Thaw, biopsy and refreeze strategy for PGT-A on previously cryopreserved embryos.

BACKGROUND: Preimplantation genetic testing for aneuploidy (PGT-A) with Next Generation Sequencing technology is a low-cost and powerful technology for the analysis of embryo quality. However, PGT-A requires freezing of embryos, suggesting that previously cryopreserved embryos cannot be tested. Here, we test whether use of the vitrification technique permits the refreezing of embryos, thus permitting PGT-A testing of cryopreserved embryos. METHODS: The results are a retrospective analysis of cases performed at Create fertility between 2016 and 2017. Results obtained after traditional PGT-A are compared with results after the thaw biopsy and refreeze (TBR) procedure. A total of 220 patients were treated with PGT-A and 54 patients with the TBR procedure. RESULTS: Maternal ages were not significantly different between the groups. The proportion of PGT-A normal embryos was not significantly different between the two groups. A clinical pregnancy rate of 61.5% was achieved with the PGT-A group and 52.4% with the TBR group. These results were not statistically significant. The efficiency of the thaw, biopsy and refreeze technique was not significantly different to that of fresh cycles for rates of survival, results obtained and aneuploidy incidence. Clinical pregnancy rates are not significantly different after the biopsy of fresh and previously cryopreserved embryos. CONCLUSION: The data shows that the TBR procedure has an equivalent success rate to that of classical PGT-A procedures.

The pharmacokinetics of liposomal encapsulated daunorubicin are not modified by HAART in patients with HIV-associated Kaposi's sarcoma.

PURPOSE: To investigate the pharmacokinetics of liposomal daunorubicin (DaunoXome) administered alone or in combination with antiviral therapy including protease inhibitors (PI) to HIV-positive patients affected by Kaposi's sarcoma (KS). PATIENTS AND METHODS: A group of 18 patients with extensive or rapidly progressing AIDS-related KS received DaunoXome at a dose of 40 mg/m2 alone or in association with a triple combination therapy consisting of one PI plus two nucleoside reverse transcriptase inhibitors (NRTI). Daunorubicin pharmacokinetics were determined in a total of 23 cycles, 6 with DaunoXome alone, 9 in combination with indinavir, 6 with ritonavir and 2 with saquinavir. Plasma samples were obtained at different times during the 72 h after DaunoXome administration. Daunorubicin and daunorubicinol plasma levels were determined by high-performance liquid chromatography. RESULTS: After the DaunoXome infusion, daunorubicin was rapidly cleared from the body following, in most cases, a one-compartment open kinetic model. The daunorubicin peak concentrations, clearances and elimination half-lives were (means +/- SD): 16.3 +/- 2.8 microg/ml, 0.3 +/- 0.1 l/h per m2 and 5.6 +/- 2.6 h after DaunoXome alone; 15.1 +/- 4.9 microg/ml, 0.5 +/- 0.3 l/ h per m2 and 5.8 +/- 2.1 h after the combination with indinavir; and 14.5 +/- 2.8 microg/ml, 0.4 +/- 0.2 l/h per m2 and 6.5 +/- 3.9 h after the combination with ritonavir. In all groups, daunorubicinol plasma levels were approximately 25-30 times lower than those of the parent drug. CONCLUSION: Our data show that there are no significant differences in the pharmacokinetic parameters of daunorubicin in patients receiving DaunoXome in combination with indinavir and ritonavir compared with those in patients not receiving PIs. Therefore in patients affected by AIDS-related KS treated with Highly Active AntiRetroviral Therapy (HAART) there is no pharmacokinetic justification for reducing the doses of DaunoXome.

Distribution of daunorubicin and daunorubicinol in human glioma tumors after administration of liposomal daunorubicin.

DaunoXome is a liposome formulation containing daunorubicin (DM). Encapsulation of the drug in liposomes presents the advantage of low-level systemic exposure and better drug penetration into the tumor. We studied the distribution of DM and its 13-dihydro metabolite, daunorubicinol (DMol), in surgical biopsies from different parts of glioblastomas. The study was performed in eight patients with recurrent glioblastoma, all of whom had previously undergone surgery and been treated with radiotherapy and chemotherapy, who received 50 mg of DaunoXome as a 1-h infusion. Surgery was performed at 24 and 48 h after the infusion in seven cases and one case, respectively. Biopsies were divided into three parts: the central area of the tumor, peripheral tumor tissue, and brain-adjacent tumor (BAT) tissue. A complete plasma pharmacokinetics study was conducted in seven cases, with samples being taken for up to 48 h after the end of the infusion. DM and DMol were determined in plasma and tissue by high-performance liquid chromatography with fluorescence detection after solvent extraction. At 24 h, concentrations of DM and DMol in the central part of the tumor ranged between < 0.005 and 0.80 microg/g and between 0.005 and 1.58 microg/g, respectively. Concentrations were similar in the peripheral tumor and in BAT tissue. From the data obtained on the patient who underwent surgery at 48 h it appears that DM and DMol remain in tumor tissue for a long time, the concentrations being 0.4 and 2.8 microg/g, respectively. DaunoXome was rapidly cleared from the body, with the plasma levels of DM and DMol determined at 48 h lying in the range of < 5-50 and < 5-20 ng/ml, respectively. The mean (+/-SD) half-life and plasmatic clearance of DM were 4.8+/-1.0 h and 0.2+/-0.06 l h(-1) m(-2). In conclusion, DaunoXome achieved and maintained potentially cytotoxic levels of both DM and DMol in glioblastoma for a long time in association with low-level systemic exposure. Further studies are therefore warranted. Although only preliminary and obtained in previously treated patients, these data suggest that DaunoXome merits investigation in CNS tumors.

Pharmacokinetic interactions of paclitaxel, docetaxel and their vehicles with doxorubicin.

BACKGROUND: The combination of doxorubicin (Dx) with paclitaxel or docetaxel is clinically effective but there are concerns regarding the higher incidence of cardiotoxicity of the combination compared with Dx alone. The mechanism of the increased toxicity is still unclear. PURPOSE: To assess whether there is a pharmacokinetic interaction between paclitaxel, docetaxel or their vehicles and Dx in mice. MATERIALS AND METHODS: CDF1 male mice were treated with Dx either alone (10 mg/kg i.v.) or in combination with paclitaxel (25 mg/kg) or docetaxel (25 mg/kg) or their vehicles, i.e., cremophor-ethanol-glucose (cremophor) or polysorbate80-ethanol-glucose (polysorbate). Four mice were killed 4, 8 or 24 hours after Dx in each experimental group and Dx was assayed in serum and in heart, liver, kidney and spleen by HPLC. RESULTS: Four hours after treatment the concentrations of Dx in heart, liver and kidney were much higher in mice concomitantly treated with paclitaxel, docetaxel (dissolved in either cremophor or polysorbate) and cremophor. At subsequent times the differences were modest and only reached statistical significance in a few cases. Dx metabolites were modified by concomitant treatment with taxanes or their vehicles. In particular, the levels of Dx aglycone in liver and kidney were significantly lower in mice treated with the combination than in mice given Dx alone. CONCLUSIONS: Paclitaxel, docetaxel and cremophor when given together with Dx modify its distribution and metabolism, increasing Dx levels in many tissues including the heart. This might have some bearing on the toxicity of regimens in which Dx is combined with taxanes.

[Localization of the LDHA-GST3-ESA4 synthetic group on human chromosome 11. Analyses of the classic man-rodent hybrids and of a new type (not adhering to the wall)]. / Localisation du groupe synténique LDHA-GST3-ESA4 sur le chromosome 11 chez l'homme. Analyses des hybrides homme-rongeur classiques et d'un type nouveau (non adhérents à la paroi).

Analysis of different human tissues showed that glutathione S-transferase (GST) of fibroblasts and leucocytes is GST3, an enzyme found in liver extracts by Board (1981). Consequently, the GST localized on human chromosome 11 by Silberstein and Shows (1981, 1982) is GST3. Analysis of tissue extracts showed a new GST band, very intense in muscle extracts and very weak or absent in other tissues, and called GSTM. Analysis of man-rodent hydrids showed synteny between LDHA, GST3, ESA4 and localization of this synteny group on chromosome 11. Analysis of discordant percentages and discordant types between markers favored the following assignments: LDHA on 11p12, GST3 on 11q13 leads to 11qter, ESA on 11q13 leads to 11q22. The present study suggests assignment of GST3 to 11q13 leads to 11q22. Different types of man-rodent hybrids are useful for human gene mapping. A new type hybrid, as used here (nonadhering to glass or plastic surfaces), appears useful because of rapid proliferation and growth in suspension, the latter feature facilitating culture and harvesting.