Papers to Appear in Forthcoming Issues

Purification of Bovine S100A12 from RecombinantEscherichia coli. Kayoko Yamashita, Yuhta Oyama, Tsuyoshi Shishibori, Osamu Matsushita, Akinobu Okabe, and Ryoji Kobayashi.Kinetic Properties of Human Dopamine Sulfotransferase (SULT1A3) Expressed in Prokaryotic and Eukaryotic Systems: Comparison with the Recombinant Enzyme Purified fromEscherichia-coli. Rana Dajani, Sheila Sharp, Steven Graham, Susanne S. Bethell, Robert M. Cooke, Derek J. Jamieson, and Michael W. H. Coughtrie.Bacterial Expression and Purification of the Fab Fragment of a Monoclonal Antibody Specific for the Low-Density Lipoprotein Receptor-Binding Site of Human Apolipoprotein E. Robert Raffai, Jelena Vukmirica, Karl H. Weisgraber, Eric Rassart, Thomas L. Innerarity, and Ross Milne.Expression, Purification, and Characterization of the Recombinant Calcium-Binding Equine Lysozyme Secreted by the Filamentous FungusAspergillus niger:Comparisons with the Production of Hen and Human Lysozymes. Andrew Spencer, Ludmilla A. Morozov-Roche, Wim Noppe, Donald A. Mackenzie, David J. Jeenes, Marcel Joniau, Christopher M. Dobson, and David B. Archer.Stable, High-Level Expression of a Type I Antifreeze Protein inEscherichia coli. Robert G. Solomon and Rudi Appels.pSKAP/S: An Expression Vector for the Production of Single-Chain Fv Alkaline Phosphatase Fusion Proteins. Remko A. Griep, Charlotte van Twisk, Randolf J. Kerschbaumer, Karen Harper, Lesley Torrance, Gottfried Himmler, Jan M. van der Wolf, and Arjen Schots.Soluble Expression inEscherichia coliof Murine Endogenous Retroviral Transmembrane Envelope Protein Having Immunosuppressive Activity. Kyung Soo Kim, Ki Hwan Kim, Sung E Choi, Ji-Won Yoon, and Yup Kang.Purification of Xyloglucan-Endotransglycosylase Based on Affinity Sorption of the Active Glycosyl–Enzyme Intermediate Complex to Cellulose. Zdena Sulová and Vladimír Farkas.     

Papers to Appear in Forthcoming Issues

Purification of A Multicatalytic Protease Complex from Developing Winged Bean Seeds by Indirect Immuno-affinity ChromatographyRajamma Usha and Manoranjan SinghProduction of Reagents and Optimization of Methods for Studying Calmodulin-Binding ProteinsBettina Ulbricht and Thierry SoldatiExpression, Folding, and Characterization of Small Proteins with Increasing Disulfide Complexity by a pT7-7-Derived PhagemidFrancis C. Peterson, Patricia J. Anderson, Lawrence J. Berliner, and Charles L. BrooksDisulfide Bond Formation and Folding of Plant Peroxidases Expressed as Inclusion Body Protein inEscherichia coliThioredoxin Reductase Negative StrainsKaare Teilum, Lars Østergaard, and Karen G. WelinderOptimized Heterologous Expression of Glutathione Reductase from CyanobacteriumAnabaenaPCC 7120 and Characterization of the Recombinant ProteinFanyi Jiang and Bengt Mannervik     

Handbook of Spatial Statistics edited by GELFAND,A. E., DIGGLE,P. J., FUENTES,M. and GUTTORP,P.

Handbook of Spatial Statistics (A. E. Gelfand, P. J. Diggle, M. Fuentes, and P. Guttorp, Editors) Daniel Commenges Computational Statistics (J. E. Gentle) Thomas Lumley Bayesian Missing Data Problems: EM, Data Augmentation and Noniterative Computation (M. T. Tan, G.‐L. Tian, and K. W. Ng) Alessandra Mattei A First Course in Bayesian Statistical Methods (P. D. Hoff) Dalene Stangl Environmental and Ecological Statistics with R (S. S. Qian) Andrew Finley A Modern Approach to Regression with R (S. J. Sheather) Mervyn G. Marasinghe Exercises and Solutions in Biostatistical Theory (L. L. Kupper, B. H. Neelon, and S. M. O’Brien) Scott S. Emerson ggplot2 : Elegant Graphics for Data Analysis (H. Wickham) Leland Wilkinson     

Analysis of the high- and low-spin Soret bands of horse-heart metmyoglobin complexes

Interest in the thermodynamics of the iron-binding site in hemoproteins has increased in recent years due to refinements in x-ray crystallographic studies of hemoproteins [see Deathage, J. F., Lee, R. S., Anderson, C. M. & Moffat, K. (1976) J. Mol. Biol. 104 , 687–706; Heidner, E. J., Ladner, R. C. & Perutz, M. F. (1976) J. Mol. Biol. 104 , 707–722; Deathage, J. F., Lee, R. S. & Moffat, K. (1976) J. Mol. Biol. 104 , 723–728; Ladner, R. C., Heidner, E. J. & Perutz, M. F. (1976) J. Mol. Biol. 114 , 385–414; Fermi, G. & Perutz, M. F. (1977) J. Mol. Biol. 114 , 421–431; Takano, T. (1977) J. Mol. Biol. 110 , 537–568 and 569–589], the synthesis and x-ray analysis of model heme compounds [see Scheidt, W. R. (1977) Acc. Chem. Res. 10 , 339–345; Kastner, M. E., Scheidt, W. R., Mashino, T. & Reed, C. A. (1978) J. Am. Chem. Soc. 100 , 666–667; Mashiko, T., Kastner, M. E., Spartalian, K., Scheidt, W. R. & Reed, C. A. (1978) J. Am. Chem. Soc. 100 , 6354–6362; Hill, H. A. O., Skite, P. P., Buchler, J. W., Luchr, H., Tonn, M., Gregson, A. K. & Pellizer, G. (1979) Chem. Commun. 4 , 151–152; and Scheidt, W. R., Cohen, I. A. & Kastner, M. E. (1979) Biochemistry 18 , 3546–3556], and the numerous data on heme–protein interactions that account for the differences observed in ligand binding between the various species of animals. Numerous probes have been used and provide information about the structure and thermodynamics of the binding site, but no single probe can provide the complete picture [see Iizuka, T. & Yonetani, T. (1970) Adv. Biophys. 1 , 157–182; Smith, D. W. & Williams, R. J. P. (1970) Struct. Bond. 7 , 1–45; and Spiro, T. G. (1975) Biochim. Biophys. Acta 416 , 169–189].     

A Primer of Ecology with R by STEVENS,M. H. H.

A Primer of Ecology with R (M. H. H. Stevens) Péter Sólymos Handbook on Analyzing Human Genetic Data: Computational Approaches and Software (S. Lin and H. Zhao, Editors) Peter M. Visscher From Finite Sample to Asymptotic Methods in Statistics (P. K. Sen, J. M. Singer, and A. C. Pedroso de Lima) Miodrag Lovric Dynamic Linear Models with R (G. Petris, S. Petrone, and P. Campagnoli) Helio S. Migon Functional Data Analysis with R and Matlab (J. O. Ramsay, G. Hooker, and S. Graves) Hervé Cardot Continuous Bivariate Distributions, 2nd edition (N. Balakrishnan and C.‐D. Lai) Márcia D'Elia Branco Brief Reports by the Editor The Elements of Statistical Learning: Data Mining, Inference, and Prediction, 2nd edition. (T. Hastie, R. Tibshirani, and J. Friedman) Gene Expression Studies Using Affymetrix Microarrays (H. Göhlmann and W. Talloen)     

Papers to Appear in Forthcoming Issues

Preparation of Recombinant Bovine, Porcine, and Porcine W4R/R5K Leptins and Comparison of Their Activity and Immunoreactivity with Ovine, Chicken, and Human LeptinsNina Raver, Eugene E. Gussakovsky, Duane H. Keisler, Radha Krishna, Jehangir Mistry, and Arieh GertlerPurification of Histidine-Tagged Mitochondrial ADP/ATP Carrier: Influence of the Conformational States of the C-Terminal RegionChristelle Fiore, Véronique Trézéguet, Pierre Roux, Agnès Le Saux, Florence Noël, Christine Schwimmer, Delphine Arlot, Anne-Christine Dianoux, Guy J.-M. Lauquin, and Gérard BrandolinExpression and Purification of Recombinant Human Indoleamine 2,3-DioxygenaseTamantha K. Littlejohn, Osamu Takikawa, Daniel Skylas, Joanne F. Jamie, Mark J. Walker, and Roger J. W. TruscottComparative Characterization of Two Forms of Recombinant Human SPC1 Secreted from Schneider 2 CellsJean-Bernard Denault, Claude Lazure, Robert Day, and Richard LeducFunctional and Immunological Analysis of Recombinant Mouse H and L Ferritins from E. coliPaolo Santambrogio, Anna Cozzi, Sonia Levi, Ermanna Rovida, Fulvio Magni, Alberto Albertini, and Paolo ArosioExpression and Purification of Soluble and Inactive Mutant Forms of Membrane Type-1 Matrix MetalloproteinaseHeli Valtanen, Kaisa Lehti, Jouko Lohi, and Jorma Keski-OjaFunctional Human Insulin-Degrading Enzyme Can Be Expressed in BacteriaValérie Chesneau and Marsha Rich RosnerHeterologous Expression in Pseudomonas aeruginosa and Purification of the 9.2 kDa c-Type Cytochrome Subunit of p-Cresol MethylhydroxylaseCiarán N. Cronin and William S. McIntire     

Identification of a uniquely immunodominant, cross-reacting site in the human immunodeficiency virus endonuclease protein.

One of the features of the life cycle of retroviruses is insertion of the proviral DNA into host chromosomes. A protein encoded by the 3' end of the pol gene of the virus genome has been shown to possess endonuclease activity (D. P. Grandgenett, A. C. Vora, and R. D. Schiff, Virology 89:119-132, 1978), which is necessary for DNA integration. Sera from the majority of human immunodeficiency virus (HIV)-infected individuals react with endonuclease protein p31 in serological tests (J. S. Allan, J. E. Coligan, T.-H. Lee, F. Barin, P. J. Kanki, S. M'Boup, M. F. McLane, J. E. Groopman, and M. Essex, Blood 69:331-333, 1987; E. F. Lillehoj, F. H. R. Salazar, R. J. Mervis, M. G. Raum, H. W. Chan, N. Ahmad, and S. Venkatesan, J. Virol. 62:3053-3058, 1988; K. S. Steimer, K. W. Higgins, M. A. Powers, J. C. Stephans, A. Gyenes, G. George-Nascimento, P. A. Liciw, P. J. Barr, R. A. Hallewell, and R. Sanchez-Pescador, J. Virol. 58:9-16, 1986). It is not known, however, which part of the protein represents the target(s) for antibody response. To study this, we synthesized peptides and used them in an enzyme-linked immunosorbent assay system to map the reactivity of human immunodeficiency virus type 1 (HIV-1) antibody-positive sera to the different regions of the HIV endonuclease. A uniquely antigenic, HIV-1- and HIV-2-cross-reacting site was identified in the central part of this protein from Phe-663 to Trp-670.     

The Statistical Analysis of Recurrent Events by COOK, R. J. and LAWLESS, J. F.

The Statistical Analysis of Recurrent Events (R. J. Cook and J. F. Lawless) Chiung‐Yu Huang and Xianghua Luo Introduction to Empirical Processes and Semiparametric Inference (M. R. Kosorok) Changyong Feng Uncertain Judgements: Eliciting Experts' Probabilities (A. O'Hagan, C. E. Buck, A. Daneshkhah, J. R. Eiser, P. H. Garthwaite, D. J. Jenkinson, J. E. Oakley, and T. Rakow) James M. Dickey Models for Intensive Longitudinal Data (T. A. Walls and J. L. Schafer, editors) Daniel Farewell Quantitative Environmental Risk Analysis for Human Health (R. A. Fjeld, N. A. Eisenberg, and K. L. Compton) Scott M. Bartell Reliable Reasoning: Induction and Statistical Learning Theory (G. Harman and S. Kulkarni) Oliver Schulte Modeling Infectious Diseases in Humans and Animals (M. J. Keeling and P. Rohani) Carol Y. Lin Cluster and Classification Techniques for the Biosciences (A. H. Fielding) Morven Leese Modern Applied U‐Statistics (J. Kowalski and X. M. Tu) Jesse Frey Handbook of Statistical Genetics, 3rd edition (D. J. Balding, M. Bishop, and C. Cannings, editors) Karin Bammann, Ronja Foraita, Frauke Günther Quality of Life: The Assessment, Analysis, and Interpretation of Patient‐Reported Outcomes, 2nd edition (P. M. Fayers and D. Machin) Jeff A. Sloan, Amylou Dueck, Rui Qin, Wenting Wu, Pamela J. Atherton, Paul Novotny, Heshan Liu, Kelli N. Burger, Angelina D. Tan, Daniel Szydlo, Victor M. Johnson, Sara J. Felten, Xinghua Zhao, and Brent Diekmann Advanced Distance Sampling: Estimating Abundance of Biological Populations (S. T. Buckland, D. R. Anderson, K. P. Burnham, J. L. Laake, C. L. Borchers, and L. Thomas) Carl James Schwarz Brief Reports by the Editor The Construction of Optimal Stated Choice Experiments: Theory and Methods (D. J. Street and L. Burgess) Design and Analysis of Experiments, Volume 1: Introduction to Experimental Design, 2nd edition (K. Hinkelmann and O. Kempthorne) Introduction to Bayesian Statistics, 2nd edition (W. M. Bolstad) Asymptotic Theory of Statistics and Probability (A. Dasgupta) Bootstrap Methods: A Guide for Practitioners and Researchers, 2nd edition (M. R. Chernick)     

Semiparametric Theory and Missing Data by TSIATIS, A. A.

Semiparametric Theory and Missing Data (A. A. Tsiatis) Andrea Rotnitzky Missing Data in Longitudinal Studies: Strategies for Bayesian Modeling and Sensitivity Analysis (M. J. Daniels and J. W. Hogan) Daniel F. Heitjan Bayesian Biostatistics and Diagnostic Medicine (L. D. Broemeling) Paul Gustafson Statistics in the Pharmaceutical Industry, 3rd edition (C. R. Buncher and J.‐Y. Tsay, Editors) Ralph B. D'Agostino Jr. Introduction to Machine Learning and Bioinformatics (S. Mitra, S. Datta, T. Perkins, and G. Michailidis) Yulan Liang The Statistics of Gene Mapping (D. Siegmund and B. Yakir) Hongyu Zhao DNA Methylation Microarrays: Experimental Design and Statistical Analysis (S.‐C. Wang and A. Petronis) Kimberly D. Siegmund Multiple Testing Procedures with Applications to Genomics (S. Dudoit and M. J. van der Laan) Ruth Heller The Statistical Analysis of Functional MRI Data (N. A. Lazar) Wesley K. Thompson Simulation and Inference for Stochastic Differential Equations with R Examples (S. M. Iacus) Dave Campbell Nonparametric Analysis of Univariate Heavy‐Tailed Data: Research and Practice (N. Markovich) M. Ivette Gomes Time Series Analysis with Applications in R, 2nd edition (J. D. Cryer and K.‐S. Chan) Timothy D. Johnson Brief Reports by the Editor Analysis of Variance and Covariance: How to Choose and Construct Models for the Life Sciences (C. P. Doncaster and A. J. H. Davey) Computational Statistics Handbook with MATLAB ^®, 2nd edition (W. L. Martinez and A. R. Martinez) Models for Probability and Statistical Inference: Theory and Applications (J. H. Stapleton) Medical Biostatistics, 2nd edition (A. Indrayan) Computational Methods in Biomedical Research (R. Khattree and D. N. Naik, Editors)     

Structural and functional recovery of electropermeabilized skeletal muscle in-vivo after treatment with surfactant poloxamer 188

A critical requirement for cell survival after trauma is sealing of breaks in the cell membrane [M. Bier, S.M. Hammer, D.J. Canaday, R.C Lee, Kinetics of sealing for transient electropores in isolated mammalian skeletal muscle cells, Bioelectromagnetics 20 (1999) 194-201; R.C. Lee, D.C. Gaylor, D. Bhatt, D.A. Israel, Role of cell membrane rupture in the pathogenesis of electrical trauma, J. Surg. Res. 44 (1988) 709-719; R.C. Lee, J.F. Burke, E.G. Cravalho (Eds.), Electrical Trauma: The Pathophysiology, Manifestations, and Clinical Management, Cambridge University Press, 1992; B.I. Tropea, R.C. Lee, Thermal injury kinetics in electrical trauma, J. Biomech. Engr. 114 (1992) 241-250; F. Despa, D.P. Orgill, J. Newalder, R.C Lee, The relative thermal stability of tissue macromolecules and cellular structure in burn injury, Burns 31 (2005) 568-577; T.A. Block, J.N. Aarsvold, K.L. Matthews II, R.A. Mintzer, L.P. River, M. Capelli-Schellpfeffer, R.L. Wollman, S. Tripathi, C.T. Chen, R.C. Lee, The 1995 Lindberg Award. Nonthermally mediated muscle injury and necrosis in electrical trauma, J. Burn Care and Rehabil. 16 (1995) 581-588; K. Miyake, P.L. McNeil, Mechanical injury and repair of cells, Crit. Care Med. 31 (2003) S496-S501; R.C. Lee, L.P. River, F.S. Pan, R.L. Wollmann, Surfactant-induced sealing of electropermeabilized skeletal muscle membranes in vivo, Proc. Natl. Acad. Sci. 89 (1992) 4524-4528; J.D. Marks, C.Y. Pan, T. Bushell, W. Cromie, R.C. Lee, Amphiphilic, tri-block copolymers provide potent membrane-targeted neuroprotection, FASEB J. 15 (2001) 1107-1109; B. Greenebaum, K. Blossfield, J. Hannig, C.S. Carrillo, M.A. Beckett, R.R. Weichselbaum, R.C. Lee, Poloxamer 188 prevents acute necrosis of adult skeletal muscle cells following high-dose irradiation, Burns 30 (2004) 539-547; G. Serbest, J. Horwitz, K. Barbee, The effect of poloxamer-188 on neuronal cell recovery from mechanical injury, J. Neurotrauma 22 (2005) 119-132]. The triblock copolymer surfactant Poloxamer 188 (P188) is known to increase the cell survival after membrane electroporation [R.C. Lee, L.P. River, F.S. Pan, R.L. Wollmann, Surfactant-induced sealing of electropermeabilized skeletal muscle membranes in vivo, Proc. Natl. Acad. Sci. 89 (1992) 4524-4528; Z. Ababneh, H. Beloeil, C.B. Berde, G. Gambarota, S.E. Maier, R.V. Mulkern, Biexponential parametrization of T2 and diffusion decay curves in a rat muscle edema model: Decay curve components and water compartments, Magn. Reson. Med. 54 (2005) 524-531]. Here, we use a rat hind-limb model of electroporation injury to determine if the intravenous administration of P188 improves the recovery of the muscle function. Rat hind-limbs received a sequence of either 0, 3, 6, 9, or 12 electrical current pulses (2 A, 4 ms duration, 10 s duty cycle). Magnetic resonance imaging (MRI) analysis, muscle water content and compound muscle action potential (CMAP) amplitudes were compared. Electroporation injury manifested edema formation and depression of the CMAP amplitudes. P188 (one bolus of 1 mg/ml of blood) was administrated 30 or 60 min after injury. Animals receiving P188 exhibited reduced tissue edema (p < 0.05) and increased CMAP amplitudes (p < 0.03). By comparison, treatment with 10 kDa neutral dextran, which produces similar serum osmotic effects as P188, had no effect on post-electroporation recovery. Noteworthy, the present results suggest that a single intravenous dose of P188 is effective to restore the structural integrity of damaged tissues with intact circulation.     

Structural and functional recovery of electropermeabilized skeletal muscle in-vivo after treatment with surfactant poloxamer 188

A critical requirement for cell survival after trauma is sealing of breaks in the cell membrane [M. Bier, S.M. Hammer, D.J. Canaday, R.C Lee, Kinetics of sealing for transient electropores in isolated mammalian skeletal muscle cells, Bioelectromagnetics 20 (1999) 194-201; R.C. Lee, D.C. Gaylor, D. Bhatt, D.A. Israel, Role of cell membrane rupture in the pathogenesis of electrical trauma, J. Surg. Res. 44 (1988) 709-719; R.C. Lee, J.F. Burke, E.G. Cravalho (Eds.), Electrical Trauma: The Pathophysiology, Manifestations, and Clinical Management, Cambridge University Press, 1992; B.I. Tropea, R.C. Lee, Thermal injury kinetics in electrical trauma, J. Biomech. Engr. 114 (1992) 241-250; F. Despa, D.P. Orgill, J. Newalder, R.C Lee, The relative thermal stability of tissue macromolecules and cellular structure in burn injury, Burns 31 (2005) 568-577; T.A. Block, J.N. Aarsvold, K.L. Matthews II, R.A. Mintzer, L.P. River, M. Capelli-Schellpfeffer, R.L. Wollman, S. Tripathi, C.T. Chen, R.C. Lee, The 1995 Lindberg Award. Nonthermally mediated muscle injury and necrosis in electrical trauma, J. Burn Care and Rehabil. 16 (1995) 581-588; K. Miyake, P.L. McNeil, Mechanical injury and repair of cells, Crit. Care Med. 31 (2003) S496-S501; R.C. Lee, L.P. River, F.S. Pan, R.L. Wollmann, Surfactant-induced sealing of electropermeabilized skeletal muscle membranes in vivo, Proc. Natl. Acad. Sci. 89 (1992) 4524-4528; J.D. Marks, C.Y. Pan, T. Bushell, W. Cromie, R.C. Lee, Amphiphilic, tri-block copolymers provide potent membrane-targeted neuroprotection, FASEB J. 15 (2001) 1107-1109; B. Greenebaum, K. Blossfield, J. Hannig, C.S. Carrillo, M.A. Beckett, R.R. Weichselbaum, R.C. Lee, Poloxamer 188 prevents acute necrosis of adult skeletal muscle cells following high-dose irradiation, Burns 30 (2004) 539-547; G. Serbest, J. Horwitz, K. Barbee, The effect of poloxamer-188 on neuronal cell recovery from mechanical injury, J. Neurotrauma 22 (2005) 119-132]. The triblock copolymer surfactant Poloxamer 188 (P188) is known to increase the cell survival after membrane electroporation [R.C. Lee, L.P. River, F.S. Pan, R.L. Wollmann, Surfactant-induced sealing of electropermeabilized skeletal muscle membranes in vivo, Proc. Natl. Acad. Sci. 89 (1992) 4524-4528; Z. Ababneh, H. Beloeil, C.B. Berde, G. Gambarota, S.E. Maier, R.V. Mulkern, Biexponential parametrization of T2 and diffusion decay curves in a rat muscle edema model: Decay curve components and water compartments, Magn. Reson. Med. 54 (2005) 524-531]. Here, we use a rat hind-limb model of electroporation injury to determine if the intravenous administration of P188 improves the recovery of the muscle function. Rat hind-limbs received a sequence of either 0, 3, 6, 9, or 12 electrical current pulses (2 A, 4 ms duration, 10 s duty cycle). Magnetic resonance imaging (MRI) analysis, muscle water content and compound muscle action potential (CMAP) amplitudes were compared. Electroporation injury manifested edema formation and depression of the CMAP amplitudes. P188 (one bolus of 1 mg/ml of blood) was administrated 30 or 60 min after injury. Animals receiving P188 exhibited reduced tissue edema (p<0.05) and increased CMAP amplitudes (p<0.03). By comparison, treatment with 10 kDa neutral dextran, which produces similar serum osmotic effects as P188, had no effect on post-electroporation recovery. Noteworthy, the present results suggest that a single intravenous dose of P188 is effective to restore the structural integrity of damaged tissues with intact circulation.     

Myxoma virus M11L blocks apoptosis through inhibition of conformational activation of Bax at the mitochondria

Many viruses inhibit or retard apoptosis, a strategy that subverts one of the most ancient antiviral mechanisms. M11L, a myxoma virus-encoded antiapoptotic protein, has been previously shown to localize to mitochondria and block apoptosis of virus-infected cells (H. Everett, M. Barry, S. F. Lee, X. J. Sun, K. Graham, J. Stone, R. C. Bleackley, and G. McFadden, J. Exp. Med. 191:1487-1498, 2000; H. Everett, M. Barry, X. Sun, S. F. Lee, C. Frantz, L. G. Berthiaume, G. McFadden, and R. C. Bleackley, J. Exp. Med. 196:1127-1139, 2002; and G. Wang, J. W. Barrett, S. H. Nazarian, H. Everett, X. Gao, C. Bleackley, K. Colwill, M. F. Moran, and G. McFadden, J. Virol. 78:7097-7111, 2004). This protection from apoptosis involves constitutive-forming inhibitory complexes with the peripheral benzodiazepine receptor and Bak on the outer mitochondrial membrane. Here, we extend the study to investigate the interference of M11L with Bax activation during the process of apoptosis. Myxoma virus infection triggers an early apoptotic signal that induces rapid Bax translocation from cytoplasm to mitochondria, despite the existence of various viral antiapoptotic proteins. However, in the presence of M11L, the structural activation of Bax at the mitochondrial membrane, which is characterized by the occurrence of a Bax conformational change, is blocked in both M11L-expressing myxoma-infected cells and M11L-transfected cells under apoptotic stimulation. In addition, inducible binding of M11L to the mitochondrially localized Bax is detected in myxoma virus-infected cells and in M11L/Bax-cotransfected cells as measured by immunoprecipitation and tandem affinity purification analysis, respectively. Importantly, this inducible Bax/M11L interaction is independent of Bak, demonstrated by the complete block of Bax-mediated apoptosis in myxoma-infected cells that lack Bak expression. Our findings reveal that myxoma M11L modulates apoptosis by multiple independent strategies which all contribute to the blockade of apoptosis at the mitochondrial checkpoint.     

Expression and functional analysis of Euglena Gracilis chloroplast initiation factor 3

A portion of a cDNA predicted to encode the mature form of Euglena gracilis chloroplast translational initiation factor 3 (IF-3_chlM, molecular mass, 46402) and the portion of this factor homologous to bacterial IF-3 (IF-3_chlH, molecular mass 22829) have been cloned and expressed in Escherichia coli as histidine-tagged proteins. The homology domain can be expressed in reasonable levels in E. coli. However, IF-3_chlM is quite toxic and can only be produced in small amounts. Both forms of the chloroplast factor are associated with E. coli ribosomes. Purification procedures have been developed for both IF-3_chlM and IF-3_chlH using Ni-NTA affinity chromatography followed by ion exchange chromatography. IF-3_chlM and IF-3_chlH are active in promoting ribosome dissociation and in promoting the binding of fMet-tRNA to E. coli ribosomes. However, IF-3_chlH has at least 5-fold more activity than either native IF-3_chl or IF-3_chlM in promoting initiation complex formation on chloroplast 30S ribosomal subunits in the presence of a mRNA carrying a natural translational initiation signal. This observation suggests that regions of IF-3_chl lying outside of the homology domain may down-regulate the activity of this factor.This work was supported in part by National Institutes of Health Grant GM24963.     

Restriction and modification of phage 47 and lambda by R factors

S. panama 47 (antibiotic-sensitive, phage pattern A) was infected with R factors from a number of field strains of Enterobacteriaceae (Salmonella, Escherichia coli, Citrobacter, Enterobacter, Klebsiella andProteus) isolated from human and animal sources. These R factors could be grouped into 11 types i.e. R1 R11 on the basis of induced changes in the phage type of the recipient.R8 and R11 renderS. panama resistant to the phages A H:S. panama 47 (R3) and 47 (R6) adsorb the phages A F, but there is no phage multiplication: phages G and H are considered to be restricted and modified in these strains. The R factors R5 and R7 also exert restriction and modification on a number of the typing phages A H. The nature of the changes in phage pattern brought about by R4, R9 and R10 is not understood. R2 does not exert restriction (i.e. no change in phage pattern). The R factors were also investigated for the fi (fertility inhibition) and spp (restriction of phage ) markers.The R factors R3 R11 readily segregate, in the sense that the restriction and modification loci, and occasionally the Resistance Transfer Factor as a whole was frequently lost after R transfer. These 9 types of R factors were encountered infrequently in the present material.Resistance to tetracycline inS.panama is nearly always due to R factors of type R1. In other members of Enterobacteriaceae, notably inE.coli, R1 is less frequently found than R2.     

Applied Spatial Data Analysis with R. by BIVAND, R. S., PEBESMA, E. J., and GOMEZ-RUBIO, V.

Applied Spatial Data Analysis with R (R. S. Bivand, E. J. Pebesma, and V. Gomez‐Rubio) Jay M. Ver Hoef Bayesian Disease Mapping: Hierarchical Modeling in Spatial Epidemiology (A. B. Lawson) J. Law Disease Surveillance: A Public Health Informatics Approach (J. S. Lombardo and D. L. Buckeridge, editors) Iris Pigeot Survival Analysis for Epidemiologic and Medical Research (S. Selvin) M. G. Valsecchi Survival and Event History Analysis: A Process Point of View (O. O. Aalen, O. Borgan, and H. K. Gjessing) Patricia Grambsch Nonlinear Dimensionality Reduction (J. A. Lee and M. Verleysen) Haonan Wang Model Selection and Model Averaging (G. Claeskens and N. L. Hjort) Thomas M. Loughin Meta‐Analysis of Binary Data Using Profile Likelihood (D. Böhning, R. Kuhnert, and S. Rattanasiri) Eloise Kaizar Wavelet Methods in Statistics with R (G. P. Nason) Jeffrey S. Morris Adaptive Design Theory and Implementation Using SAS and R (M. Chang) Feifang Hu Ecological Models and Data in R (B. M. Bolker) Rachel M. Fewster Applied Multiway Data Analysis (P. M. Kroonenberg) Renato Coppi Brief Reports by the Editor Sampling of Populations: Methods and Applications, 4th edition (P. S. Levy and S. Lemeshow) Applied Survival Analysis: Regression Modeling of Time‐to‐Event Data, 2nd edition (D. W. Hosmer, S. Lemeshow, and S. May) SAS for Data Analysis: Intermediate Statistical Methods (M. G. Marasinghe and W. J. Kennedy) Advances in Mathematical and Statistical Modeling (B. C. Arnold, N. Balakrishnan, J. M. Sarabia, and R. Mínguez, editors) An Introduction to Generalized Linear Models, 3rd edition (A. J. Dobson and A. G. Barnett) Design and Analysis of Bioavailability and Bioequivalence Studies, 3rd edition (S.‐C. Chow and J.‐P. Liu)     

M-like,immunoglobulin-binding protein ofStreptococcus pyogenes type M15

An M-like protein fromStreptococcus pyogenes type M15 strain EF1949 (EMML15) was cloned inEscherichia coli and sequenced. Recombinant EMML15 protein revealed a unique binding pattern for human IgG subclasses not described previously. Comparative analysis of the EMML15 amino acid sequence with those of other M-like proteins of opacity factor positive (OF⁺) serotypes and protein H, an IgG receptor from OF^– serotype M1, showed that IgG-binding proteins with common binding of IgG3 were closely related and distinct from streptococcal IgG receptors not binding IgG3. Thus, the Ig-binding proteins fromS. pyogenes were subdivided into two main categories according to binding pattern, protein structure, and gene location.     

Studies on the effects ofin vitro methyiation on aminoacylation of transfer RNA

In vitro methyiation ofEscherichia coli transfer ribonucleic acid by cell free extracts ofMycobacterium smegmatis leads exclusively to the formation of 1-methyl adenine [Vani, B. R., Ramakrishnan, T., Taya, Y., Noguchi, S., Yamaiuzumi, Z. and Nishimura, S. (1978)J. Bact., 137, 1085]. We have studied the effect of this modification on aminoacylationof Escherichia coli tRNA by mycobacterial enzymes. Aminoacylation with total algal protein hydrolysate as well as several individual aminoacids like methionine, valine, tyrosine, aspartic acid and lysine were monitored. In all the cases methyiation had a positive effect on the extent of aminoacylation by mycobacterial enzymes. Decreased aminoacylationin vitro was observed when hypomethylated transfer RNA from ethionine treated cells was used as the substrate for aminoacylation     

Lineage and Genogroup-Defining Single Nucleotide Polymorphisms of Escherichia coli O157:H7

Escherichia coli O157:H7 is a zoonotic human pathogen for which cattle are an important reservoir host. Using both previously published and new sequencing data, a 48-locus single nucleotide polymorphism (SNP)-based typing panel was developed that redundantly identified 11 genogroups that span six of the eight lineages recently described for E. coli O157:H7 (J. L. Bono, T. P. Smith, J. E. Keen, G. P. Harhay, T. G. McDaneld, R. E. Mandrell, W. K. Jung, T. E. Besser, P. Gerner-Smidt, M. Bielaszewska, H. Karch, M. L. Clawson, Mol. Biol. Evol. 29:2047–2062, 2012) and additionally defined subgroups within four of those lineages. This assay was applied to 530 isolates from human and bovine sources. The SNP-based lineage groups were concordant with previously identified E. coli O157:H7 genotypes identified by other methods and were strongly associated with carriage of specific Stx genes. Two SNP lineages (Ia and Vb) were disproportionately represented among cattle isolates, and three others (IIa, Ib, and IIb) were disproportionately represented among human clinical isolates. This 48-plex SNP assay efficiently and economically identifies biologically relevant lineages within E. coli O157:H7.     

Translation enhancement by optimized downstream box sequences in <Emphasis Type="Italic">Escherichia coli </Emphasis>and<Emphasis Type="Italic"> Mycobacterium smegmatis</Emphasis>

The effect on translation of downstream box sequences optimized for binding to Mycobacterium smegmatis and Escherichia coli 16S rRNA in the absence of a Shine--Dalgarno (SD) region was investigated. The relative translational efficiency of each construct in either M. smegmatis or E. coli was determined. Eradication of the SD region in the absence of a downstream box abolished the translation activity. In contrast, optimized downstream box constructs resulted in a 13- and 18-fold increase in protein synthesis, relative to non-optimized DB controls in E. coli and M. smegmatis, respectively.