Individual bovine in vitro embryo production and cumulus cell transcriptomic analysis to distinguish cumulus-oocyte complexes with high or low developmental potential.

Studying cumulus cell (CC) transcriptome is of great interest as it could provide a noninvasive method to assess oocyte quality. In cattle, the search for quality markers has not been done with cumulus-oocyte complexes (COCs) cultured individually from maturation to blastocyst stage. Here, differences between high- and low-potential COCs were examined by transcriptomic analysis of CC biopsies obtained from COCs of 2 to 6 mm follicles (n = 249; eight replicates) before individual in vitro maturation, fertilization, and culture until Day 8 after fertilization. Each COC was individually tracked and categorized based on his fate: embryo at blastocyst stage (CC-Blast) or embryo arrested at 2- to 8-cell stage (CC-2-8-cells). Average blastocyst rates were 27.7% for individual culture and 31.2% for group control (not significantly different). For transcriptomic analysis, five cumulus biopsies per replicate were pooled for each fate. Three CC replicates underwent transcriptomic analysis using RNA microarray assay. Some clear differences in gene expression between the CC-Blast and the CC-2-8-cell groups were identified. Considering a 1.5-fold change (P < 0.05), 68 genes were differentially expressed between the CC-Blast and CC-2-8-cells. Quantitative reverse transcription-polymerase chain reaction validations were performed for 12 selected genes: six upregulated genes for each COC fate. Higher expression of 1-acylglycerol-3-phosphate O-acyltransferase 9 (AGPAT9) (lipid metabolism), Chloride intracellular channel 3 (CLIC3), Keratin 8 (KRT8), and Lumican (LUM) (molecular transport) was observed in CC-2-8-cells (P < 0.05). The CC-Blast fate analysis revealed a significantly higher expression of Glycine amidinotransferase (L-arginine:glycine amidinotransferase) (GATM) (posttranslational modification, amino acid metabolism, and free radical scavenging). This newly identified set of genes could provide new markers to distinguish COCs associated with good quality embryos from COCs with limited developmental potential.

Contribution of PET in the diagnosis of recurrent colorectal cancer: comparison with conventional imaging.

The clinical value of total body PET with FDG was evaluated in 76 patients presenting with or suspected of recurrent local or distant colorectal cancer. PET results were compared to those of routine imaging (CT pelvis, CT/US liver and CXR). The accuracy of PET for local disease was 95% which was superior to CT-pelvis (accuracy 65%). PET accuracy for liver metastases (98%) compared favourably to CT/US-liver accuracy (93%). Unexpected extra-hepatic mestastases were detected by PET in 14 locations in 10 patients. Also, a primary breast cancer was found in one patient. The main value of PET appeared an improved staging of apparently resectable, local or distant recurrent disease. Thereby, a more adequate indication of major secondary surgery could be attained.

Effect of an equal volume replacement with 500 mL 6% hydroxyethyl starch on the blood and plasma volume of healthy volunteers.

The effect of an equal volume replacement with 6% low molecular weight (200,000) hydroxyethyl starch (substitution ratio 0.60-0.66) (Elohaes) on the blood volume and plasma volume of eight healthy volunteers was investigated with special attention to volume overshooting. After replacement of 500 mL whole blood, blood volume was measured repeatedly for 8 1/2 h using chromium-51 labelled erythrocytes. Red cell volume and plasma volume were calculated from standard formulae. After replacement, blood and plasma volumes were unchanged from control (4754 +/- 630 mL and 2900 +/- 450 mL, respectively). The haematocrit decreased from 43.7 +/- 2.5% to 39.0 +/- 1.4% (1 h after starch infusion) and 41.1 +/- 1.9% (7.5 h after starch infusion). There were no significant changes in haemodynamic variables (systolic and diastolic blood pressure; heart rate), liver function (bilirubin; transaminases), renal function (serum creatinine; serum sodium) or coagulation (activated partial thromboplastin time). Alpha-amylase was significantly increased, which is a well known effect of starches. The replacement of 500 mL whole blood by hydroxyethyl starch maintains isovolaemia for at least 8 h in normal volunteers without any significant volume overshooting; this is not what was seen in earlier studies. This should increase safety in patients with cardiac disease.

Effect of ticlopidine on blood loss, platelet turnover and platelet deposition on prosthetic surfaces in patients undergoing aorto-femoral bypass grafting.

Sixty patients in whom a prosthetic aorto-femoral bypass graft was inserted were pretreated with ticlopidine (250 mg b.i.d.) or placebo in a double-blind study. Blood loss and transfusion need were higher in ticlopidine treated patients. Platelet survival was partially corrected and platelet deposition on prosthetic surfaces was inhibited with ticlopidine. The study suggests that ticlopidine does not increase excessively the bleeding risk during peripheral vascular surgery and that the antiplatelet drug may have beneficial effects in the early phase after insertion of prosthetic vascular graft.