A mammalian 2-5A system functions as an antiviral pathway in transgenic plants.

Resistance to virus infections in higher vertebrates is mediated in part through catalysis of RNA decay by the, interferon-regulated 2-5A system. A functional 2-5A system requires two enzymes, a 2-5A synthetase that produces 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A) in response to double-stranded RNA, and the 2-5A-dependent RNase L. We have coexpressed these human enzymes in transgenic tobacco plants by using a single plasmid containing the cDNAs for both human RNase L and a low molecular weight form of human 2-5A synthetase under control of different, constitutive promoters. Expression of the human cDNAs in the transgenic plants was demonstrated from Northern blots, by specific enzyme assays, and by immunodetection (for RNase L). Infection of leaves, detached or in planta, of the coexpressing transgenic plants by tobacco mosaic virus, alfalfa [correction of alfafa] mosaic virus, or tobacco etch virus resulted in necrotic lesions. In contrast, leaves expressing 2-5A synthetase or RNase L alone and leaves containing the plasmid vector alone produced typical systemic infections. While alfalfa mosaic virus produced lesions only in the inoculated leaves regardless of the concentration of virus in the inoculum, high, but not low, levels of tobacco etch virus inoculum resulted in escape of virus to uninoculated leaves. Nevertheless, there was a substantial reduction of tobacco etch virus yield as measured by ELISA assay in the coexpressing transgenic plants. These results indicate that expression of a mammalian 2-5A system in plants provides resistance to virus infections.

Structure determination of murine mitochondrial carbonic anhydrase V at 2.45-A resolution: implications for catalytic proton transfer and inhibitor design.

The three-dimensional structure of murine mitochondrial carbonic anhydrase V has been determined and refined at 2.45-A resolution (crystallographic R factor = 0.187). Significant structural differences unique to the active site of carbonic anhydrase V are responsible for differences in the mechanism of catalytic proton transfer as compared with other carbonic anhydrase isozymes. In the prototypical isozyme, carbonic anhydrase II, catalytic proton transfer occurs via the shuttle group His-64; carbonic anhydrase V has Tyr-64, which is not an efficient proton shuttle due in part to the bulky adjacent side chain of Phe-65. Based on analysis of the structure of carbonic anhydrase V, we speculate that Tyr-131 may participate in proton transfer due to its proximity to zinc-bound solvent, its solvent accessibility, and its electrostatic environment in the protein structure. Finally, the design of isozyme-specific inhibitors is discussed in view of the complex between carbonic anhydrase V and acetazolamide, a transition-state analogue. Such inhibitors may be physiologically important in the regulation of blood glucose levels.

A serine proteinase inhibitor locus at 18q21.3 contains a tandem duplication of the human squamous cell carcinoma antigen gene.

The squamous cell carcinoma antigen (SCCA) is a member of the ovalbumin family of serine proteinase inhibitors (serpins). A neutral form of the protein is found in normal and some malignant squamous cells, whereas an acidic form is detected exclusively in tumor cells and in the circulation of patients with squamous cell tumors. In this report, we describe the cloning of the SCCA gene from normal genomic DNA. Surprisingly, two genes were found. They were tandemly arrayed and flanked by two other closely related serpins, plasminogen activator inhibitor type 2 (PAI2) and maspin at 18q21.3. The genomic structure of the two genes, SCCA1 and SCCA2, was highly conserved. The predicted amino acid sequences were 92% identical and suggested that the neutral form of the protein was encoded by SCCA1 and the acidic form was encoded by SCCA2. Further characterization of the region should determine whether the differential expression of the SCCA genes plays a causal role in development of more aggressive squamous cell carcinomas.

Human autoantibody recognition of DNA.

Combinatorial IgG Fab phage display libraries prepared from a systemic lupus erythematosus (SLE) donor and a healthy donor were affinity selected against human placental DNA. Human monoclonal antibody Fab fragments specific for DNA were isolated from both libraries, although Fabs of the highest affinity were isolated only from the lupus library. Generally, apparent affinities of the Fabs for human placental DNA, purified double-stranded DNA, and denatured DNA were approximately equivalent. Surface plasmon resonance indicated Fab binding constants for a double-stranded oligodeoxynucleotide of 0.2-1.3 x 10(8) M-1. The higher-affinity Fabs, as ranked by binding to human placental DNA or to the oligonucleotide probe, tested positive in the Crithidia luciliae assay commonly used in the diagnosis of SLE, and interestingly the genes encoding the heavy-chain variable regions of these antibodies displayed evidence of only minimal somatic hypermutation. The heavy chains of the SLE Fabs were characterized by a predominance of basic residues toward the N terminus of complementarity-determining region 3 (CDR3). The crucial role of heavy-chain CDR3 (HCDR3) in high-affinity DNA recognition was suggested by the creation of DNA binding in an unrelated antibody by HCDR3 transplantation from SLE antibodies. We propose that high-affinity DNA-binding antibodies can arise in SLE without extensive somatic hypermutation in the variable-region genes because of the expression of inappropriate HCDR3s.

(Table 2) Radiocarbon age determination from the polar planktonic foraminifera Neogloboquadrina pachyderma s for sediment core HU90-013-029

An additional Heinrich ice-rafting event is identified between Heinrich events 5 and 6 in eight cores from the Labrador Sea and the northwest Atlantic Ocean. It is characterized by sediment rich in detrital carbonate (40% CaCO3) with high concentration of floating dropstones, high coarse-fraction (% > 150 µm) content, and has a sharp contact with the underlying but grades into the overlying hemipelagic sediment. It also shows lighter d18ONpl values, indicating freshening due to iceberg rafting and/or meltwater discharge. This event is correlated with Dansgaard-Oeschger event 14 and interpreted as an additional Heinrich event, H5a. The thickness of H5a in the Labrador Sea reaches up to 220 cm. This additional Heinrich event has also been reported in cores PS2644 and SO82-5 from the northern North Atlantic. With the recognition of H5a the temporal spacing between Heinrich events 1 to 6 becomes more uniform (~7 ka).

AID assistance to local government : experience and issues /

"PN-AAL-026"--Cover.

UM Men's Swimming Team, 1988

Front Row: swimming coach Jon Urbanchek, Rob Silverman, Mike Bayerl, Bill Hayes, Lee Michaud, Alec Campbell, Jan-Erick Olsen, Tim Petsche, Eric Kitchen, Martin Moran, Mike Barrowman, diving coach Dick Kimball

2nd Row: trainer Dave Tomchek, Eric Wise, D, Mott, Rick Reising, M. Smith, Alex Alvizuri, Zeb Esselstyn, Tato Ceresa, D. McNear, M. Curry, E. Gotting

3rd Row: David Parrish, Dan Dewhirst, Sean Gallagher, Steve Pancratz, Scott Van Appledorn, Dave Goch, Bill Kopas, G. Williams, Ron Howard, Rick Wilkening, A. DeLuca

Back Row: assistant coach Joe Parker, Scott Ryan, Bjoern Warland, Jarret Winter, Ron Melnyk, L. Aspenwall, Jeff Heydanek, Mats Nygren, Mike Creaser, graduate assistant Dan Kerska

UM Men's Swimming Team, 1992

Front Row: swimming coach Jon Urbanchek, Steve Duttenhofer, Steve Bigelow, Jim Hume, Noel Strauss, Greg Gooch, Brian Gunn, diving coach Dick Kimball

Middle Row: Gustavo Borges, Tom Hay, Rodney VanTassell, Matt Jaffe, Steve West, Tim Bower Chris Veber

Back Row: Bill Pettit, Dan Abruzzi, Thomas Blake, Kevin Glass, Bill Kopas, Kent Tschannen, trainer J. Petter. Eric Lesser

Not Pictured: Eric Bailey, Alex Bogaerts, Jayson Field, R. Genova, Drew Hansz, Jeff Jozwiak, John Kiani, Brad Lambert, Chad McCloud, asst. coach Mark Noetzel, Scott Pjesky, Abel Sanchez, Rob Silverman, Dan Valdes

The use of context cues in teaching machines /

"Technical report: NAVTRADEVCEN 507-1."

Special you; a story of adoption.

Mode of access: Internet.

Mystery of Edwin Drood; Master Humphrey's clock, and Sketches by "Boz."

The mystery of Edwin Drood.--Master Humphrey's clock, and other pieces.--Sketches by Boz.--Holiday romance.--George Silverman's explanation.

The mystery of Edwin Drood, and other stories.

On cover: Standard edition.

Supplemental report of the Special Prosecutor /

"September 10, 1982."

School of Nursing Class of 1975

Top Row: Kris Aasvved, Phyllis Askew, Stephanie Babboni, Carolyn Backus, Carol Bockeloo, Veronica Banks, Patte Barland, Sally Barling, Rowena Beebe, Ginger Behr, Bobbi Bergmooser, Clary Bestor, Terry A. Bilinski, Debbie Blauer, Kathleen Bly, Lois K. Boer, Aurelia boyer, Polly Bradley, Sue Brenkert, Sherry Brezina

Row 2: Andrea Brown, Phyllis Buchholz, Michele Bujak, Barbara Burcham, Carol Burg, Mary Ann Campbell, Nancy Cartwright, Sally Chin, Kathleen Christmas, Barbara Clark, Marlene Clarkson, Alma Cole, Judy Coltson, Donna Craig, Janet L. Davies, Catherine Davidson, Sandra Detrisac, Toni Doherty, Kathleen Dumas, Deretha Eddings

Row 3: Marcia Ferrand, Karen Finger, Carol Fischer, Susan Fischer, Suzanne M. Fleszar, Barbara Fritz, Lola Garland, Susan Goldstein, Pam Goltz, Diane Gorman, Debby Goudreau, Diane Greenfield, Debbie Gross, Joan Hamman, Cheryl Hauch, Michelle Hays, Betty Henderson, Christena Henson, Constance Hill, Linda Hill

Row 4: Pamela Hill, Marilyn Holland, Patricia Horvath, Lois Huissen, Nance J. Huston, Phyllis Isackson, Angela Janik, Kim Johnson, Marjorie Kelsey, Wanda Kent, Eugenie Kimura, Lesley Kinnard, Kathleen Klute, Peggy Koskela, Linda Ksiazkiewicaz, Barbara Lang, Karen C. Carson, Kathryn Linder, Kathleen Lipinski, Janie Locke</p>

Row 5: Nancy Luth, Denise Lyons, Susan Malkewitz, Diane Mannino, Nancy Marsh, Denise M. McCann, Carol McVannel, Vicky Melancon, Darlene Mikolajczak, Jane Monroe, Pam Morris, Cari Mulholland, Sandra Muller, Jacqueline Murphy, Terri Murtland, Colleen Nash, Debbie Nichols, Nancy Nowacek, Denise D. O'Brien, Sue Olejniczak

Row 6: Susan Panozzo, Marty Parmelee, Nancy Parr, Alexandra Paul, Pam Pennington, Patricia Phelps, Helen Piggush, Jan Pinkham, Molly Power, Janet Primeau, Ilona Proskie, Gretel Quitmeyer, Vicki Jo Ray, Josephine Reed, Ruth Riley, Norine Rowe, Beata Rudnik, Pat Rutowski, Linda Sanders, Patricia Saran

Row 7: Judy Sayles, Janis Schlicker, Janice Schmidt, Janiece Selecky, Deborah Silverman, Susan K. Smith, Theresa Sobanski, Marcia Sosnowski, Joyce Stein, Cathie Stepien, Pam Stoeffler, Sharon Swann, Susan Truchan, Susan Turke, Susan Valentine, Delores Vander Wal, Mary Jane VanLoon, Pamela A. Van Riper, Jeanne M. Wade, Karen Warner

Row 8: Deborah White, Rebecca E. Wildgen, Karen Williams, Sharon Williams, Debra Wilson, KEn Wilson, Nancy Wiltz, Maribeth Wooldridge, Martha Zawacki, JoAnn Zlotnick

Row 9: Julie Sochalski, Norma Shumaker, Kristin Brawner, Susan Archambault, Lauralee Hess, Rita M. Gibes, Barbara Terrien, Laurie Cushman, Mary Markey