Proteases,actinidin, papain and trypsin reduce oral biofilm on the tongue in elderly subjects and in vitro

ObjectiveDental plaque is a causative factor for oral disease and a potential reservoir for respiratory infection in the elderly. Therefore, there is a critical need for the development of effective methods to remove oral biofilm. The objective of this study was to investigate the effect of proteases on oral biofilm formation andremoval.DesignThe in vivo effect of actinidin, a cysteine protease, on the removal of tongue coating was assessed after orally taking a protease tablet. Effects of the proteases trypsin, papain and actinidin on Actinomyces monospecies biofilm and multispecies biofilm that was reconstructed using a plaque sample from the tongue coating were investigated using the microtiter plate method. Antimicrobial tests and limited proteolysis of fimbrial shaft proteins were also performed to clarify underlying mechanisms of oral biofilm removal.ResultsTablets containing actinidin removed tongue coating in elderly subjects. Oral Actinomyces biofilm was significantly reduced by the proteases papain, actinidin and trypsin. Papain and trypsin effectively digested the major fimbrial proteins, FimP and FimA, from Actinomyces. Actinidin, papain and trypsin reduced multispecies biofilm that was reconstructed in vitro. Papain and trypsin inhibited formation of multispecies biofilm in vitro.ConclusionsThis study shows that proteases reduced oral biofilm in vivo in elderly subjects and in vitro, and suggests that protease digests fimbriae and inhibits biofilm formation.     

不同来源木瓜蛋白酶合成Aspartame前体能力的研究

研究了Ｓｉｇｍａ公司、Ｍｅｒｃｋ公司和广州远天酶制剂厂生产的木瓜蛋白酶和从木瓜青果中提取的木瓜蛋白酶在水－乙酸乙酯两相反应系统中合成二肽甜味剂Ａｓｐｅｒｔａｍｅ前体（Ｐ－ＡＰＭ）的能力，并以ＨＰＬＣ（流动相为乙腈－水＝４５：５５，检测波长２６０ｎｍ，流速１ｍｌ／ｍｉｎ）和ＴＬＣ（硅胶ＧＦ＿（２５４）板，展开剂为正丁醇－醋酸－水＝３：１：１，显色剂为１％茚三酮丙酮液）对Ｐ－ＡＰＭ进行检测，实验结果表明，Ｐ－ＡＰＭ与以嗜热蛋白酶催化合成的Ａｓｐａｒｔａｍｅ中间产物极为相似。不同来源的木瓜蛋白酶由于其酶活力不同，对Ｐ－ＡＰＭ的合成和水解能力也有差异，选择国产的木瓜蛋白酶合成Ａｓｐａｒｔａｍｅ，为酶法合成二肽甜味剂提供了可探讨的途径。     

不同浓度木瓜蛋白酶建立兔膝骨关节炎模型的比较研究

目的:比较不同浓度木瓜蛋白酶建立的兔膝骨关节炎模型,为研究骨关节炎病理及诊疗方法提供数据支持。方法:将60只新西兰大耳白兔随机分为4组,分别在1、3、5d右膝关节腔注射2%(低剂量组)、5%(中剂量组)、10%(高剂量组)(w/v)木瓜蛋白酶和0.03mol/L的L-半胱氨酸混合溶液0.1ml/kg(实验组),以及等量生理盐水(对照组)。整个实验过程观察兔状态并记录体重和膝关节宽度。首次注射后第2、4、6周将兔分批处死,取股骨髁、胫骨平台及滑膜做大体及组织病理学观察,并进行评分。结果:实验组均出现关节宽度增大、关节软骨变薄、基质分解破坏、滑膜增生等炎症及退行性改变,严重程度随浓度增高而增高,也随时间延长而发展;对照组无上述改变,二者比较差异有统计学意义。结论:兔膝骨关节腔注射2%、5%、10%木瓜蛋白酶和0.03mol/LL-半胱氨酸混合溶液0.1ml/kg可制备不同严重程度的骨关节炎模型,周期短,可重复且成功率高。     

Effects of nerve growth factor and basic fibroblast growth factor on survival of cultured septal cholinergic neurons from adult rats

We have established a primary culture technique for neuronal cells from rat basal forebrain from postnatal day 58 (P58) to study the effects of neurotrophic factors on the neurons. The survival of acetylcholinesterase (AChE)-positive neurons of 2-week-old rat septum has already been reported to be strongly supported by nerve growth factor (NGF) in culture. In this culture study of neurons from adult rat brains, the survival of AChE-positive neurons from P58 rat septum was slightly improved by NGF, although low affinity NGF receptor expression was also observed on cultured P58 rat septum neurons as well as on those from 2-week-old rats. The addition of basic fibroblast growth factor (bFGF) improved the survival of AChE-positive neurons cultured from P58 rat septum, but did not promote the survival of neurons from P12 rat septum. These results suggest that NGF changes to a maintenance factor in adult rat brain from a survival factor in postnatal 2-week-old rats. The survival of cholinergic neurons in culture of adult rat septum might be supported by factor(s) other than NGF, such as bFGF.     

木瓜蛋白酶及胰蛋白酶所致大鼠肺气肿模型的差异及对比

目的研究木瓜蛋白酶及胰蛋白酶所致大鼠肺气肿模型的差异。方法采用猪胰蛋白酶和木瓜蛋白酶气管内滴注的方法,将体质量170～220 g的60只SPF级SD大鼠随机分为正常对照组(A组),猪胰蛋白酶组（B组）,木瓜蛋白酶组（C组）,各20只,B组、C组气管内分别滴入猪胰蛋白酶1 U/g、5％木瓜蛋白酶0.001 ml/kg造模...     

Nutritional composition of Rainbow papaya, the first commercialized transgenic fruit crop

Rainbow papaya (Carica papaya L.) is a genetically engineered (GE) cultivar with resistance to papaya ringspot virus (PRSV). This cultivar currently accounts for about 70% of Hawaii's papaya acreage. The nutritional composition of Rainbow papaya and a non-transgenic control were analyzed to address GE food safety concerns regarding the potential for altered nutritional composition and altered expression of inherent allergens and toxic proteins. Rainbow papaya fruit were analyzed at three ripening stages and the data compared to that of a non-transgenic papaya which shares a similar pedigree. No differences were observed between GE and non-GE papaya for 36 nutrients at any of the tested fruit ripeness stages. However, vitamin A was higher and calcium levels were lower in the GE fruit. The GE fruit showed higher levels of protein and papain at the earliest stage of ripening (color break), but in ripened fruit these differences were insignificant. Benzyl isothiocyanate (BITC) levels were very low and similar for both Rainbow and the non-transgenic control fruit at all ripeness stages. Our data show that the contents of nutrients, BITC and papain of GE Rainbow papaya are within the range of those of non-GE papaya and that the Rainbow cultivar is substantially similar to the non-GE cultivar.     

血型不规则抗体检测的方法学评价

目的寻找简便、快速、敏感的用于红细胞不规则抗体筛选的试验方法。方法收集6例临床疑难病例样本和1份IgG型试剂,采用盐水法、木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法平行检测患者血清中不规则抗体的效价,比较5种方法的敏感性、特异性。结果盐水法仅检出IgM类不规则抗体;木瓜酶法、凝聚胺法、抗人球蛋白法、微柱凝胶法在检测IgG类抗体效价时差异不大,木瓜酶法的敏感性略低,微柱凝胶法对弱阳性IgG抗体检测的敏感性最高,但4种方法间差异无统计学意义（P〉0.05）,相关性较好（r〉0.99）。结论盐水法不能检出IgG抗体;木瓜酶法检测IgG抗体具有一定的局限性;凝聚胺法简便、快速,但为手工操作,不易标准化;抗人球蛋白法需要反复洗涤红细胞,费时,不便于开展大规模筛查;微柱凝胶法操作简便迅速、敏感性高、重复性好、易于标准化,适于临床输血前和孕妇产前大批量抗体筛选,是检测IgG抗体最为敏感的方法。     

木瓜酶造成软骨损伤的实验研究

本文观察了通过不同方法注射木瓜酶所引起的软骨损伤。软骨损伤的部位及程度因给药方式的差别而不同。在描述了各自特点的同时,本文着重观察了腹腔注射木瓜酶后骺板软骨损伤和修复过程。初步认为,这种方法效果稳定且简便易行,在某些骨、软骨疾病研究中具有一定的实用价值。     

Thiirancarboxamides as inhibitors of papain

Derivatives of the thiirancarboxylic acid building-block containing a peptide bond were synthesised and screened against the model cysteine protease papain. The most active of the series showed a second-order rate constant of inactivation comparable to that of the parent compound. The insertion of a peptide moiety seems to compensate the lack of a free carboxylate interacting with the histidinium ion at the enzyme's active site.     

木瓜蛋白酶对补骨脂有效成分煎出的影响

利用木瓜蛋白酶预处理补骨脂粉后煎提，与普通煎煮法比较，煎液总氮量提高了２倍，磷脂提高了４倍；补骨脂素、异补骨脂素提高了近１倍。结果提示，蛋白酶水解蛋白质的同时，促进了磷脂的溶出，发挥增溶作用，最终导致各种有效成分煎出量增加。