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    • 76. 发明申请
    • PREVENTING ARRHYTHMIAS ASSOCIATED WITH CELL TRANSPLANTATION
    • 预防与细胞迁移相关的ARRHYTHMIAS
    • WO2005092033A2
    • 2005-10-06
    • PCT/US2005/009358
    • 2005-03-22
    • THE JOHNS HOPKINS UNIVERSITYMARBAN, EduardoABRAHAM, Maria, Roselle
    • MARBAN, EduardoABRAHAM, Maria, Roselle
    • A61K48/00
    • A61K48/005C12N5/0657C12N5/0658C12N15/86C12N2501/11C12N2740/16043
    • Skeletal myoblasts are an attractive cell type for transplantation since they are autologous and resistant to ischemia. However, clinical trials of myoblasts transplantation in heart failure have been plagued by ventricular tachy-arrhythmias and sudden cardiac death. The pathogenesis of these arrhythmias is poorly understood, but may be related to the fact that skeletal muscle cells, unlike heart cells, are electrically isolated by the absence of gap junctions. An in vitro model of myoblasts transplantation into cardiomyocyte monolayers can be used to investigate the mechanisms of transplant-associated arrhythmias. Co-cultures of human skeletal myoblasts and rat cardiomyocytes result in reentrant arrhythmias (spiral waves) that reproduce the features of ventricular tachycardia seen in patients receiving myoblasts transplants. These arrhythmias can be terminated by nitrendipine, an L-type calcium channel Mocker, but not by the Na channel blocker lidocaine. Genetic modification of myoblasts to stably express the gap junction protein connexin 43 decreases arrhythmogenicity in co-cultures. It similarly can be used to increase the safety of myoblasts transplantation in patients.
    • 骨骼肌成肌细胞是一种有吸引力的移植细胞类型,因为它们是自体的,对局部缺血有抗性。 然而,成肌细胞移植在心力衰竭中的临床试验一直困扰着心室心律失常和心源性猝死。 这些心律失常的发病机制很少被理解,但可能与骨髓肌细胞不同,心肌细胞不通过间隙连接而被电隔离的事实有关。 将成肌细胞移植到心肌细胞单层中的体外模型可用于研究移植相关性心律失常的机制。 人类骨骼肌成肌细胞和大鼠心肌细胞的共培养物导致再现心律失常(螺旋波),其再现了接受成肌细胞移植的患者中观察到的室性心动过速的特征。 这些心律失常可以由尼群地平,一种L型钙通道Mocker,而不是Na通道阻断剂利多卡因终止。 成肌细胞的遗传修饰以稳定表达间隙连接蛋白连接蛋白43降低共培养物中的心律失常。 它同样可以用于增加患者成肌细胞移植的安全性。
    • 78. 发明申请
    • MULTI-COLOR IN VITRO TRANSLATION
    • 多色翻译
    • WO2005017182A2
    • 2005-02-24
    • PCT/US2004/019097
    • 2004-07-21
    • THE JOHNS HOPINKS UNIVERSITY SCHOOL OF MEDICINETRAVERSO, Carlo, GiovanniDIEHL, FrankKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • TRAVERSO, Carlo, GiovanniDIEHL, FrankKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • C12Q
    • G01N33/6845G01N33/542
    • In vitro translation is a widely used tool for both analytical and preparative purposes. For analytical purposes, small amounts of proteins are synthesized and visualized through labeled amino acids incorporated during translation. The radioactively labeled amino acids originally used, such as [ 35 S]methionine or [ 14 C]leucine, have been superceded by the addition of antigenic tags or the incorporation of biotin-labeled or fluorescently-labeled amino acids. Such non-radioactive tags are simpler to visualize following translation and do not pose a radiation hazard. Among the non-radioactive tags, fluorescently labeled-lysine is the most convenient, as proteins that have incorporated this amino acid can be directly visualized following gel electrophoresis. Multiple fluorophores introduced into proteins significantly extend their utility, particularly for the comparison of in vitro translated proteins from related sources. This methodology can be employed for several purposes, including the simplified detection of rare truncating mutations in clinical samples from cancer patients.
    • 体外翻译是用于分析和制备目的的广泛使用的工具。 为了分析的目的,通过在翻译过程中并入的标记氨基酸合成少量蛋白质并使其可视化。 最初使用的放射性标记的氨基酸,例如[35 S]甲硫氨酸或[[14 C]亮氨酸)已经通过加入抗原标签或引入生物素标记或荧光标记的氨基酸而被替代。 这种非放射性标签更容易可视化翻译,并且不会造成辐射危害。 在非放射性标签中,荧光标记的赖氨酸是最方便的,因为结合了该氨基酸的蛋白质可以在凝胶电泳后直接显现。 引入蛋白质的多种荧光团显着扩展其效用,特别是用于比较来自相关来源的体外翻译蛋白质。 该方法可用于多种目的,包括简化检测癌症患者临床样品中罕见的截短突变。
    • 79. 发明申请
    • METHOD AND COMPOSITIONS FOR DETECTION AND ENUMERATION OF GENETIC VARIATIONS
    • 用于检测和产生遗传变异的方法和组合物
    • WO2005010145A2
    • 2005-02-03
    • PCT/US2004/015587
    • 2004-06-09
    • THE JOHNS HOPKINS UNIVERSITYDRESSMAN, DevinYAN, HaiKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • DRESSMAN, DevinYAN, HaiKINZLER, Kenneth, W.VOGELSTEIN, Bert
    • C12N
    • C12Q1/6858C07H21/04C12N15/1075C12Q1/686C12Q2565/537C12Q2563/149C12Q2563/143
    • Many areas of biomedical research depend on the analysis of uncommon variations in individual genes or transcripts. Here we describe a method that can quantify such variation at a scale and ease heretofore unattainable. Each DNA molecule in a collection of such molecules is converted into a single particle to which thousands of copies of DNA identical in sequence to the original are bound. This population of beads then corresponds to a one-to-one representation of the starting DNA molecules. Variation within the original population of DNA molecules can then be simply assessed by counting fluorescently-labeled particles via flow cytometry. Millions of individual DNA molecules can be assessed in this fashion with standard laboratory equipment. Moreover, specific variants can be isolated by flow sorting and employed for further experimentation. This approach can be used for the identification and quantification of rare mutations as well as to study variations in gene sequences or transcripts in specific populations or tissues.
    • 生物医学研究的许多领域取决于对个别基因或转录本中不常见变异的分析。 在这里,我们描述一种可以量化这种变化的方法,其规模和容易度迄今为止是无法实现的。 将这样的分子的集合中的每个DNA分子转化成单个颗粒,其中与原始序列顺序相同的数千个拷贝的DNA被结合。 这种珠子群对应于起始DNA分子的一对一表示。 然后可以通过流式细胞术对荧光标记的颗粒进行计数,简单地评估原始DNA分子群体内的变异。 可以用标准实验室设备以这种方式评估数百万个单独的DNA分子。 此外,可以通过流动分选分离特定的变体并用于进一步的实验。 该方法可用于鉴定和定量稀有突变,并研究特定群体或组织中基因序列或转录本的变异。