Multiple Ab Research Articles

High-density IgE recognition of the major grass pollen allergen Phl p 1 revealed with single-chain IgE antibody fragments obtained by combinatorial cloning.

The timothy grass pollen allergen Phl p 1 belongs to the group 1 of highly cross-reactive grass pollen allergens with a molecular mass of ∼25-30 kDa. Group 1 allergens are recognized by >95% of grass pollen allergic patients. We investigated the IgE recognition of Phl p 1 using allergen-specific IgE-derived single-chain variable Ab fragments (IgE-ScFvs) isolated from a combinatorial library constructed from PBMCs of a grass pollen-allergic patient. IgE-ScFvs reacted with recombinant Phl p 1 and natural group 1 grass pollen allergens. Using synthetic Phl p 1-derived peptides, the binding sites of two ScFvs were mapped to the N terminus of the allergen. In surface plasmon resonance experiments they showed comparable high-affinity binding to Phl p 1 as a complete human IgE-derived Ab recognizing the allergens' C terminus. In a set of surface plasmon resonance experiments simultaneous allergen recognition of all three binders was demonstrated. Even in the presence of the three binders, allergic patients' polyclonal IgE reacted with Phl p 1, indicating high-density IgE recognition of the Phl p 1 allergen. Our results show that multiple IgE Abs can bind with high density to Phl p 1, which may explain the high allergenic activity and sensitizing capacity of this allergen.

P018 : IDENTIFICATION OF UNACCEPTABLE HLA ANTIGENS WHEN PRESENT IN TRANSPLANT PATIENT SERA AT AN UNDILUTED OR NEAT TITER BUT NOT PRESENT AT A LOW TITER (⩽1:16)

Characteristics identifying clinically deleterious HLA antibodies (Ab) may include specificity, mean fluorescence intensity (MFI) and C1q reactivity. These factors are also used when identifying unacceptable HLA antigens (Ags). However, if unacceptable HLA Ab specificities are present in undiluted sera but not present when sera are diluted 1:16 then these Ab specificities may be ignored when deciding on which unacceptable Ab specificities to report. This reduction in unacceptable Ags would allow for easier matching of highly sensitized patients (with high numbers of HLA Ab specificities) to an appropriate donor. Thirty-nine cI/cII high PRA sera were evaluated for the presence of HLA Abs and their Ag specificities at neat and at a 1:16 titer. The number of Ab specificities were compared at the two dilutions. One Lambda, Inc. reagents were used per the manufacturers instructions. The HLA cI/cII PRAs were 77 ± 25% and 84 ± 21% respectively for the 39 sera. The total number of HLA cI/cII Ags identified in undiluted sera were 46 ± 22 and 34 ± 22 respectively per patient. In addition, there were 7 ± 8 cI and 5 ± 4 cII falsely identified HLA Ags respectively per patient. When sera were titrated to a 1:16 dilution 71 ± 25% of cI and 51 ± 28% of cII unacceptable Ags were eliminated. The total number of cI Ags eliminated were 21 ± 10 and 15 ± 9 cII Ags. When false positive Ags were included in the evaluation the frequency for Ag elimination at a 1:16 dilution for cI became 85 ± 14% and 66 ± 24% for cII respectively. Fifty-four patients with donor specific antibody (DSA) titers of ⩽1:16 were identified. 81% (44/54) had both negative cytotoxic (AHG) and flow cytometric (FCXM) donor specific crossmatches whereas 10 patients (19%, 10/54) displayed a negative AHG but a positive FCXM. When transplanted, the FCXM negative patients had a two year graft survival of 91% vs. 60% for the FCXM positive patients ( p These data suggest that titration studies of high PRA, multiple Ab specificity presenting sera, can be used when identifying unacceptable Ags. Moreover, while the presence of DSA in low concentration (⩽1:16) may falsely suggest a positive virtual crossmatch it may not result in a real time positive crossmatch. HLA Ab specificities that are eliminated at a low titer (1:16) may allow for easier matching of highly sensitized patients to donors for transplant.

Anion recognition by uranyl-salophen derivatives as probed by infrared multiple photon dissociation spectroscopy and ab initio modeling.

The vibrational features and molecular structures of complexes formed by a series of uranyl-salophen receptors with simple anions, such as Cl(-) , H(-) , and HCOO(-) , have been investigated in the gas phase. Spectra of the anionic complexes were studied in the $\tilde \nu $=800-1800 cm(-1) range by mass-selective infrared multiple photon dissociation (IRMPD) spectroscopy with a continuously tunable free-electron laser. The gas-phase decarboxylation of the formate adducts produces uranyl-salophen monohydride anions, which have been characterized for the first time and reveal a strong UH bond, the nature of which has been elucidated theoretically. The spectra are in excellent agreement with the results obtained from high-quality ab initio calculations, which provided the structure and binding features of the anion-receptor complexes.

Polymorphism in paracetamol: evidence of additional forms IV and V at high pressure.

The structural phase stability of N-(4-hydroxyphenyl) acetamide (paracetamol) has been studied at ambient temperature up to 23 GPa using Raman spectroscopy. Spectral changes have provided further evidence for a highly kinetically driven Form I → II transition that occurs as a mixed phase from 4.8 to 6.5 GPa, and might complete as early as 7 GPa. Upon further compression to 8.1 GPa, a drastic shift in spectral signature was observed providing the first evidence for a previously undiscovered Form IV of paracetamol. Additional shifts in mode intensities were observed near 11 GPa indicating a potential restructuring of the hydrogen bonding network and/or structural modification to a potentially new Form V. Phase boundaries at 7 and 8 GPa were confirmed under hydrostatic conditions using Raman spectroscopy. Spectral changes indicate that the transition Form IV → V occurs near 11 GPa. Multiple ab initio harmonic frequency calculations at different levels of theory were performed with a B3LYP/6-31G** being used to provide a more robust mode assignment to our experimentally obtained Raman modes. High pressure X-ray diffraction (XRD) was performed up to 21 GPa, which provided further evidence for a highly kinetically driven Form I → II transition in agreement with our Raman measurements. In addition, the XRD provided further evidence for the existence of Form IV near 8 GPa and Form V near 11 GPa with Form V persisting up to 21 GPa.

Elimination of Unacceptable HLA Antigens When Present in Transplant Patient Sera at an Undiluted or Neat Titer But Not Present at a Low Titer (≤ 1:16).

HLA antibodies (Ab) are routinely identified in transplant recipient sera. Characteristics determining clinically deleterious Abs include specificity, MFI and C1q. These factors are also used when identifying unacceptable HLA antigens (Ags). However, if unacceptable HLA Ab specificities are present in undiluted sera but not present when the sera is diluted 1:16 then these Ab specificities can possibly be ignored when deciding on which unacceptable Ab specificities to report. This reduction in unacceptable antigens would allow for an easier matching of highly sensitized patients to an appropriate donor for transplant. Thirty-nine cI/cII high PRA sera were evaluated for the presence of HLA Abs and their antigen specificities at neat and at a 1:16 titer. The number of Ab specificities were compared at the two dilutions. One Lambda, Inc. reagents were used per the manufacturers instructions. The HLA cI/cII PRAs were 77±25% and 84±21% respectively for the 39 sera. The total number of HLA cI/cII Ags identified in undiluted sera were 46±22 and 34±22 respectively per patient. In addition, there were 7±8 cI and 5±4 cII falsely identified HLA Ags respectively per patient. When sera were titrated to a 1:16 dilution 71±25% of cI and 51±28% of cII unacceptable Ags were absent. The total number of cI Ags eliminated were 21±10 and cII Ags were 15±9. When false positive Ags were included in the evaluation the frequency for Ag elimination at a 1:16 dilution for cI became 85±14% and 66±24% for cII respectively. Fifty-four patients with donor specific antibody (DSA) titers of ≤ 1:16 were identified. 81% (44/54) had both negative cytotoxic (AHG) and flow cytometric (FCXM) donor specific crossmatches whereas 10 patients (19%, 10/54) displayed a negative AHG but a positive FCXM. When transplanted, the FCXM negative patients had a two year graft survival of 91% vs. 60% for the FCXM positive patients (p < 0.01). These data suggest that titration studies of high PRA, multiple Ab specificity presenting sera can be used when identifying unacceptable Ags. Moreover, the presence of DSA in low concentration (≤ 1:16) while falsely suggesting a positive virtual crossmatch may not result in a real time positive crossmatch. HLA Ab specificities that are eliminated at a low titer (1:16) may allow for easier matching of highly sensitized patients to donors for transplant.

Transient Antibody-Mucin Interactions Produce a Dynamic Molecular Shield against Viral Invasion

Given the difficulty in finding a cure for HIV/AIDS, a promising prevention strategy to reduce HIV transmission is to directly block infection at the portal of entry. The recent Thai RV144 trial offered the first evidence that an antibody-based vaccine may block heterosexual HIV transmission. Unfortunately, the underlying mechanism(s) for protection remain unclear. Here we theoretically examine a hypothesis that builds on our recent laboratory observation: virus-specific antibodies (Ab) can trap individual virions in cervicovaginal mucus (CVM), thereby reducing infection in vivo. Ab are known to have a weak—previously considered inconsequential—binding affinity with the mucin fibers that constitute CVM. However, multiple Ab can bind to the same virion at the same time, which markedly increases the overall Ab-mucin binding avidity, and creates an inheritable virion-mucin affinity. Our model takes into account biologically relevant length and timescales, while incorporating known HIV-Ab affinity and the respective diffusivities of viruses and Ab in semen and CVM. The model predicts that HIV-specific Ab in CVM leads to rapid formation and persistence of an HIV concentration front near the semen/CVM interface, far from the vaginal epithelium. Such an HIV concentration front minimizes the flux of HIV virions reaching target cells, and maximizes their elimination upon drainage of genital secretions. The robustness of the result implies that even exceedingly weak Ab-mucin affinity can markedly reduce the flux of virions reaching target cells. Beyond this specific application, the model developed here is adaptable to other pathogens, mucosal barriers, and geometries, as well as kinetic and diffusional effects, providing a tool for hypothesis testing and producing quantitative insights into the dynamics of immune-mediated protection.

Multiwavelength Observations of AB Doradus

AbstractWe have observed the bright, magnetically active multiple star AB Doradus in a multiwavelength campaign centring around two large facility allocations in November 2006 and January, 2007. Our observations have covered at least three large flares. These flares were observed to produce significant hardening of the X-ray spectra during their very initial stages. We monitored flare-related effects using the Suzaku X-ray satellite and the Australia Telescope Compact Array (ATCA) at 3.6 and 6 cm. Observations at 11 and 21 cm were also included, but they were compromised by interference. Optical monitoring was also provided by broadbandBandVphotometry and some high-dispersion spectrograms. From this multiwavelength coverage we find that the observed flare effects can be mainly associated with a large active region near longitude zero. The second major X-ray and microwave flare of Jan 8, 2007 was observed with a favourable geometry that allowed its initial high-energy impulsive phase to be observed in the higher frequency range of Suzaku’s XIS detectors. The fractional circular polarisation (Stokes V/I) was measured in the uv data for the complete runs, for 25 min integrations and, at 4.80 GHz, for 5 min integrations, using the radio data of Nov 21 2006 and Jan 08 2007. Most of the full data sets showed V/I fractions from AB Dor B that were significant at greater than the 3σ level. In several of the 5 min integrations at 4.80 and 8.64 GHz this fraction reached a significance level between 3 and 9σ. Lack of angular resolution prevented identification of these high V/I values with one or other of the two low-mass red-dwarf components of AB Dor B.

Comparação entre medidas da Classe de Respostas “Separar o lixo adequadamente”

As medidas mais usadas para o estudo da classe de respostas “separação adequada do lixo” em pesquisas na análise do com¬portamento são a pesagem e a contagem de itens recicláveis depositados nas lixeiras. Embora diferentes medidas sejam utilizadas, não parece haver na literatura qualquer discussão da adequação relativa das diferentes medidas à variável dependente. O presente estudo teve como objetivo realizar uma comparação entre as medidas citadas e a observação direta do comportamento dos participantes. Para realizar a comparação entre medidas, o estudo foi delineado como replicação sistemática do estudo de Austin et al. (1993) sobre com¬portamento de reciclagem. Os participantes foram membros de dois prédios de uma universidade e eventuais transeuntes das proximi¬dades. Foi utilizado delineamento AB de linha de base múltipla. específicos foram introduzidos como variável independente. O índice de correlação entre a observação direta e a contagem de itens mostrou-se superior ao índice de correlação entre a observação direta e a pesagem. Os resultados obtidos pela pesquisa original não foram replicados. Os resultados do presente estudo parecem sugerir que pesquisas futuras devam atribuir maior grau de confiança à contagem de itens.

Passenger Lymphocyte Syndrome Following Solid Organ Transplantation: Graft Source, Incidence, Specificity, Duration, and Severity Of Hemolysis

BackgroundPassenger Lymphocyte Syndrome (PLS) is a rare complication of solid organ transplantation (SOT) and is marked by production of donor-derived antibodies towards host red blood cell (RBC) antigens. At Canada's largest SOT program, >400 transplants are conducted annually. The affiliated Transfusion Laboratory detects possible cases of PLS when previously seronegative hosts develop (delayed, non-transfusion-attributable) post-transplant antibodies (Ab) bearing unexpected, autoreactive (rather than alloreactive) RBC specificities. Donor attributability is established by discovery of the same specificity (on serologic lookback) and/or by the antithetical (seroconversion risk) phenotype. The incidence, duration, and severity of PLS remain unknown due to the infrequency of reported cases and the general absence of active surveillance. MethodBetween 1/10/2006 – 30/06/2013, PLS cases were discovered by the Blood Bank through pre-transfusion specimens. The specificities, phase (direct or indirect), correlative markers of hemolysis, and temporal profile of the Ab were subsequently followed according to patient and clinician interest. The severity of hemolysis (grade 0 to 5) at the time of nadir hemoglobin was judged according to the tiers defined by the Canadian Blood Services/IVIG Hemolysis Pharmacovigilance Group (CL#2011-34). Implicated organ and donor review (for relevant serology, and outcomes in any parallel organ distributions) were also attempted. ResultsOver 88 months, 2772 transplants occurred (992 kidney, 187 pancreas [with/after kidney in 113/74 respectively], 909 liver, 529 lung [437 double, 83 single, 9 +heart], 151 heart, 4 bowel). PLS was found in 14 or 0.5% (sex: 5F: 9M; mean age 56 years), with the implicated organ being lung or liver (9 [2%] and 5 [0.6%] respectively). All had a negative pre-transplant DAT with a pre-sensitized donor. PLS Ab specificities included ABO (8: 5A, 3B), RH (5 with ≥1 target [C:3, D:5, E:2, V:1]), and others (KIDD: Jka in 2; MNS: N in 1; unidentified in 1). Multiple Ab occurred in 5 (36%), usually within the same system (3 RH cases: C,D,E in 2; C,D,V in 1), but otherwise towards ABO + ≥1 target (B,Jka,N; B, unidentified). The time to the first detectable expression of an Ab (as signaled by a +DAT and/or screen) was 4-120 days (mean 24, median 15, n=14). Clinically significant hemolysis occurred in 11 (79%), with six grade 3 and five grade 4; the duration of hemolysis was 0-776 days (mean 148, median 38, n=11). The duration of Ab detectability (or the point of “last positivity”) was 12-851 days (mean 196, median 78, n=10), while the point of disappearance (“subsequent sustained negativity”) occurred between day 12-1288 (mean 298, median 45, n=10). Lookbacks were feasible with 5 implicated donors, and in only 1 case (of a contralateral lung's transplantation) was PLS detected in another recipient. PLS did not account directly for any deaths in the cohort, although hemolytic morbidity required treatment in 2 cases. Transfusion support with antigen-negative blood was provided to 11 patients for anemia that may have been due to hemolysis and/or other causes while the Ab was still evident. In one case, severe hemolysis provoked rate-uncontrolled atrial fibrillation with a supply and demand ischemia which culminated in irreversible congestive heart failure and cardiac death 2 months after PLS resolution. [Display omitted] Discussion/ConclusionsThis is the largest series of PLS reported to date. Clinician-driven submissions of blood for transfusion compatibility testing, and the discovery of discrepant serologies, represented an unforced but prospective means by which to identify PLS in roughly 1 in 200 transplants. Several cases transcended the limits of previously described temporal profiles (with the most delayed onset at 120 days, and the most prolonged duration at 2.3 years). The duration of hemolysis was consistently shorter than the duration of seropositivity, and was rarely life-threatening, while the most severe grades associated with RH rather than ABO or other targets. The surveillance achieved in this cohort revealed a longer-than-previously-appreciated duration of Ab productivity, and raised the question of whether or not such prolongation is a function of observation bias or the current-day armamentarium of anti-rejection therapeutics, which may be more permissive to donor-derived chimerism. Disclosures:No relevant conflicts of interest to declare.

Valence shell direct double photodetachment in polyanions

A valence shell study of electrosprayed insulin protein polyanion photodetachment was carried out on a vacuum ultra-violet synchrotron radiation beamline coupled to a radiofrequency ion trap, for both close- and open-shell species. A two-electron photodetachment is observed, which arises from two different mechanisms that are disentangled: a sequential multi-photon absorption and a direct one-photon two-electron process. The threshold for the direct double-electron ejection is measured at 11.4 eV and corresponds to electronic excitation in the valence shell, which makes it the first observation of direct double photodetachment in the valence shell. The results are discussed in the light of previous knowledge from multiple photoionization and ab initio calculations on model polyanions. Double photodetachment appears to be a relaxation mechanism that leads to oxidized anions of striking stability, a feature of high relevance in radiobiology.

3-P

Aim Luminex based analysis of HLA antibody concentration in serum (represneted as mean fluorescence intensity-MFI) is increasingly used for patient management. However, different factors contribute to the variation in these MFI values including a real biological condition, type of antigen, disease or assay, drug treatment and technical variation. Here, we evaluated MFI variation for HLA class I and II single antigen assay when performed by different technologist for sera known to have multiple HLA abs of varied strength. Methods A total of 6 highly sensitized samples (3 samples each for HLA class I and II) were tested by three technologists using labscreen ® Single antigen assay. To ensure uniformity, all experiments were performed according to the manufacturer’s guidelines on the same day using same reagents, centrifuges & Luminex analyzer and reactions were performed on the same position of the tray. Mean, standard deviation (STD), and coefficient variance (CV) was calculated. Results Significant technical variation was observed, where average CV was negatively correlated with the strength of antibody. Higher CV was observed for abs with MFI of 10,000 or more. Differences were significant for HLA Class I abs and showed a trend of significance for HLA class II abs.[ Table 1 ]A total of 60 HLA class I and 83 HLA class II abs were detected. Significantly lower CV was observed in case of HLA class I than HLA class II abs (avg CV = 8.3% and 16.5% respectively), though this was primarily due to the variation in quantification of HLA-DP abs (avg CV = 29%). Conclusions Technical variation in MFI values for different HLA abs was inversely proportional to the strength of the antibody. Our results suggest that contribution from technical variation should be considered in interpreting single antigen assay results especially when a longitudinal study for patient serum is being performed.

The 2012 ICC Rules of Arbitration – An Accelerated Procedure and Substantial Changes

The 2012 International Chamber of Commerce (ICC) Rules of Arbitration pave the way for more expeditious arbitral proceedings since under the added requirements, the parties are to provide more extensive information during the initial stages of the proceedings. The delays caused by further submissions are thereby avoided. A wider approach towards the case, may enable the joinder of additional parties, to address claims between multiple parties and multiple contracts ab initio, and to consolidate arbitration proceedings. The new Rules establish an emergency arbitrator procedure for urgent interim or conservatory measures, as well as case management techniques to reduce the costs. The work of the members of the ICC Commission of Arbitration, the ICC National Committees, the International Court of the Arbitration, the representatives of the National Committees, and selected counsels and arbitrators, over a two-year period bore results: in June 2011 in Mexico City, a revised version of the 1998 ICC Rules of Arbitration was adopted by The World Council of the ICC (hereinafter 'the 2012 ICC Rules' and/or 'the new Rules').

Structural Basis for the 14-3-3 Protein-dependent Inhibition of the Regulator of G Protein Signaling 3 (RGS3) Function

Regulator of G protein signaling (RGS) proteins function as GTPase-activating proteins for the α-subunit of heterotrimeric G proteins. The function of certain RGS proteins is negatively regulated by 14-3-3 proteins, a family of highly conserved regulatory molecules expressed in all eukaryotes. In this study, we provide a structural mechanism for 14-3-3-dependent inhibition of RGS3-Gα interaction. We have used small angle x-ray scattering, hydrogen/deuterium exchange kinetics, and Förster resonance energy transfer measurements to determine the low-resolution solution structure of the 14-3-3ζ·RGS3 complex. The structure shows the RGS domain of RGS3 bound to the 14-3-3ζ dimer in an as-yet-unrecognized manner interacting with less conserved regions on the outer surface of the 14-3-3 dimer outside its central channel. Our results suggest that the 14-3-3 protein binding affects the structure of the Gα interaction portion of RGS3 as well as sterically blocks the interaction between the RGS domain and the Gα subunit of heterotrimeric G proteins.

Tight and efficient enclosure of matrix multiplication by using optimized BLAS

AbstractThis paper is concerned with the tight enclosure of matrix multiplication AB for two floating‐point matrices A and B. The aim of this paper is to compute component‐wise upper and lower bounds of the exact result C of the matrix multiplication AB by floating‐point arithmetic. Namely, an interval matrix enclosing C is obtained. In this paper, new algorithms for enclosing C are proposed. The proposed algorithms are designed to mainly exploit the level 3 operations in BLAS. Although the proposed algorithms take around twice as much costs as a standard algorithm promoted by Oishi and Rump, the accuracy of the result by the proposed algorithms is better than that of the standard algorithm. At the end of this paper, we present numerical examples showing the efficiency of the proposed algorithms. Copyright © 2010 John Wiley &amp; Sons, Ltd.

CXCR7 Protein Is Not Expressed on Human or Mouse Leukocytes

Since the discovery that CXCR7 binds to CXCL12/SDF-1α, the role of CXCR7 in CXCL12-mediated biological processes has been under intensive scrutiny. However, there is no consensus in the literature on the expression of CXCR7 protein by peripheral blood cells. In this study we analyzed human and mouse leukocytes and erythrocytes for CXCR7 protein expression, using a competitive CXCL12 binding assay as well as by flow cytometry and immunohistochemistry using multiple CXCR7 Abs. CXCR7(-/-) mice were used as negative controls. Together, these methods indicate that CXCR7 protein is not expressed by human peripheral blood T cells, B cells, NK cells, or monocytes, or by mouse peripheral blood leukocytes. CXCR7 protein is, however, expressed on mouse primitive erythroid cells, which supply oxygen to the embryo during early stages of development. These studies therefore suggest that, whereas CXCR7 protein is expressed by primitive RBCs during murine embryonic development, in adult mammals CXCR7 protein is not expressed by normal peripheral blood cells.

Support in everyday activities with a home-based electronic memory aid for persons with memory impairments

Purpose. The purposes of this study was to examine the possibilities of a home-based electronic memory aid with sensors for persons with memory impairments, as support to carry out everyday activities in their own home environments.Method. The method involved a single-subject study with a multiple baseline AB design. Five participants identified three activities each that they usually forget to carry out. An electronic memory aid with individually spoken reminders was installed in the participant's home. There were automatic computer registrations of completed activities during the study phase of 12 weeks. Assessments of functioning and quality of life (QoL) were conducted before and after the intervention and at follow-up after 2 months.Results. Four participants improved in completing most of the self-chosen activities when the electronic memory aid was used. Performance and satisfaction with performance and QoL improved, but there was no memory function improvement. There were technical problems with the aid, which had a negative effect for users.Conclusions. Electronic memory aids have a large potential for supporting persons with cognitive impairments. It is important to conduct follow-up afterwards, because the use of an aid and the need of support change over time and put high demands on technical reliability of the electronic memory aid.

Isolation of Cross-Clade HIV-Neutralizing Human Antibodies from Memory B Cell Repertoires Using Short Term Culture and High-Throughput Primary Neutralization Screens (38.17)

Abstract Background. We have recently isolated 2 broadly neutralizing monoclonal antibodies (bNAbs), PG9 and PG16, from an HIV-infected subject in an African Protocol G cohort that are more potent than the bNAbs discovered in the past decade. The discovery highlights our human antibody (Ab) discovery approach based on short-term memory B cell culture, high-throughput primary microneutralization screening, and recombinant monoclonal antibody (mAb) reconstitution (Walker LM, et al, 2009, Science, 326:285-9). The epitope specificities of PG9 and PG16 reveal a novel target for designing vaccine immunogens that may elicit broad anti-HIV protection. To identify additional epitopes for vaccine design, we extended the bNAb discovery to more HIV-infected subjects. Method. We selected several HIV-infected subjects with broad neutralization activity in serum. Memory B cells from these elite neutralizers were activated and the IgG-containing culture media were screened for neutralizing activity against multiple pseudotyped viruses. mAbs were rescued from selected B cell culture wells by RT-PCR and subcloning. Results. From an HIV-infected elite neutralizer, we have identified in the primary screen multiple human Abs that cross-neutralize MGRM-C-26 of clade C, and 93BR020 and JR-CSF of clade B. Antibodies that neutralize either MGRM-C-26 alone or 94UG103 of clade A alone were also detected. Recombinant mAbs are currently being rescued for further characterization. (Supported by IAVI and USAID)

Study of AB 2 alloy electrodes for Ni/MH battery prepared by centrifugal casting and gas atomization

Centrifugal casting and gas atomization processes were applied on multiple phase AB 2 alloys and compared to the conventional melt-and-cast. Four different compositions were chosen for this study. The roles of Zr, Mn, Cr, and Ni in various battery aspects are identified. Cooling speed was found to be crucial for the C14 and C15 phase abundances. As the cooling speed increased from 10 2 to 10 4 degrees per second, a higher percentage of C15 was found. The centrifugal casting process provided better cycle life and low temperature performance with the only trade-off being slower activation. The gas atomization process can achieve lower production cost due to the elimination of a grinding procedure and extended cycle life, but suffers from higher bulk oxygen content and thicker surface oxide, and thus inferior in all battery performance characteristics other than cycle life and charge retention.

Development of the Releasable Asbestos Field Sampler

The releasable asbestos field sampler (RAFS) was developed as an alternative to activity-based sampling (ABS; personal breathing zone sampling during a simulated activity). The RAFS utilizes a raking motion to provide the energy that releases particulate material from the soil and aerosolizes the asbestos fibers. A gentle airflow laterally transports the generated aerosol inside of a tunnel to one end where filter sampling cassettes or real-time instruments are used to measure asbestos and particulate release. The RAFS was tested in a series of laboratory experiments to validate its performance and then was deployed for field trials in asbestos-contaminated soil at multiple geographical locations. Laboratory data showed the RAFS generated repeatable and representative aerosol particulate concentrations. Field tests showed the RAFS aerosolized asbestos concentrations were statistically correlated with total particle concentrations. Field tests also showed the RAFS aerosolized asbestos concentrations were statistically correlated with asbestos concentrations measured by multiple ABS tests with different activities, different soil/environmental conditions, and at different geographical locations. RAFS provides a direct measurement of asbestos emission from soil in situ without consideration of meteorology and personal activity on the asbestos transport to the breathing zone.

Internal validation of 29 autosomal SNP multiplex using a ABI 310 genetic analyser

In the last few years genetic identification and paternity testing have begun to make increasing use of autosomal SNP (Single Nucleotide Polymorphism) typing as a supplement or alternative to STR analysis. With the improvement in detection technology SNP analysis is likely to be easier and more sensitive, with the generation of new methods and multiplex systems for a growing array of SNP markers. SNPforID consortium developed 52 SNP PCR multiplex for human identification purposes detected with 23 plex and 29 plex single base extension reactions (Auto1 and 2 respectively). In this study, internal validation for the 29 SNPs of Auto2 was carried out by performing a 29 plex PCR and single base extension reaction on control samples and previously analyzed forensic casework and subsequent detection with an AB 310 Genetic Analyzer. We tested the accuracy, precision, sensitivity and reproducibility of the Auto2 multiplex with this instrument in our laboratory. We used 9947A control DNA samples of the AmpFℓSTR Identifiler™ kit to test the validation parameters together with non-probative DNA samples from whole blood and buccal swab samples of 29 healthy donors from different parts of Istanbul. Good results were obtained but interpretation of the peak patterns obtained on the AB 310 requires care and thorough optimization before they can be readily compard to those obtained from multiple capillary AB 31xx Analyzers. We succesfully optimized and validated the SNP forID Auto2 multiplex system for identification analyses in our laboratory.