Simultaneous HPLC analysis of hydrophobic prodrugs and their hydrophilic metabolites in biological media without sample preparation. Application to the biotransformation studies of pronucleotides derived from 5-fluorouracil

Improvements of an "on-line cleaning" HPLC method for analysis of biological samples are presented: (i) the use of cleaning precolumns filled with hydrophobic stationary phases instead of the hydrophilic ones previously used to eliminate the biological matrix: (ii) the combination in the mobile phase of anionic and cationic pairing reagents in order to retain on the precolumn all the metabolites, whatever their hydrophilicity and ionicity are. Such modifications allowed to study the biotransformation of prodrugs of 5-Fluorouracil, designed to act as antitumoral pronucleotides.     

Does Competition for Clients Increase Service Quality in Cleaning Gobies?

In a biological market, members of one trading class try to outbid each other to gain access to the most valuable partners. Competition within class can thus force individuals to trade goods or services more cheaply, ultimately resulting in conflict (e.g. cheating) over the value of commodities. Cleaning symbioses among fish appear to be good examples of biological markets. However, the existence and effect of outbidding competition among either types of traders (cleaners or clients) have never been tested. We examined whether increasing competition among cleaning gobies ( Elacatinus spp.) for access to clients results in outbidding in the form of provision of a better cleaning service. On reefs where fish clients visited cleaning stations less frequently, and thus competition among cleaners was higher, cleaning gobies ingested fewer scales relative to the number of ingested parasites, i.e. they cleaned more honestly. This shift in cleaner behaviour towards greater honesty is consistent with a greater market value of access to clients in the face of competition among cleaners. However, this pattern could have also arisen as a result of differences in ectoparasite availability across reefs and therefore in value of the commodity offered by clients. Experimental manipulations will be required to determine whether cleaning service quality by cleaning gobies was enhanced solely because of competitive outbidding.     

Effect of two cleaning processes for bone allografts on gentamicin impregnation and in vitro antibiotic release

Bone allografts are a useful and sometimes indispensable tool for the surgeon to repair bone defects. Microbial contamination is a major reason for discarding allografts from bone banks. To improve the number of safe allografts, we suggest chemical cleaning of the grafts followed by antibiotic impregnation. Comparison of two chemical cleaning processes for bone allografts aiming for antibiotic impregnation and consequently delivery rates in vitro. Bone chips of 5–10 mm were prepared from human femoral heads. Two cleaning methods (cleaning A and cleaning B) based on solutions containing hydrogen peroxide, paracetic acid, ethanol and biological detergent were carried out and compared. After the cleaning processes, the bone chips were impregnated with gentamicin. Bacillus subtilis bioassay was used to determine the gentamicin release after intervals of 1–7 days. Differences were compared with non-parametric Mann–Whitney U tests. The zones of inhibition obtained from the bone grafts cleaned with both cleaning processes were similar between the groups. The concentration of the released antibiotic was decreasing gradually over time, following a similar pattern for both groups. The cleaning procedure A as well as the cleaning procedure B for bone allografts allowed the impregnation with gentamicin powder in the same concentrations in both groups. The delivery of gentamicin was similar for both groups. Both cleaning procedures were easy to be carried out, making them suitable for routine use at the bone banks.     

Advantages of using microbial technology over traditional chemical technology in removal of black crusts from stone surfaces of historical monuments

This study compares two cleaning methods, one involving an ammonium carbonate-EDTA mixture and the other involving the sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris ATCC 29579, for the removal of black crust (containing gypsum) on marble of the Milan Cathedral (Italy). In contrast to the chemical cleaning method, the biological procedure resulted in more homogeneous removal of the surface deposits and preserved the patina noble under the black crust. Whereas both of the treatments converted gypsum to calcite, allowing consolidation, the chemical treatment also formed undesirable sodium sulfate.     

稗草主要生物学特性及其防除

本文概述了稗草(Echinochlon spp．)的生物学特性及其防除效果。试验证明稗草萌发的最低温度为12℃,稗草种子在充足的水分条件下,当有效积温达33.8—49.5℃(生物零度为10℃)时,出现田间稗草的发生高峰期。提出消除稗草要从秧田入手,并应掌握在稗草3叶期以前、化学防除应以药效期相吻合,才能达到预期的效果。通过试验对9种除草剂进行了综合评价,实践证明化学防除配套技术和农业措施协调应相结合,能有效地控制稻田稗草的危害。     

A1M,an extravascular tissue cleaning and housekeeping protein

Alpha-1-microglobulin (A1M) is a small protein found intra- and extracellularly in all tissues of vertebrates. The protein was discovered 40 years ago and its physiological role remained unknown for a long time. A series of recent publications have demonstrated that A1M is a vital part of tissue housekeeping. A strongly electronegative free thiol group forms the structural basis of heme-binding, reductase, and radical-trapping properties. A rapid flow of liver-produced A1M through blood and extravascular compartments ensures clearing of biological fluids from heme and free radicals and repair of oxidative lesions. After binding, both the radicals and the A1M are electroneutral and therefore do not present any further oxidative stress to tissues. The biological cleaning cycle is completed by glomerular filtration, renal degradation, and urinary excretion of A1M heavily modified by covalently linked radicals and heme groups. Based on its role as a tissue housekeeping cleaning factor, A1M constitutes a potential therapeutic drug candidate in treatment or prophylaxis of diseases or conditions that are associated with pathological oxidative stress elements.     

Advantages of Using Microbial Technology over Traditional Chemical Technology in Removal of Black Crusts from Stone Surfaces of Historical Monuments

This study compares two cleaning methods, one involving an ammonium carbonate-EDTA mixture and the other involving the sulfate-reducing bacterium Desulfovibrio vulgaris subsp. vulgaris ATCC 29579, for the removal of black crust (containing gypsum) on marble of the Milan Cathedral (Italy). In contrast to the chemical cleaning method, the biological procedure resulted in more homogeneous removal of the surface deposits and preserved the patina noble under the black crust. Whereas both of the treatments converted gypsum to calcite, allowing consolidation, the chemical treatment also formed undesirable sodium sulfate.     

Flux enhancement for membrane filtration of bacterial suspensions using high-frequency backpulsing

A promising method for reducing membrane fouling during crossflow microfiltration of biological suspensions is backpulsing. Very short backpulses (0.1-1.0 s) have been used to increase the net flux for washed bacterial suspensions and whole bacterial fermentation broths. The net fluxes under optimum backpulsing conditions for the washed bacteria are approximately 10-fold higher than those obtained during normal crossflow microfiltration operation, whereas only a 2-fold improvement in the net flux is achieved for the fermentation broths. A theory is presented that is based on external fouling during forward filtration and nonuniform cleaning of the membrane during reverse filtration. The model contains an adjustable parameter which is a measure of the cleaning efficiency during backpulsing; the cleaning efficiency found by fitting the model to the experiments increases with increasing frequency and duration of the backpulses. The theory predicts an optimum backpulsing frequency, as was observed experimentally. An economic analysis shows that crossflow microfiltration with backpulsing has lower costs than centrifugation, rotary vacuum filtration, and crossflow microfiltration without backpulsing.     

Quantifying the cleanliness of glass capillaries

I used capillary rise methods to investigate the lumenal surface properties of quartz (fused silica, Amersil T-08), borosilicate (Corning 7800), and high-lead glass (Corning 0010) capillaries commonly used to make patch pipets. I calculated the capillary rise and contact angle for water and methanol from weight measurements. The capillary rise was compared with the theoretical maximum value calculated by assuming each fluid perfectly wetted the lumenal surface of the glass (i.e., zero contact angle, which reflects the absence of surface contamination). For borosilicate, high-lead, and quartz capillaries, the rise for water was substantially less than the theoretical maximum rise. Exposure of the borosilicate, lead, and quartz capillaries to several cleaning methods resulted in substantially better—but not perfect—agreement between the theoretical maximum rise and calculated capillary rise. By contrast, the capillary rise for methanol was almost identical in untreated and cleaned capillaries, but less than its theoretical maximum rise. The residual discrepancy between the observed and theoretical rise for water could not be improved on by trying a variety of cleaning procedures, but some cleaning methods were superior to others. The water solubility of the surface contaminants, deduced from the effectiveness of repeated rinsing, was different for each of the three types of capillaries examined: Corning 7800>quartz>Corning 0010. A surface film was also detected in quartz tubing with an internal filament. I conclude that these borosilicate, quartz, and high-lead glass capillaries have a film on the lumenal surface, which can be removed using appropriate cleaning methods. The surface contaminants may be unique to each type of capillary and may also be hydrophobic. Two simple methods are presented to quantitate the cleanliness of glass capillary tubing commonly used to make pipets for studies of biological membranes. It is not known if the surface film is of importance in electrophysiological studies of biological membranes.     

Characterization of biosurfactants and their use in pollution removal – State of the Art. (Review)

Surface-active compound of biological origin (biosurfactants) have only been described in the past few decades. With the advantage of biodegradability and production on renewable resources, biosurfactants have been gaining prominence and their applications are becoming wider. So far, literature contains mixed reports on the successes of the applications of biosurfactants and their economical viability. They remain compounds which are not very well understood, yet, with several important applications. The target industries for biosurfactant use are the petroleum remediation industries and environmental conservation agencies. These industries, however, seem reluctant to use them for fear of dealing with microbes or microbial products. This includes cleaning up oil spills from the environment, remediation of metal-contaminated soils or waste streams, mobilizing heavy oil sludge and enhanced oil recovery. The importance of biosurfactants, their production, characteristics and limited successes and applications in oil pollution remediation and oil storage tank cleaning are discussed.     

Enzyme‐enabled responsive surfaces for anti‐contamination materials

Many real‐life stains have origins from biological matters including proteins, lipids, and carbohydrates that act as gluing agents binding along with other particulates or microbes to exposed surfaces of automobiles, furniture, and fabrics. Mimicking naturally occurring self‐defensive processes, we demonstrate in this work that a solid surface carrying partially exposed enzyme granules protected the surface in situ from contamination by biological stains and fingerprints. Attributed to the activities of enzymes which can be made compatible with a wide range of materials, such anti‐contamination and self‐cleaning functionalities are highly selective and efficient toward sticky chemicals. This observation promises a new mechanism in developing smart materials with desired anti‐microbial, self‐reporting, self‐cleaning, or self‐healing functions. Biotechnol. Bioeng. 2013; 110: 1805–1810. © 2013 Wiley Periodicals, Inc.     

Oviposition behavior of Phymastichus coffea LaSalle (Hymenoptera: Eulophidae) on its natural host

Phymastichus coffea LaSalle is an African endoparasitoid of adults of the coffee berry borer, Hypothenemus hampei (Ferrari) that was introduced into Mexico for biological control purposes. The objective of this study was to investigate the oviposition behavior of P. coffea on its host under laboratory conditions. The results indicate that P. coffea females exhibited 12 behavioral acts during and after oviposition, such as, walking, cleaning, flying, surface landing, resting, host landing, host antennation, touching the host with forelegs, host recognition, walking on the host, oviposition and cleaning after oviposition. The sequence of each behavior is described in an ethogram.     

Efficacy of Conventional Endoscopic Disinfection and Sterilization Methods Against Helicobacter pylori Contamination

Background. Iatrogenic transmission of Helicobacter pylori via contaminated endoscopic devices is well documented. Despite the prevalence of this infectious agent, few controlled studies have investigated the major factors that impact on reprocessing of endoscopes contaminated with H. pylori. Materials and Methods. An endoscope (Pentax) was contaminated with 10⁸ cfu/ml of H. pylori in 5% bovine calf serum as standardized inoculum. The endoscope then was passed through one of eight arms (five repetitions per arm = 40 total runs), as follows: 1, recovery control (no cleaning or disinfection); 2, manual cleaning alone; 3–5, manual precleaning followed by either 10-, 20-, or 45-minute exposure to 2% glutaraldehyde and ethanol (ETOH) drying; 6, manual cleaning followed by automated reprocessing by STERIS System; 7 and 8, automated reprocessing by STERIS with and without active peracetic acid sterilant (wash-off control). Suction-biopsy channels and air-water channels were harvested for microbiological culture. Results. Control runs recovered more than 1 × 10⁶ cfu per site, confirming the viability of the test organism and the adequacy of the biological burden for challenge. When instruments underwent manual cleaning alone (without subsequent disinfection), test organisms remained in 40% of runs at the air-water site. Manual cleaning followed by 10-, 20-, or 45-minute glutaraldehyde exposure and ETOH drying removed all test organisms from all sites in all runs (i.e., 100% disinfection). The automated STERIS system with or without active peracetic acid sterilant also removed all test organisms from all sites in all runs, as did manual cleaning followed by STERIS use. Conclusion. Manual cleaning alone does not effectively remove H. pylori from an endoscope. Current joint association recommendations for minimal disinfection (manual cleaning followed by at least 20 minutes of immersion in glutaraldehyde and ETOH drying) are effective in preventing cross-transmission of H. pylori. Reprocessing using the automated STERIS system according to manufacturer's recommendations also is highly effective in sterilizing endoscopes contaminated with H. pylori.     

石首鱼科鱼肌肉胆碱酯酶对有机磷化合物反应灵敏度的研究

胆碱酯酶对有机磷化合物的反应灵敏度高,特异性强,但不同来源的生物材料的胆碱酯酶在灵敏度上差异甚大。鲈形目石首鱼科的小黄鱼、大黄鱼、白姑鱼、鳘和黄姑鱼肌肉胆碱酯酶对沙林和ＶＸ均相当敏感,两种化合物对它们的胆碱酯酶抑制的ＰＩ５０值几乎均在７．０以上。进一步研究,这些胆碱酯酶可能成为分析有磷化合物和农药的有效试剂,对筛选生物活性物质有一定的指导意义。     

BACTERIOLOGICAL STUDIES OF IMMERSION CLEANING OF MILKING EQUIPMENT ON COMMERCIAL FARMS

SUMMARY: A study has been made of the immersion cleaning technique as applied on commercial farms. In an initial survey bacteriological results for rinses and swabs of the milking equipment were satisfactory on most farms, but on one farm the results were very disappointing. Further investigations at this farm showed that the poor results were caused partly by sporing bacteria which survived the caustic soda treatment, presumably in the spore state, and partly by non-sporing organisms, chiefly chromobacteria and micrococci.
These non-sporing organisms are found in abundance on the surface of dirty wash troughs, and under certain conditions re-contamination of the milking equipment can occur during the rinsing operation. Suggested modifications to the original immersion cleaning technique include the use of hot water for the before-milking hypochlorite rinse, brushing the equipment in the after-milking rinse to remove extraneous dirt, regular cleansing of the wash trough and the monthly cleansing of the basket and bin.     

肌萎缩性侧索硬化症相关的Cu,Zn-SOD

超氧化物歧化酶（superoxide dismutase,SOD）被称为生物体内自由基的清洁剂,其主要形式Cu,Zn-SOD称SOD1. SOD1突变体可引起致死性运动神经元疾病肌萎缩性侧索硬化症(ALS).但是,SOD1的毒性机理尚未完全清楚.本文概述了SOD1、Cu分子伴侣（copper chaperone for SOD1,CCS）的分子结构和CCS活化SOD1的机理,重点分析了突变体SOD1构象变化的原因及其在ALS中的可能致病机制的最新研究进展.     

An AFM determination of the effects on surface roughness caused by cleaning of fused silica and glass substrates in the process of optical biosensor preparation

The covalent attachment of organic films and of biological molecules to fused silica and glass substrates is important for many applications. For applications such as biosensor development, it is desired that the immobilised molecules be assembled in a uniform layer on the surface so as to provide for reproducibility and speed of surface interactions. For optimal derivatisation the surface must be appropriately cleaned to remove contamination, to create surface attachment sites such as hydroxyl groups, and to control surface roughness. The irregularity of the surface can be significant in defining the integrity and density of immobilised films. Numerous cleaning methods exist for fused silica and glass substrates and these include gas plasmas, and combinations of acids, bases and organic solvents that are allowed to react at varying temperatures. For many years, we have used a well established method based on a combination of washing with basic peroxide followed by acidic peroxide to clean and hydroxylate the surface of fused silica and glass substrates before oligonucleotide immobilisation. Atomic force microscopy (AFM) has been used to evaluate the effect of cleaning on surface roughness for various fused silica and glass samples. The results indicate that surface roughness remains substantial after use of this common cleaning routine, and can provide a surface area that is more than 10% but less than 30% larger than anticipated from geometric considerations of a planar surface.     

生态工程中食物链组合的环分析

本文研究了一个用于治理水体生态系统富营养化的生态工程,应用环分析方法对工程中的食物链的组合进行了分析。在这里,贝类被用于清除过量的藻类;种植水生植物改变营养的流转途径并且放养食植性鱼类,通过捕捞鱼和贝,使水体中的有机物和无机盐沿着从藻类到贝类和从水生植物到食植性鱼类两条途径由水体生态系统中输出,应用环分析的手段,可以调整生态工程中食物链的组合结构,使整个生态工程具有某种特定的功能。     

Cleaning characterization of protein drug products using UV-vis spectroscopy

This study uses on-line absorbance monitoring to evaluate cleanability of protein drug products. Characterization and validation of equipment cleanliness is a key requirement for a biopharmaceutical facility. A manufacturing-scale cleaning cycle has to be developed and validated for its ability to clean all of the equipment parts for a given soil. Cleaning validation in a multiproduct fill-finish facility could benefit from using a worst-case-based approach that involves validating the cleaning process for the most difficult to clean product. Such an approach minimizes the number of required validation runs. Scaled-down cleaning evaluations can provide helpful information for evaluating multiple products and determine the worst case. This study presents a simple and rapid technique for bench-scale characterization of cleanability of protein drug products. On-line A280 (UV absorbance at 280 nm) measurements are performed using a fiber optic probe, and the data are used to establish the dynamics of protein dissolution in cleaning solution. The model not only helps to estimate cleaning time of different formulated proteins (and peptides) but also provides insights into the kinetics of cleaning under different thermal and chemical conditions. Protein product degradation during cleaning is also evaluated through gel electrophoresis. Such information is useful in designing new cleaning cycles. While the study is performed using drug products, the model as well as the findings are also applicable for characterization of final purified bulk soils relevant to bulk drug manufacturing.     

国外环境生物技术的发展和展望

环境生物技术在社会的发展中起着越来越重要的积极作用,相关产业也随之迅速发展起来,环境治理要依赖对生物,尤其是微生物及其生理,生化特性的了解和认识,从而可以对其生理,生化和遗传方面的性能加以利用。在这方面,分离解毒微生物及阐明毒物降解过程和机理仍是现在研究的焦点,与此同时,对遗传基因方面的研究和利用还有许多研究工作待进行,监测和跟踪微生物已进入到了分子水平,而传感监测方面已有一些可喜的成果正在向生产转化,生物修复作为消除污染的手段正治理中发挥巨大的作用,但环境保护的根本应是在于进行无污染产生的生产,也称之为绿色生产,未来的社会中,环境生物技术仍对将社会产生巨大的影响,对环境保护起到重要的作用,特别是在新型生物能源的开发和探索方面,本文同时也对世界不同地区在环境生物技术的发展及其特点进行了综述。