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991.
Comparison of ethanol versus formalin fixation on preservation of histology and RNA in laser capture microdissected brain tissues 总被引:1,自引:0,他引:1
Su JM Perlaky L Li XN Leung HC Antalffy B Armstrong D Lau CC 《Brain pathology (Zurich, Switzerland)》2004,14(2):175-182
Although RNA can be retrieved from formalin-fixed, paraffin-embedded (FFPE) tissues, the yield is low, and the RNA is fragmented. Recent advances in gene expression profiling underscore the importance of identifying a fixative that preserves histology and mRNA. We demonstrated that, for immersion fixation of brains, 70% ethanol is superior to formalin for mRNA preservation. RNA yield from ethanol-fixed tissues was 70% of the yield from fresh frozen specimens, but only a negligible quantity was recovered from formalin-fixed tissues. RNA from ethanol-fixed brains showed integrity comparable to RNA from fresh frozen tissues, and RT-PCR using RNA from ethanol-fixed tissues was consistently successful. RNA from FFPE tissues composed of low-molecular weight fragments, and their use in RT-PCR failed repeatedly. The yield and quality of RNA from ethanol-fixed brains were unaffected after immersion at 4 degrees C for 2 weeks. In a blinded comparison to FFPE tissues, ethanol-fixed specimens were judged to show comparable histology and superior immunostaining. After laser capture microdissection (LCM), we failed to recover mRNA from FFPE tissues but retrieved mRNA from ethanol-fixed tissues for RT-PCR and cDNA microarray analysis. We conclude that 70% ethanol preserves RNA integrity and is suitable for expression profiling of brain tissues by LCM and cDNA microarray. 相似文献
992.
SARS病人外周血T淋巴细胞的动态变化及其在发病进程和发病机理中的意义 总被引:10,自引:0,他引:10
目的 :通过研究严重急性呼吸综合征 (SevereAcuteRespiratorySyndrome,SARS)患者外周血免疫细胞的动态变化 ,探讨外周血免疫细胞在SARS发病进程中的意义 ,初步探讨SARS的发病机理 ,并为SARS的诊断和治疗提供有力的实验室依据。方法 :回顾性动态观察我院收治的已治愈SARS病例和SARS死亡病例外周血CD4 + 和CD8+ T淋巴细胞 ,评估外周血免疫细胞在SARS发病进程及预后中的作用。实验方法为流式细胞术检测和分析免疫细胞表面特异性荧光抗体标记 ,T细胞表面标志组合为CD3 CD8 CD4 5 CD4。结果 :所有治愈的SARS病人的CD4 + 和CD8+ T淋巴细胞都有不同程度的可逆性下降 ,而所有的死亡病例有不可逆性显著下降 ,直至死亡。T细胞下降程度和维持的时间与病情密切相关。普通型SARS病例最低CD4 + T淋巴细胞数为 30 5± 15 0cells μl(P <0 .0 0 1)、重型为 139± 6 9cells μl(P <0 .0 0 1) ,普通型SARS病例最低CD8+ T淋巴细胞数为 2 2 3± 89cells μl(P <0 .0 0 1)、重型为 171± 92cells μl(P <0 .0 0 1)。所有普通型病例和大部分重型病例T淋巴细胞恢复正常 ,个别重型病例低于正常或接近正常 ,恢复后普通型平均为 991± 2 86cells μl,重型平均为 5 4 5± 2 2 5cells μl。恢复时间有所不同 ,普通型平均为 17± 5天 相似文献
993.
Summary Venezuelan equine encephalitis virus was passaged in KB cell cultures. The virus lost its mouse pathogenicity following subcutaneous inoculation during KB cell passage; the attenuated strain also produced smaller plaques than the pathogenic strain. Both strains grew to the same extent inAedes aegypti (L.) mosquitoes after intrathoracic inoculation. If any reversions to pathogenicity occur during development of the attenuated virus in mosquitoes, then the mutation frequency per duplication per particle must be smaller than 3.5×10–6. 相似文献
994.
Septins: a ring to part mother and daughter 总被引:15,自引:0,他引:15
The septins are well conserved GTPases found in animals and fungi. In yeast, they are required for the formation of 10-nm filaments, with which they co-localize at the bud neck. Therefore, septins have been proposed to be components of the neck filaments and to have polymerization properties. In support of this hypothesis, septin complexes purified from yeast and flies form filaments in vitro. However, recent studies have questioned the relevance of septin filament formation for septin function. Particularly, septin polymerization may not be required for their function in cytokinesis. New septin functions have also been recently uncovered: in budding yeast, the septin ring is required for the maintenance of cell polarity. It forms a cortical barrier that prevents lateral diffusion of membrane-associated proteins through the bud neck. Here, we review the most recent functional and biochemical data, to discuss whether there is a link between septin polymerization properties and septin function. 相似文献
995.
Genetic Conservation of Hemagglutinin Gene of H9 Influenza Virus in Chicken Population in Mainland China 总被引:1,自引:0,他引:1
The hemagglutinin (HA) genes of 12 H9N2 influenza virus strains isolated from chickens in Mainland China during the period 1995–2002 were genetically analyzed. All the isolates possessed the same amino acid motif -R-S-S-R/G-L- at the cleavage site of HA. Except for the conserved amino acids, as is the case in the other avian influenza viruses, located in the receptor binding site, all of the 12 isolates possessed N at amino acid position 183; A, T, or V at position 190; K at position 137, whereas the representative strains of the other lineage (except Dk/HK/Y280/97-like lineage) virus of H9N2 viruses had H, E, and R at these positions respectively. These could be considered as the partial molecular markers of the H9 viruses isolated from chickens in Mainland China. Phylogenetic analyses showed HA genes of these isolates belonged to that of A/duck/Hong Kong/Y280/97-like virus lineage. No A/quail/Hong Kong/Gl/97-like virus was found in chicken, population since the outbreak of H9N2 influenza in Mainland China in 1992. The available evidence indicates that HA genes of H9 influenza virus circulating in Mainland China during the past years were well conserved. 相似文献
996.
Charles R. Bursey Stephen R. Goldberg 《Acta parasitologica / Witold Stefański Institute of Parasitology, Warszawa, Poland》2007,52(4):368-370
Aplectana krausi sp. nov. (Ascaridida, Cosmocercidae) from the intestines of Platymantis boulengeri (Anura, Ceratobatrachidae) is described and illustrated. Aplectana krausi represents the 42nd species assigned to the genus, the 4th species reported from the Australo-Papuan region. It is easily
separated from the three species previously reported from the region by the distribution pattern of male caudal papillae:
A. macintoshii and A. novaezelandiae have irregular patterns; A. zweifeli and A. krausi have defined patterns. Aplectana zweifeli has 8–10 precloacal, no adcloacal, and 9 postcloacal pairs of papillae, there is a single median papillae just anterior to
the cloaca; A. krausi has 5 precloacal, 1 adcloacal, and 5 postcloacal pairs of papillae, a median papillae is absent. 相似文献
997.
Francesca Fernandez Robert P Curtain Natalie J Colson Micky Ovcaric John MacMillan Lyn R Griffiths 《BMC medical genetics》2007,8(1):57
Background
Migraine with aura (MA) is a subtype of typical migraine. Migraine with aura (MA) also encompasses a rare severe subtype Familial Hemiplegic Migraine (FHM) with several known genetic loci. The type 2 FHM (FHM-2) susceptibility locus maps to chromosome 1q23 and mutations in the ATP1A2 gene at this site have recently been implicated. We have previously provided evidence of linkage of typical migraine (predominantly MA) to microsatellite markers on chromosome 1, in the 1q31 and 1q23 regions. In this study, we have undertaken a large genomic investigation involving candidate genes that lie within the chromosome 1q23 and 1q31 regions using an association analysis approach. 相似文献998.
999.
Francesca Fernandez Teresa Esposito Rod A Lea Natalie J Colson Alfredo Ciccodicola Fernando Gianfrancesco Lyn R Griffiths 《BMC medical genetics》2008,9(1):109
Background
Migraine is a neurological disorder characterized by recurrent attacks of severe headache, affecting around 12% of Caucasian populations. It is well known that migraine has a strong genetic component, although the number and type of genes involved is still unclear. Prior linkage studies have reported mapping of a migraine gene to chromosome Xq 24–28, a region containing a cluster of genes for GABA A receptors (GABRE, GABRA3, GABRQ), which are potential candidate genes for migraine. The GABA neurotransmitter has been implicated in migraine pathophysiology previously; however its exact role has not yet been established, although GABA receptors agonists have been the target of therapeutic developments. The aim of the present research is to investigate the role of the potential candidate genes reported on chromosome Xq 24–28 region in migraine susceptibility. In this study, we have focused on the subunit GABA A receptors type ε (GABRE) and type θ (GABRQ) genes and their involvement in migraine. 相似文献1000.
Mechanisms of Neuronal Death in Alzheimer's Disease 总被引:9,自引:0,他引:9
Recent data in cell culture has shown that brain neurons are particularly vulnerable to degeneration by apoptosis. Further the inducers that activate the program (e.g. β-amyloid, oxidatative damage, low energy metabolism) correspond to conditions present in the Alzheimer's disease (AD) brain. This suggests the possibility that apoptosis may be one of the mechanisms contributing to neuronal loss in this disease. Indeed, some neurons in vulnerable regions of the AD brain show evidence of DNA damage, nuclear apoptotic bodies, chromatin condensation, and the induction of select genes characteristic of apoptosis in cell culture and animal models. This suggests the existence of apoptosis in the AD brain, a hypothesis also consistent with evolving research in one of the regulatory functions of the presenilin genes. On the other hand, DNA damage is present in the majority of neurons in vulnerable regions in early and mild cases. In most tissues, cells in fully activated apoptosis degenerate and are removed within hours to days and thus it seems all DNA damage is unlikely to signify terminal apoptosis. The presence of extensive DNA damage suggests an acceleration of damage, faulty repair process, loss of protective mechanisms, or an activation and arrest of aspects of the apoptotic program. DNA damage is unlikely to be an artifact of postmortem delay or agonal state. The existence of protective mechanisms for neurons may exist as these cells are nondividing and essential. In this context it is interesting that Bcl-2 is upregulated in most neurons with DNA damage. Further, at least one DNA repair enzyme is also upregulated. Thus it appears as if neurons are in a struggle between degeneration and repair. As research advances it is critical to reduce the stimuli that cause the neuronal damage and discover the key intervention points to assist neurons in the repair processes. 相似文献