Swine testis (ST) cell lines producing a murine recombinant retrovirus (RRV) were established in order to transfer the bacterial lacZ gene fused to a nuclear location signal (nlslacZ) into animal cells. ST cells were infected with the supernatant of the cat G355.5LacZ2 cell line which produces amphotropic and xenotropic MMuLVSVnlslacZ-defective RRV and wild amphotropic and xenotropic MMuLV. Expression of the nlslacZ reporter gene was under the transcriptional control of both the SV40 early promoter and the retroviral LTR. ST cells expressing the reporter gene were sorted and cloned by limiting dilutions. Fourteen STLacZ-cell lines were isolated and subsequently tested for virus production. Depending on the host range of the retroviruses, two cell lines (STBF11 and STAA3) produced both a xenotropic recombinant pseudotype and wild retroviruses; another (STAB 10) produced both an amphotropic recombinant pseudotype and wild retroviruses. Southern blot analysis of the producer cell lines was carried out to verify proviral integration. The efficiency of the different pseudotypes in the transfer of the nlslacZ reporter gene to cultured animal cells, including porcine cells, was compared to the pseudotyped RRV produced by cat lines. Our results showed that the xenotropic RRV produced by the porcine STBF 11 cell line has a high titre for cells from different species and led to a higher number of porcine endothelial and lymphoblastoid cells expressing the reporter gene than did RRV produced by the cat packaging cell lines.

Animals↗

PubMedDate not supplied

[Pharmacokinetics of methotrexate and clinical response associated in the medical treatment of ectopic pregnancies].

Methotrexate (MTX) is used in the medical treatment of unruptured ectopic pregnancy. This drug is administered by intramuscular (IM) or intrasaccular (IS) injection under sonographic control. No data are available concerning the pharmacokinetics of MTX in this new indication. This study compares the two administration routes in 12 patients (21 to 37 years) taken as their own control and presenting with ectopic pregnancy (51 +/- 12 days of amenorrhea). Our aim was to compare the pharmacokinetic profile of MTX for each route to facilitate its use in the future. The initial level of hCG was 4.474 +/- 4.184 mIU/ml. Each patient firstly received 1 mg/kg of MTX intrasaccularly under vaginal sonography. The same dose was injected intramuscularly 48 hours later. The pharmacokinetic profiles of MTX after IS and IM administrations were determined after both injections during 48 hours. MTX serum levels were measured by Fluorescence Polarization Immuno Assay. Data were analyzed by model independent methods and compared by a Wilcoxon T test (p 0.01 was considered as significant). All the unruptured ectopic pregnancy were cured and the hCG serum levels were normalized (10 mIU/ml) in 37 +/- 18 days. After IM administration, AUC0-infinity is significantly (p 0.01) increased i.e., 15.1 +/- 4.1 mumol.h/l versus 11.2 +/- 4.8 after IS injection. T1/2 lambda z and MRT remained unchanged whatever the route is i.e., 11.3 +/- 4.9 h and 8.6 +/- 3.9 h (IS) versus 12.1 +/- 5.9 h and 7.3 +/- 1.8 h (IM). The decrement of AUC0-infinity 8 determined after IS injection might be the consequence of the capture of the MTX by the trophoblastic cells (target cells).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗