BACKGROUND & AIMS: :Polymer-associated infections are a major problem in implanted or intravascular devices. Among others, microorganisms of the staphylococcal family have been identified as the most important culprit. Prevention of bacterial adhesion and colonization of polymeric surfaces by release of antimicrobial agents incorporated into the polymers itself are currently under study. We have developed a novel method for the functionalization of a polymeric surface which is based on the deposition of covalently coupled lipid structures from antibiotic loaded vesicles. We have found that such process significantly reduces the bacterial growth on polystyrene material. In this work, lipid coverage obtained from multilamellar (MLVs) and extruded unilamellar (LUVs) vesicles were analyzed with respect to their adhesion efficiency on three types of polystyrene (PS) well-plates. Two methods of lipid deposition were characterized and compared in terms of surface lipid density and time stability: deposition of cationic vesicles on negatively charged surfaces and formation of covalent linkages between functionalized lipids and amines enriched surfaces. In order to study the antibiotic encapsulation efficiency we measured how the rifampicin (RIF) loading was affected by changes of liposome charge upon introduction of various amounts of stearylamine (SA), distearoyl-trimethylammonium propane (DSTAP) or dioleoyloxypropyl-trimethylammonium chloride (DOTAP) into the liposomal formulation. RIF-coated polymeric surfaces were also tested against a Staphylococcus epidermidis strain to evaluate their efficacy in vitro, showing that only approximately 2% of such bacteria inoculated on MLV-treated PS substrate were able to proliferate. Covalently immobilized lipid films showed about a tenfold increase in time stability compared to electrostatically bonded lipid films. Furthermore, substrates covalently modified with RIF-loaded MLVs retained an antibacterial activity for up to 12 days when aged in buffer at 37 degrees C. Such antimicrobial polymer coatings show promise for their use as antibacterial barrier for the prevention of catheter-related infections.

背景与目标： ：与聚合物相关的感染是植入式或血管内装置的主要问题。其中，葡萄球菌家族的微生物已被确定为最重要的罪魁祸首。目前正在研究通过释放掺入到聚合物本身中的抗微生物剂来防止细菌粘附和聚合物表面定居。我们已经开发了一种用于聚合物表面功能化的新方法，该方法基于从载有抗生素的囊泡中共价偶联的脂质结构的沉积。我们已经发现，这种方法显着减少了细菌在聚苯乙烯材料上的生长。在这项工作中，分析了从多层（MLV）和单层挤出（LUV）囊泡获得的脂质覆盖度，分析了它们在三种类型的聚苯乙烯（PS）孔板上的粘附效率。表征了两种脂质沉积的方法，并根据表面脂质密度和时间稳定性进行了比较：阳离子小泡在带负电荷的表面上的沉积以及功能化脂质和富含胺的表面之间形成共价键。为了研究抗生素的包封效率，我们测量了引入各种量的硬脂胺（SA），二硬脂酰三甲基铵丙烷（DSTAP）或二油酰氧基丙基三甲基氯化铵（DOTAP）时脂质体电荷的变化如何影响利福平（RIF）的负载量进入脂质体制剂。还用RIF涂层的聚合物表面针对表皮葡萄球菌菌株进行了测试，以在体外评估其功效，结果表明，仅约2％的此类细菌接种在MLV处理的PS底物上能够繁殖。与静电结合的脂质膜相比，共价固定的脂质膜显示出约10倍的时间稳定性。此外，当在37摄氏度的缓冲液中老化时，用RIF负载的MLV共价修饰的底物在长达12天的时间内仍保留了抗菌活性。此类抗菌聚合物涂层有望将其用作预防导管相关感染的抗菌屏障。

BACKGROUND & AIMS: :Klebsiella oxytoca (NRRL-B199), although able to produce 2, 3-butanediol from glucose, converted lactose mainly into acetic acid. By addition of a preparation of lactase (beta-galactosidase, EC 3.2.1.23), the fermentation of lactose in a stirred vessel was three-times faster and resulted in a high concentration of 2, 3-butanediol. The lactase confined in dead cells of Kluyveromyces lactis (CBS 683) was prepared by permeabilization with solvents and fixation with glutaraldehyde. The cells were coimmobilized by adhesion to glass wool after treatment of the latter with chitosan, which ensured cell-support electrostatic attraction. The cell loading (dry weight) was ca. 9 gL(-1) for the yeast and ca. 2 gL(-1) for the bacteria. In the presence of culture medium, the adhesion of both cells was stable and the bacteria tended to form biofilms. The stability of the coimmobilized cells was demonstrated by the continous conversion of lactose into 2, 3-butanediol at 30oC during 25 days. The coimmobilization system gave output concentrations (14 gL(-1)) and rate of production (1 gL(-1) h(-1)) of 2, 3-butanediol from lactose, similar to those obtained in the literature with immobilized cells and glucose. Compared to the literature data on direct conversion of lactose using pure cultures, the present results showed higher butanediol concentrations and 10 to 100 times higher rates of production.

背景与目标： 产酸克雷伯氏菌（KRLbsiella oxytoca）（NRRL-B199）尽管能够从葡萄糖中产生2,3-丁二醇，但主要将乳糖转化为乙酸。通过添加乳糖酶制剂（β-半乳糖苷酶，EC 3.2.1.23），在搅拌容器中进行乳糖发酵的速度加快了三倍，并产生了高浓度的2，3-丁二醇。限制在乳酸克鲁维酵母（CBS 683）死细胞中的乳糖酶是通过溶剂渗透和戊二醛固定制备的。在用壳聚糖处理玻璃棉后，通过将其粘附到玻璃棉上来共同固定细胞，从而确保细胞支持静电吸引。电池负荷（干重）约为1。酵母约9 gL（-1），细菌2 gL（​​-1）。在培养基的存在下，两个细胞的粘附是稳定的，细菌倾向于形成生物膜。共固定细胞的稳定性通过将乳糖在30oC下连续25天连续转化为2，3-丁二醇来证明。共固定系统产生了乳糖2,3,3-丁二醇的输出浓度（14 gL（-1））和产生速率（1 gL（-1）h（-1）），与文献中固定细胞获得的相似和葡萄糖。与使用纯培养物直接转化乳糖的文献数据相比，目前的结果显示出更高的丁二醇浓度和更高的10至100倍的生产率。

BACKGROUND & AIMS: :Fermentation rates and intracellular compositions have been determined for alginate-entrapped Saccharomyces cerevisiae and for identical cells in suspension. Glucose uptake and ethanol and glycerol production are approximately two times faster in immobilized cells than in suspended cells. Phosphorus-31 nuclear magnetic resonance (NMR) spectroscopy of fermenting immobilized and suspended cells shows differences in intermediate metabolite levels such as fructose-1,6 diphosphate, glucose-6-phosphate, and 3-phosphoglycerate and in internal pH. Carbon-13 NMR shows an increase in polysaccharide production. These data suggest that immobilization has accelerated the rate of glucose transport or of glucose phosphorylation. These effects of immobilization upon cell metabolism are observed in a very short period of time under conditions in which negligible DNA, RNA, or protein synthesis takes place.

背景与目标： ：已确定了藻酸盐捕获的酿酒酵母和悬浮细胞中相同细胞的发酵速率和细胞内组成。固定细胞中的葡萄糖吸收以及乙醇和甘油的产生比悬浮细胞中快约两倍。发酵固定化和悬浮细胞的磷31核磁共振波谱显示中间代谢物水平（如果糖1,6二磷酸，葡萄糖6磷酸和3-磷酸甘油酸）和内部pH的差异。碳13 NMR显示多糖产量增加。这些数据表明固定化加速了葡萄糖转运或葡萄糖磷酸化的速率。固定化对细胞代谢的这些作用在很短的时间内，在发生可忽略的DNA，RNA或蛋白质合成的条件下观察到。

BACKGROUND & AIMS: :The enzyme penicillin G acylase (PGA) is not adsorbed at pH 7 on DEAE- or PEI-coated supports, neither is it adsorbed on carboxymethyl (CM)- or dextran sulfate (DS)-coated supports. The surface of the enzyme was chemically modified under controlled conditions: chemical amination of the protein surface of carboxylic groups (using soluble carbodiimide and ethylendiamine) and chemical succinylation (using succinic anhydride) of amino groups. The full chemical modification produced some negative effects on enzyme stability and activity, although partial modification (mainly succinylation) presented negligible effects on both enzyme features. The chemical amination of the protein surface permitted the immobilization of the enzyme on CM- and DS-coated support, while the chemical succinylation permitted the enzyme immobilization on DEAE- and PEI-coated supports. Immobilization was very strong on these supports, mainly in the polymeric ones, and dependent on the degree of modification, although the enzymes still can be desorbed after inactivation by incubation under drastic conditions. Moreover, the immobilization on ionic polymeric beds allowed a significant increase in enzyme stability against the inactivation and inhibitory effects of organic solvents, very likely by the promotion of a certain partition of the organic solvent out of the enzyme environment. These results suggest that the enrichment of the surface of proteins with ionic groups may be a good strategy to take advantage of the immobilization of industrial enzymes via ionic exchange on ionic polymeric beds.

背景与目标： ：青霉素G酰基转移酶（PGA）在pH 7时不吸附在DEAE或PEI包被的载体上，也不吸附在羧甲基（CM）或硫酸葡聚糖上（DS）的载体上。酶的表面在受控条件下进行了化学修饰：羧基的蛋白质表面化学胺化（使用可溶性碳二亚胺和乙二胺）和氨基的化学琥珀酰化（使用琥珀酸酐）。完全的化学修饰对酶的稳定性和活性产生了一些负面影响，尽管部分修饰（主要是琥珀酰化）对两种酶的特征影响可忽略不计。蛋白质表面的化学胺化使酶固定在CM和DS涂层的支持物上，而化学琥珀酰化使酶固定在DEAE和PEI涂层的支持物上。这些酶的固定化非常强，主要是在聚合的载体上，并且取决于修饰的程度，尽管在剧烈条件下孵育灭活后酶仍可以解吸。而且，固定在离子聚合物床上使得抵抗有机溶剂的失活和抑制作用的酶稳定性显着增加，这很可能是由于有机溶剂的一定分配的促进。这些结果表明，利用离子基团富集蛋白质表面可能是利用工业酶通过离子聚合床上的离子交换固定化酶的一种好策略。

BACKGROUND & AIMS: :Chondroitin-4-sulphate (ChS A) was immobilized by matrix assisted pulsed laser evaporation (MAPLE) with the aid of a UV KrF* excimer laser source. Distilled water was used as solvent for the preparation of the frozen composite MAPLE targets. The surface morphology, chemical structure and functional properties of laser transferred ChS A were investigated as a function of laser processing conditions. The results indicate that the amount of laser immobilized material, structure, and functional properties can be controlled by the laser fluence value used for the irradiation of the MAPLE targets. Under selected irradiation conditions besides the molecular structure, the functional properties of the laser processed ChS A molecules can be maintained.

背景与目标： ：借助紫外线KrF *受激准分子激光源，通过基质辅助脉冲激光蒸发（MAPLE）固定软骨素4-硫酸盐（ChS A）。蒸馏水用作制备冷冻复合材料MAPLE靶的溶剂。研究了激光转移的ChS A的表面形态，化学结构和功能特性随激光加工条件的变化。结果表明，激光固定材料的数量，结构和功能特性可以通过用于辐照MAPLE目标的激光通量值来控制。在选定的辐射条件下，除了分子结构外，还可以保持经过激光处理的ChS A分子的功能特性。

BACKGROUND & AIMS: :The remediation of heavy metal-contaminated soils has attracted increased attention worldwide. The immobilization of metals to prevent their uptake by plants is an efficient way to remediate contaminated soils. This work aimed to seek the immobilization of cadmium in contaminated soils via a combination method. Flask experiments were performed to investigate the effects of hydroxyapatite (HAP) and the Cupriavidus sp. strain ZSK on soil pH and DTPA-extractable cadmium. Pot experiments were carried out to study the effects of the combined amendment on three plant species. The results showed that HAP has no obvious influence on the growth of the strain. With increasing concentrations of HAP, the soil pH increased, and the DTPA-extractable Cd decreased. Via the combined amendment of the strain and HAP (SH), the DTPA-extractable Cd in the soil decreased by 58.2%. With the combined amendment of the SH, the cadmium accumulation in ramie, dandelion, and daisy decreased by 44.9%, 51.0%, and 38.7%, respectively. Moreover, the combined amendment somewhat benefitted the growth of the three plant species and significantly decreased the biosorption of cadmium. These results suggest that the immobilization by the SH combination is a potential method to decrease the available cadmium in the soil and the cadmium accumulation in plants.

背景与目标： ：重金属污染土壤的修复在全世界引起了越来越多的关注。固定金属以防止植物吸收金属是修复污染土壤的有效方法。这项工作旨在通过组合方法寻求将镉固定在受污染的土壤中。进行烧瓶实验以研究羟基磷灰石（HAP）和Cupriavidus sp。的作用。 ZSK菌株对土壤pH和DTPA可萃取镉的影响。进行盆栽试验以研究组合的改良剂对三种植物的影响。结果表明，HAP对菌株的生长没有明显的影响。随着HAP浓度的增加，土壤pH值升高，而DTPA可萃取的Cd降低。通过对菌株和HAP（SH）的联合改良，土壤中DTPA可萃取的Cd降低了58.2％。通过对SH的综合修正，麻，蒲公英和雏菊中的镉积累量分别下降了44.9％，51.0％和38.7％。此外，组合的改良剂在某种程度上有益于三种植物的生长，并显着降低了镉的生物吸收。这些结果表明，通过SH组合固定化是减少土壤中有效镉和植物中镉积累的潜在方法。

BACKGROUND & AIMS: :In this study, three pentachlorophenol (PCP) laboratory-spiked and one field-contaminated soil were amended with 2.0% char, humic acid (HA) and peat, respectively. The amended soils were aged for either 7 or 250 days. After amendment, CaCl(2) extractability of PCP was significantly decreased. Desorption kinetics indicated that the proposed amendment could lead to a strong binding and slow desorption of PCP in soils. Amendment with char reduced the bioaccumulation factor (BAF) of PCP most significantly for earthworms (Eisenia fetida) in all soils studied. The results of both physicochemical and biological tests suggested that amendment reduced PCP bioavailability quickly and enduringly, implying that carbonaceous material amendment, especially char amendment, was a potentially attractive in situ remediation method for sequestration of PCP in contaminated soil.

背景与目标： ：在这项研究中，分别用2.0％的炭，腐殖酸（HA）和泥炭对3种五氯酚（PCP）实验室加标的土壤和1种被田地污染的土壤进行了修正。修改过的土壤要老化7天或250天。修改后，PCP的CaCl（2）可萃取性显着降低。解吸动力学表明，拟议的修正可能导致土壤中五氯苯酚的牢固结合和缓慢解吸。在所有研究的土壤中，对炭黑的修正最大程度地降低了（（Eisenia fetida）的五氯苯酚的生物积累因子（BAF）。物理化学和生物学测试的结果均表明，改性剂快速而持久地降低了PCP的生物利用度，这意味着碳质材料改性剂（尤其是炭化剂改性剂）是一种潜在的有吸引力的原位修复方法，用于隔离受污染土壤中的PCP。

BACKGROUND & AIMS: :Herein, the zwitterionic material poly (carboxybetaine acrylamide) was grafted onto iron oxide to obtain biocompatible magnetic nanoparticles Fe3O4-pCBAA which were employed to immobilize enzymes. The nanocomplxes Fe3O4-pCBAA were characterized using scanning electron microscopy (SEM), dynamic light scattering (DLS), zeta potential, Fourier transform-infrared (FT-IR) spectra and energy dispersive X-ray spectrometry (EDX). The urease as a model enzyme was immobilized with the novel supports and the properties of immobilized urease were further investigated in comparison with the free urease counterpart. The immobilized urease exhibited excellent thermodynamic and chemical stability. Particularly, 60% of initial activity was remained after being stored at 70 °C for 2 h while the free urease only remained 30%. Besides, the relative activity of immobilized enzyme was 1.7 times that of free ones after disposed in ethanol and 2-propanol for 2 h, and 7 times in DMF. Moreover, immobilized urease retained >80% of its initial activity after 5 cycles. In addition, the immobilization carrier Fe3O4-pCBAA displayed famous biocompatibility, and the immobilized urease performed better in complex biological samples, which were >85% and <60% of its initial activity for the immobilized and dissociative urease, respectively, in 20% and 25% of serum. These results confirm that the nanoparticles Fe3O4-pCBAA are biofriendly and efficient supports for enzyme immobilization and potential for practical applications in bio-microenvironments.

背景与目标： ：在此，将两性离子材料聚（羧基甜菜碱丙烯酰胺）接枝到氧化铁上，获得生物相容性磁性纳米粒子Fe3O4-pCBAA，用于固定酶。使用扫描电子显微镜（SEM），动态光散射（DLS），ζ电势，傅立叶变换红外（FT-IR）光谱和能量色散X射线光谱（EDX）对Fe3O4-pCBAA纳米复合材料进行了表征。用新型载体固定了作为模型酶的脲酶，并与游离脲酶的对应物进行了比较，进一步研究了固定脲酶的性质。固定的脲酶表现出优异的热力学和化学稳定性。特别是，在70°C下保存2小时后，保留了60％的初始活性，而游离脲酶仅保留了30％。此外，固定化酶在乙醇和2-丙醇中放置2h后的相对活性是游离酶的1.7倍，在DMF中则为7倍。此外，固定的脲酶在5个循环后保留了其初始活性的80％以上。此外，固定化载体Fe3O4-pCBAA表现出著名的生物相容性，固定化脲酶在复杂的生物样品中表现更好，分别占固定化和解离脲酶初始活性的> 85％和<60％，分别为20％和70％。血清的25％。这些结果证实纳米颗粒Fe 3 O 4 -pCBAA是酶固定的生物友好且有效的载体，并且在生物微环境中的实际应用具有潜力。

BACKGROUND & AIMS: :In the present study, a facile functionalization of magnetic nanoparticles has been described for the immobilization of enzyme that offers many advantages for reuse and excellent efficiencies. The magnetic gold nanocomposites have been fabricated for the successful immobilization of an industrially important enzyme "papain". For immobilization of papain on magnetic gold nanocomposites, magnetic nanoparticles were modified with 3-(mercaptopropyl) trimethoxy silane (MPTS). Further, the citrate stabilized gold nanoparticles were chemisorbed on these thiol coated magnetic nanoparticles to fabricate the desired magnetic gold nanocomposites. Papain containing net positive charge (isoelectric point of papain=8.75) in PBS buffer (pH 7.4) has immobilized on the surface of the negatively charged magnetic gold nanocomposites through the ionic or electrostatic interaction. The Michaelis-Menten kinetic constant (K(m)) and maximum reaction velocity (V(max)) for free papain were 0.236×10(5) g ml(-1) and 4.08 g ml(-1)/s respectively whereas for immobilized papain, K(m) and V(max) values were 0.308×10(5) g ml(-1) and 5.4 g ml(-1)/s respectively. The loading amount of papain on magnetic gold nanocomposites was 54 mg/g support and the activity recovery of the immobilized papain reached to 47 (±5)% compared to native papain. The main advantage of this papain nanobiocatalyst is the easy isolation of enzyme from the reaction medium.

背景与目标： ：在本研究中，已描述了磁性纳米粒子的简便功能化，用于固定化酶，该酶具有许多可重复使用的优点和出色的效率。已经制造出磁性金纳米复合材料，以成功固定工业上重要的酶“木瓜蛋白酶”。为了将木瓜蛋白酶固定在磁性金纳米复合材料上，用3-（巯基丙基）三甲氧基硅烷（MPTS）修饰了磁性纳米粒子。此外，将柠檬酸盐稳定的金纳米颗粒化学吸附在这些硫醇涂覆的磁性纳米颗粒上，以制造所需的磁性金纳米复合材料。在PBS缓冲液（pH 7.4）中含有净正电荷的木瓜蛋白酶（木瓜蛋白酶的等电点= 8.75）已通过离子或静电相互作用固定在带负电荷的磁性金纳米复合材料的表面上。游离木瓜蛋白酶的Michaelis-Menten动力学常数（K（m））和最大反应速度（V（max））分别为0.236×10（5）g ml（-1）和4.08 g ml（-1）/ s固定化木瓜蛋白酶的K（m）和V（max）值分别为0.308×10（5）g ml（-1）和5.4 g ml（-1）/ s。木瓜蛋白酶在磁性金纳米复合材料上的负载量为54 mg / g载体，与天然木瓜蛋白酶相比，固定化木瓜蛋白酶的活性回收率达到47（±5）％。这种木瓜蛋白酶纳米生物催化剂的主要优点是易于从反应介质中分离出酶。

BACKGROUND & AIMS: :Immobilization reduces muscle performance, and despite these performance losses being associated with neural impairments little is known regarding adaptations in cortical properties. We utilized transcranial magnetic stimulation to assess changes in flexor carpi radialis (FCR) intracortical facilitation (ICF), and short- and long-interval intracortical inhibition (SICI and LICI) in healthy humans undergoing 3 weeks of immobilization. Measurements were obtained at rest and during contraction (15% intensity). Central activation and the Hoffman reflex (H-reflex) were also assessed. Strength decreased 43.2% +/- 6.1% following immobilization, and central activation also decreased (97.5% +/- 2.4% to 73.2% +/- 8.3%). No changes in ICF, SICI, or LICI were observed at rest; however, LICI was increased during contraction (67.5% +/- 6.9% to 53.1% +/- 6.7% of unconditioned response). The increase in LICI correlated with the loss of strength (r = -0.63). The H-reflex increased following immobilization. These findings suggest that immobilization increases intracortical inhibition during contraction, and this increase is primarily mediated by GABA(B) receptors.

背景与目标： ：固定会降低肌肉性能，尽管这些性能下降与神经损伤相关，但对皮层特性的适应性知之甚少。我们利用经颅磁刺激来评估接受固定3周的健康人的radial屈腕（FCR）皮质内促进作用（ICF）以及短时间隔和长时间隔皮质内抑制作用（SICI和LICI）的变化。在静止和收缩期间（15％强度）获得测量值。还评估了中枢激活和霍夫曼反射（H-反射）。固定后强度降低了43.2％/-6.1％，中枢激活也降低了（97.5％/-2.4％至73.2％/-8.3％）。静止时未观察到ICF，SICI或LICI的变化；但是，收缩期的LICI升高（无条件反应的67.5％/-6.9％至53.1％/-6.7％）。 LICI的增加与强度下降相关（r = -0.63）。固定后H反射增加。这些发现表明固定化在收缩过程中增加了皮层内抑制，并且这种增加主要是由GABA（B）受体介导的。

BACKGROUND & AIMS: :The aim of this work is to determine the effect of chronic immobilization stress on kinetics and dosimetry of 67Ga in a mouse model. A control group (CG) and a stress group (SG), each with 15 mice, were included in the study, and the latter group was subjected to a chronic immobilization stress model 2 h daily for 14 consecutive days. At day 13, 67Ga-citrate was administered intraperitoneally (11.24 ± 0.44 MBq) to each mouse. Then, sets of three mice were obtained sequentially at 24, 36, 48, 60 and 72 h, in which the radionuclide activity was measured with an activity counter. The 67Ga biokinetic data showed a fast blood clearance in the SG, with a mean residence time of 0.06 h. The calculated mean radiation absorbed doses were: liver (2.45 × 10-03 Gy), heart (3.17 × 10-04 Gy) and kidney (1.88 × 10-04 Gy) in the SG. The results show that stress reduced weight gain by approximately 13% and also increased adrenal gland weight by 26%. On the other hand, chronic stress accelerates 67Ga clearance after 24 h compared to normal conditions. It is concluded that murine organisms under chronic immobilization stress have higher gallium-67 clearance rates, decreasing the cumulated activity and absorbed dose in all organs.

背景与目标： ：这项工作的目的是确定长期固定应力对小鼠模型中67Ga动力学和剂量学的影响。对照组（CG）和压力组（SG）各自有15只小鼠，并且该组每天2h接受慢性固定应激模型，连续14天。在第13天，向每只小鼠腹膜内给予67Ga柠檬酸（11.24±0.44MBq）。然后，在24、36、48、60和72h依次获得三只小鼠的集合，其中用活性计数器测量放射性核素的活性。 67Ga的生物动力学数据显示SG中血液快速清除，平均停留时间为0.06 h。计算得出的平均辐射吸收剂量为：SG中的肝脏（2.45××10-03 Gy），心脏（3.17××10-04 Gy）和肾脏（1.88××10-04 Gy）。结果表明，压力使体重增加减少了约13％，肾上腺重量也增加了26％。另一方面，与正常情况相比，慢性应激会在24小时后加速67Ga的清除。结论是，在慢性固定压力下的鼠类生物具有较高的镓67清除率，从而降低了所有器官的累积活性和吸收剂量。

BACKGROUND & AIMS: :Thermophilic fungi produce thermostable enzymes which have a number of applications, mainly in biotechnological processes. In this work, we describe the characterization of a protease produced in solidstate (SSF) and submerged (SmF) fermentations by a newly isolated thermophilic fungus identified as a putative new species in the genus Myceliophthora. Enzyme-production rate was evaluated for both fermentation processes, and in SSF, using a medium composed of a mixture of wheat bran and casein, the proteolytic output was 4.5-fold larger than that obtained in SmF. Additionally, the peak of proteolytic activity was obtained after 3 days for SSF whereas for SmF it was after 4 days. The crude enzyme obtained by both SSF and SmF displayed similar optimum temperature at 50 degrees C, but the optimum pH shifted from 7 (SmF) to 9(SSF). The alkaline protease produced through solid-state fermentation (SSF), was immobilized on beads of calcium alginate, allowing comparative analyses of free and immobilized proteases to be carried out. It was observed that both optimum temperature and thermal stability of the immobilized enzyme were higher than for the free enzyme. Moreover, the immobilized enzyme showed considerable stability for up to 7 reuses.

背景与目标： ：嗜热真菌产生的热稳定酶主要在生物技术过程中具有许多应用。在这项工作中，我们描述了新分离的嗜热真菌在固态（SSF）和淹没（SmF）发酵中产生的蛋白酶的特性，该真菌被鉴定为Myceliophthora属中假定的新物种。在两个发酵过程中以及在SSF中，使用由麦麸和酪蛋白的混合物组成的培养基，评估了酶的产生速率，其蛋白水解产量是SmF的4.5倍。另外，对于SSF，蛋白水解活性的峰值在3天后获得，而对于SmF，蛋白水解活性的峰值是在4天之后。通过SSF和SmF获得的粗酶在50摄氏度时显示出相似的最佳温度，但最佳pH从7（SmF）变为9（SSF）。将通过固态发酵（SSF）产生的碱性蛋白酶固定在藻酸钙珠上，从而可以对游离的和固定的蛋白酶进行比较分析。观察到固定化酶的最佳温度和热稳定性均高于游离酶。此外，固定化酶显示出相当高的稳定性，最多可重复使用7次。

BACKGROUND & AIMS: OBJECTIVE:This study used a unilateral knee joint immobilization model in adult guinea pigs to test the hypothesis that retrograde degeneration of motor neurons in the spinal cord is the result of attenuation of knee joint activities. METHODS:A total of 32 adult guinea pigs were used and divided into 8 groups based on the duration of knee joint immobilization. Light microscopic studies of Nissl, nitric oxide synthase immunohistochemistry, horseradish peroxidase, and fast blue were carried out to examine the neurons in the spinal cord. Electron microscopy was also performed to examine the neurons and axons. RESULTS:After various periods of knee joint immobilization, a variety of features of motor neuronal degeneration were observed. Specific characteristics included gradual increases in the expressions of neuronal nitric oxide synthase and ultrastructural changes in affected motor neurons including reduction of cell organelles, indentation of the nuclear envelop, and small compact clumps of chromatin in the nuclei. Observation of the peripheral nerve (femoral nerve) also revealed demyelination alterations in some axons innervating the muscles of the knee joint. Interestingly, motor neuronal degenerative changes and demyelination were reversible after the knee joint immobilization was removed and knee joint activity was restored. These findings may assist in further development of models for spinal dysfunction such as the chiropractic subluxation complex. CONCLUSION:We conclude that motor neuronal degeneration in the spinal cord and axons in this study was the result of knee joint immobilization. Increases in motor neuronal nitric oxide-mediated oxidative stress level after reduction of target tissue activity may contribute to the mechanism for degenerative changes in the motor neurons in adult spinal cord of the guinea pig.

背景与目标： 目的：本研究使用成年豚鼠单侧膝关节固定模型来验证以下假设：脊髓中运动神经元的逆行变性是膝关节活动减弱的结果。
方法：共使用32只成年豚鼠，根据固定膝关节的时间分为8组。进行了Nissl，一氧化氮合酶免疫组织化学，辣根过氧化物酶和耐晒蓝的光学显微镜研究，以检查脊髓中的神经元。还进行电子显微镜检查以检查神经元和轴突。
结果：膝关节固定不同时期后，观察到运动神经元变性的各种特征。具体特征包括神经元一氧化氮合酶表达的逐渐增加和受影响的运动神经元的超微结构变化，包括细胞器的减少，核包膜的压痕以及核中染色质的小块状团块。对周围神经（股神经）的观察还发现，在支配膝关节肌肉的某些轴突中脱髓鞘改变。有趣的是，去除固定膝关节并恢复膝关节活动后，运动神经元变性和脱髓鞘是可逆的。这些发现可能有助于脊柱功能障碍模型的进一步开发，例如整脊半脱位复合体。
结论：我们得出结论，本研究中脊髓和轴突的运动神经元变性是膝关节固定的结果。靶组织活性降低后，运动神经元一氧化氮介导的氧化应激水平升高可能是豚鼠成年脊髓运动神经元退行性变化的机制。

BACKGROUND & AIMS: :The synthesis of cyclohexadiene and maleimide derivatives and their use for the functionalization of oligonucleotides and the coating of glass surfaces is reported. A method for the covalent attachment of diene or maleimide modified oligonucleotides to the coated glass surfaces via aqueous Diels-Alder reactions is presented.

背景与目标： ：报道了环己二烯和马来酰亚胺衍生物的合成及其在寡核苷酸的功能化和玻璃表面涂层中的用途。提出了一种通过水的Diels-Alder反应将二烯或马来酰亚胺修饰的寡核苷酸共价附接到涂覆的玻璃表面的方法。

BACKGROUND & AIMS: :A novel cell-free translation system is described in which template-mRNA molecules were captured onto solid surfaces to simultaneously synthesize and immobilize proteins in a more native-state form. This technology comprises a novel solid-phase approach to cell-free translation and RNA-protein fusion techniques. A newly constructed biotinylated linker-DNA which enables puromycin-assisted RNA-protein fusion is ligated to the 3' ends of the mRNA molecules to attach the mRNA-template on a streptavidin-coated surface and further to enable the subsequent reactions of translation and RNA-protein fusion on surface. The protein products are therefore directly immobilized onto solid surfaces and furthermore were discovered to adopt a more native state with proper protein folding and superior biological activity compared with conventional liquid-phase approaches. We further validate this approach via the production of immobilized green fluorescent protein (GFP) on microbeads and by the production and assay of aldehyde reductase (ALR) enzyme with 4-fold or more activity. The approach developed in this study may enable to embrace the concept of the transformation of 'RNA chip-to-protein chip' using a solid-phase cell-free translation system and thus to the development of high-throughput microarray platform in the field of functional genomics and in vitro evolution.

背景与目标： ：描述了一种新颖的无细胞翻译系统，其中模板mRNA分子被捕获到固体表面上，以更天然的形式同时合成和固定蛋白质。该技术包括用于无细胞翻译和RNA蛋白质融合技术的新型固相方法。将能够使嘌呤霉素辅助的RNA-蛋白质融合的新构建的生物素化的接头DNA连接到mRNA分子的3'末端，以将mRNA模板附着在链霉亲和素包被的表面上，并进一步实现翻译和RNA的后续反应-蛋白质在表面融合。因此，将蛋白质产物直接固定在固体表面上，并且与常规液相方法相比，还发现其具有更天然的状态，具有适当的蛋白质折叠和优异的生物学活性。我们通过在微珠上生产固定的绿色荧光蛋白（GFP）以及通过生产和测定具有4倍或更多活性的醛还原酶（ALR）酶来进一步验证这种方法。这项研究中开发的方法可能能够包含使用固相无细胞翻译系统将“ RNA芯片转化为蛋白质芯片”的概念，从而促进高通量微阵列平台的发展。功能基因组学和体外进化。