Histamine Transmission Modulates the Phenotype of Murine Narcolepsy Caused by Orexin Neuron Deficiency

Narcolepsy type 1 is associated with loss of orexin neurons, sleep-wake derangements, cataplexy, and a wide spectrum of alterations in other physiological functions, including energy balance, cardiovascular, and respiratory control. It is unclear which narcolepsy signs are directly related to the lack of orexin neurons or are instead modulated by dysfunction of other neurotransmitter systems physiologically controlled by orexin neurons, such as the histamine system. To address this question, we tested whether some of narcolepsy signs would be detected in mice lacking histamine signaling (HDC-KO). Moreover, we studied double-mutant mice lacking both histamine signaling and orexin neurons (DM) to evaluate whether the absence of histamine signaling would modulate narcolepsy symptoms produced by orexin deficiency. Mice were instrumented with electrodes for recording the electroencephalogram and electromyogram and a telemetric arterial pressure transducer. Sleep attacks fragmenting wakefulness, cataplexy, excess rapid-eye-movement sleep (R) during the activity period, and enhanced increase of arterial pressure during R, which are hallmarks of narcolepsy in mice, did not occur in HDC-KO, whereas they were observed in DM mice. Thus, these narcolepsy signs are neither caused nor abrogated by the absence of histamine. Conversely, the lack of histamine produced obesity in HDC-KO and to a greater extent also in DM. Moreover, the regularity of breath duration during R was significantly increased in either HDC-KO or DM relative to that in congenic wild-type mice. Defects of histamine transmission may thus modulate the metabolic and respiratory phenotype of murine narcolepsy.     

Covalent enzyme immobilization by poly(ethylene glycol) diglycidyl ether (PEGDE) for microelectrode biosensor preparation

Poly(ethylene glycol) diglycidyl ether (PEGDE) is widely used as an additive for cross-linking polymers bearing amine, hydroxyl, or carboxyl groups. However, the idea of using PEGDE alone for immobilizing proteins on biosensors has never been thoroughly explored. We report the successful fabrication of microelectrode biosensors based on glucose oxidase, d-amino acid oxidase, and glutamate oxidase immobilized using PEGDE. We found that biosensors made with PEGDE exhibited high sensitivity and a response time on the order of seconds, which is sufficient for observing biological processes in vivo. The enzymatic activity on these biosensors was highly stable over several months when they were stored at 4 °C, and over at least 3d at 37 °C. Glucose microelectrode biosensors implanted in the central nervous system of anesthetized rats reliably monitored changes in brain glucose levels induced by sequential administration of insulin and glucose. PEGDE provides a simple, low cost, non-toxic alternative for the preparation of in vivo microelectrode biosensors.     

Altered cholesterol metabolism in Niemann-Pick type C1 mouse brains affects mitochondrial function

Niemann-Pick type C1 (NPC1) disease is a fatal hereditary disorder characterized by a defect in cholesterol trafficking and progressive neurodegeneration. Although the NPC1 gene has been identified, the molecular mechanism responsible for neuronal dysfunction in brains of patients with NPC1 disease remains unknown. This study demonstrates that the amount of cholesterol within mitochondria membranes is significantly elevated in NPC1 mouse brains and neural cells. In addition, the mitochondrial membrane potential, the activity of ATP synthase, and henceforth the level of ATP are markedly decreased in NPC1 mouse brains and neurons. Importantly, reducing the level of cholesterol within mitochondrial membranes using methyl-beta-cyclodextrin can restore the activity of ATP synthase. Finally, NPC1 neurons show an impaired neurite outgrowth, which can be rescued by exogenous ATP. These results suggest that mitochondrial dysfunctions and subsequent ATP deficiency, which are induced by altered cholesterol metabolism in mitochondria, may be responsible for neuronal impairment in NPC1 disease.     

Sequence and pH effects of LNA-containing triple helix-forming oligonucleotides: physical chemistry, biochemistry, and modeling studies

Triple helix-forming oligonucleotides (TFOs) have been demonstrated to be capable of interfering with gene expression and modifying genomic DNA in a sequence-specific manner. Partial incorporation of 2'-O,4'-C-methylene linked locked nucleic acid (LNA) residues in TFOs has been shown to enhance significantly triple helix formation, whereas the full-length LNA TFO failed to form a stable triplex. This work is aimed at understanding the triple helix-forming properties of LNA-containing TFOs and at optimally designing their sequences. Both DNA thermal melting, gel retardation, and restriction enzyme experiments as well as modeling studies by molecular mechanics were carried out to investigate the base composition/sequence and pH-dependence effects of LNA-containing TFOs, as well as their structural features underlying triple helix formation. Alternating LNA substitution every 2-3 nucleotides in TFOs is mandatory, whereas the use of thymine LNA residues should be favored under neutral pH conditions. A rule for designing optimal LNA-containing TFOs is proposed. In addition, alternative LNA and 2'-O-methyl residues in TFOs do not significantly improve triple helix formation.     

大肠杆菌F18菌毛及其亚型的PCR鉴定

F18菌毛是产肠毒素大肠杆菌 (ETEC)与产vero细胞毒素大肠杆菌 (VTEC)的重要致病因子 ,可介导细菌对小肠细胞的黏附 ,并具有F18ab和F18ac 2个抗原亚型。根据已发表的F18ab菌毛A亚单位 (FedA ab)的基因 (fedA ab)设计 3条引物 ,建立了 2种聚合酶链式反应 (PCR)扩增方法。通过对F18ab 大肠杆菌、F18ac 大肠杆菌、K88 大肠杆菌、K99 大肠杆菌、987P 大肠杆菌、F4 1 大肠杆菌的试验 ,结果表明所建立的PCR方法可特异性鉴定F18 大肠杆菌并区别其亚型F18ab与F18ac     

植物体内的鞘氨醇-1-磷酸(S1P)

鞘氨醇-1-磷酸（S1P）在植物体内也可作为一种信号分子参与ABA介导的保卫细胞信号转导过程,这一途径和异三聚体G蛋白相耦联。文章对此问题的研究进展作了介绍。     

Distribution and radiosensitizing effect of cholesterol-coupled Dbait molecule in rat model of glioblastoma

Background

Glioma is the most aggressive tumor of the brain and the most efficient treatments are based on radiotherapy. However, tumors are often resistant to radiotherapy due to an enhanced DNA repair activity. Short and stabilized DNA molecules (Dbait) have recently been proposed as an efficient strategy to inhibit DNA repair in tumor.

Methodology/Principal Findings

The distribution of three formulations of Dbait, (i) Dbait alone, (ii) Dbait associated with polyethylenimine, and (iii) Dbait linked with cholesterol (coDbait), was evaluated one day after intratumoral delivery in an RG2 rat glioma model. Dbait molecule distribution was assessed in the whole organ with 2D-FRI and in brain sections. CoDbait was chosen for further studies given its good retention in the brain, cellular localization, and efficacy in inducing the activation of DNA repair effectors. The radiosensitizing effect of coDbait was studied in four groups of rats bearing RG2-glioma: no treatment, radiotherapy only, coDbait alone, and CoDbait with radiotherapy. Treatment started 7 days after tumor inoculation and consisted of two series of treatment in two weeks: coDbait injection followed by a selective 6-Gy irradiation of the head. We evaluated the radiosensitizing effect using animal survival, tumor volume, cell proliferation, and vasculature characteristics with multiparametric MRI. CoDbait with radiotherapy improved the survival of rats bearing RG2-glioma by reducing tumor growth and cell proliferation without altering tumor vasculature.

Conclusion/Significance

coDbait is therefore a promising molecular therapy to sensitize glioma to radiotherapy.     

略论东方白鹳的繁殖分布区域的扩展

东方白鹳(Ciconia boyciana)为远东特有鸟种.体长100～115 cm,体羽白色,初级和次级飞羽黑色,嘴长而厚重强直,脚红色,曾被视为白鹳(Ciconia ciconia)下之一亚种,后分立[1-6].东方白鹳与白鹳的主要区别在于体型明显为大,眼周裸出皮肤粉红色,嘴灰黑色至黑色而非红色.作者介绍何芬奇,男;研究方向:鸟类学.     

携带鼠源基因的沙门氏菌对肿瘤的预防作用

探讨减毒鼠伤寒沙门氏菌作为口服基因治疗载体的可行性.通过电转化法将真核表达载体pCMVmIL-12、pCMVmGM-CSF、EGFPN1导入减毒鼠伤寒沙门氏菌SL3261中,经由胃管饲于BALB/c和C57BL/6小鼠.6周后分别用4T₁乳腺癌细胞和Lewis肺癌细胞进行攻击.通过流式细胞仪、共聚焦显微镜检测绿色荧光蛋白在小鼠各组织中的表达,通过PCR和ELISA的方法检测mIL-12、mGM-CSF基因的整合和表达情况.并考察肿瘤的受抑情况和小鼠的生存期.结果表明：在小鼠的肝、脾、小肠、肾脏和肿瘤中可检测到绿色荧光蛋白的表达和相应细胞因子基因的整合.血清中相应的细胞因子水平较对照组明显升高（P＜0.05）,生存期远远超过对照组小鼠（P＜0.05）.减毒沙门氏菌可作为口服基因治疗载体,为肿瘤的预防和治疗提供一条简便、安全、有效的途径.     

基于支持向量机的细菌基因组水平转移基因预测

随着各种生物基因组序列测定工作的完成,大量的DNA序列数据涌现出来,为研究在基因组中寻找水平转移基因提供了极大的便利.将基因序列特征分析和支持向量机技术结合起来,通过分析基因序列的特征差异发现水平转移基因.依据以前研究工作的基础,选取了绝对密码子使用频率(FCU)作为序列特征,主要因为它既包含了基因密码子使用偏性的信息,也包含了基因所编码蛋白的氨基酸组成信息,支持向量机利用这些信息进行水平转移基因分析和预测,可以提高预测的准确性.另外,提出了基于分链的水平转移基因预测新方法,即将细菌基因组前导链和滞后链上的基因区别对待,分别进行水平转移基因预测.结果显示,基本预测方法要优于目前预测结果最好的Tsirigos等提出的基于八联核苷酸频率的打分算法,命中率的相对提高率最高达31.47%,而基于分链的方法对水平转移基因的预测取得了更好的结果.