Quinone-based antibody labeling reagent for enzyme-free chemiluminescent immunoassays. Application to avidin and biotinylated anti-rabbit IgG labeling

Chemiluminescence-enzyme immunoassays make it possible to measure trace components with high sensitivity and selectivity due to the high specificity of the antigen-antibody reaction and the high sensitivity of chemiluminescence assays. However, using an enzyme-labeled antibody suffers from many problems such as low reproducibility due to the instability of the enzyme and inhibition of antigen-antibody reaction due to its steric effect. Therefore, herein we report an innovative non-enzymatic chemiluminescence immunoassays labeling reagent through using quinone as a signal-generating tag coupled with biotin as a binder, to overcome enzymatic labeling problems. Biotinylated-1,4-naphthoquinone (biotin-NQ) was synthesized and characterized and it could produce long-lasting chemiluminescence upon mixing with dithiothreitol and luminol based on the redox cycle of quinone. Biotin-NQ showed exceptional stability towards different stress factors that may be encountered during performing the immunoassay such as high temperatures, highly acidic and alkaline conditions, and repeated freeze-thaw cycles. On the other hand, all these conditions lead to decreased labeling enzyme reactivity due to possible denaturation of its protein structure. Finally, the measurement of the biotin-labeled antibody was successfully performed using biotin-NQ and avidin. As a result, the antibody could be detected down to 25.7 nM which is 2.5 times sensitive than biotin-HRP chemiluminescence-enzyme immunoassays. Moreover, our method was applied successfully for determination of avidin using immobilized biotinylated antibody and biotin-NQ, which simulates immunoassays. Avidin could be detected down to 23.4 nM with excellent linearity (r = 0.996). Accordingly, our developed reagent, biotin-NQ, could be used as a universal highly stable, cost-effective, and steric free non-enzymatic label for immunoassays

A Crowdsourcing Approach for Finding Misidentifications of Bibliographic Records

Because there is no perfect technique for automatic identification of bibliographic records, cleaning the identification results manually is indispensable. However, to recruit human resources for the task is often difficult. This paper discusses a microtask-based crowdsourcing approach to the problem. An important issue is to design a good strategy for generating tasks to be assigned to workers, maintaining the quality and reducing the number of tasks. In this study, we explore a design space defined by two criteria to reduce the number of assigned microtasks for finding misidentifications caused by automatic identification techniques. We compare four task-generation strategies using bibliographic records of the National Diet Library. One of the strategies reduced 55.7% of tasks from the baseline strategy and statistic analysis showed that the quality of its result is comparable to those of the other three strategies.published or submitted for publicationis peer reviewe

A New Device Facilitating Intracorporeal Purse-string Suture during Endoscopic Surgery

Standard laparoscopic colorectal surgery requires additional incision or enlargement of the trocar incision for the retrieval of the surgical specimen. A natural orifice specimen extraction （NOSE） procedure, in which the specimen is retrieved through the anus or vagina without any additional skin incision, requires purse-string suture （PSS） of the rostral intestinal segment in order to fix the anvil head of the stapler and perform extracorporeal mechanical anastomosis. Colorectal surgery has a limited NOSE in cases where the end of the rostral segment could be pulled through the anus. Broader application of NOSE depends on intracorporeal PSS. We developed a new forceps for intracorporeal PSS during NOSE and evaluated its efficacy. The PSS instrument was refined to pass through a 12-mm trocar in an intracorporeal PSS and achieve anastomosis using double stapling. In trials utilizing an endoscopic practice box, regular spacing of stitches during PSS were consistent （n＝10）, and tight intracorporeal anastomosis of the porcine colon was successfully performed （n＝2）. We then confirmed efficacy through an operation on a pig. Our novel PSS device will help us perform NOSE not only in laparoscopic colorectal surgery but also in any operation requiring intracorporeal PSS, which should contribute to further advances in endoscopic digestive surgery

品川区「市民科」教科書の政治学的分析

第1節序論 1シティズンシップへの関心の高まり 2 品川区「市民科」の取り組み 3 問題設定 第2節品川区「市民科」教科書の量的分析 1 1・2年生 2 3・4年生 3 5・6・7年生 4 8・9年生 5 「市民科」カリキュラムの流れ 6 各指導領域が想定する「市民性」 第3節中間考察 第4節品川区「市民科」教科書の質的分析 1 自治的活動能力 2 道徳実践能力 3 社会的判断・行動能力 4 自治的活動領域で育成される能力 5 非政治性の政治性 第5節結論法政研究　別

三叉神経運動枝除神経モデルにおけるラットとマウスのアポトーシス関連分子活性の差異

Introduction / Materials and Methods / Results / Discussion / ReferencesSubmitted by 真弓 小柳 ([email protected]) on 2012-05-08T07:56:29Z No. of bitstreams: 1 dent559.pdf: 1119414 bytes, checksum: f217eae29f9f04b92ef0e4babf8e4dbf (MD5)Made available in DSpace on 2012-05-08T07:56:29Z (GMT). No. of bitstreams: 1 dent559.pdf: 1119414 bytes, checksum: f217eae29f9f04b92ef0e4babf8e4dbf (MD5) Previous issue date: 2012-03-27歯学府_歯学[Background] We previously developed a rat trigeminal motor neuron axotomy model involving masseter and temporal muscle resection to study pathological changes of the central nucleus after peripheral nerve injury caused by oral surgery. Because motor neurons are reported to be more vulnerable to axotomy in mice than rats, we compared the degeneration process of the trigeminal motor nucleus in the rat model with a similar mouse model. [Methods] We removed masseter and temporal muscles of adult mice or rats. Animals were sacrificed at 3, 7, 14, 28, 42, and 56 days post-operation, and the trigeminal motor nuclei were histologically analyzed. [Results] Size reduction, but no neuronal loss, was seen in the trigeminal motor nuclei in both mice and rats. Time-dependent Noxa expression, starting at 1 week post-operation (wpo), was seen in the mouse model. By 8 wpo, mice expressed a higher level of Noxa than rats. Additionally, we noted persistent expression of cleaved caspase 3 in mice but not rats. Conversely, apoptosis-inducing factor (AIF), which executes DNA fragmentation in the nucleus, was not translocated to the nucleus in either model. [Conclusions] Our findings indicate differential activation of motor neuron apoptosis pathways after axotomy in mice and rats. Lack of activation of caspase-independent pathways and distal-end denervation in our model might be related to the survival of motor neurons after axonal injury. These findings could be relevant to future neuroprotective strategies for peripheral nerve injury caused by oral surgeries