf birth weights among treatments. The alternate-day feeding strategy reduced hay and total intake, altered concentrations of serum urea and NEFA, and altered feeding behavior compared with other supplementation methods.

Technical note: physiological response of beef heifers after receiving a reused controlled internal drug release insert processed with different heat-treating methods.

Eighty-one prepubertal beef heifers were used to evaluate effects of used controlled internal drug release (CIDR) insert heating methods on concentrations of progesterone after CIDR insert reinsertion. Heifers were stratified by weight and birth date and then assigned to receive a new CIDR insert (New; n = 10) or 1 of 8 used (7 d prior use) CIDR insert treatments: 1) no processing (Used; n = 10), 2) autoclaved (Autoclaved; n = 8), 3) processed in dishwasher (Dishwasher; n = 8), 4) processed in microwave for 30 s (Microwave; n = 10), 5) processed in toaster oven (Oven; n = 9), 6) processed in clothes dryer (Dryer; n = 10), 7) processed in boiling water (Boiled; n = 8), or 8) stored outdoors for 60 d (Outside; n = 8). Used CIDR inserts were processed at 121°C for 30 min for autoclaved and oven treatments, at 121°C for boiled treatment, and for 30 min for dryer and dishwasher treatments. Blood samples were collected on d -10, immediately before CIDR insert insertion (d 0), 3 h after CIDR insert insertion (3 h), daily while CIDR insert was in place (d 1 to 11), and 24 h after CIDR insert removal (d 12) for analysis of concentrations of progesterone. Subjective color scores (1 = bright white to 5 = completely stained yellow/red) were assigned to each CIDR insert after d 11. A treatment × time interaction (P < 0.0001) was present for concentrations of progesterone. Concentrations of progesterone were similar (P > 0.10) for heifers receiving a used CIDR insert compared with heifers receiving CIDR inserts processed in a dishwasher, microwave, oven, dryer, or boiling water (collectively reported as "Processed"). However, heifers receiving autoclaved CIDR inserts had greater (P < 0.05) concentrations of progesterone from h 3 to d 3 but similar (P > 0.10) concentrations of progesterone from d 4 to d 11 compared with heifers receiving used or processed CIDR inserts. From d 1 to 11 heifers receiving outside CIDR inserts had decreased (P < 0.05) concentrations of progesterone compared with all other treatments. Heifers receiving autoclaved CIDR inserts had greater (P < 0.05) concentrations of progesterone compared with all other treatments at 3 h and 1 d, whereas heifers receiving new CIDR inserts had greater (P < 0.05) concentrations of progesterone from d 6 to 11 compared with all other treatments. Outside CIDR inserts were more discolored (P < 0.001) compared with all other treatments. Processing used CIDR inserts with a dishwasher, microwave, oven, clothes dryer, boiling water, or full environmental exposure did not result in a pattern of concentrations of progesterone similar to that of autoclaved or new CIDR inserts.

Effects of temporary calf removal before fixed-time artificial insemination on pregnancy rates and subsequent calf performance in suckled beef cows.

Two experiments were conducted to determine the effect of calf removal (CR) on pregnancy rate (PR) and calf performance in suckled beef cows. Cows in both experiments were synchronized with the 7-d CO-Synch + CIDR protocol [i.e., 100-µg injection of GnRH at controlled internal drug release (CIDR) device insertion (d -7) with 25-mg injection of PGF2α at CIDR removal (d 0), followed by injection of GnRH and timed AI (TAI) on d 3]. Cows were blocked by location (6 locations), stratified by days postpartum (DPP) and parity, and assigned to 1 of 2 treatments in Exp. 1: 1) control (Control; n = 156); 2) calves were separated from their dams between d 0 and 3 (CR72; n = 168); and 1 of 4 treatments in Exp. 2: 1) Control (n = 103); 2) CR72 (n = 104); 3) calves were separated from their dams between d 0 and 2 (CR48A; n = 95); and 4) similar to CR48A but CR between d 1 and 3 (CR48B; n = 53). Transrectal ultrasonography of ovarian structures was performed on d 0, 1, 2, 3, 4, and 10 (in a subset of cows) to determine pregnancy status on d 33. Blood samples were collected on d -14, -7, 0, 3, and 10 (in a subset of cows) to determine concentrations of progesterone (P4) and estradiol (E2). Calves were blocked by age as young (25 to 59 d), medium (60 to 79 d), and old (≥80 d), and were weighed on d 0, 3, 33, and 63. Overall PR did not differ among treatments and averaged 50%. Follicle growth rate from d 0 to 3 tended (P = 0.06) to be greater for CR72 (0.42 ± 0.15 mm/d) compared with Control (0.02 ± 0.15 mm/d). Young (-3.9 ± 0.3%) and old (-3.1 ± 0.4%) calves lost a greater (P < 0.001) percent of BW (PBW) during CR than medium-age (-1.6 ± 0.3%) calves exposed to CR72. In Exp. 2, PR were similar among all 3 locations (49%; P = 0.15). Young (-4.8 ± 0.6%) and medium (-3.0 ± 0.5%) calves lost greater (P < 0.01) percent body weight (PBW) during CR than old (-1.4 ± 0.6%) calves within the CR72 treatment. Calves exposed to CR48 (-2.2 ± 0.6%, -1.1 ± 0.6%, and -2.4 ± 0.6% PBW change for young, medium, and old, respectively) lost more BW than calves in the Control group (-3.7 ± 0.4%, -1.7 ± 0.5%, and -2.1 ± 0.5% PBW change for young, medium, and old, respectively). Subsequent calf weights on d 33 and 63 were greater (P < 0.05) in Controls than cows exposed to CR48 or CR72 treatments. We conclude that CR stimulated follicle growth but failed to enhance PR to TAI. However, CR had a negative impact on subsequent calf performance, which differed, depending on the duration and age of the calf when exposed to CR.

Modeling a system of phosphated cross-linked high amylose for controlled drug release. Part 2: physical parameters, cross-linking degrees and drug delivery relationships.

High amylose cross-linked to different degrees with sodium trimetaphosphate by varying base strength (2% or 4%) and contact time (0.5-4h) was evaluated as non-compacted systems for sodium diclophenac controlled release. The physical properties and the performance of these products for sodium diclophenac controlled release from non-compacted systems were related to the structures generated at each cross-linking degree. For samples at 2% until 2h the swelling ability, G' and eta* values increased with the cross-linking degree, because the longer polymer chains became progressively more entangled and linked. This increases water uptake and holding, favoring the swelling and resulting in systems with higher viscosities. Additionally, the increase of cross-linking degree should contribute for a more elastic structure. The shorter chains with more inter-linkages formed at higher cross-linking degrees (2%4h and 4%) make water caption and holding difficult, decreasing the swelling, viscosity and elasticity. For 2% samples, the longer drug release time exhibited for 2%4h sample indicates that the increase of swelling and viscosity contribute for a more sustained drug release, but the mesh size of the polymeric network seems to be determinant for the attachment of drug molecules. For the 4% samples, smaller meshes size should determine less sustained release of drug.

Superiority of enoxaparin over heparin in combination with a GPIIb/IIIa receptor antagonist during coronary thrombolysis in dogs.

It is known that a low-molecular-weight heparin (LMWH) is more effective than unfractionated heparin in unstable angina/non-Q-wave myocardial infarction (UA/NQMI) and the platelet GPIIb/IIIa receptors play an important role in acute myocardial infarction (AMI). Therefore, enoxaparin might have a similar advantage over heparin when used with a GPIIb/IIIa antagonist (RPR109891) in coronary thrombolysis. After induction of coronary thrombosis in anesthetized dogs, infusion of saline, enoxaparin, heparin, RPR109891, enoxaparin+RPR109891, or heparin+RPR109891 was initiated followed 15 min later by recombinant tissue plasminogen activator (rt-PA). The incidence of reperfusion in the enoxaparin+RPR109891- and the heparin+RPR109891-treated groups was similar, but time to reperfusion tended to be shorter for enoxaparin versus heparin. Only 43% of the vessels reoccluded in the enoxaparin+RPR109891 group, compared to 100% vessels in the heparin+RPR109891 group. Enoxaparin+RPR109891 maintained flow for a significantly longer time compared to saline, enoxaparin, heparin, and heparin+RPR109891. Enoxaparin+RPR109891 and heparin+RPR109891 increased the template bleeding time by 2- and 3-fold and activated partial thromboplastin time (APTT) by 1.3- and 3-fold, respectively. These data suggest that enoxaparin is more effective and potentially safer than heparin when combined with a GPIIb/IIIa receptor antagonist during rt-PA-induced coronary thrombolysis.

Crystal structures of human factor Xa complexed with potent inhibitors.

Involved in the coagulation cascade, factor Xa (FXa) is a serine protease which has received great interest as a potential target for the development of new antithrombotics. Although there is a great wealth of structural data on thrombin complexes, few structures of ligand/FXa complexes have been reported, presumably because of the difficulty in growing crystals. Reproducible crystallization conditions for human des-Gla1-45 coagulation FXa have been found. This has led to an improvement in the diffraction quality of the crystals (about 2.1 A) when compared to the previously reported forms (2.3-2.8 A) thus providing a suitable platform for a structure-based drug design approach. A series of crystal structures of noncovalent inhibitors complexed with FXa have been determined, three of which are presented herein. These include compounds containing the benzamidine moiety and surrogates of the basic group. The benzamidine-containing compound binds in a canonical fashion typical of synthetic serine protease inhibitors. On the contrary, molecules that contain surrogates of the benzamidine group do not make direct hydrogen-bonding interactions with the carboxylate of Asp189 at the bottom of the S1 pocket. The structural data provide a likely explanation for the specificity of these inhibitors and a great aid in the design of bioavailable potent FXa inhibitors.

Growth and differentiation of bone marrow hematopoietic cells in mice bearing Ehrlich ascite tumor and treated with Dicyclopentadienildichlorotitanium (IV).

In this work we have investigated the growth and differentiation of bone marrow stem cells in mice bearing Ehrlich ascites tumor and treated with three dose-regimens of Dicyclopentadienyldichlorotitanium (IV) (DDCT). We also studied the presence of colony stimulating factors in the serum of DDCT-treated animals, as well as the effects of the drug on the survival of the tumor-bearing mice. The results demonstrated that the myelosuppression developed in the tumor-bearing animals is prevented by the administration of 1, 2 or 3 doses of 15 mg/kg DDCT. In the treatment with three doses, however, 23% of the animals died. Moreover, DDCT treatment in normal animals resulted in increased numbers of CFU-GM. We observed the presence of stimulating factors in the serum of drug-treated animals which induced the growth and differentiation of bone marrow progenitor cells from normal animals in vitro. On the other hand, in vitro addition of the drug to these cultures had no effect. Thus, we conclude that the drug protects against the myelosuppression induced by the tumor and that this protection may be related to an indirect action of the drug.

Design of a new class of orally active fibrinogen receptor antagonists.

The integrin receptor recognition sequence Arg-Gly-Asp was successfully used as a template from which to develop a series of potent, selective, orally active, peptide-based fibrinogen receptor antagonists with a long duration of action. Simple modifications centered on the Arg and Gly residues quickly led to a modified peptide (1) with significantly enhanced ability to inhibit in vitro platelet aggregation. Substitution of the guanidino group in 1 by piperidine provided 3, which showed not only a further increase in potency but also a modest degree of oral efficacy. Finally, exploration of the nature of the C-terminal amino acid, with respect to its side-chain functionality and the carboxy terminus, yielded a group of molecules that showed excellent in vitro potency for inhibiting platelet aggregation, excellent integrin selectivity, a high level of oral efficacy, and an extended duration of action.

Identification and initial structure-activity relationships of a novel class of nonpeptide inhibitors of blood coagulation factor Xa.

The discovery and some of the basic structure-activity relationships of a series of novel nonpeptide inhibitors of blood coagulation Factor Xa is described. These inhibitors are functionalized beta-alanines, exemplified by 2a. Docking experiments placing 2a in the active site of Factor Xa implied that the most expeditious route to enhancing in vitro potency was to modify the group occupying the S3 site of the enzyme. Increasing the hydrophobic contacts between the inhibitor and the enzyme in this region led to 8, which has served as the prototype for this series. In addition, an enantioselective synthesis of these substituted beta-alanines was also developed.

Anti-thrombotic activity of RG13965, a novel platelet fibrinogen receptor antagonist.

RG13965, a pseudotetrapeptide analogue of Arg-Gly-Asp (RGD), inhibited collagen-induced dog, monkey, human, hamster, mouse, and pig platelet aggregation in vitro with IC50 values of 3.7, 4.6, 6.3, 126, 136 and 1600 microM, respectively. RG13965 (3, 10, and 30 mg/kg, i.v.) decreased the incidence of collagen/epinephrine-induced thrombosis in mice from 90% in untreated animals to 63, 37, and 0%, respectively. In hamsters, RG13965 (10 and 30 mg/kg, i.v.) prolonged the time required for formation of a hemostatic plug in severed mesenteric arteries by 1.6- and 3.6-fold, respectively. In a canine model of repetitive platelet thrombus formation in the coronary artery, RG13965 (0.1, 0.3, and 1 mg/kg, i.v.) reversibly inhibited cyclic flow reductions (CFRs) and inhibited ADP-induced ex vivo platelet aggregation by 29, 57, and 77%, respectively. RG13965 (1 mg/kg) completely inhibited CFRs for at least 40 min. Platelet count was not altered at any dose and template bleeding time was prolonged modestly (1.8-fold) at only the highest dose. RG13965 dose-dependently and reversibly inhibited thrombus formation at doses which did not completely inhibit ex vivo platelet aggregation and only modestly prolonged template bleeding time.