Polyethylene Oxide (PEO) and Polyethylene Glycol (PEG) Polymer Sieving Matrix for RNA Capillary Electrophoresis

The selection of sieving polymer for RNA fragments separation by capillary electrophoresis is imperative. We investigated the separation of RNA fragments ranged from 100 to 10,000 nt in polyethylene glycol (PEG) and polyethylene oxide (PEO) solutions with different molecular weight and different concentration. We found that the separation performance of the small RNA fragments (<1000 nt) was improved with the increase of polymer concentration, whereas the separation performance for the large ones (>4000 nt) deteriorated in PEG/PEO solutions when the concentration was above 1.0%/0.6%, respectively. By double logarithmic plot of mobility and RNA fragment size, we revealed three migration regimes for RNA in PEG (300-500k) and PEO (4,000k). Moreover, we calculated the smallest resolvable nucleotide length (N_min) from the resolution length analysis.     

Branched Polyethylene Glycol (Bpeg) Conjugated Antisense Oligonucleotides

Abstract

The synthesis of new oligonucleotide conjugates bearing an high molecular weight, branched polyethylene glycol (bPEG) chain is reported.     

聚对苯二甲酸乙二醇酯(PET)降解酶的研究进展

聚对苯二甲酸乙二醇酯(Polyethylene terephthalate,PET)因其优越的物理化学性质,在各个领域尤其在包装产业得到了广泛的应用.然而,由于使用后的PET处置不当,对生态环境造成了严重威胁.目前生物降解尤其是酶促降解已成为极具可行性且环境友好的PET处理方式.本文集中梳理和总结了近年来已报道的PET...     

Polyethylene glycol (PEG)-induced mouse model of choroidal neovascularization

In this study, we describe a new method for inducing choroidal neovascularization (CNV) in C57BL/6 mice, an animal model of wet age-related macular degeneration (AMD). AMD is a disease that causes central blindness in humans. We injected PEG-8 subretinally in different doses (0.125-2 mg) to induce CNV. After PEG-8 injection, we examined CNV at several time points (days 3-42). We also used Western blotting, immunohistochemistry, and ELISA to examine the complement component C3 split products, C9, VEGF, TGF-β2, and basic FGF. As early as day 1 after treatment, we found that a single subretinal injection of 1 mg of PEG-8 increased the C3 split products and the C9, TGF-β2, and basic FGF levels in the retinal pigment epithelium-choroid tissue. By day 3 after PEG-8 injection, the intraocular activation of the complement system caused induction and progression of CNV, including new vessels penetrating the Bruch's membrane. At day 5 after PEG-8 injection, we observed a fully developed CNV and retinal degeneration. Thus, in this study, we present a new, inexpensive, and accelerated mouse model of CNV that may be useful to study AMD.     

Uses of a Polyethylene Foam (Ethafoam) in Cryostat Operation

Round plugs of Ethafoam (Dow Chemical Co.) prevent water vapor condensation on the port sleeves; a wedge-shaped plug satisfactorily replaces an ineffectively sealing access door. A dropping bottle insulated by encasing it in a block of Ethafoam keeps water from freezing for 3-4 hr. A block of Ethafoam is usuable as a Coplin jar filler for processing single slides with a minimum volume of reagent, and for making a single-slide staining receptacle of 7-8 ml capacity. A cryostat section, covered with a piece of aluminum foil can be flattened by brushing with a pad of Ethafoam.     

Robust Identification of Polyethylene Terephthalate (PET) Plastics through Bayesian Decision

Recycling is one of the most efficient methods for environmental friendly waste management. Among municipal wastes, plastics are the most common material that can be easily recycled and polyethylene terephthalate (PET) is one of its major types. PET material is used in consumer goods packaging such as drinking bottles, toiletry containers, food packaging and many more. Usually, a recycling process is tailored to a specific material for optimal purification and decontamination to obtain high grade recyclable material. The quantity and quality of the sorting process are limited by the capacity of human workers that suffer from fatigue and boredom. Several automated sorting systems have been proposed in the literature that include using chemical, proximity and vision sensors. The main advantages of vision based sensors are its environmentally friendly approach, non-intrusive detection and capability of high throughput. However, the existing methods rely heavily on deterministic approaches that make them less accurate as the variations in PET plastic waste appearance are too high. We proposed a probabilistic approach of modeling the PET material by analyzing the reflection region and its surrounding. Three parameters are modeled by Gaussian and exponential distributions: color, size and distance of the reflection region. The final classification is made through a supervised training method of likelihood ratio test. The main novelty of the proposed method is the probabilistic approach in integrating various PET material signatures that are contaminated by stains under constant lighting changes. The system is evaluated by using four performance metrics: precision, recall, accuracy and error. Our system performed the best in all evaluation metrics compared to the benchmark methods. The system can be further improved by fusing all neighborhood information in decision making and by implementing the system in a graphics processing unit for faster processing speed.     

聚乙二醇对栝楼种子萌发的影响

针对栝楼籽萌发困难的问题,本试验采用聚乙二醇（PEG）对栝楼种子进行处理,结果表明：15％PEG浸种24h,可有效调节渗透水势,改变种子生理生化反应特性,并保持淀粉酶活性连续上升,使可溶性蛋白质含量明显提高,可见PEG在加速大分子动员和提高代谢能力方面发挥了作用,从而促进植物体的组织器官形态建成和生长发育;同时,PEG处理提高了过氧化物酶（POD）活性,降低了丙二醛（MDA）含量,减少了自由基积累,减轻了浸种吸涨对膜系统的损伤,降低了不利因素的影响,提高了种子活力。     

Efficient Polyethylene Glycol (PEG) Mediated Transformation of the Moss Physcomitrella patens

A simple and efficient method to transform Physcomitrella pantens protoplasts is described. This method is adapted from protocols for Physocmitrella protonemal protoplast and Arabidopsis mesophyll protoplast transformation¹. Due to its capacity to undergo efficient mitotic homologous recombination, Physcomitrella patens has emerged as an important model system in recent years². This capacity allows high frequencies of gene targeting^3-9, which is not seen in other model plants such as Arabidopsis. To take full advantage of this system, we need an effective and easy method to deliver DNA into moss cells. The most common ways to transform this moss are particle bombardment¹⁰ and PEG-mediated DNA uptake¹¹. Although particle bombardment can produce a high transformation efficiency¹², gene guns are not readily available to many laboratories and the protocol is difficult to standardize. On the other hand, PEG mediated transformation does not require specialized equipments, and can be performed in any laboratory with a sterile hood. Here, we show a simple and highly efficient method for transformation of moss protoplasts. This method can generate more than 120 transient transformants per microgram of DNA, which is an improvement from the most efficient protocol previously reported¹³. Because of its simplicity, efficiency, and reproducibility, this method can be applied to projects requiring large number of transformants as well as for routine transformation.     

聚乙二醇修饰超氧化物歧化酶的初步研究

目的:对玉米超氧化物歧化酶(SOD)进行共价修饰,以提高其稳定性.方法:以分子量为6000Da的活化聚乙二醇(PEG)为修饰剂,对玉米超氧化物歧化酶(SOD)进行共价修饰,并确定最佳反应条件.将天然SOD与PEG-SOD分别进行热稳定性、酸碱稳定性及抗蛋白酶稳定性的比较实验.结果:在最佳反应时间10h、最佳反应温度4℃时PEG与SOD反应获得的PEG-SOD比天然SOD在热、酸、碱及抗酶解三方面的稳定性均有不同程度的提高.结论:玉米超氧化物歧化酶经PEG修饰后稳定性显著提高.     

Purification and Crystallization of d-Arabinose (l-Fucose) Isomerase from Aerobacter aerogenes by Polyethylene Glycol

d-Arabinose(l-fucose) isomerase (d-arabinose ketol-isomerase, EC 5.3.1.3) was purified from the extracts of d-arabinose-grown cells of Aerobacter aerogenes, strain M-7 by the procedure of repeated fractional precipitation with polyethylene glycol 6000 and isolating the crystalline state. The crystalline enzyme was homogeneous in ultracentrifugal analysis and polyacrylamide gel electrophoresis. Sedimentation constant obtained was 15.4s and the molecular weight was estimated as being approximately 2.5 × 10⁵ by gel filtration on Sephadex G-200.

Optimum pH for isomerization of d-arabinose and of l-fucose was identical at pH 9.3, and the Michaelis constants were 51 mm for l-fucose and 160 mm for d-arabinose. Both of these activities decreased at the same rate with thermal inactivation at 45 and 50°C. All four pentitols inhibited two pentose isomerase activities competitively with same Ki values: 1.3–1.5 mm for d-arabitol, 2.2–2.7 mm for ribitol, 2.9–3.2 mm for l-arabitol, and 10–10.5 mm for xylitol. It is confirmed that the single enzyme is responsible for the isomerization of d-arabinose and l-fucose.     

聚乙二醇对羟基丁酸和羟基己酸共聚物的共混改性研究

本研究以聚羟基脂肪酸酯家族中的新成员羟基丁酸和羟基己酸共聚物（PHBHHx）为基础,采用与聚乙二醇（PEG）共混的方法对其进行改性,研究结果证实：PHBHHx与PEG 共混物中比例为3:1及2:1时,两者完全物理相容。而PEG在共混物中比例升高时则导致相分离,成为部分相容体系。PEG掺入显著提高材料亲水性及表面自由能,使血管平滑肌细胞（RaSMCs）及血管内皮细胞（HUVECs）的细胞粘附及增殖大幅度提高,并且均具有一定的PEG含量依赖性。其中对RaSMCs的作用最为明显,RaSMCs能在PEG/PHBHHx比例为1:1的共混膜（E1X1）上持续增殖至融合,而HUVECs则呈粘附较差的类球形形貌,证实E1X1可以潜在应用于复合血管组织工程支架中的近内膜基材。     

Involvement of a Polyethylene glycol (PEG)-oxidizing Enzyme in the Bacterial Metabolism of PEG

Eighty-seven strains of acetic acid bacteria were surveyed for plasmids by CsCl-ethidium bromide equilibrium centrifugation of cell lysates. Twenty-seven of the 33 strains of Acetobacter were found to harbor plasmid DNA and most strains contained multiple species of low-molecular-weight plasmids. On the other hand, plasmid DNAs were detected in 23 of the 36 strains of Gluconobacter and most of them had molecular weights of more than 5 megadaltons. Of the 18 strains newly isolated from a vinegar factory, some of which were examined taxonomically, 17 contained plasmid DNA. The molecular weights of plasmids detected in this study were in the range of about one to over 17 megadaltons. The plasmids with molecular weights of less than 5 megadaltons were characterized with restriction endonucleases. The physical maps of two plasmids designated as pMV1O1 and pMV102, which were found in the isolated strains, were constructed.     

Polyethylene barrier impregnated with lambda-cyhalothrin for exclusion of subterranean termites (Isoptera: Rhinotermitidae) from structures

Polyethylene film impregnated with lambda-cyhalothrin was placed over a sand plot and covered with a concrete slab to allow insecticide movement into the sand for a period of 5.5 yr. Discs of polyethylene film and sand beneath them were sampled annually for 5 yr and at 5.5 yr for bioassay with the Formosan and eastern subterranean termite. Results demonstrated that sufficient quantities of lambda-cyhalothrin were released from the impregnated polyethylene film into adjacent sand to prevent termite penetration. The impregnated film has less environmental impact than conventional liquid termiticides because the insecticide is held in the polymer. Other advantages include its dual function as a construction moisture barrier and ease in verifying its proper installation.     

Preparation of Long-Acting Superoxide Dismutase Using High Molecular Weight Polyethylene Glycol (41,000-72,000 Daltons)

Superoxide dismutase (SOD) has demonstrated therapeutic potential for treating a variety of conditions including radiation injury, oxygen toxicity, reperfusion injury, and inflammation, especially arthritis. However, the native enzyme's short half-life in plasma (6 minutes in mice. 25 minutes in man) limits the enzyme's effectiveness in many applications, or requires infusion of large doses. High doses of SOD derived from either natural or rDNA sources may increase the potential for immunologic sensitization. One effective use of native SOD is intra particular administration for treatment of arthritis, where injection of SOD into joints retards elimination (15 hour terminal half-life), allowing the effective use of lower doses.

To overcome the limitations resulting from rapid clearance. various researchers have increased the persistence of SOD by cross-linking SOD or by attaching polymeric substances, including dextrans, albumin, Ficoll, polyvinyl alcohol or polyethylene glycol (PEG). PEG is relatively safe; however. the amount of modification by PEG. is the MW range 1.900–5.000 daltons, which is necessary to optimally increase serum persistence and reduce immunogenicity, results in the loss of much of the enzymatic activity.

In this report we describe the preparation of SOD adducts containing I to 4 strands of high MW PEG (41,000–72,000 daltons). The MW range of these adducts, measured by steric exclusion HPLC based on protein standards, is 200,000 to over 1,100,000 daltons. The number of PEG strands attached per SOD dimer (32,000 daltons) was measured by HPLC. Because of the low degree of protein modification required to produce very high MW products, these PEG-SODS retain 90%-100% of the SOD activity of the native enzyme. Additionally, these very large adducts demonstrate longer persistence and lower immunogenicity and antigenicity compared to the more highly modified PEG-SODS containing low MW PEG (i.e., 7–16 strands of 5.000 dalton methoxy-PEG).     

Preparation of Long-Acting Superoxide Dismutase Using High Molecular Weight Polyethylene Glycol (41,000-72,000 Daltons)

Superoxide dismutase (SOD) has demonstrated therapeutic potential for treating a variety of conditions including radiation injury, oxygen toxicity, reperfusion injury, and inflammation, especially arthritis. However, the native enzyme's short half-life in plasma (6 minutes in mice. 25 minutes in man) limits the enzyme's effectiveness in many applications, or requires infusion of large doses. High doses of SOD derived from either natural or rDNA sources may increase the potential for immunologic sensitization. One effective use of native SOD is intra particular administration for treatment of arthritis, where injection of SOD into joints retards elimination (15 hour terminal half-life), allowing the effective use of lower doses.

To overcome the limitations resulting from rapid clearance. various researchers have increased the persistence of SOD by cross-linking SOD or by attaching polymeric substances, including dextrans, albumin, Ficoll, polyvinyl alcohol or polyethylene glycol (PEG). PEG is relatively safe; however. the amount of modification by PEG. is the MW range 1.900-5.000 daltons, which is necessary to optimally increase serum persistence and reduce immunogenicity, results in the loss of much of the enzymatic activity.

In this report we describe the preparation of SOD adducts containing I to 4 strands of high MW PEG (41,000-72,000 daltons). The MW range of these adducts, measured by steric exclusion HPLC based on protein standards, is 200,000 to over 1,100,000 daltons. The number of PEG strands attached per SOD dimer (32,000 daltons) was measured by HPLC. Because of the low degree of protein modification required to produce very high MW products, these PEG-SODS retain 90%-100% of the SOD activity of the native enzyme. Additionally, these very large adducts demonstrate longer persistence and lower immunogenicity and antigenicity compared to the more highly modified PEG-SODS containing low MW PEG (i.e., 7-16 strands of 5.000 dalton methoxy-PEG).     

IgY Technology: Extraction of Chicken Antibodies from Egg Yolk by Polyethylene Glycol (PEG) Precipitation

Hens can be immunized by means of i.m. vaccination (Musculus pectoralis, left and right, injection volume 0.5-1.0 ml) or by means of Gene-Gun plasmid-immunization. Dependent on the immunogenicity of the antigen, high antibody-titres (up to 1:100,000 - 1:1,000,000) can be achieved after only one or 3 - 4 boost immunizations. Normally, a hen lays eggs continuously for about 72 weeks, thereafter the laying capacity decreases. This protocol describes the extraction of total IgY from egg yolk by means of a precipitation procedure (PEG. Polson et al. 1980). The method involves two important steps. The first one is the removal of lipids and the second is the precipitation of total IgY from the supernatant of step one. After dialysis against a buffer (normally PBS) the IgY-extract can be stored at -20°C for more than a year. The purity of the extract is around 80 %, the total IgY per egg varies from 40-80 mg, dependent on the age of the laying hen. The total IgY content increases with the age of the hen from around 40 mg/egg up to 80 mg/egg (concerning PEG precipitation). The laying capacity of a hen per year is around 325 eggs. That means a total potential harvest of 20 g total IgY/year based on a mean IgY content of 60 mg total IgY/egg (see Table 1).     

Effect of Drug Solubility on Polymer Hydration and Drug Dissolution from Polyethylene Oxide (PEO) Matrix Tablets

The purpose of the study was to investigate the effect of drug solubility on polymer hydration and drug dissolution from modified release matrix tablets of polyethylene oxide (PEO). Different PEO matrix tablets were prepared using acetaminophen (ACE) and ibuprofen (IBU) as study compounds and Polyox WSR301 (PEO) as primary hydrophilic matrix polymer. Tablet dissolution was tested using the USP Apparatus II, and the hydration of PEO polymer during dissolution was recorded using a texture analyzer. Drug dissolution from the preparations was dependent upon drug solubility, hydrogel formation and polymer proportion in the preparation. Delayed drug release was attributed to the formation of hydrogel layer on the surface of the tablet and the penetration of water into matrix core through drug dissolution and diffusion. A multiple linear regression model could be used to describe the relationship among drug dissolution, polymer ratio, hydrogel formation and drug solubility; the mathematical correlation was also proven to be valid and adaptable to a series of study compounds. The developed methodology would be beneficial to formulation scientists in dosage form design and optimization.     

Polyethylene glycol (PEG)-mediated transient gene expression in a red alga, Cyanidioschyzon merolae 10D

DNA introduction into cells is an essential technique for molecular genetic analysis. Here, we show that DNA is easily introduced into cells of the unicellular red alga Cyanidioschyzon merolae by a polyethylene glycol (PEG)-mediated protocol. In this study, the beta-tubulin gene of C. merolae was cloned on a plasmid and a hemagglutinin (HA) tag then added at the C-terminus. This plasmid was then introduced into C. merolae cells by a PEG-mediated transformation protocol. At 24 h after PEG-mediated transformation, intracellular localization of the tagged protein was detected by anti-HA immunocytochemistry, indicating the utility of this transient expression system for molecular genetic analyses.