Biologically relevant effects of mRNA amplification on gene expression profiles

Background  

Gene expression microarray technology permits the analysis of global gene expression profiles. The amount of sample needed limits the use of small excision biopsies and/or needle biopsies from human or animal tissues. Linear amplification techniques have been developed to increase the amount of sample derived cDNA. These amplified samples can be hybridised on microarrays. However, little information is available whether microarrays based on amplified and unamplified material yield comparable results.     

黄地老虎核型多角体病毒的一些特性

黄地老虎核型多角体病毒(Agrotis segetum Nulear Polyhedrosis Virus简称AsNPV)的国内分离株(AsNPV^C),多角体呈六边形,大小1.7—2.6μm,为多粒包埋类型.每个病毒束内有2—7个核衣壳,大小约52nm×308nm.感染烟青虫(Heliothis assttlta)后分离到的多角体(As-HaNPV)其形状不规则,大小0.7—2.6μm,亦为多粒包埋类型.核衣壳2—6个不等,大小约40nm×300nm.EcoR1和HindⅢ限制性内切酶电泳图谱分析表明,AsNPV^CDNA和As-HaNPV DNA的EcoRI、HindIII酶切图谱一致,两者与HaNPV DNA的EcoRI,HindⅢ酶切图谱存在明显差异,AsNPV^C DNA的EcoRI酶切图谱共有15个片段,分子量在12.74×10⁶—1.18×10⁶道尔顿之间,总分子量约88.6×10⁶道尔顿,相当于134.25kbp.HaNPV DNA的EcoRI酶切图谱共有19个片段,分子量在13.89×10⁶—1.10×10⁶道尔顿之间,总分子量约93.86×10⁶道尔顿,相当于142.25kbp.AsNPV对黄地老虎2龄和4龄幼虫以及对烟青虫4龄幼虫的LD_{50分别为:1.4×10⁵pIB、7.4×10⁴PIB和2.61×10⁴PIB.}     

Photosynthetic induction times in shade-tolerant species with long and short-lived leaves

In the understory of a tropical rainforest, light flecks can contribute 10–80% of the total light flux. We investigated the capacity of eight shade-tolerant species to use light flecks by examining the time required for full induction of photosynthesis during an artificial light fleck. CO₂ fixation rates were measured with a portable LiCor gas-exchange system for plants growing in the field on Barro Colorado Island, Panama. In all species induction to 50% of maximum CO₂ fixation occurred quickly, from 1 to 3 min. In species with short leaf lifetimes (1 year), induction to 90% of maximum also occurred quickly, in 3–6 min. In contrast, the species with longer lived leaves (>4 years) required 11–36 min for induction to 90% of maximum. Induction times for leaves from gap and understory plants of the same species were indistinguishable. Elevated CO₂ did not eliminate the slow induction phase of long-lived leaves. This suggests that slow induction did not result from stomatal limitation. O₂ evolution, measured on excised leaf disks, induced in 3–4 min in species with short-lived leaves, and 4–8 min in species with long-lived leaves. The rapid induction of O₂ evolution indicates that the slower induction of CO₂ fixation in long-lived leaves was not caused by a delay in the induction of electron transport. Activation of rubisco may be the major factor limiting response times in species with long-lived leaves. Species from Panama with short-lived leaves had remarkably rapid induction times that are comparable to those of algae or higher plant chloroplasts. We also found that understory forest plants induced two to seven times more quickly than did potted plants.     

可切除的食管胃结合部腺癌的联合治疗

近年来胃癌的发病率有所下降,相比之下胃食管结合部腺癌的发病率却快速增长。手术治疗仍然是早期食管胃结合部腺癌的标准治疗方法,同时手术联合化疗、放化疗治疗食管胃结合部腺癌也逐渐得到国际认可。尽管在手术治疗、放疗和化疗治疗技术得到完善和改进,但食管癌和食管胃结合部腺癌的预后仍然较差。目前有数个大型临床随机对照试验数据支持对食管下端和食管胃交界部腺癌使用术前联合化疗,但辅助治疗的贡献仍不能确定。最近有meta分析表明手术联合化疗、放化疗可以提高胃食管结合部腺癌患者术后存活率,但也有一些临床随机试验的数据表明手术联合化疗、放化疗并无明显好处。本文通过总结最新的临床试验及meta分析结果,阐述不同的可切除的胃食管结合部腺癌的联合治疗方法。     

Desiccation tolerance of five tropical seedlings in panama. Relationship to a field assessment of drought performance

Studies of the desiccation tolerance of the seedlings of five tropical trees were made on potted plants growing in a greenhouse. Pots were watered to field capacity and then dehydrated for 3 to 9 weeks to reach various visual wilting stages, from slightly wilted to dead. Saturated root hydraulic conductance was measured with a high-pressure flowmeter, and whole-stem hydraulic conductance was measured by a vacuum chamber method. Leaf punches (5.6-mm diameter) were harvested for measurement of leaf water potential by a thermocouple psychrometer method and for measurement of fresh and dry weight. In a parallel study, the same five species were studied in a field experiment in the understory of a tropical forest, where these species frequently germinate. Control seedlings were maintained in irrigated plots during a dry season, and experimental plants were grown in similar plots with rain exclusion shelters. Every 2 to 4 weeks, the seedlings were scored for wilt state and survivorship. After a 22-week drought, the dry plots were irrigated for several weeks to verify visual symptoms of death. The field trials were used to rank drought performance of species, and the greenhouse desiccation studies were used to determine the conditions of moribund plants. Our conclusion is that the desiccation tolerance of moribund plants correlated with field assessment of drought-performance for the five species (r(2) > 0.94).     

Depletion of CD4+ T cells during immunization with nonviable Listeria monocytogenes causes enhanced CD8+ T cell-mediated protection against listeriosis

Immunization of mice with nonviable Listeria monocytogenes generates an insufficient CD8(+) T cell response and consequently only limited protection against subsequent L. monocytogenes infection. We have recently demonstrated that depletion of regulatory CD4(+) T cells during immunization significantly enhances CD8(+) T cell responses. In the present study, we determined the impact of CD4(+) T cell depletion on the CD8(+) T cell response against heat-killed LISTERIA: Treatment of mice with anti-CD4 mAb during boost immunization with heat-killed Listeria significantly increased numbers of Listeria-specific CD8(+) T cells and improved protection against subsequent infection with L. monocytogenes. During challenge infection, numbers of Listeria-specific CD8(+) T cells were enhanced, and these cells expressed effector functions in terms of IFN-gamma production. In summary, we demonstrate that combining nonviable L. monocytogenes vaccination and CD4(+) T cell depletion improves generation of long-lasting and functional Listeria-specific CD8(+) memory T cells.     

Cinnamoyl glucosides of catechin and dimeric procyanidins from young leaves of Inga umbellifera (Fabaceae)

The rapidly growing, nearly achlorophyllous, young leaves of Inga umbellifera express high concentrations of mono and dimeric 3-O-gluco-cinnamoyl catechin/epicatechin, rare forms of substituted flavan-3-ols. Here we present structures for five novel compounds in this class: three monomers [catechin-3-O-beta-D-gluco(2-cinnamoyl)pyranoside, catechin-3-O-beta-D-gluco(6-cinnamoyl) pyranoside, catechin-3-O-beta-D-gluco(2,6-biscinnamoyl)pyranoside] and two dimeric procyanidins [catechin-3-O-beta-D-glucopyrano-(4alpha-->8)-catechin-3-O-beta-D-gluco(2-cinnamoyl)pyranoside and catechin-3-O-beta-D-glucopyrano-(4alpha-->8)-epicatechin-3-O-beta-D-gluco(6-cinnamoyl)pyranoside]. The young leaves of Inga umbellifera express high concentrations of 3-O-(cinnamoyl)glucosides of catechin and epicatechin.     

Comparison of dot chromosome sequences from D. melanogaster and D. virilis reveals an enrichment of DNA transposon sequences in heterochromatic domains

Background

Chromosome four of Drosophila melanogaster, known as the dot chromosome, is largely heterochromatic, as shown by immunofluorescent staining with antibodies to heterochromatin protein 1 (HP1) and histone H3K9me. In contrast, the absence of HP1 and H3K9me from the dot chromosome in D. virilis suggests that this region is euchromatic. D. virilis diverged from D. melanogaster 40 to 60 million years ago.

Results

Here we describe finished sequencing and analysis of 11 fosmids hybridizing to the dot chromosome of D. virilis (372,650 base-pairs) and seven fosmids from major euchromatic chromosome arms (273,110 base-pairs). Most genes from the dot chromosome of D. melanogaster remain on the dot chromosome in D. virilis, but many inversions have occurred. The dot chromosomes of both species are similar to the major chromosome arms in gene density and coding density, but the dot chromosome genes of both species have larger introns. The D. virilis dot chromosome fosmids have a high repeat density (22.8%), similar to homologous regions of D. melanogaster (26.5%). There are, however, major differences in the representation of repetitive elements. Remnants of DNA transposons make up only 6.3% of the D. virilis dot chromosome fosmids, but 18.4% of the homologous regions from D. melanogaster; DINE-1 and 1360 elements are particularly enriched in D. melanogaster. Euchromatic domains on the major chromosomes in both species have very few DNA transposons (less than 0.4 %).

Conclusion

Combining these results with recent findings about RNAi, we suggest that specific repetitive elements, as well as density, play a role in determining higher-order chromatin packaging.