Continuous Monitoring of Post Mortem Temperature Changes in the Human Brain

Recent developments in neurochemistry research on the post mortem human brain require a detailed understanding of the post mortem changes in the human brain, including the correlation between time related temperature changes and alterations in biochemical parameters. As an initial step towards our deeper insight into the intricate relationships between post mortem time, temperature and neurochemical processes, in the present study we set out to monitor continuously temperature changes in the post mortem human brain in eight cadavers for a period of up to 24 h after death under 'standard' clinical conditions at a neurosurgery clinic. A main objective of the study was to find a simple and reliable mathematical formula, requiring only time and an easily obtainable body temperature measurement parameter, with the help of which the superficial and deep brain temperatures can be obtained without invasive interactions. With a portable thermoprobe data logger system superficial (4 cm from skull surface) and deep (8 cm) brain temperatures, the temperature of the liver and that of the forehead skin, as well as the ambient temperature of the room were measured at regular time intervals (every 1 or 5 min). Various mathematical models were fitted to the data in order to create a simple model capable to predict brain temperatures from easily accessible measurements, such as that of the forehead skin. On the basis of the tested models we propose that with simple polynomial equations the deep and superficial brain temperatures can be described reliably as T (br4) ( degrees C)=T (fh)-0.001t (3)+0.0541t (2)-1.0622t+7.5933 and T (br8) ( degrees C)=T (fh)-0.0003t (3)+0.0201t (2)-0.619t+7.9036, respectively, where T (br4) is the superficial (4 cm) brain temperature, T (br8) is the deep (8 cm) brain temperature, T (fh) is the forehead temperature and t is the time from death. These measurements can, in combination with further neurochemical studies, contribute to our better understanding of the human brain's time- and temperature-related post mortem biochemical changes.     

mRNA选择性剪接的分子机制

真核细胞mRNA前体经过剪接成为成熟的mRNA,而mRNA前体的选择性剪接极大地增加了蛋白质的多样性和基因表达的复杂程度,剪接位点的识别可以以跨越内含子的机制(内含子限定)或跨越外显子的机制(外显子限定)进行。选择性剪接有多种剪接形式：选择不同的剪接位点,选择不同的剪接末端,外显子的不同组合及内含子的剪接与否等。选择性剪接过程受到许多顺式元件和反式因子的调控,并与基本剪接过程紧密联系,剪接体中的一些剪接因子也参与了对选择性剪接的调控。选择性剪接也是1个伴随转录发生的过程,不同的启动子可调控产生不同的剪接产物。mRNA的选择性剪接机制多种多样,已发现RNA编辑和反式剪接也可参与选择性剪接过程。     

基于PCR 技术的人脑基因可变剪接差异性研究

目的:对人脑的基因可变剪接位点差异性进行分析。方法:通过RNA-Seq得到的转录组数据,分析推测出在不同年龄的7对样本的大脑前额叶皮层(prefrontal cortex,PFC)表达的某些基因具有可变剪接位点的差异性;为验证此推测,设计7对引物,提取不同年龄样本的RNA,通过反转录合成cDNA,采用PCR技术扩增目的序列。结果:在已剪接和(或)未剪接产物预期片段处有明显的条带。结论:通过Quantity One软件分析后,确定在不同年龄间存在可变剪接差异。基于PCR技术的基因可变剪接分析对RNA-seq数据集的预测结果进行验证是一种有效新颖的PCR新应用。     

mRNA选择性剪切在植物发育中的作用

mRNA选择性剪切（altemativesplicing）是生物体基因转录调控的基本方式之一,随着新一代测序技术的广泛应用,越来越多的基因的选择性剪切现象被揭示。在植物发育的不同阶段及其应对外界逆境胁迫的过程中,许多基因发生了选择性剪切,产生植物各个发育阶段所需的特定蛋白质来完成不同的发育过程和形成不同应答因子以适应外界环境的变化。本文从种子发育、器官形态发育、开花时间、生物钟调控、环境胁迫等方面综述选择性剪切在植物发育中的研究进展。     

Post Mortem DNA Degradation of Human Tissue Experimentally Mummified in Salt

Mummified human tissues are of great interest in forensics and biomolecular archaeology. The aim of this study was to analyse post mortem DNA alterations in soft tissues in order to improve our knowledge of the patterns of DNA degradation that occur during salt mummification. In this study, the lower limb of a female human donor was amputated within 24 h post mortem and mummified using a process designed to simulate the salt dehydration phase of natural or artificial mummification. Skin and skeletal muscle were sampled at multiple time points over a period of 322 days and subjected to genetic analysis. Patterns of genomic fragmentation, miscoding lesions, and overall DNA degradation in both nuclear and mitochondrial DNA was assessed by different methods: gel electrophoresis, multiplex comparative autosomal STR length amplification, cloning and sequence analysis, and PCR amplification of different fragment sizes using a damage sensitive recombinant polymerase. The study outcome reveals a very good level of DNA preservation in salt mummified tissues over the course of the experiment, with an overall slower rate of DNA fragmentation in skin compared to muscle.     

Alternative Splicing,Expression and Cellular Localization of Calneuron-1 in the Rat and Human Brain

Calneuron-1 and -2 are members of the neuronal calcium-binding protein family (nCaBP). They are transmembrane Calmodulin-like EF-hand Ca²⁺-sensors, and a function in the control of Golgi-to-plasma membrane vesicle trafficking has been assigned to both proteins. In this paper, we describe the distribution of Calneuron-1 in rat and human brains. We show that Calneuron-1 is ubiquitously expressed in all brain regions examined. The protein is most abundant in Purkinje cells of the cerebellum and principal neurons of the cortex and limbic brain whereas no expression in glial cells is apparent. In addition, we identify two novel splice isoforms of Calneuron-1 with extended N-termini. These isoforms are particular abundant in the cerebellum. Taken together, these data set grounds for a better understanding of the cellular function of Calneurons.     

Splicing Pattern of Gsα mRNA in Human and Rat Brain

The enzyme tyrosine hydroxylase catalyzes the first step in the biosynthesis of dopamine, norepinephrine, and epinephrine. Tyrosine hydroxylase is a substrate for cyclic AMP-dependent protein kinase as well as other protein kinases. We determined the Km and Vmax of rat pheochromocytoma tyrosine hydroxylase for cyclic AMP-dependent protein kinase and obtained values of 136 microM and 7.1 mumol/min/mg of catalytic subunit, respectively. These values were not appreciably affected by the substrates for tyrosine hydroxylase (tyrosine and tetrahydrobiopterin) or by feedback inhibitors (dopamine and norepinephrine). The high Km of tyrosine hydroxylase correlates with the high content of tyrosine hydroxylase in catecholaminergic cells. We also determined the kinetic constants for peptides modeled after actual or potential tyrosine hydroxylase phosphorylation sites. We found that the best substrates for cyclic AMP-dependent protein kinase were those peptides corresponding to serine 40. Tyrosine hydroxylase (36-46), for example, exhibited a Km of 108 microM and a Vmax of 6.93 mumol/min/mg of catalytic subunit. The next best substrate was the peptide corresponding to serine 153. The peptide containing the sequence conforming to serine 19 was a very poor substrate, and that conforming to serine 172 was not phosphorylated to any significant extent. The primary structure of the actual or potential phosphorylation sites is sufficient to explain the substrate behavior of the native enzyme.     

鉴定9个新的RHD基因mRNA可变剪接体

为了研究各种RHD基因mRNA可变剪接体的基因结构, 应用逆转录聚合酶链反应(RT-PCR)检测正常人脐血样本RHD mRNA, 对RHD cDNA进行TA克隆和序列分析, 对各可变剪接体的剪接位点进行DNA序列分析, 并将RHD mRNA进行表达序列标签(ESTs)分析。结果在28个阳性克隆中, 除全长RHD cDNA外, 共检测到12种(包括9种新的)RHD可变剪接体, 发现外显子遗漏、5′和3′剪接位点变异3种剪接形式, 涉及外显子2~9, 其中6种新的剪接体同时存在RHD和RHCE基因同源杂交现象。ESTs分析还检索到内含子保留形式的剪接体。研究表明, RHD基因mRNA存在复杂的可变剪接机制, 除已报道的剪接体外, 检测到9种新的RHD可变剪接体, 并发现了可变剪接和同源杂交并存现象。