Aberrant placenta gene expression pattern in bovine pregnancies established after transfer of cloned or in vitro produced embryos.

In the present study, we used the global transcriptome profile approach to identify dysregulated genes, molecular pathways, and molecular functional alterations in bovine placentas derived from somatic cell nuclear transfer (SCNT) and in vitro embryo production (IVP) pregnancies compared with their artificial insemination (AI) counterparts at day 50 of gestation. For this, day 7 blastocysts derived from AI, IVP, or SCNT were transferred to oestrus-synchronized cows. The pregnant animals were slaughtered at day 50 of gestation, and the placentas were then recovered and used for transcriptome analysis using Affymetrix GeneChip bovine genome array. Results showed the SCNT placenta to be different from its AI counterpart in the expression of 1,196 transcripts. These genes were found to be associated with alterations in key biological processes and molecular pathways in SCNT placenta, and the dysregulation of 9% (n = 110) of these genes was due to transcriptional reprogramming error. IVP placenta also displayed alterations in the expression of 72 genes, of which 58 were common to SCNT placenta. Gene enrichment analysis revealed that the expression of genes involved in organ development, blood vessel development, extracellular matrix organization, and the immune system was affected in both SCNT and IVP placentas. However, 96% of the affected genes in SCNT were not significantly altered in IVP groups. Thus, the higher transcriptome dysregulation in SCNT placenta followed by IVP would reflect the degree of placental abnormality in SCNT and IVP pregnancies at day 50 of the gestation, which may have a profound effect on subsequent fetal development and health of the offspring.

Molecular mechanisms and pathways involved in bovine embryonic genome activation and their regulation by alternative in vivo and in vitro culture conditions.

Understanding gene expression patterns in response to altered environmental conditions at different time points of the preimplantation period would improve our knowledge on regulation of embryonic development. Here we aimed to examine the effect of alternative in vivo and in vitro culture conditions at the time of major embryonic genome activation (EGA) on the development and transcriptome profile of bovine blastocysts. Four different blastocyst groups were produced under alternative in vivo and in vitro culture conditions before or after major EGA. Completely in vitro- and in vivo-produced blastocysts were used as controls. We compared gene expression patterns between each blastocyst group and in vivo blastocyst control group using EmbryoGENE's bovine microarray. The data showed that changing culture conditions from in vivo to in vitro or vice versa, either before or after the time of major EGA, had no effect on the developmental rates; however, in vitro conditions during that time critically influenced the transcriptome of the blastocysts produced. The source of oocyte had a critical effect on developmental rates and the ability of the embryo to react to changing culture conditions. Ontological classification highlighted a marked contrast in expression patterns for lipid metabolism and oxidative stress response between blastocysts generated in vivo versus in vitro, with opposite trends. Molecular mechanisms and pathways that are influenced by altered culture conditions during EGA were defined. These results will help in the development of new strategies to modify culture conditions at this critical stage to enhance the development of competent blastocysts.

Survival and prognostic factors in surgically resected synchronous multiple primary lung cancers.

OBJECTIVE: The presence of synchronous multiple primary non-small-cell lung cancers (SMPLC) is a rare condition and the optimal treatment remains unclear. In this study, the survival of surgically treated SMPLC patients and the factors affecting survival were analyzed. METHODS: Between 2001 and 2008, 26 consecutive patients diagnosed with SMPLC, who had all of their tumors resected, were retrospectively evaluated. Patients, who had bronchoalveolar carcinoma or carcinoid tumors and satellite nodules, were excluded. Prognostic factors were analyzed using univariate and multivariate analyses. RESULTS: The tumors were unilateral in 14 and bilateral in 12 patients. In total, 38 procedures were performed. A complete resection was achieved in 35 (92.1%) procedures. The in-hospital mortality rate was 7.6% (two patients). The overall 5-year survival rate was 49.7%, and the median survival time was 40 months. The 5-year survival rate was 40.6% for unilateral and 62.8% for bilateral SMPLC patients (p = 0.47). Histopathologic tumor type, N1 nodal disease, tumor (T) status, and older age did not influence survival. There was no survival disadvantage for patients, upon whom a sublobar resection had been performed. There was a trend toward poorer survival in patients upon whom a pneumonectomy had been performed (p = 0.12). The 3-year survival rate for patients, who received adjuvant chemotherapy and/or radiotherapy (66.7%), was better than other patients (56.3%). In the multivariate analysis, we found a trend toward poor survival in patients, who received a pneumonectomy, and a trend toward better survival in patients, who received adjuvant therapy (p = 0.05 and p = 0.06). CONCLUSIONS: The survival of SMPLC patients, who were treated surgically, was satisfactory. Pneumonectomy was a poor prognostic factor, whereas adjuvant therapy was a good prognostic factor.