Aberrant DNA methylation profiles in the premature aging disorders Hutchinson-Gilford Progeria and Werner syndrome

DNA methylation gradiently changes with age and is likely to be involved in aging-related processes with subsequent phenotype changes and increased susceptibility to certain diseases. The Hutchinson-Gilford Progeria (HGP) and Werner Syndrome (WS) are two premature aging diseases showing features of common natural aging early in life. Mutations in the LMNA and WRN genes were associated to disease onset; however, for a subset of patients the underlying causative mechanisms remain elusive. We aimed to evaluate the role of epigenetic alteration on premature aging diseases by performing comprehensive DNA methylation profiling of HGP and WS patients. We observed profound changes in the DNA methylation landscapes of WRN and LMNA mutant patients, which were narrowed down to a set of aging related genes and processes. Although of low overall variance, non-mutant patients revealed differential DNA methylation at distinct loci. Hence, we propose DNA methylation to have an impact on premature aging diseases.     

人类新基因C17orf32的电子克隆和编码区序列RT-PCR验证

利用生物信息学与实验验证的技术路线,成功地克隆了人类新基因C17orf32的cDNA(GenBank登记号：AY074907和TPA: BK000260),发现C17orf32的完整开放阅读框架(ORF,31～657 bp）cDNA（627 bp）与人类假定基因LOC124919 ORF(25～807 bp)的25～651位只有一个碱基不同.经RT-PCR验证并cDNA测序、人类表达序列标签(EST)数据库的BLAST检索和基因组成规律分析三方面的结果,均支持C17orf32的序列,而不支持LOC124919的编码序列.C17orf32基因组序列全长4.610 kb,含有6个外显子和5个内含子,cDNA序列全长1 679 bp, ORF横跨全部6个外显子.该基因ORF翻译起始处符合Kozak规则,ORF起始码上游同一相位有终止码,ORF后有2个加尾信号和PolyA尾.C17orf32基因的成功克隆表明,NCBI GENOME Annotation Project在2001年12月预测的人类假定蛋白XP-058865编码基因LOC124919的模式参考序列XM-058865中存在偏差,即在C17orf32基因cDNA的406与407位碱基之间错误插入一个碱基G, 从而导致在插入位点后,ORF编码125位氨基酸以后蛋白质序列的改变,出现260个氨基酸的多肽.因此,应慎重看待计算机注释的人类基因组编码序列.建立的技术路线有助于发现更多新的人类功能基因.     

Potential impacts of climate warming on active soil organic carbon contents along natural altitudinal forest transect of Changbai Mountain

Understanding soil carbon fractions and their responses to the global warming is important for improving soil carbon management of natural altitudinal forest ecosystem. In this study, the contents of soil total organic carbon (SOC), soil labile organic carbon (LOC), and microbial biomass carbon (MBC) in soil upper layers (0–20 cm) were measured along a natural altitudinal transect in the north slope of Changbai Mountain. The results showed that under natural conditions the contents of SOC and LOC were largest in Betula ermanii forest (altitude 1996 m), moderate in spruce-fir forest (altitude 1350 m), and smallest in Korean pine mixed broad-leaf tree forest (altitude 740 m). MBC contents in different forest ecosystems decreased in the order of Betula ermanii forest, Korean pine mixed broad-leaf tree forest, and dark coniferous forest. In addition, the responses of SOC, LOC, and MBC to soil warming were conducted by relocating intact soil cores from high- to low-elevation forests for one year. As expected, the soil core relocation caused significant increase in soil temperature but made no significant effect on soil moisture. After one year incubation, soil relocation significantly decreased SOC contents, whereas the contents of LOC, MBC, and the ratios of LOC to SOC and MBC to SOC increased.     

Double-tailed lipid modification as a promising candidate for oligonucleotide delivery in mammalian cells

Background

The potential use of nucleic acids as therapeutic drugs has triggered the quest for oligonucleotide conjugates with enhanced cellular permeability. To this end, the biophysical aspects of previously reported potential lipid oligodeoxyribonucleotide conjugates were studied including its membrane-binding properties and cellular uptake.

Methods

These conjugates were fully characterized by MALDI-TOF mass spectrometry and HPLC chromatography. Their ability to insert into lipid model membrane systems was evaluated by Langmuir balance and confocal microscopy followed by the study of the internalization of a lipid oligodeoxyribonucleotide conjugate bearing a double-tail lipid modification (C₂₈) into different cell lines by confocal microscopy and flow cytometry. This compound was also compared with other lipid containing conjugates and with the classical lipoplex formulation using Transfectin as transfection reagent.

Results

This double-tail lipid modification showed better incorporation into both lipid model membranes and cell systems. Indeed, this lipid conjugation was capable of inserting the oligodeoxyribonucleotide into both liquid-disordered and liquid-ordered domains of model lipid bilayer systems and produced an enhancement of oligodeoxyribonucleotide uptake in cells, even better than the effect caused by lipoplexes. In addition, in β₂ integrin (CR3) expressing cells this receptor was directly involved in the enhanced internalization of this compound.

Conclusions

All these features confirm that the dual lipid modification (C₂₈) is an excellent modification for enhancing nucleic acid delivery without altering their binding properties.

General significance

Compared to the commercial lipoplex approach, oligodeoxyribonucleotide conjugation with C₂₈ dual lipid modification seems to be promising to improve oligonucleotide delivery in mammalian cells.