Enhanced thrombin generation in plasma of severe thrombocytopenic patients due to rFVIIa.

UNLABELLED: RFVIIa-enhanced thrombin generation has been shown to be dependent on platelets. In previous work we have shown that addition of monocytes and rFVIIa to microparticle free plasma causes a distinct thrombin generation. The aim of our study has been to examine whether there is enough surface provided by microparticles in thrombocytopenic plasma to allow an effect of rFVIIa. PATIENTS, METHODS: Thrombin generation was measured in platelet rich plasma (PRP) and microparticle free plasma (MFP) of thrombocytopenic haemato-oncological patients with and without addition of rVIIa by means of calibrated automated thrombography. Microparticles were analyzed in PRP by FACS flow cytometry. RESULTS: Microparticle free plasma showed no thrombin generation with or without addition of rFVIIa. Addition of rFVIIa to PRP of thrombocytopenic patients led to a significant shortening of lag time and time to peak in thrombin generation, while ETP and peak remained unchanged. CONCLUSION: Our results show that even in plasma of severe thrombocytopenic patients enough surface may be provided by microparticles to allow an enhancement of thrombin generation by rFVIIa.

cDNA cloning and analysis of tissue-specific expression of mouse peroxisomal straight-chain acyl-CoA oxidase.

Straight-chain acyl-CoA oxidase is the first and rate limiting enzyme in the peroxisomal beta-oxidation pathway catalysing the desaturation of acyl-CoAs to 2-trans-enoyl-CoAs, thereby producing H2O2. To study peroxisomal beta-oxidation we cloned and characterized the cDNA of mouse peroxisomal acyl-CoA oxidase. It consists of 3778 bp, including a 1983-bp ORF encoding a polypeptide of 661 amino-acid residues. Like the rat and human homologue the C-terminus contains an SKL motif, an import signal present in several peroxisomal matrix proteins. Sequence analysis revealed high amino-acid homology with rat (96%) and human (87%) acyl-CoA oxidase in addition to minor homology ( approximately 40%) with other related proteins, such as rabbit trihydroxy-cholestanoyl-CoA oxidase, human branched chain acyl-CoA oxidase and rat trihydroxycoprostanoyl-CoA oxidase. Acyl-CoA oxidase mRNA and protein expression were most abundant in liver followed by kidney, brain and adipose tissue. During mouse brain development acyl-CoA oxidase mRNA expression was highest during the suckling period indicating that peroxisomal beta-oxidation is most critical during this developmental stage. Comparing tissue mRNA levels of peroxisome proliferator-activated receptor alpha and acyl-CoA oxidase, we noticed a constant relationship in all tissues investigated, except heart and adipose tissue in which much more, and respectively, much less, peroxisome proliferator-activated receptor alpha mRNA in proportion to acyl-CoA oxidase mRNA was found. Our data show that acyl-CoA oxidase is an evolutionary highly conserved enzyme with a distinct pattern of expression and indicate an important role in lipid metabolism.

Pathologic features of placentas from singleton pregnancies obtained by in vitro fertilization and embryo transfer.

Fifty placentas were collected at term from singleton pregnancies resulting from in vitro fertilization (IVF) and intrauterine embryo transfer. Their pathologic features were compared with those of a control group composed of 50 placentas obtained from spontaneous singleton pregnancies. The mean maternal age, mean gestational age, mean fetal weight, sex ratio, and rate of pregnancy complications did not differ. There was also no significant difference between the groups in the mean placental weight and in the incidence of placental pathologic lesions, including extended infarcts, massive perivillous fibrin depositions, chorioangiomas, and placental inflammatory lesions. The incidence of abnormal placental shapes was significantly (P less than .05) greater in the IVF group (22%) compared with the control group (6%). A significant (P less than .025) difference was observed between the groups in the distribution of umbilical cord insertions. The mean distance between the cord insertion and the closest placental margin was significantly (P less than .005) shorter in the group conceived by IVF (3.23 +/- 1.91 cm) than in the control group (4.54 +/- 2.42 cm). A relationship between these placental morphologic features and the superficial implantation and/or inadequate orientation of the blastocyst after IVF and intrauterine embryo transfer is proposed.

Amyloidosis of the joints: evidence that human hip capsules have a unique predisposition for amyloid of the senile systemic type.

Amyloid-containing tissues were sampled from 19 capsules and 10 cartilages from hip, knee and sternoclavicular joints of 16 elderly patients. Heart ventricles from 3 of these patients were also studied. Histochemistry and immunofluorescence were performed and amyloid fibrils extracted from one hip capsule. The results indicate that 9 of 13 hip capsules contained prealbumin-like amyloid fibril protein ASc1, a finding characteristic of senile systemic amyloidosis. The amyloid of 16 out of 17 other joint structures showed no reaction with anti-ASc1 or antisera to 3 other systemic forms of amyloid, indicating that they represent localized forms of amyloid.