Preparation of Polymer-Conjugated Bilirubin Oxidase and Treatment of Jaundice

本研究旨在阐明细菌蛋白酶在流感感染中的作用，详细分析流感感染的致病过程。所使用的蛋白酶是从粘质沙雷氏菌中获得的，作为细菌感染的模型。当小鼠感染流感病毒时，与未添加蛋白酶的小鼠相比，添加蛋白酶的小鼠肺部病毒显著增加（约100倍）。这在小鼠的生存中被发现是一个协同结果；所有的老鼠都以更快的速度死亡，其中大约40%的老鼠在没有蛋白酶的情况下死亡。下一个问题是流感感染中的致病分子是什么；病毒本身或其他间接后果。我们关注了细支气管肺泡灌洗液（BALF）中自由基的产生。令我们惊讶的是，黄嘌呤氧化酶（OX）的水平比对照高了400倍。XO酶是一种以低氧自由基或黄嘌呤为底物产生O_2^-和OH自由基的酶。另一种O_2^-生成源NADPH氧化酶水平在BALF中最多升高7倍。然后，我们研究了次黄嘌呤或黄嘌呤生成的上游级联；它是由腺苷脱氨酶将腺苷脱氨后产生的肌苷。未感染小鼠的腺苷脱氨酶活性升高20倍；这是增加XO燃料供应的确凿证据。然后，我们测试了去除O_2^-的效果，治愈90%的病毒感染小鼠，否则全部死亡。别嘌呤醇对XO的抑制有利于小鼠的存活，这一事实进一步证实了这一发现。黄嘌呤脱氢酶产生XO是通过丝氨酸蛋白酶介导的，丝氨酸蛋白酶可以被大豆胰蛋白酶抑制剂抑制。这种抑制剂对病毒感染小鼠的生存也有效。该抑制剂可能还能有效抑制BALF中丝氨酸蛋白酶对病毒的激活，并抑制缓激肽的产生，缓激肽是血管通透性和炎症的主要原因。少

The purpose of the present investigation is to clarify the role of bacterial protease in the influenza infection and the detailed analysis pathogenic process in influenza infection. The protease used was obtained from Serratia marcescens as a model of bacterial infection. When mice were infected witt influenza virus, addition of the protease yielded remarkable increase (about 100 fold) of virus in the lung of the infected mice compared with those without protease. This was found as a synergistic outcome in the survival of mice ; all mice died at much accelerated rate, where about 40% died without the protease. Next question is that what is the pathogenic molecules in influenza infection ; virus itself or other indirect consequence. We focused free radical generation in the Bronchiol Alveolar Lavage Fluid (BALF). To our surprise the level of xanthiee oxidase (OX) was elevated 400 fold higher than that of control. XO is an enzyme which generate O_2^- as well as OH radicals using hypoxant … More hine or xanthine as its substrate. The level of NADPH oxidase, another source of O_2^- generation was elevated at most 7 fold in BALF. Then, we investigated upstream cascadeof hypoxanthine or xanthine generation ; which is inosine derived after deamination of adenosine by adenosine deaminase.The adenosine deaminase activity was elevated 20 fold of uninfected mice ; a solid evidence of enhanced supply of fuel for XO. Then, we tested effect of removal of O_2^- cure 90% of virus infected mice otherwise all died. This finding was further strengthened by the fact that inhibition of XO by allopurinol was beneficial to the survival of mice. Generation of XO from xanthine dehydrogenase is mediated via serine protease which can be inhibited by soybean trypsin inhibitor. The effect of this inhibitor was found also effective to the viral infected mice for their survival. This inhibitor may be effective also inhibiting activation of virus by serine protease in BALF and also inhibiting kallikrein which generate bradykinin a major cause of vascular permeability and inflammation. Less