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Screening System (screening + system)
Selected AbstractsInvasion Possibility and Potential Effects of Rhus typhina on Beijing MunicipalityJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 5 2008Guangmei Wang Abstract Rhus typhina, an alien species introduced from North America, was identified as a main afforestation species in Beijing municipality. However, its invasiveness is still at odds. To clarify this problem, we applied the North American Screening System and the Australian Screening System to preliminarily predict its invasion possibility. Both screening systems gave the same recommendation to "reject". The geographical distribution was surveyed, with the population features of R. typhina against the native plant communities being assessed. With anthropogenic assistance, R. typhina has been scattered on almost all habitats from downtown to mountains, including roadsides, farmlands and protected areas. As a clonal shrub, R. typhina possessed a high spreading rate, varying from 6.3 m/3 years at sterile habitats to 6.7 m/3 years at fertile ones. Significantly lower species richness, individual density and diversity were observed in the R. typhina community than those of the native Vitex negundo Linn.var. heterophylla (Franch.) Rehd. community at both sterile and fertile habitats. Continual wide plantation of R. typhina may further foster its population expansion, which helps the species to overcome spatial isolation. The fact that each root fragment can develop into a new individual makes R. typhina very difficult to be eradicated once established. From a biological point of view, we believe that R. typhina is a plant invader in Beijing. We therefore suggest the government should remove the name of R. typhina from the main tree species list in afforesting Beijing. [source] Cell Detachment Model for an Antibody-Based Microfluidic Cancer Screening SystemBIOTECHNOLOGY PROGRESS, Issue 5 2006Swapnil P. Wankhede We consider cells bound to the floor of a microfluidic channel and present a model of their flow-induced detachment. We approximate hydrodynamic force and cell elastic response using static finite-element simulation of a single cell. Detachment is assumed to occur when hydrodynamic and adhesive forces are roughly equal. The result is extended to multiple cells at the device level using a sigmoidal curve fit. The model is applied to a microfluidic cancer-screening device that discriminates between normal epithelial cells and cells infected with human papillomavirus (HPV), on the basis of increased expression of the transmembrane protein ,6 integrin in the latter. Here, the cells to be tested are bound to a microchannel floor coated with anti ,6 integrin antibodies. In an appropriate flow rate range, normal cells are washed away while HPV-infected cells remain bound. The model allows interpolation between data points to choose the optimal flow rate and provides insight into interaction of cell mechanical properties and the flow-induced detachment mechanism. Notably, the results suggest a significant influence of cell elastic response on detachment. [source] European Standard Series patch test results from a contact dermatitis clinic in Israel during the 7-year period from 1998 to 2004CONTACT DERMATITIS, Issue 2 2006Aneta Lazarov The results of a 7-year retrospective study (1998,2004) from patch testing with the European Standard Series (ESS) establishing the frequency of sensitization in a contact dermatitis clinic in Israel are presented. 23 allergens were patch tested on 2156 patients, 1462 females (67.8%) and 694 males (32.2%). Atopy and asthma were present in 21.9% of the patients. One or more allergic reactions were observed in 937 patients (43.5%). The highest yield of patch test positives from the 1076 positive reactions were obtained from nickel sulfate (13.9%), fragrance mix (7.1%), potassium dichromate (3.8%), Balsam of Peru (3.6%), CL + Me-isothiazolinone (3.4%) and cobalt chloride (3.4%). Allergens which produced the least amount of positive results were primin and clioquinol. Allergic contact dermatitis (ACD) was established in 32.8%, whereas occupationally related allergic (8.0) and irritant contact dermatitis (5.6%) affected a total of 13.6% of the cases studied. The most common clinical forms of dermatitis were chronic dermatitis (47.7%) followed by acute dermatitis (22.8%), and lichenification and hyperkeratosis (7.9%). The hands (30.7%), face and neck (23.9%) and extremities (11.3%) were the most frequently affected areas. Four allergens in our study differed from the top 10 allergens in Europe namely: Cl + Me-isothiazolinone, formaldehyde, 4-tert-butylphenol formaldehyde resin and sesquiterpene lactone mix reflecting an existing difference in environmental exposure. Our study is the first to provide data on the frequency of sensitization and important allergens in the aetiology of ACD in Israel. In spite of the existing differences with Europe, we conclude that ESS is an appropriate screening system for the diagnosis of ACD in Israel. [source] Functional interaction of general transcription initiation factor TFIIE with general chromatin factor SPT16/CDC68GENES TO CELLS, Issue 4 2000Seung-Woo Kang Background Transcriptional initiation of class II genes is one of the major targets for the regulation of gene expression and is carried out by RNA polymerase II and many auxiliary factors, which include general transcription initiation factors (GTFs). TFIIE, one of the GTFs, functions at the later stage of transcription initiation. As recent studies indicated the possibility that TFIIE may have a role in chromatin transcriptional regulation, we isolated TFIIE-interacting factors which have chromatin-related functions. Results Using the yeast two-hybrid screening system, we isolated the C-terminal part of the human homologue of Saccharomyces cerevisiae (y) Spt16p/Cdc68p, a general chromatin factor. The C-terminal part of human SPT16/CDC68 directly interacts with TFIIE, and ySpt16p/Cdc68p also interacts with yTFIIE (Tfa1p/Tfa2p), thus indicating the existence of an evolutionarily conserved interaction between TFIIE and SPT16/CDC68. Functional interaction of yTFIIE and ySpt16p/Cdc68p was examined using a conditional yTFIIE-, mutant strain. Over-expression of ySpt16p/Cdc68p suppressed the phenotype of cold sensitivity of the yTFIIE-,- cs mutant strain, and in vitro binding assays revealed that yTFIIE-,- cs mutant protein showed diminished binding affinity to ySpt16p/Cdc68p. Conclusions These observations indicate that general transcription initiation factor TFIIE functionally interacts with general chromatin factor SPT16/CDC68, a finding which provides new insight into the involvement of TFIIE in chromatin transcription. This may well lead to a breakthrough in relationships between the transcription initiation process and structural changes in chromatin. [source] Usefulness of EGFR mutation screening in pleural fluid to predict the clinical outcome of gefitinib treated patients with lung cancerINTERNATIONAL JOURNAL OF CANCER, Issue 10 2006Junichi Soh Abstract The importance of epidermal growth factor receptor (EGFR) gene mutation has been recognized in nonsmall cell lung cancer (NSCLC), requiring the standardization of mutation screening system including the kind of samples. Here, we examined the EGFR mutation status in 61 pleural fluid samples from NSCLC cases using direct sequencing, nonenriched PCR, mutant-enriched PCR and peptide nucleic acid-locked nucleic acid (PNA-LNA) PCR clamp assay. The mutant-enriched PCR assay detected 16 mutant cases. Among them, the nonenriched PCR assay failed to detect 3 mutant cases. Regarding the discrepancy between mutant-enriched PCR and PNA-LNA PCR clamp assays, 3 cases of exon19-deletions were detected only by mutant-enriched PCR assay and no difference at the L858R mutation. There was no difference in results between direct sequencing and nonenriched PCR assay. We also correlated the EGFR mutation with clinical outcome of gefitinib-treated 29 cases. EGFR mutations were present in 10 cases, revealing 7 partial response and 3 no change (NC). In EGFR wild-type cases, 10 revealed NC and 9 progressive disease. The responders were significantly more frequent among the EGFR mutant cases than among the wild-type (p < 0.0001). Overall survival (p = 0.0092) and progression-free survival (p = 0.018) were significantly longer among the EGFR mutant cases than among the wild-type. In summary, we evaluated the utility of EGFR mutation screening in pleural fluid using 4 assays that showed some discrepancies arising from the designs of the assays. As clinical importance, the EGFR mutation status in pleural fluid can be a biomarker for the favorable outcome of gefitinib-treated NSCLC cases. © 2006 Wiley-Liss, Inc. [source] Comparison of different methods of bacterial detection in blood componentsISBT SCIENCE SERIES: THE INTERNATIONAL JOURNAL OF INTRACELLULAR TRANSPORT, Issue 1 2009M. Schmidt Background, Over the last two decades, the residual risk of acquiring a transfusion-transmitted viral infection has been reduced to less than 1 : 1 000 000 via improvements in different techniques (e.g. donor selection, leuco-depletion, introduction of 3rd or 4th generation enzyme-linked immunosorbent assays and mini-pool nucleic acid testing (MP-NAT). In contrast, the risk for transfusion-associated bacterial infections has remained fairly stable, and is estimated to be in a range between 1 : 2000 and 1 : 3000. Platelets are at an especially higher risk for bacterial contamination, because they are stored at room temperature, which provides good culture conditions for a broad range of bacterial strains. To improve bacterial safety of blood products, different detection systems have been developed that can be divided into culture systems like BacT/ALERT or Pall eBDS, rapid detection systems like NAT systems, immunoassays and systems based on the FACS technique. Culture systems are used for routine bacterial screening of platelets in many countries, whereas rapid detection systems so far are mainly used in experimental spiking studies. Nevertheless, pathogen-reduction systems are currently available for platelet concentrates and plasma, and are under investigation for erythrocytes. Methods, In this review, the functional principles of the different assays are described and discussed with regard to their analytical sensitivity, analytical specificity, diagnostic sensitivity, diagnostic specificity and clinical efficiency. The detection methods were clustered into three groups: (i) detection systems currently used for routine screening of blood products, (ii) experimental detection systems ready to use for routine screening of blood products, and (iii) new experimental detection systems that need to be investigated in additional spiking studies and clinical trials. Results, A recent International Society of Blood Transfusion international forum reported on bacterial detection methods in 12 countries. Eight countries have implemented BacT/ALERT into blood donor screening, whereas in three countries only quality controls were done by culture methods. In one country, shelf-life was reduced to 3 days, so no bacterial screening was implemented. Screening data with culture methods can be used to investigate the prevalence of bacterial contamination in platelets. Differing results between the countries could be explained by different test definitions and different test strategies. Nevertheless, false-negative results causing severe transfusion-related septic reactions have been reported all over the world due to a residual risk of sample errors. Rapid screening systems NAT and FACS assays have improved over the last few years and are now ready to be implemented in routine screening. Non-specific amplification in NAT can be prevented by pre-treatment with Sau3AI, filtration of NAT reagents, or reduction of the number of polymerase chain reaction cycles. FACS systems offer easy fully automated handling and a handling time of only 5 min, which could be an option for re-testing day-5 platelets. New screening approaches like immunoassays, detection of bacterial adenosine triphosphate, or detection of esterase activity need to be investigated in additional studies. Conclusion, Bacterial screening of blood products, especially platelets, can be done with a broad range of technologies. The ideal system should be able to detect one colony-forming unit per blood bag without a delay in the release process. Currently, we are far away from such an ideal screening system. Nevertheless, pathogen-inactivation systems are available, but a system for all blood components will not be expected in the next few years. Therefore, existing culture systems should be complemented by rapid systems like NAT or FACS especially for day-5 platelets. [source] Development of an Antibody Hapten-Chip System for Detecting the Residues of Multiple Antibiotic Drugs,JOURNAL OF FORENSIC SCIENCES, Issue 4 2009Ailiang Chen M.Sc. Abstract:, The abuse of antibiotic drugs during animal production remains a worldwide problem and the subsequent detection of the residues of various drugs present at low concentrations in complex biological matrices poses significant analytical challenges. The present study outlines a practical biochip assay system to identify antibiotic residues in different animal tissue extracts. The system uses a simple but efficient multiresidue sample extraction procedure to isolate the antibiotic residues which were then identified directly using high-affinity monoclonal antibodies presented in a competitive immunoassay with conjugated antibiotic hapten-chips. The hapten-chip can analyze six samples each for eight antibiotics on a single chip within 3 h. The analytical results with both artificial positive standard samples and the incurred samples show that the antibody hapten-chip system has a comparable accuracy and a similar sensitivity to a standard ultra performance liquid chromatography,mass spectrometry (MS)/MS assay. In conclusion, an effective analytical screening system based on antibody hapten-chip was developed for detecting multiple antibiotic residues from multiple samples. [source] Biological, pharmaceutical, and analytical considerations with respect to the transport media used in the absorption screening system, Caco-2JOURNAL OF PHARMACEUTICAL SCIENCES, Issue 8 2003Françoise M. Ingels Abstract During the evaluation and selection of drug candidates, the Caco-2 cell culture system is commonly used for the determination of intestinal transport characteristics and to anticipate permeability limited drug absorption. Although classic HBSS-like buffered salt solutions are commonly used to perform Caco-2 transport experiments, different shortcomings (e.g., adsorption and low solubility) have been associated with the use of plain aqueous buffers. As transport experiments performed with unoptimized conditions may compromize the value of the Caco-2 model as a permeation screening tool, many efforts have been made to optimize the experimental conditions of Caco-2 transport assays. In this minireview, the hurdles associated with the use of saline aqueous buffers in Caco-2 transport experiments are summarized and the different options, which have been proposed to overcome these issues, are reviewed and discussed. Biologically, pharmaceutically, as well as analytically relevant media affecting the outcome of the transport experiments are described. Unfortunately, up to now, no systematic studies comparing the different experimental conditions have been performed, jeopardizing the possibility to define a (single) optimal solution to overcome the different issues associated with the use of saline aqueous buffers. Based on the reported options it can be proposed to use DMSO (,1%) in standard screening procedures for the ranking of compounds based on their apical to basolateral transport. If compounds are not soluble in DMSO 1%, dimethylacetamide (3%) or N -1-methyl-pyrrolidone (2.5%) are good alternatives. However, these options do not imitate the in vivo situation. If one wants to take into account the physiological relevance of the media, the use of a biologically relevant apical medium (e.g., FASSIF) in combination with an analytically friendly, sink condition creating basolateral solvent (e.g., containing a micelle forming agent) can be suggested. © 2003 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 92:1545,1558, 2003 [source] Further validation of the Systematic Screening for Behavior Disorders in middle and junior high school,PSYCHOLOGY IN THE SCHOOLS, Issue 7 2009Michael J. Richardson The Systematic Screening for Behavior Disorders (SSBD), a screening system to identify elementary students at risk for emotional and behavioral disorders, was evaluated for use in middle and junior high schools. Teachers completed SSBD Stages One and Two on students in grades 6 to 8 who had characteristics of internalizing or externalizing disorders. Teacher, parent, and self-rating forms of the Achenbach System of Empirically Based Assessment (ASEBA) and the Social Skills Rating System (SSRS) were also completed on 66 students nominated via the SSBD as at risk for internalizing and externalizing problems. Office discipline referrals and grade point averages, for students nominated at SSBD Stage One, were compared with nonnominated students resulting in medium to large effect sizes. Small to moderate correlations were also found between SSBD Stage Two scores and ASEBA and SSRS scores, including several from the parent and student forms. © 2009 Wiley Periodicals, Inc. [source] Downstream utrophin enhancer is required for expression of utrophin in skeletal muscleTHE JOURNAL OF GENE MEDICINE, Issue 6 2008Jun Tanihata Abstract Background Duchenne muscular dystrophy is caused by the absence of the muscle cytoskeletal protein dystrophin. Utrophin is an autosomal homologue of dystrophin, and overexpression of utrophin is expected to compensate for the dystrophin deficit. We previously reported that the 5.4-kb 5,-flanking region of the utrophin gene containing the A-utrophin core promoter did not drive transgene expression in heart and skeletal muscle. To clarify the regulatory mechanism of utrophin expression, we generated a nuclear localization signal-tagged LacZ transgenic (Tg) mouse, in which the LacZ gene was driven by the 129-bp downstream utrophin enhancer (DUE) and the 5.4-kb 5,-flanking region of the utrophin promoter. Methods Two Tg lines were established. The levels of transgene mRNA expression in several tissues were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and quantitative RT-PCR. Cryosections of several tissues were stained with haematoxylin and eosin and X-gal. Results The transgene expression patterns were consistent with endogenous utrophin in several tissues including heart and skeletal muscle. Transgene expression was also up-regulated more in regenerating muscle than in nonregenerating muscle. Moreover, utrophin expression was augmented in the skeletal muscle of DUE Tg/dystrophin-deficient mdx mice through cross-breeding experiments. We finally established cultures of primary myogenic cells from this Tg mouse and found that utrophin up-regulation during muscle differentiation depends on the DUE motif. Conclusions Our results showed that DUE is indispensable for utrophin expression in skeletal muscle and heart, and primary myogenic cells from this Tg mice provide a high through-put screening system for drugs that up-regulate utrophin expression. Copyright © 2008 John Wiley & Sons, Ltd. [source] Construction and evaluation of a porcine bacterial artificial chromosome libraryANIMAL GENETICS, Issue 1 2000K Suzuki Summary A porcine bacterial artificial chromosome (BAC) library consisting of 103 488 clones has been constructed. The average insert size in the BAC vector was calculated to be 133 kb based on the examination of 189 randomly selected clones, indicating that the library contained 4.4 genome equivalents. The library can be screened by two-step PCR. The first screening step is performed on 22 superpools, each containing 4704 clones (49×96 well plates). In the second screening step, 49 plates comprising a superpool are arrayed in a 7×7 matrix and 4D-PCR is performed. Screening of the library superpools by PCR for 125 marker sequences selected from different regions of swine genome revealed 123 sequences, indicating that the library is not biased. Subsequent screenings (4D-PCR) were successfully applied for identification of clones containing each marker sequence. This porcine BAC library and the PCR screening system are useful for isolation of genomic DNA fragments containing desired sequences. [source] Synthesis of Monomeric and Dimeric Acridine Compounds as Potential Therapeutics in Alzheimer and Prion DiseasesARCHIV DER PHARMAZIE, Issue 12 2009René Csuk Abstract Starting from substituted 9-chloroacridines, a series of quinacrine and spacered dimeric acridine compounds was prepared. Their ability to interrupt the protein association of prion- and Alzheimer-specific proteins and Ab peptides was explored using a fast screening system based on FACS analysis. The bis-acridines displayed a higher activity than the corresponding monomers. Among these derivatives, best results were obtained with the 2,4-dimethoxy-6-nitro compound 7h for A,-peptides and the 2-methoxy-6-nitro compound 7f for PrP. [source] Synthesis of Dimeric Quinazolin-2-one, 1,4-Benzodiazepin-2-one, and Isoalloxazine Compounds as Inhibitors of Amyloid Peptides AssociationARCHIV DER PHARMAZIE, Issue 8 2009Alexander Barthel Abstract The synthesis of dimeric compounds derived from quinazolin-2-one and 1,4-benzodiazepin-2-one possessing a piperazine or homopiperazine spacer was investigated. In addition, a piperazine spacered bis-isoalloxazine and a bis-riboflavin compound were prepared and their ability to interrupt the association of prion proteins and Alzheimer-specific A, peptides was investigated using a fast screening system based on flow cytometry. The bis-isoalloxazine 14 was identified as a new lead structure. [source] In this issue: Biotechnology Journal 8/2010BIOTECHNOLOGY JOURNAL, Issue 8 2010Article first published online: 12 AUG 2010 Biocatalyst microemulsions Pavlidis et al., Biotechnol. J. 2010, 5, 805,812 Enzymes maintain their catalytic activity when hosted in aqueous nanodroplets like reverse micelles. Researchers from Ioannina, Greece, propose the use of water-in-ionic liquid microemulsionbased organogels (w/IL MBGs) as novel supports for the immobilization of lipase B from Candida antarctica and lipase from Chromobacterium viscosum. These novel lipase-containing w/IL MBGs can be effectively used as solid phase biocatalysts in various polar and non-polar organic solvents or ILs, exhibiting up to 4.4-fold higher esterification activity compared to water-in-oil microemulsion-based organogels. The immobilized lipases retain their activity for several hours at 70°C, while their half life time is up to 25-fold higher compared to that observed in w/IL microemulsions Biocatalyst cryogelation Bieler et al., Biotechnol. J. 2010, 5, 881,885 Entrapment of biocatalysts in hydrogel beads allows stable operation in otherwise deteriorating solvents. Doing this by cryogelation is a gentle method to extend the scope of biocatalysis. To foster the use of this versatile method, researchers from Aachen, Germany, devised an automated injector for the production of PVA/PEG-enzyme immobilisates. The device consists of a thermostated reservoir connected to a programmable injector nozzle and an agitated receiving bath for the droplets. This lab-scale production unit yields up to 1500 beads with immobilized enzyme per minute with a narrow size distribution and good roundness. Biocatalyst membrane reactor Lyagin et al., Biotechnol. J. 2010, 5, 813,821 Screening of biocatalysts, substrates or conditions in the early stages of bioprocess development requires an enormous number of experiments and is a tedious, expensive and time-consuming task. Currently available screening systems can only be operated in batch or fed-batch mode, which can lead to severe misinterpretations of screening results. Researchers from Berlin, Germany, now developed a novel screening system that enables continuous feeding of substrates and continuous removal of products. A prototype based on the membrane reactor concept was designed and operated for a model reaction, the hydrolysis of cellulose. [source] Continuous screening system for inhibited enzyme catalysis: A membrane reactor approachBIOTECHNOLOGY JOURNAL, Issue 8 2010Evgenij Lyagin Abstract The screening of catalysts, substrates or conditions in the early stages of bioprocess development requires an enormous number of experiments and is a tedious, expensive and time-consuming task. Currently available screening systems can only be operated in batch or fed-batch mode, which can lead to severe misinterpretations of screening results. For example, catalysts that are inhibited by substrates or accumulating products will be excluded from further investigations in the early stages of process development despite the fact that they might be superior to other candidates in a different operational mode. Important and advantageous properties such as turnover stability can also be overshadowed by product inhibition. The aim of this study was to develop a novel screening system that enables continuous feeding of substrates and continuous removal of products. A prototype based on the membrane reactor concept was designed and operated for a model reaction, the hydrolysis of cellulose. [source] High-throughput protein refolding screening method using zeoliteBIOTECHNOLOGY PROGRESS, Issue 4 2009Takayuki Y. Nara Abstract We established a 96-well-plate-based refolding screening system using zeolite. In this system, protein denatured and solubilized with 6 M guanidine hydrochloride is adsorbed onto zeolite placed in a 96-well plate. The refolding conditions can be tested by incubating the samples with refolding buffers under various conditions of pH, salts, and additives. In this study, we chose green fluorescent protein as the model protein. Green fluorescent protein was expressed as inclusion bodies, and we tested the effects of four pH conditions and six additives on its refolding. The results demonstrate that green fluorescent protein was more efficiently refolded with zeolite than with the conventional dilution method. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source] Measurement of Homogeneous Kinase Activity for Cell Lysates Based on the Aggregation of Gold NanoparticlesCHEMBIOCHEM, Issue 8 2007Jun Oishi When the charge sticks. A kinase activity screening system based on the aggregation of gold nanoparticles induced by cationic substrate peptides as coagulants has been constructed. The system can conduct and monitor kinase reactions under homogeneous conditions and has been successfully applied to the kinase assay of cell lysates. [source] Adolescent Satisfaction with Computer-Assisted Behavioural Risk Screening in Primary CareCHILD AND ADOLESCENT MENTAL HEALTH, Issue 4 2008Deena J. Chisolm Background:, This study measures patient satisfaction with a computerised mental health and risk-behaviour screening tool and predictors of satisfaction. Method:, Youth, aged 11,20, were recruited to use a laptop-based screening system in nine primary care clinics. The study assessed correlations between satisfaction with the system and selected predictors. Results:, Most users were satisfied with their experience. Multivariate logistic regression found perceived ease of use, perceived usefulness, and trust to be significantly associated with high satisfaction. Satisfaction was not related to computer experience or risk behaviour status. Conclusions:, Adolescent patients, even those at risk, accept computer-assisted screening in primary care. [source] Engineering Chiral Catalysts through Asymmetric Activation and Super High Throughput Screening (SHTS)CHINESE JOURNAL OF CHEMISTRY, Issue 6 2001Koichi Mkami Abstract A conceptually new strategy for asymmetric catalysis, namely asymmetric activation, in which a chiral activator selectively activates one enantiomer of a racemic chiral catalyst, and a highly efficient screening system for finding the most effective catalysts, namely super high throughput screening (SHTS), by which the reaction can be conducted in parallel and the ee% of the product is allowed to determine within minutes, are summarized in the present account. It is reasonable to believe that SHTS technique combined with asymmetric activation or deactivation principle will provide a very powerful methodology for finding the new catalysts and the best catalyst tuning for asymmetric reactions. [source] A European interlaboratory testing of three well-known procedures for immunocytochemical detection of epithelial cells in bone marrow.CYTOMETRY, Issue 6 2006Results from analysis of normal bone marrow Abstract Background: This investigation intended to study the unspecific background to be expected in normal bone marrow (BM), comparing three well recognized protocols for immunocytochemical detection of disseminated carcinoma cells. The interlaboratory variation in screening and evaluation of stained cells was analyzed and different screening methods were compared. Methods: BM mononuclear cells (BM MNC) from 48 normal BMs were immunostained in parallel by three participating laboratories. The protocols, based on three different anti-cytokeratin antibodies, have all been in common use for detection of disseminated carcinoma cells: the A45-B/B3 protocol (Hamburg), the CK2 protocol (Augsburg) and the AE1AE3 protocol (Oslo). For all protocols, the immunostained cells were visualized by the same alkaline-phosphatase (AP) detection system (APAAP) followed by detection of the cells by manual screening and by two different automated screening systems (ACIS from Chromavision and MDS1 from Applied Imaging). Detected AP-visualized cells were morphologically classified into unambiguous hematopoietic (Uhc) and questionable cells (Qc, potentially interpreted as tumor cells). Results: Seven of 48 BMs (15%) harbored ,1 AP-visualized cell(s) among 1 × 106 BM MNC, both for the A45-B/B3- and for the AE1AE3 protocol, while for CK2 a higher proportion of BMs (21 BMs; 44%) harbored AP-visualized cells (P < 0.01, McNemar's test). The number of Qc was, for all protocols, 1 log lower than the total number of AP-visualized cells. On average, the frequency of Qc was 0.04, 0.08, and 0.02 per 106 BM MNC with A45-B/B3, CK2 and AE1AE3, respectively, and the number of Qc-positive BMs 1, 4, and 1. The MDS1 screening sensitivity was similar to manual screening, while ACIS detected fewer cells (P < 0.001, McNemar's test). Conclusions: All protocols resulted in AP-visualization of occasional hematopoietic cells. However, morphological classification brings the specificity to a satisfactory high level. Approximately 10% of AP-visualized cells were categorized "questionable". The CK2 protocol turned out less specific than the A45-B/B3 and AE1AE3 protocols. © 2006 International Society for Analytical Cytology. [source] Invasion Possibility and Potential Effects of Rhus typhina on Beijing MunicipalityJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 5 2008Guangmei Wang Abstract Rhus typhina, an alien species introduced from North America, was identified as a main afforestation species in Beijing municipality. However, its invasiveness is still at odds. To clarify this problem, we applied the North American Screening System and the Australian Screening System to preliminarily predict its invasion possibility. Both screening systems gave the same recommendation to "reject". The geographical distribution was surveyed, with the population features of R. typhina against the native plant communities being assessed. With anthropogenic assistance, R. typhina has been scattered on almost all habitats from downtown to mountains, including roadsides, farmlands and protected areas. As a clonal shrub, R. typhina possessed a high spreading rate, varying from 6.3 m/3 years at sterile habitats to 6.7 m/3 years at fertile ones. Significantly lower species richness, individual density and diversity were observed in the R. typhina community than those of the native Vitex negundo Linn.var. heterophylla (Franch.) Rehd. community at both sterile and fertile habitats. Continual wide plantation of R. typhina may further foster its population expansion, which helps the species to overcome spatial isolation. The fact that each root fragment can develop into a new individual makes R. typhina very difficult to be eradicated once established. From a biological point of view, we believe that R. typhina is a plant invader in Beijing. We therefore suggest the government should remove the name of R. typhina from the main tree species list in afforesting Beijing. [source] Advances in membrane receptor screening and analysisJOURNAL OF MOLECULAR RECOGNITION, Issue 4 2004Matthew A. Cooper Abstract During the last decade there has been significant progress in the development of analytical techniques for the screening of ligand binding to membranes and membrane receptors. This review focuses on developments using label-free assays that facilitate ligand,membrane,receptor screening without the need for chemical-, biological- or radiological-labelled reagents. These assays include acoustic, optical surface plasmon resonance biosensing, sedimentation (analytical ultracentrifugation), chromatographic assays, isothermal titration calorimetry and differential scanning calorimetry. The merits and applications of cell-based screening systems and of different model membrane systems, including planar supported lipid layers, bead-supported membranes and lipid micro-arrays, are discussed. Recent advances involving more established techniques including intrinsic fluorescence, FRET spectroscopy, scintillation proximity assays and automated patch clamping are presented along with applications to peripheral membrane proteins, ion channels and G protein-coupled receptors. Novel high-throughput assays for determination of drug- and protein-partitioning in membranes are also highlighted. To aid the experimenter, a brief synopsis of the techniques commonly employed to purify and reconstitute membranes and membrane receptors is included. Copyright © 2004 John Wiley & Sons, Ltd [source] Quick prediction of the retention of solutes in 13 thin layer chromatographic screening systems on silica gel by classification and regression treesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15 2008ukasz Komsta Abstract The use of classification and regression trees (CART) was studied in a quantitative structure,retention relationship (QSRR) context to predict the retention in 13 thin layer chromatographic screening systems on a silica gel, where large datasets of interlaboratory determined retention are available. The response (dependent variable) was the rate mobility (RM) factor, while a set of atomic contributions and functional substituent counts was used as an explanatory dataset. The trees were investigated against optimal complexity (number of the leaves) by external validation and internal crossvalidation. Their predictive performance is slightly lower than full atomic contribution model, but the main advantage is the simplicity. The retention prediction with the proposed trees can be done without computer or even pocket calculator. [source] In this issue: Biotechnology Journal 8/2010BIOTECHNOLOGY JOURNAL, Issue 8 2010Article first published online: 12 AUG 2010 Biocatalyst microemulsions Pavlidis et al., Biotechnol. J. 2010, 5, 805,812 Enzymes maintain their catalytic activity when hosted in aqueous nanodroplets like reverse micelles. Researchers from Ioannina, Greece, propose the use of water-in-ionic liquid microemulsionbased organogels (w/IL MBGs) as novel supports for the immobilization of lipase B from Candida antarctica and lipase from Chromobacterium viscosum. These novel lipase-containing w/IL MBGs can be effectively used as solid phase biocatalysts in various polar and non-polar organic solvents or ILs, exhibiting up to 4.4-fold higher esterification activity compared to water-in-oil microemulsion-based organogels. The immobilized lipases retain their activity for several hours at 70°C, while their half life time is up to 25-fold higher compared to that observed in w/IL microemulsions Biocatalyst cryogelation Bieler et al., Biotechnol. J. 2010, 5, 881,885 Entrapment of biocatalysts in hydrogel beads allows stable operation in otherwise deteriorating solvents. Doing this by cryogelation is a gentle method to extend the scope of biocatalysis. To foster the use of this versatile method, researchers from Aachen, Germany, devised an automated injector for the production of PVA/PEG-enzyme immobilisates. The device consists of a thermostated reservoir connected to a programmable injector nozzle and an agitated receiving bath for the droplets. This lab-scale production unit yields up to 1500 beads with immobilized enzyme per minute with a narrow size distribution and good roundness. Biocatalyst membrane reactor Lyagin et al., Biotechnol. J. 2010, 5, 813,821 Screening of biocatalysts, substrates or conditions in the early stages of bioprocess development requires an enormous number of experiments and is a tedious, expensive and time-consuming task. Currently available screening systems can only be operated in batch or fed-batch mode, which can lead to severe misinterpretations of screening results. Researchers from Berlin, Germany, now developed a novel screening system that enables continuous feeding of substrates and continuous removal of products. A prototype based on the membrane reactor concept was designed and operated for a model reaction, the hydrolysis of cellulose. [source] Continuous screening system for inhibited enzyme catalysis: A membrane reactor approachBIOTECHNOLOGY JOURNAL, Issue 8 2010Evgenij Lyagin Abstract The screening of catalysts, substrates or conditions in the early stages of bioprocess development requires an enormous number of experiments and is a tedious, expensive and time-consuming task. Currently available screening systems can only be operated in batch or fed-batch mode, which can lead to severe misinterpretations of screening results. For example, catalysts that are inhibited by substrates or accumulating products will be excluded from further investigations in the early stages of process development despite the fact that they might be superior to other candidates in a different operational mode. Important and advantageous properties such as turnover stability can also be overshadowed by product inhibition. The aim of this study was to develop a novel screening system that enables continuous feeding of substrates and continuous removal of products. A prototype based on the membrane reactor concept was designed and operated for a model reaction, the hydrolysis of cellulose. [source] | |||||||||||||||||||||||||||||||