The influence of warm ischemia elimination on kidney injury during transplantation - Clinical and molecular study

Kidney surface cooling was used during implantation to assess the effect of warm ischemia elimination on allograft function, histological changes and immune-related gene expression. 23 recipients were randomly assigned to a group operated on with kidney surface cooling during implantation (ice bag technique, IBT group), and the other 23 recipients receiving the contralateral kidney from the same donor were operated on with a standard technique. Three consecutive kidney core biopsies were obtained during the transplantation procedure: after organ recovery, after cold ischemia and after reperfusion. Gene expression levels were determined using low-density arrays (Format 32, TaqMan). The IBT group showed a significantly lower rate of detrimental events (delayed graft function and/or acute rejection, p = 0.015) as well as higher glomerular filtration rate on day 14 (p = 0.026). A greater decrease of MMP9 and LCN2 gene expression was seen in the IBT group during total ischemia (p = 0.003 and p = 0.018). Elimination of second warm ischemia reduced the number of detrimental events after kidney transplantation, and thus had influence on the short-term but not long-term allograft function. Surface cooling of the kidney during vascular anastomosis may reduce some detrimental effects of immune activation resulting from both brain death and ischemia-reperfusion injury

Merkel cell carcinoma of skin-current controversies and recommendations

The review covers the current recommendations for Merkel cell carcinoma (MCC), with detailed discussion of many controversies. The 2010 AJCC staging system is more in-line with other skin malignancies although more complicated to use. The changes in staging system over time make comparison of studies difficult. A wide excision with margins of 2.5–3 cm is generally recommended. Even for primary </= 1 cm, there is a significant risk of nodal and distant metastases and hence sentinel node biopsy should be done if possible; otherwise adjuvant radiotherapy to the primary and nodal region should be given. Difficulties of setting up trials owing to the rarity of the disease and the mean age of the patient population result in infrequent reports of adjuvant or concurrent chemotherapy in the literature. The benefit, if any, is not great from published studies so far. However, there may be a subgroup of patients with high-risk features, e.g. node-positive and excellent performance status, for whom adjuvant or concurrent chemotherapy may be considered. Since local recurrence and metastases generally occur within 2 years of the initial diagnosis, patients should be followed more frequently in the first 2 years. However delayed recurrence can still occur in a small proportion of patients and long-term follow-up by a specialist is recommended provided that the general condition of the patient allows it. In summary, physician judgment in individual cases of MCC is advisable, to balance the risk of recurrence versus the complications of treatment

Comparative transcriptomics of a complex of four European pine species

Background: Pinus sylvestris, P. mugo, P. uliginosa and P. uncinata are closely related but phenotypically and ecologically very distinct European pine species providing an excellent study system for analysis of the genetic basis of adaptive variation and speciation. For comparative genomic analysis of the species, transcriptome sequence was generated for 17 samples collected across the European distribution range using Illumina paired-end sequencing technology. Results: De novo transcriptome assembly of a reference sample of P. sylvestris contained 40968 unigenes, of which fewer than 0.5% were identified as putative retrotransposon sequences. Based on gene annotation approaches, 19659 contigs were identified and assigned to unique genes covering a broad range of gene ontology categories. About 80% of the reads from each sample were successfully mapped to the reference transcriptome of P. sylvestris. Single nucleotide polymorphisms were identified in 22041-24096 of the unigenes providing a set of ~220-262 k SNPs identified for each species. Very similar levels of nucleotide polymorphism were observed across species (π=0.0044-0.0053) and highest pairwise nucleotide divergence (0.006) was found between P. mugo and P. sylvestris at a common set of unigenes. Conclusions: The study provides whole transcriptome sequence and a large set of SNPs to advance population and association genetic studies in pines. Our study demonstrates that transcriptome sequencing can be a very useful approach for development of novel genomic resources in species with large and complex genomes

Population genetic structure of Juniperus phoenicea (Cupressaceae) in the western Mediterranean Basin: gradient of diversity on a broad geographical scale

Introduction, Material and Methods: The genetic structure and diversity of ten natural populations of Juniperus phoenicea L. from the western part of the species range have been studied using random amplified polymorphic DNA (RAPD) markers. Results and discussion: Among 10 analyzed primers only 3 reproduced consistently across successful PCR reactions and gave 45 loci. The percentage of polymorphic loci (P) and Nei’s heterozygosity (H e ) have average values of 64.9% and 0.177. The average expected heterozygosity of particular populations positively correlate with latitude and negatively with altitude (τ = 0.556, P = 0.025; τ = −0.494, P = 0.047, respectively). The proportion of genetic variation contributed by the differences between populations was low (G ST  =  0.056). The gene flow (N m) has an average value of 4.2, and was higher in subsp. turbinata (7.3) than in subsp. phoenicea (4.1). Significant proportion of the variation (ΦST = 0.106) was attributable to differences among populations, as revealed in analysis of molecular variance analysis of pair-wise RAPD distances. No evidence for isolation by distance was detected in Mantel test on genetic (ΦST) and geographic distances. European populations differed at a higher level from the African, subsp. phoenicea from turbinata (3.97% and 3.14% of total variance, respectively). The significant level of differences between European and African populations can result from (1) the earlier divergence and considerably low level of gene flow between them, or (2) a different mutation rate within population of different continent. Conclusion: The results suggest rather local forest economy with J. phoenicea, without seed exchange on large distance.The study was partly sponsored by the Polish Committee for Scientific Research, grant no. 2P04C 030 26 and partly by the Institute of Dendrology. The collection of plant material was made possible thanks to the cooperation between CSIC (Spanish Research Council) and PAS (Polish Academy of Sciences).Peer Reviewe