Poor reproductive performance of dairy cattle has a major negative impact on farm profitability. This is in part attributed to the use of semen from bulls of below-average fertility and to early embryonic mortality. Osteopontin (OPN) is an acidic glycoprotein that has been detected in the epithelium of the ampulla, seminal vesicles, and seminal fluid of high-fertility Holstein bulls. The objective of this study was to determine whether freezing semen with purified bovine milk OPN would improve fertilization and embryonic development in vitro. In a first step, frozen semen from 6 bulls was used with different concentrations of OPN (0, 1, 10, and 100 g mL-1) to evaluate fertilization, and in a second step, semen from 2 of the 6 bulls was frozen with OPN (0, 5, 10, 20, 40, and 80 g mL-1) to evaluate its effect on in vitro fertilization and embryonic development. In vitro-matured bovine oocytes were inseminated with 1 105 frozen–thawed spermatozoa per 10 oocytes. After 18 h (39C, 5% CO2 in air), oocytes designated for evaluation of fertilization were fixed, stained, and observed to determine the presence of pronuclei. Oocytes destined for embryo culture were placed in 4-well dishes containing 500 L of synthetic oviduct fluid per well at 5% O2, 5% CO2, and 90% N2 (v/v). Each experiment was repeated 4 times, and data from each experiment were pooled. Analysis of variance using a general linear model was performed using a weighted mean based on the number of oocytes per treatment. Significantly more (P < 0.05) oocytes were fertilized when frozen semen was used with 10 g mL-1 of OPN (85 4.0%, 78 4.0%) from 2 of the 6 bulls than when 0 g mL-1 was used (75 4.0%, 69 4.0%). Semen from those bulls resultewd in more fertilized oocytes when 20 g mL-1 (86 3.5%, 79 3.4%) and 40 g mL-1 (88 3.4%, 81 3.9%) of OPN were used during the second phase of the experiment. Oocytes inseminated with 10 g mL-1 (84 2.5%, 77 2.3%), 20 g mL-1 (87 2.8%, 79 1.9%), or 40 g mL-1 (89 1.9%, 81 2.4%) of OPN in the frozen semen had increased cleavage rates at Day 4 compared with those inseminated with 0 g mL-1 (75 3.5%, 69 3.4%). At Day 8, blastocyst development was greater for 10 g mL-1 (40 1.8%, 37 1.6%), 20 g mL-1 (42 2.1%, 38 2.9%), and 40 g mL-1 (45 2.9%, 40 2.5%) of OPN than for the semen with 0 g mL-1 (33 2.3%, 29 2.8%). We conclude that semen from some bulls that was frozen with purified bovine milk OPN increases IVF, cleavage, and embryonic development, suggesting a facilitative role for OPN in some reproductive technologies.

This study was designed to investigate the effects of pre-incubating cattle spermatozoa or matured oocytes with purified osteopontin (OPN) from cattle milk on fertilization in cattle and embryonic development in vitro. There were two different experiments, semen from six mature Holstein bulls (Bos Taurus) was frozen with different concentrations of OPN (0, 1, 10, 100 microg/mL). Matured cattle oocytes were also pre-treated with OPN (0, 10, 100 microg/mL). In both experiments, pre-treated oocytes or frozen semen, was processed for in vitro fertilization and embryo development. Significantly more oocytes were fertilized when using frozen semen with 10 microg/mL OPN (bull 2=85+/-4% and bull 5=78+/-4%) than without OPN (bull 2=75+/-4% and bull 5=69+/-4%). Those bulls also had increase in cleavage and embryo development (bull 2=85+/-3%, 41+/-1.9%; bull 5=76+/-2%, 37+/-1.8%) compared with control (bull 2=75+/-3%, 30+/-2%; bull 5=68+/-2%, 29+/-2%). Incubating matured oocytes in 10 microg/mL OPN (87+/-3%) and 100 microg/mL OPN (88+/-3%) significantly increased fertilization than control (73+/-3%). OPN also improve cleavage, and embryo development in treatments with 10 microg/mL OPN (82.7+/-1.3%; 31.7+/-1.4%) and 100 microg/mL OPN (85.8+/-1.3%; 33.8+/-1.5%) when compared with control (74.1+/-1.3%; 24.2+/-1.2%). These data suggest that both, spermatozoa from some bulls and oocytes may associate with OPN, suggesting a facilitory role on in vitro fertilization and embryo development.

The concentration-time profiles of Doxorubicin (DOXO) from day 0 to day 21 after i.v. infusion of 25 or 30 mg/m2 doxorubicin HCl stealth liposomes (Caelyx®) were investigated in 9 patients receiving combination polychemotherapy with cyclophosphamide, vinorelbine and prednisone. Peak serum concentrations occurred from 0.04 to 4.0 days after infusion (mean tmax=1.79±1.55 d) with a mean cmax of 4595±2849 ng/ml. A total amount of 12.84±2.47 mg liposomal DOXO in the plasma volume (Vp=2794+537 ml) could be estimated at tmax (=27% of the mean dose of 47.6 mg). Stealth liposomes were eliminated slowly from the blood with a mean t1/2el of 1.9+0.5 days (MRT was 4.6+2.5 days).

BACKGROUND:Cytoprotection of healthy cells represents a new approach in cancer chemotherapy, but a pharmacokinetic drug interaction between the cytostatic and the cytoprotectant is undesired.METHODS:The purpose was to evaluate the clinical pharmacokinetics (PHK) of paclitaxel (PACLI) and its metabolites under cytoprotection in patients suffering from breast cancer. PACLI was administered alone and in a second cycle in combination with amifostine (AMI) as a paired cross over.RESULTS:In both treatment schedules the steady state of PACLI occurred after 3 hours, the tmax of metabolites between 3 and 4 hours. The mean steady-state concentration was cmax = 5432 +/- 1238 ng/ml in the control group and cmax = 5140 +/- 2407 ng/ml in the AMI group. For the serum metabolites, the findings were very similar: 6-OH-PACLI: cmax 413 +/- 153 ng/ml versus 432 +/- 304 ng/ml, 3"-OH-PACLI: cmax 99 +/- 103 ng/ml versus 123 +/- 98 ng/ml, 3",6-DiOH-PACLI: cmax 43 +/- 55 ng/ml versus 75 +/- 85 ng/ml. AUC values of metabolites were slightly higher in the AMI group, but PHK seemed equal.CONCLUSION:The results gave evidence, that cytoprotection with AMI has no clinical consequences on PACLI pharmacokinetics and biotransformation.