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Applications of murine and humanized chimaeric monoclonal antibodies for red cell phenotyping.

The limited supply of reagent human polyclonal antibodies to high prevalence antigens, like Js(b), is driving the search for alternative reagents. Murine immunoglobulin G (IgG) monoclonal antibodies (Mabs) and their humanized chimaeric IgM isoforms can now be used for typing patients and screening donors. Antigen typing of red blood cells (RBC) with a positive direct antiglobulin test (DAT) is also possible using these antibodies. Blood from patients with sickle cell disease and African donors were tested with reagent anti-Js(b), murine Mab IgG anti-Js(b) [murine immunochemistry monoclonal antibody-8 (MIMA-8)], and humanized chimaeric IgM anti-Js(b) [human immunochemistry monoclonal antibody-8 (HIMA-8)] by haemagglutination and gel cards. RBC samples that were DAT positive were used to evaluate the humanized chimaeric IgM monoclonal anti-Fy(a) (HIMA-19). RBC samples (n = 243) of known Js(b) type were tested in parallel with MIMA-8 and reagent anti-Js(b), and 132 samples were tested with MIMA-8 in gel cards and HIMA-8 by direct tube testing. No discrepant results were obtained. DAT-positive RBC samples (n = 27) were correctly phenotyped using HIMA-19. We conclude that MIMA-8 is suitable for screening donors and typing patient RBCs. Testing MIMA-8 with gel cards containing anti-mouse IgG enables the screening of donors by automated methods. Humanized chimaeric IgM anti-Js(b) and anti-Fy(a) are suitable as typing reagents by direct agglutination methods.

Anemia, Sickle Cell↗

A hand infection from A. calcoaceticus (M. polymorpha).

Acinetobacter calcoaceticus (Mima polymorpha) is not a well known organism, and its identification can be confusing due to its variable morphology and sensitivity pattern. In a joint infection, delay in identification or appropriate treatment can result in added morbidity. An A. calcoaceticus (Mima) infection in the hand seems not to have been previously reported. The present case involving a finger joint infection illustrates the pathogenicity of A. calcoaceticus (Mima) and problems of treatment. Minocycline is recommended as the drug of choice in subsequent A. calcoaceticus (Mima) and (Herella) infections.

Acinetobacter↗

Combined chemotherapy with carmustine, doxorubicin, etoposide, vincristine, and cyclophosphamide plus mitoxantrone, cytarabine and methotrexate with citrovorum factor for the treatment of aggressive non-Hodgkin lymphoma: a long-term follow-up study.

BACKGROUND: The standard treatment for patients with aggressive non-Hodgkin lymphoma (NHL) is cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP). Since 1989, the authors have used a new chemotherapy regimen with combined carmustine, doxorubicin, etoposide, vincristine, and cyclophosphamide plus mitoxantrone, cytarabine and methotrexate with citrovorum factor called BAVEC-MiMA. The objective of the current study was to explore, after a long follow-up period, the impact of this third-generation regimen for the treatment of aggressive NHL. METHODS: One hundred and one consecutive patients (median age, 41 years) with either B-cell (n=94 patients) or non-B-cell (n=7 patients), aggressive lymphoma were diagnosed and treated between 1989 and 1999 with the BAVEC-MiMA regimen. RESULTS: The complete response rate was 74%, and the overall response rate was 89%. Eleven patients with refractory disease died rapidly after a median period of 5 months. The major toxicity was Grade 4 neutropenia (according to World Health Organization criteria), which was observed in 15 patients (15%). There were four toxicity-related deaths. The overall survival rate was 63% at 9 years. In multivariate analysis, factors that were associated with advantage in overall survival were response to induction therapy, bulky disease, and high score on the International Prognostic Index (IPI). The disease-free survival rate was 77% at 9 years. In multivariate analysis, the IPI was the most important variable for the definition of disease-free survival. CONCLUSIONS: The BAVEC-MiMA regimen was feasible on an outpatient basis, it was tolerated well, and it showed a low toxicity-related mortality. The long follow-up in patients with NHL, which is a rapidly fatal disease, led the authors to observe that, with this regimen, a cure was obtained in > 50% of patients who had low-risk or low-to-intermediate-risk, aggressive NHL.

Adolescent↗

Comparative biokinetics of plutonium and americium after inhalation of PuO2 and mixed oxides (U, Pu)O2 in rat.

PURPOSE: To compare the biokinetics of Pu and Am in rat after inhalation of PuO2 and two (U, Pu) mixed oxides (MOX), referred to as MIMAS and SOLGEL. MATERIALS AND METHODS: Lung clearance was measured in vivo by X-and y-ray spectrometry. Retention of Pu and Am in femurs, liver and kidneys was measured by alpha-spectrometry. RESULTS: Observed lung clearance was in the same range for all three powders. Extra-pulmonary transfers were expressed as the percent of the initial deep lung deposit (IDLD) measured 7 days after inhalation. After PuO2 exposure, bone retention remained nearly constant throughout the 270-day experiment. It was approximately 0.7% of the IDLD for Pu and Am. By contrast, a gradual increase was observed for the two MOX. After 7 days, bone retention of Pu and Am was respectively 0.05 and 0.08% for MIMAS, and 0.2 and 0.6% for SOLGEL. The retention reached maximal values between 180 and 270 days post-exposure, which were 0.2 and 0.3% for MIMAS, and 1.2 and 2.8% for SOLGEL for Pu and Am respectively. CONCLUSIONS: Different transfer rates of Pu and Am from the lung were observed depending on the chemical composition of the oxides and/or the method of their preparation.

Administration, Inhalation↗

Herellea (Acinetobacter) and Pseudomonas ovalis (P. putida) from frozen foods.

Seventeen strains of Herellea vaginicola (Acinetobacter antitratus) and 8 of Pseudomonas ovalis (P. putida), isolated from 23 (6.3%) of 364 samples of frozen, foil-pack foods, were identified and characterized morphologically and biochemically. Herellea was isolated from 17 foods (4.7%), P. ovalis from 6 (1.6%). No Mima were found. The food samples included precooked frozen meats, precooked and uncooked frozen vegetables, and uncooked frozen desserts. The bacteria were detected in the food with a procedure used generally for the detection of salmonellae. The pseudomonad simulated the characteristics of Herellea on Sellers differential agar, except for the fact that it fluoresced. From consideration of the habitat and pathogenicity of Herellea and Mima, it is concluded that, although the presence of these bacteria may not be desirable, their significance in food remains unanswered.

Acinetobacter↗

Evaluation of media for differentiating nonfermenting gram-negative bacteria of medical significance.

An evaluation was made of media and tests used for differentiating nonfermenting gram-negative bacteria encountered in medical bacteriology in order to determine those diagnostic procedures most useful in identifying these bacteria. The organisms examined included Alcaligenes faecalis, A. odorans var. viridans, Moraxella duplex (Mima polymorpha var. oxidans), Acinetobacter anitratum (Herellea vaginicola), A. lwoffi (Mima polymorpha), Pseudomonas fluorescens, P. putida, P. maltophilia, P. pseudomallei, P. stutzeri, P. alcaligenes, and atypical strains of P. aeruginosa. The media and tests evaluated included Sellers' medium; Hugh and Leifson's OF medium; acid production from 10% lactose infusion agar; gluconate oxidation; starch, aesculin, and Tween 80 hydrolysis; lysine decarboxylase, arginine dihydrolase, deoxyribonuclease, and tyrosinase activity; tolerance to triphenyl tetrazolium chloride, cetrimide, cadmium sulfate, 2.5% and 6.5% sodium chloride, and pH 5.6; utilization of glucose, acetamide, and malonate.

Alcaligenes↗

Deoxyribonucleic acid homologies among species of the genus Neisseria.

Eleven aerobic species of Neisseria, a Mima sp., and a Herellea sp. were tested for deoxyribonucleic acid (DNA) homology in direct hybridization experiments. DNA labeled with either (14)C or (32)P was prepared from five species of Neisseria. Unlabeled DNA from the various microorganisms was immobilized on membrane filters, which, after pretreatment, were incubated with labeled DNA (4,000 counts per min per filter) for 14 hr at 67 C. The measure of relatedness was expressed as the relative percentage of direct binding compared to that obtained with homologous DNA. All serological types of N. meningitidis, including the newly proposed types, were homologous to the standard strain of N. meningitidis with one possible exception, type Z. The genus Neisseria is heterogeneous in nature, forming at least three distinct groups: first, N. meningitidis and N. gonorrhoeae; second, N. perflava, N. subflava, N. sicca, N. flavescens, and N. flava; third, N. catarrhalis and N. caviae. Mima and Herellea species show no significant homology with the Neisseria.

Bacteria↗

Bovine infectious keratoconjunctivitis: serological aspects of Moraxella bovis infection.

A modification of a gel diffusion precipitin test (GDPT) was used to detect antibodies for Moraxella bovis (M. bovis) in the sera of cattle affected with bovine infectious keratoconjunctivitis (BIK). The test was also used for the detection of sequential antibody development in cattle vaccinated with cultures of M. bovis. Also, strains of M. bovis isolated from cattle herds affected with BIK were characterized serologically as a part of an identification scheme using the test.A comparison of the antigenic properties of various strains of M. bovis and M. bovis-like organisms was conducted using the test. The results indicated that there might be antigenic relationships between M. bovisand M. bovis-like organisms such as Moraxella liquefaciens, Moraxella nonliquefaciens, an unidentified hemolytic diplococcus, Mima polymorpha, Mima polymorpha var. oxidans and Herellea vaginicolaThe authors suggest that the GDPT can be used for serological studies of BIK, and the identification and antigenic analysis of M. bovis. They indicate, however, that a more definitive study is needed to evaluate the reliability of the test for quantitative work.

Animals↗

Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids.

DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli, seems to be regulated through its binding to acidic phospholipids, such as cardiolipin. In our previous paper (Hase, M., Yoshimi, T., Ishikawa, Y., Ohba, A., Guo, L., Mima, S., Makise, M., Yamaguchi, Y., Tsuchiya, T., and Mizushima, T. (1998) J. Biol. Chem. 273, 28651-28656), we found that mutant DnaA protein (DnaA431), in which three basic amino acids (Arg(360), Arg(364), and Lys(372)) were mutated to acidic amino acids showed a decreased ability to interact with cardiolipin in vitro, suggesting that DnaA protein binds to cardiolipin through an ionic interaction. In this study, we construct three mutant dnaA genes each with a single mutation and examined the function of the mutant proteins in vitro and in vivo. All mutant proteins maintained activities for DNA replication and ATP binding. A mutant protein in which Lys(372) was mutated to Glu showed the weakest interaction with cardiolipin among these three mutant proteins. Thus, Lys(372) seems to play an important role in the interaction between DnaA protein and acidic phospholipids. Plasmid complementation analyses revealed that all these mutant proteins, including DnaA431 could function as an initiator for chromosomal DNA replication in vivo.

Adenosine Diphosphate↗

MEDIUM FOR DIFFERENTIATING THE GRAM-NEGATIVE, NONFERMENTING BACILLI OF MEDICAL INTEREST.

Sellers, Walter (U.S. Air Force School of Aerospace Medicine, Brooks Air Force Base, Tex.). Medium for differentiating the gram-negative, nonfermenting bacilli of medical interest. J. Bacteriol. 87:46-48. 1964.-An agar-slant medium is described for differentiating gram-negative, nonfermenting bacilli of medical interest. Differences in the ability of bacilli to grow anaerobically in the presence of nitrate, to produce pH changes, to produce N(2) from nitrite and nitrate (singly and in combination), to fluoresce, and to oxidize a drop of glucose solution to acid in the presence of high peptone concentrations were used in the development of the medium. Organisms differentiated by the medium include Pseudomonas aeruginosa, Bacterium anitratum, Mima polymorpha, and Vibrio alcaligenes or Alcaligenes faecalis.

Acinetobacter↗

Approaches to understanding the searching behavior of CrossFire users.

Due to the high costs of purchasing, supporting and training users of desktop chemical information systems, it is important to understand users' behavior in order that deficiencies in their search efficiency and effectiveness can be identified and addressed. CrossFire generates comprehensive log files that can be examined to determine the nature of search activity. At GSK (GlaxoSmithKline) a Log File Parser, CrossParse, has been developed in Visual Basic that enables analysis by individual user name, groups of users or the whole user population. Log files can be analyzed for occasions when specific structural features are built, specific types of search are done and how the results are manipulated. CrossParse produces output that can be saved and analyzed within Microsoft Excel. It also allows determination of numbers of active concurrent users on the CrossFire system. CrossParse has been used at GSK (ex-SmithKline Beecham sites) to examine the search behavior of medicinal and synthetic chemists. Additionally, it has been used by MIMAS (Manchester Information and Associated Services) to compare the search behavior of trained and untrained users in the higher education community and to identify any areas where improvements to training can be made. Use of CrossParse in both organizations has allowed identification of areas where users may have difficulties using CrossFire. This will provide valuable feedback to MDL Inc., the authors of the CrossFire application, and guide them in enhancing CrossFire.

Journal Article↗

In situ multi-satellite detection of coherent vortices as a manifestation of Alfvénic turbulence.

Turbulence in fluids and plasmas is a ubiquitous phenomenon driven by a variety of sources-currents, sheared flows, gradients in density and temperature, and so on. Turbulence involves fluctuations of physical properties on many different scales, which interact nonlinearly to produce self-organized structures in the form of vortices. Vortex motion in fluids and magnetized plasmas is typically governed by nonlinear equations, examples of which include the Navier-Stokes equation, the Charney-Hasegawa-Mima equations and their numerous generalizations. These nonlinear equations admit solutions in the form of different types of vortices that are frequently observed in a variety of contexts: in atmospheres, in oceans and planetary systems, in the heliosphere, in the Earth's ionosphere and magnetosphere, and in laboratory plasma experiments. Here we report the discovery by the Cluster satellites of a distinct class of vortex motion-short-scale drift-kinetic Alfvén (DKA) vortices-in the Earth's magnetospheric cusp region. As is the case for the larger Kelvin-Helmholtz vortices observed previously, these dynamic structures should provide a channel for transporting plasma particles and energy through the magnetospheric boundary layers.

Journal Article↗

The fate of bacteria in frozen red cells.

Units of blood were intentionally contaiminated with suspensions of either Aerobacter aerogenes, Escherichia coli, Bacillus subtilis, Enterobacter cloacae, Pseudomonas aeruginosa, Sarcina lutea, Seratia marcesens, Staphylococcus epidermidis, Streptococcus faecalis, Paracolabacterum aerogenoides (Enterobacter hafriae), Mima polymorpha or Acinetobacter- calcoaceticus. When inoculation was made prior to glycerolization, the subsequent glycerolization, freezing, thawing, and deglycerolization resulted in roughly a two log reduction in the number of bacteria. When inoculation was made with a final concentration of between 10(1) and 10(5) organisms per milliliter, immediately following deglycerolization or following washing without glycerolization and freezing, no increase in the number of bacteria was seen after 72 hours storage at 4 C. Three of the 12 organisms studied decreased in number during 72 hours of storage. These data suggest that the current 24-hour limit on the post-thaw storage of frozen red blood cells may be unnecessarily restrictive.

Bacteria↗

Immunization of transgenic mice for production of MoAbs directed at polymorphic blood group antigens.

BACKGROUND: Antibodies of human origin for blood typing are increasingly difficult to obtain, and, despite aggressive efforts, MoAbs with specificities to several blood group polymorphisms have eluded production. As an approach for the generation of MoAbs with defined specificities, the feasibility of immunizing mice that are transgenic for the target polymorphism, Fy(a)/Fy(b) of the Duffy blood group system, was tested with a source of the antithetical antigen. STUDY DESIGN AND METHODS: Nontransgenic mice were immunized with recombinant Fy(b), and transgenic mice expressing human Fy(b) were immunized with recombinant Fy(a). RESULTS: Immunization of the nontransgenic mice resulted in the production of MoAbs to the Duffy protein, but not to the Fy(a)/Fy(b) blood group polymorphism. However, immunization of the transgenic mice resulted in production of the first example of murine Fy(a) MoAb (MIMA-19). This antibody is being used to screen for Fy(a-) blood donors and has been evaluated by many laboratories in an international workshop. CONCLUSION: This approach provides an effective method for producing MoAbs with specificities to polymorphic epitopes. These MoAbs are needed in transfusion medicine to identify antigen-negative donors and to alleviate the critical shortage of blood bank typing reagents, which currently are available only from human-derived sources.

Animals↗

Production of recombinant murine-human chimeric IgM and IgG anti-Js(b) for use in the clinical laboratory.

BACKGROUND: Directly agglutinating MoAbs are more useful than IgG MoAbs of murine origin for typing RBCs from donors and patients. The molecular manipulation and conversion of a murine IgG MoAb into mouse- human chimeric IgM and IgG antibodies are described. STUDY DESIGN AND METHODS: cDNA encoding the variable heavy- and light-chain genes of a murine hybridoma anti-Jsb cell line (MIMA-8) were cloned into human IgM or IgG expression vectors, which were then separately stably transfected into SP2/0-Ag14 B-cells. The secreted antibodies were screened by ELISA and analyzed by flow cytometry and hemagglutination. RESULTS: Forty percent (16 of 40) of the stable clones secreted IgM and 66 percent (12 of 18) of the stable clones secreted IgG. The chimeric IgM from the highest expressing clone reacted 4+ in LISS at room temperature. The chimeric IgG from one clone reacted 4+ by the IAT, resembling the specificity of the original murine antibody. Both manipulated MoAbs reacted specifically with RBCs as assessed by flow cytometry. CONCLUSION: Human-mouse chimeric IgM and IgG from a murine IgG MoAb anti-Jsb has been successfully engineered for use in the clinical laboratory. This approach can potentially be used to manipulate other murine MoAbs to blood group antigens into more clinically useful human isotypes.

Animals↗

Acinetobacter.

Members of the genus Acinetobacter are oxidase negative, aerobic Gram-negative coccobacilli, which have evolved taxonomically from former strains of the Mima-Herrelia group. Their natural habitat is human skin and mucous membranes, water, soil, vegetation, and sewage. The most common multiresistant nosocomial pathogen among 19 genospecies is the A. calcoaceticus-baumannii complex ( A. baumannii). This species is not a common component of normal human acinetobacter colonization, and its ecological origin remains unknown. Outbreaks of nosocomial acinetobacter infection are due to spread of one or a few clones among patients, personnel, and the inanimate hospital environment. Thus, strict implementation of infection control procedures is the major technique for prevention and suppression of such outbreaks. Surveillance cultures of personnel and the environment; molecular genotyping of isolates; cohorting of colonized or infected patients and staff; and topical application of polymyxin B to colonized wounds have been used to enhance standard infection control procedures. Antimicrobial resistance to beta lactam antibiotics in Acinetobacter is due primarily to a combination of chromosomal beta lactamase production and reduced outer membrane permeability. Carbapenem resistance is an increasing phenomenon and restriction of late-generation cephalosporin and carbapenem utilization should be considered in outbreak control. Effective therapy of multiresistant Acinetobacter infection may require a variety of potentially synergistic antibiotic combinations.

Journal Article↗

Some remarks on coherent structures out of chaos in planetary atmospheres and oceans.

In the context of planetary atmospheres and oceans, it is natural to define "coherent structures" as "long-lived," or "solitary," Rossby vortices. These can be described by the generalized Charney-Obukhov equation (in fluid dynamics) or the analogous generalized Hasegawa-Mima equation (in plasma physics). These two equations contain KdV-type nonlinearities which (together with the compensating dispersive spreading) determine the formation of the coherent structures and explain the clear-cut cyclonic/anticyclonic asymmetry observed experimentally in long-lived planetary Rossby vortices. Examples are given of natural vortices which are (and which are not) coherent structures.

Journal Article↗

Quasi-two-dimensional dynamics of plasmas and fluids.

In the lowest order of approximation quasi-two-dimensional dynamics of planetary atmospheres and of plasmas in a magnetic field can be described by a common convective vortex equation, the Charney and Hasegawa-Mima (CHM) equation. In contrast to the two-dimensional Navier-Stokes equation, the CHM equation admits "shielded vortex solutions" in a homogeneous limit and linear waves ("Rossby waves" in the planetary atmosphere and "drift waves" in plasmas) in the presence of inhomogeneity. Because of these properties, the nonlinear dynamics described by the CHM equation provide rich solutions which involve turbulent, coherent and wave behaviors. Bringing in nonideal effects such as resistivity makes the plasma equation significantly different from the atmospheric equation with such new effects as instability of the drift wave driven by the resistivity and density gradient. The model equation deviates from the CHM equation and becomes coupled with Maxwell equations. This article reviews the linear and nonlinear dynamics of the quasi-two-dimensional aspect of plasmas and planetary atmosphere starting from the introduction of the ideal model equation (CHM equation) and extending into the most recent progress in plasma turbulence.

Journal Article↗