Determination of kinetic and retention properties of cartridge and disk devices for solid-phase extraction

The kinetic properties of cartridge and disk solid-phase extraction devices are determined by forced-flow liquid chromatography. Typical cartridges provide about 5–15 theoretical plates per cm of bed height and particle-loaded membranes provide about 4–9 theoretical plates for a 0.5-mm-thick membrane. It is shown that cartridge devices fail to provide their maximum trapping performance because of inadequate packing density and that the required packing density could be easily achieved in practice with particles of a standard size. The retention properties of common sorbents for extraction from water and air are characterized with the solvation parameter model. For predominantly aqueous solutions a favorable cavity term results in increased retention while polar interactions tend to reduce retention. Retention on porous polymer sorbents is more complicated because of their capacity to absorp significant amounts of the sample processing solvent resulting in solvent-dependent changes in retention properties. For trapping organic volatiles from air cavity formation and dispersion interactions are important, and in the case of Tenax its capacity for induction interactions is also significant.     

The use of a drug resistance cartridge for in vitro insertion and deletion mutagenesis of a cosmid clone

A drug-resistant cartridge was employed in the construction of families of insertion mutants of a cosmid clone. The cartridge contains a cml gene and has identical restriction enzyme sites, EcoRI, BamHI, SalI, and PstI, on both ends. The families of mutants were made by ligation of the cartridge to the cosmid, which was linearized or partially digested, followed by in vitro packaging and transduction. From these families we selected cosmid derivatives which either have a unique BamHI site at a predetermined site in the cosmid or have deletions covering different portions of the original clone. The extent of a large gene cluster cloned into the original cosmid was identified by confirming the gene function in some of the deletion mutants. The possibility for further and various uses of this cartridge is discussed.     

When consumer behavior dictates life cycle performance beyond the use phase: case study of inkjet cartridge end-of-life management

Purpose

Conventional wisdom suggests that product reuse can provide environmental savings. The purpose of this study is to first compare the environmental impacts of retail refilling and remanufactured inkjet cartridge alternatives to production of new inkjet cartridges, and then determine the extent to which consumer behavior can influence life cycle outcomes.

Methods

A life cycle inventory was developed for an inkjet cartridge with an integral print head using material composition data collected from cartridge disassembly and material processing, product manufacturing, and transportation inputs estimated from market data and the ecoinvent database in SimaPro 7.3. Although previous comparative life cycle assessment (LCA) studies for printer cartridges typically use “pages printed” or a variation thereof for the functional unit, “cartridge use cycles” is more suitable for examining reused inkjet cartridge alternatives that depend on the inkjet cartridge end-of-life (EOL) route chosen by the consumer. Since multiple reuse cycles achieved from refilling by a retailer was of specific interest, a functional unit defined in the form of “five use cycles” included the mode and manner in which consumers purchased inkjet cartridge use cycles.

Results and discussion

Cartridge refills present the lowest environmental impact, offering a 76 % savings in global warming potential (GWP) impact compared to production and purchase of a new inkjet cartridge alternative, followed by the remanufacturing case, which provided a 36 % savings in GWP impact compared to the new inkjet cartridge. However, results varied widely, even switching to favor new cartridge purchase, depending on how consumer transport was modeled, specifically the mode of travel, travel patterns (number of trips), and method of allocating impact to each trip.

Conclusions

Refilling an original equipment manufacturer (OEM) cartridge four consecutive times provides the best alternative for reducing environmental impact for those consumers that purchase inkjet cartridges one at a time. On the other hand, consumers that purchase multiple cartridges in a single trip to a retailer reduce environmental impact more by transport minimization than by refilling. Results reinforce the need for more comprehensive inclusion of consumer behavior when modeling life cycle environmental impact of product alternatives.     

Experimental comparison and cross-validation of Affymetrix HT plate and cartridge array gene expression platforms

The successful use of gene expression microarrays in basic research studies has spawned interest in the use of this technology for clinical trial and population-based studies, but cost, complexity of sample processing and tracking, and limitations of sample throughput have restricted their use for these very large-scale investigations. The Affymetrix GeneChip Plate Array System addresses these concerns and could facilitate larger studies if the data prove to be comparable to industry-standard cartridge arrays. Here we present a comparative evaluation of performance between Affymetrix GeneChip Human 133A cartridge and plate arrays with an emphasis on the assessment of systematic variation and its impact on log ratio data. This study utilized two standardized control RNAs on four independent lots of plate and cartridge arrays. We found that HT plate arrays showed improved specificity and were more reproducible over a wide intensity range, but cartridge arrays exhibit better sensitivity. Not surprisingly, artifactual changes due to positional effects were detectable on plate arrays, but were generally small in number and magnitude and in practice may be removed using standard fold-change and p-value thresholds. Overall, log ratio data between cartridges and plate arrays were remarkably concordant. We conclude that HT arrays offer significant improvements over cartridge arrays for large-scale studies.     

Microsequence analysis of peptides and proteins. V. Design and performance of a novel gas-liquid-solid phase instrument

We describe the construction and performance of a novel, automated, Edman chemistry-based microsequencer. The reagent and solvent delivery system, the reaction cartridge for coupling and cleavage, and the conversion flask are all constructed from chemically inert perfluoroelastomers. The delivery valves are of a new design incorporating the use of electromagnetically actuated solenoids and zero-dead-volume construction, and may be connected in a modular fashion resulting in multiple inputs with a single output line which can be flushed with inert gas. The bottle closures are of a new design based on an all-Teflon compression fitting. The reaction cartridge and conversion flask are thermostated by solid-state heaters in an aluminum block. The overall size of the instrument is 25 X 34 X 14 in. The chemistry utilizes 2% aqueous triethylamine as the coupling base which is delivered to the reaction cartridge via a stream of nitrogen. The "gas-phase" delivery of the coupling base and the cleavage acid (trifluoroacetic acid) is modeled after the method described by R. M. Hewick et al. (J. Biol. Chem. 256, 7990-7997,1981). The instrument has performed well over a period of 3 years in terms of low background peaks, sensitivity in the picomole range, and reliability of operation. The use of economical components, ease of construction and operation, and sensitive analytical capability make this instrument a useful tool for microsequence analysis of peptides and proteins.     

Automatic mechanical alveolar gas sampler for multiple-sample collection in field

A mechanical alveolar gas sampler using the revolver principle capable of collecting six individual expired gas samples is described. The 0.91-kg sampler collects 19-ml samples in pre-evacuated aluminum ampoules equipped with spring-loaded valves from a sampling chamber equipped with two removable one-way valves. On depression of external handles, one of six ampoules located in a removable cartridge is aligned and advanced into the sampling chamber where its valve is opened and then closed. Releasing the handles removes the ampoule from the sampling chamber and automatically rotates the cartridge through 60 degrees to position a new ampoule in preparation for the next sampling sequence. A lock-out mechanism prevents reexposure of any of the ampoules after six samples have been taken. The performance of the sampler is described including its successful use in the field to collect alveolar gas samples on the summit of Mount Everest.     

Evaluation of recalcitrance of mascara in a cartridge container system to microbial challenge

Summary A cartridge mascara system designed to minimize the exposure of mascara to the environment during use was evaluated for recalcitrance to bacteria. The design of the system contributed to low numbers of bacteria found in the mascara, but the need for preservatives was not alleviated.     

A charcoal cartridge for the removal of anionic detergent and electrophoresis stains

Instructions are given for the construction of a charcoal-containing cartridge that allows the rapid recirculation through charcoal of any fluid in which the cartridge is submerged; recirculatory flow is achieved by magnetic stirring of the fluid by a stirring bar placed under the cartridge. The device is assembled from nylon mesh and conical sections cut from polypropylene beakers. The device can be used to accelerate the destaining of electrophoresis gels and to remove SDS (sodium lauryl sulfate) from SDS gels. The removal of SDS prior to staining is essential for the staining of SDS gels with Coomassie Brilliant Blue G-250.     

Cryopreservation of the anaerobic rumen fungus Neocallimastix patriciarum

A rapid extraction and purification procedure is described for the preparation of toxic peptides from freshwater blooms and laboratory isolates of Microcystis aeruginosa . Extraction with methanol/butanol, followed by C₁₈ cartridge concentration; gel filtration and high performance liquid chromatography yields discrete toxin peaks. Elution profiles for the laboratory isolates and bloom extracts are compared and the applicability of the method for detecting cyanobacterial toxins in natural waters is discussed.     

Efficient purification of DNA fragments using a protein binding membrane

A novel and efficient method has been developed for isolation of correctly digested DNA fragments without the use of classic size-dependent electrophoretic separation methods. To achieve this, DNA fragments are end-labelled by haptens. After specific endonuclease digestion of the hapten-labelled DNA, the DNA is incubated with a protein that specifically binds to the hapten. The incubation mixture is then passed through a cartridge containing a protein-binding membrane that does not bind DNA. Undigested and partly digested DNA are retained on the membrane, while correctly digested DNA is selectively recovered for use in further downstream applications.     

An automated microfluidic-based immunoassay cartridge for allergen screening and other multiplexed assays

A microfluidic cartridge and system for multiplexed immunoassays is described. The passive microfluidic cartridge was composed of three layers of injection molded plastic sealed together using a thermal staking technique. Using this platform technology, a specific immunoglobulin E (IgE) panel assay was constructed. Allergen extract targets, positive and negative controls, and IgE calibration standards were immobilized within the cartridge as a microarray. A computer-controlled solenoid array provided the necessary actuation force for pumping reagents within the cartridge to perform an automated, chemiluminescent indirect immunoassay. A 20-target allergen extract panel was demonstrated on the device with a total analysis time of 27 min. Allergen screening results showed 84% agreement for 3 house dust mites (N = 300) compared with a commercial test and 80% agreement overall (N = 978). Average coefficients of variation (N = 80) were measured as 20.5% for low/medium levels and 20.4% for medium/high levels. The average limit of detection (N = 160) was measured at 0.535 AU, and cutoff levels of 1.0 AU were estimated at less than 1 IU/ml (2.4 ng/ml). Such a system has potential applications in decentralized allergen screening as well as in other near-patient diagnostic immunoassays where multiplexed analysis, ease of use, and short analysis time are critical.     

The role of macrofungi in environmental conservation

Abstract

The motives for the conservation of fungi are briefly outlined. The data and methods needed for successful use of fungi in environmental conservation are mentioned. A survey is given of the main habitats of threatened fungi in Western and Central Europe, including data on the significance of their mycoflora, the causes of decrease and possible measures to improve the situation. The value of macrofungi as bio-indicators for environmental quality is discussed and demonstrated with three examples: the indicator value of wood-inhabiting fungi for undisturbed forest sites; of ectomycorrhizal fungi for air pollution stress in forests and of saprotrophic fungi for the duration of undisturbed grassland use. Lists of indicator species with different indicator values for air pollution stress and grassland use are provided.     

Extraction and analysis of microcystins RR and LR in cyanobacteria using a cyano cartridge

A new method for the extraction of microcystins RR and LR in cyanobacteria was developed using a cyano cartridge. Lyophilized cells (100 mg) were extracted with 5% (v/v) acetic acid. The extract was centrifuged and then the supernatant was applied to a CN cartridge. The cartridge that contained microcystins was rinsed with 5 ml of water and 5 ml of 0.5 M acetic acid, followed by 5 ml of 5% acetonitrile in water. Microcystins were finally eluted from the CN cartridge with 70% acetonitrile in water and were determined by HPLC. Better recoveries and chromatograms were observed than with ODS cartridges.     

Anatomical identification of spectral receptor types in the retina and lamina of the Australian orchard butterfly,Papilio aegeus aegeus D.

Summary The nine receptor cells examined in each ommatidium of the butterfly Papilio aegeus aegeus can be named according to their positional orientation across the fused rhabdom. Six of them end as short visual fibres (svf) in the second stratum of the lamina, whereas the remaining three retinula cells (lvf) pass together with the lamina fibres (L-fibres) the first optic ganglion and the outer chiasma to end in the three most distal layers of the second optic ganglion, the medulla. The organization of the retinula-cell axons within the pseudocartridge and the cartridge remains almost uniform throughout the first optic ganglion. Five L-fibres, which have their origin in the fenestrated layer (FL), join each laminar cartridge before entering the neuropil of the first optic region. Four of these L-fibres (L-1, L-2, L-3 and L-4) could be definitely located and characterized using Golgi-stained light- and electron-microscopic techniques. Whereas L-1 and L-3 show a lateral branching pattern reaching only fibres of the same cartridge, L-2 and L-4 have long collaterals interconnecting several neighbouring cartridges in a characteristic pattern. Serial sections of silver-impregnated retinula-cell axons as well as L-fibres were investigated for their synaptic connectivity patterns between and within these fibres. These cellular interactions and possible information processing are discussed.     

An nptI-sacB-sacR cartridge for constructing directed, unmarked mutations in gram-negative bacteria by marker exchange-eviction mutagenesis

A technique for marker exchange-eviction mutagenesis that enables the construction of directed, unmarked mutations in Gram-negative bacteria was demonstrated in Erwinia chrysanthemi. The technique employs an nptI-sacB-sacR cartridge that is carried on a 3.8-kb BamHI fragment and confers kanamycin (Km) resistance and sucrose sensitivity (due to the production of levansucrase by sacB) in E. chrysanthemi. The cartridge was inserted into a Sau3A site in a cloned E. chrysanthemi pelC gene (encoding pectate lyase isozyme PLc) and then introduced into the Erwinia genome by gene exchange recombination. The resulting mutant was KmR, sucrose-sensitive, and PLc-deficient. The cartridge was then excised from the plasmid-borne pelC gene by PstI cleavage to leave a 28-bp frame-shifting insertion. The pelC allele containing the 28-bp insertion was exchanged for the chromosomal allele containing the nptI-sacB-sacR cartridge by selection for sucrose tolerance. The resulting E. chrysanthemi mutant was Kms and PLc-deficient. The technique permits the construction of complex strains with many directed mutations without the introduction of a corresponding number of antibiotic resistance markers and should prove useful, for example, in exploring the role of the multiple pel genes in E. chrysanthemi.     

The Choice of HLA‐Associated Peptide Enrichment and Purification Strategy Affects Peptide Yields and Creates a Bias in Detected Sequence Repertoire

Understanding the most appropriate workflow for biochemical human leukocyte antigen (HLA)‐associated peptide enrichment prior to ligand sequencing is essential to achieve optimal sensitivity in immunopeptidomics experiments. The use of different detergents for HLA solubilization as well as complementary workflows to separate HLA‐bound peptides from HLA protein complex components after their immunoprecipitation including HPLC, C18 cartridge, and 5 kDa filter are described. It is observed that all solubilization approaches tested led to similar peptide ligand identification rates; however, a higher number of peptides are identified in samples lysed with CHAPS compared with other methods. The HPLC method is superior in terms of HLA‐I peptide recovery compared with 5 kDa filter and C18 cartridge peptide purification methods. Most importantly, it is observed that both the choice of detergent and peptide purification strategy creates a significant bias for the identified peptide sequences, and that allele‐specific peptide repertoires are affected depending on the workflow of choice. The results highlight the importance of employing a suitable strategy for HLA peptide enrichment and that the obtained peptide repertoires do not necessarily reflect the true distributions of peptide sequences in the sample.     

Simple System for Isothermal DNA Amplification Coupled to Lateral Flow Detection

Infectious disease diagnosis in point-of-care settings can be greatly improved through integrated, automated nucleic acid testing devices. We have developed an early prototype for a low-cost system which executes isothermal DNA amplification coupled to nucleic acid lateral flow (NALF) detection in a mesofluidic cartridge attached to a portable instrument. Fluid handling inside the cartridge is facilitated through one-way passive valves, flexible pouches, and electrolysis-driven pumps, which promotes a compact and inexpensive instrument design. The closed-system disposable prevents workspace amplicon contamination. The cartridge design is based on standard scalable manufacturing techniques such as injection molding. Nucleic acid amplification occurs in a two-layer pouch that enables efficient heat transfer. We have demonstrated as proof of principle the amplification and detection of Mycobacterium tuberculosis (M.tb) genomic DNA in the cartridge, using either Loop Mediated Amplification (LAMP) or the Exponential Amplification Reaction (EXPAR), both coupled to NALF detection. We envision that a refined version of this cartridge, including upstream sample preparation coupled to amplification and detection, will enable fully-automated sample-in to answer-out infectious disease diagnosis in primary care settings of low-resource countries with high disease burden.     

基因枪介导真核表达质粒转染体外培养的COS-7细胞系的实验研究

目的：建立基因枪子弹制备及转染体外培养COS-7细胞系的方法。方法：以亚精氨、氯化钙沉淀法制备子弹（DNA＋金颗粒）,利用原子力显微镜观察子弹制备情况;采用基因枪方法分别将真核表达质粒pVax-Dsred-IRES-EGFP转染对照组和实验组COS-7细胞,转染后24h,利用激光扫描共聚焦显微镜观察细胞中红、绿荧光蛋白的表达。结果：制备了基因枪子弹,DNA紧密包裹在金颗粒周围;基因枪介导的pVax-Dsred-IRES-EGFP被转染入体外培养的COS-7细胞,转染后24h可检测到红、绿荧光,而对照组则没有荧光蛋白的表达。结论：国产新芝SJ-500型基因枪能够有效介导外源基因转移,基因枪转染的COS-7细胞能够有效表达报告基因。     

Method of isolating conditionally-pathogenic Gram-negative microorganisms, agents of intrahospital infections, from air

The possibility of detecting Pseudomonas aeruginosa and other Gram-negative bacteria in the air of the burn department at the Institute of Surgery was studied. The investigation of large volumes of air (0.5-1 m3) in the wards and the corridor with the use of a new bacteriological aerosol sampler, model IIAB-5, resulted in the detection of Pseudomonas aeruginosa. Besides, in a number of other rooms Klebsiella, Proteus, Citrobacter and Enterobacter were detected in the air. The possibility of the spread of Gram-negative opportunistic bacteria through the air in hospital conditions is discussed.