Life Sciences Commons^™

Intracellular Lipid-Binding Proteins And Their Genes, David A. Bernlohr, Melanie A. Simpson, Ann Vogel Hertzel, Leonard J. Banaszak

Department of Biochemistry: Faculty Publications

Intracellular lipid-binding proteins are a family of low-molecularweight single-chain polypeptides that form 1:1 complexes with fatty acids, retinoids, or other hydrophobic ligands. These proteins are products of a large multigene family of unlinked loci distributed throughout the genome. Each lipid-binding protein exhibits a distinctive pattern of tissue distribution. Transcriptional control, regulated by a combination of peroxisome proliferator activated receptors and CCAAT/enhancer-binding proteins, allows for a variety of both cell and tissue-specific expression patterns. In some cells, fatty acids increase the expression of the lipid-binding protein genes. Fatty acids, or their metabolites, are activators of the peroxisome proliferator–activated receptor family of …

Ets-Core Binding Factor: A Common Composite Motif In Antigen Receptor Gene Enhancers, Batu Erman, Marta Cortes, Barbara S. Nikolajczyk, Nancy A. Speck, Ranjan Sen

Dartmouth Scholarship

A tripartite domain of the murine immunoglobulin μ heavy-chain enhancer contains the μA and μB elements that bind ETS proteins and the μE3 element that binds leucine zipper-containing basic helix-loop-helix (bHLH-zip) factors. Analysis of the corresponding region of the human μ enhancer revealed high conservation of the μA and μB motifs but a striking absence of the μE3 element. Instead of bHLH-zip proteins, we found that the human enhancer bound core binding factor (CBF) between the μA and μB elements; CBF binding was shown to be a common feature of both murine and human enhancers. Furthermore, mutant enhancers that bound …

Probing The Intermolecular Hydrogen Bonding Of Water Molecules At The Ccl4/Water Interface In The Presence Of Charges Soluble Surfactant, D. E. Gragson, G. L. Richmond

Chemistry and Biochemistry

The molecular structure and hydrogen bonding of water molecules at the CCl₄/water interface in the presence of a charged soluble surfactant has been explored in this study using vibrational sum frequency generation. By examining OH stretching modes that are highly sensitive to the local hydrogen bonding environment it is found that water molecules at the CCl₄/water interface both in the presence and absence of a charged soluble surfactant are predominantly in a tetrahedral arrangement much like the structure ofice. Isotopic dilution studies have been employed to further characterize this icelike interfacial structure. Ablueshift ofapproximately120 cm^{-1 …}

Role Of Egr-1 Gene Expression In B Cell Receptor-Induced Apoptosis In An Immature B Cell Lymphoma, Subramanian Muthukkumar, Seong-Su Han, Sumathi Muthukkumar, Vivek M. Rangnekar, Subbarao Bondada

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Ligation of B cell receptor (BCR) on BKS-2, an immature B cell lymphoma by anti-IgM antibodies (Ab) caused apoptosis. Here we report that signaling through B cell receptor in wild type BKS-2 cells down-regulated the expression of Egr-1, a zinc finger-containing transcription factor. A reduction in the level ofEgr-1 mRNA could be demonstrated as early as 30 min after the ligation of BCR on BKS-2 cells. Immunocytochemical and Western blot analysis revealed that the expression of EGR-1 protein was also inhibited by anti-IgM treatment. Antisense oligonucleotides to Egr-1 caused growth inhibition and apoptosis in BKS-2 cells, suggesting that …

Liquid-Liquid Immiscibility In Lipid Monolayers, John P. Hagen, Harden M. Mcconnell

Chemistry and Biochemistry

Some binary lipid mixtures form coexisting liquid phases when spread at the air/water interface. This work describes the pressure–composition phase diagrams of binary mixtures of four unsaturated phosphatidylcholines with dihydrocholesterol. These four binary mixtures have critical compositions of approximately fifty mole percent, and average critical exponents of 0.25±0.07. The data can also be approximated by a regular solution thermodynamic model, yielding parameters for the non-ideality of these mixtures.

A Fluid In Contact With A Semipermeable Surface: Second-Order Integral Equation Approach, Douglas Henderson, Pawel Bryk, Stefan Sokolowski

Faculty Publications

An integral equation approach for a binary hard-sphere mixture interacting with a planar semipermeable wall (membrane) is formulated by using the second-order nonuniform or pair Ornstein–Zernike equation as well as the usual singlet Ornstein–Zernike equation. The results of the pair theory are compared with those obtained from the singlet theory and with computer simulation data. The pair approach is more accurate than the singlet theory.

Emissions From Smoldering Combustion Of Biomass Measured By Open-Path Fourier Transform Infrared Spectroscopy, Robert J. Yokelson, Ronald A. Susott, Darold E. Ward, James J. Reardon, David W. T. Griffith

Chemistry and Biochemistry Faculty Publications

Biomass samples from a diverse range of ecosystems were burned in the Intermountain Fire Sciences Laboratory open combustion facility. Midinfrared spectra of the nascent emissions were acquired at several heights above the fires with a Fourier transform infrared spectrometer (FTIR) coupled to an open multipass cell. In this report, the results from smoldering combustion during 24 fires are presented including production of carbon dioxide, carbon monoxide, methane, ethene, ethyne, propene, formaldehyde, 2-hydroxyethanal, methanol, phenol, acetic acid, formic acid, ammonia, hydrogen cyanide, and carbonyl sulfide. These were the dominant products observed, and many have significant influence on atmospheric chemistry at the …

Interferon Gamma (Ifn-Gamma) Is Necessary For The Genesis Of Acetylcholine Receptor-Induced Clinical Experimental Autoimmune Myasthenia Gravis In Mice., Balaji Balasa, Caishu Deng, Jae Lee, Linda M. Bradley, Dyanna K. Dalton, Premkumar Christadoss, Nora Sarvetnick

Journal Articles: Regenerative Medicine

Experimental autoimmune myasthenia gravis (EAMG) is an animal model of human myasthenia gravis (MG). In mice, EAMG is induced by immunization with Torpedo californica acetylcholine receptor (AChR) in complete Freund's adjuvant (CFA). However, the role of cytokines in the pathogenesis of EAMG is not clear. Because EAMG is an antibody-mediated disease, it is of the prevailing notion that Th2 but not Th1 cytokines play a role in the pathogenesis of this disease. To test the hypothesis that the Th1 cytokine, interferon (IFN)-gamma, plays a role in the development of EAMG, we immunized IFN-gamma knockout (IFN-gko) (-/-) mice and wild-type (WT) …

Ms2 Coat Protein Mutants Which Bind Qβ Rna, Marc Spingola, David Peabody

Biology Department Faculty Works

The coat proteins of the RNA phages MS2 and Qβ are structurally homologous, yet they specifically bind different RNA structures. In an effort to identify the basis of RNA binding specificity we sought to isolate mutants that convert MS2 coat protein to the RNA binding specificity of Qβ. A library of mutations was created which selectively substitutes amino acids within the RNA binding site. Genetic selection for the ability to repress translation from the Qβ translational operator led to the isolation of several MS2 mutants that acquired binding activity for Qβ RNA. Some of these also had reduced abilities to …

Distribution Of Endosomal, Lysosomal And Mhc Markers In A Monocytic Cell Line Infected With Chlamydia Psittaci, David M. Ojcius, Raymond Hellio, Alice Dautry-Varsat

All Dugoni School of Dentistry Faculty Articles

The intracellular fate of Chlamydia psittaci during infection of a monocytic cell line, THP1, was characterized. Cytochalasin D inhibited phagocytosis of latex beads but had no effect on infection by C. psittaci, and vacuoles expressed the transferrin receptor, suggesting accessibility to the endocytic pathway. Early Chlamydia-containing vacuoles expressed major histocompatibility complex (MHC) class I molecules, and most vacuoles fused with host cell lysosomes, since they expressed LAMP-1 and had acidic pHs. In cells prestimulated with gamma interferon, vacuoles also expressed MHC class II molecules, suggesting that the monocytes might effectively process Chlamydia-derived antigens for presentation by MHC class I and …

Grand Canonical Monte Carlo And Modified Singlet Integral Equations For The Density Profile Of A Yukawa Fluid Near A Planar Wall, Douglas Henderson, Wilmer Olivares-Rivas, Leo Degreve, Jacqueline Quintana

Faculty Publications

Results for the density profile for Yukawa molecules near a hard wall and an exponential attractive wall are presented for Grand Canonical Monte Carlo (GCMC) simulations, for the singlet hypernetted chain (HNC) integral equation and for a modified version of the Lovett–Mou–Buff–Wertheim (LMBW-1) which uses the exact contact value theorem. The results of the standard singlet HNC are quite poor. If the LMBW equation is modified (but still using the bulk direct correlation function) the results at high temperature become reasonable. However, the results at low temperatures, close to the bulk coexistence curve, are only a partial improvement. The contact …

Local Delivery Of Interleukin 4 By Retrovirus-Transduced T Lymphocytes Ameliorates Experimental Autoimmune Encephalomyelitis., Michael K. Shaw, James B. Lorens, Archana Dhawan, Richard Dalcanto, Harley Y. Tse, Alyssa B. Tran, Colleen Bonpane, Shanti L. Eswaran, Stefan Brocke, Nora Sarvetnick, Lawrence Steinman, Garry P. Nolan, C. Garrison Fathman

Journal Articles: Regenerative Medicine

Experimental autoimmune encephalomyelitis (EAE) is an inflammatory autoimmune disease of the central nervous system which serves as a model for the human disease multiple sclerosis. We demonstrate here that encephalitogenic T cells, transduced with a retroviral gene, construct to express interleukin 4, and can delay the onset and reduce the severity of EAE when adoptively transferred to myelin basic protein-immunized mice. Thus, T lymphocytes transduced with retroviral vectors can deliver "regulatory cytokines" in a site-specific manner and may represent a viable therapeutic strategy for the treatment of autoimmune disease.

Laser-Initiated Decomposition Products Of Indocyanine Green (Icg) And Carbon Black Sensitized Biological Tissues, John M. Kokosa, Andrzej Przyjazny, Kenneth Eugene Bartels, Massoud Motamedi, Donald J. Hayes, David B. Wallace, Christopher J. Frederickson

Chemistry & Biochemistry Publications

Organic dyes have found increasing use a s sensitizers in laser surgical procedures, due to their high optical absorbances. Little is known, however, about the nature of the degradation products formed when these dyes are irradiated with a laser. Previous work in our laboratories has shown that irradiation of polymeric and biological tissues with CO2 and Nd:YAG lasers produces a host of volatile and semivolatile by-products, some of which are known to be potential carcinogens. This work focuses on the identification of the chemical by-products formed by diode laser and Nd:YAG laser irradiation of indocyanine green (ICG) and carbon black …

Fish Primary Cultures For Toxicity And Pathological Studies, Maria Lyons Alcantara

Doctoral

In this study, epithelial primary cultures from the rainbow trout (Oncorhynchus mykiss) were developed as a model for environmental effects assessment. Cadmium was used as representative of heavy metals. The possible use of this system for pathological studies was also evaluated. The differential cytotoxic effects of cadmium on fish and mammalian epithelial cells in established and primary culture wee assessed by looking at the reduction of colony-forming ability and the reduction in the extent of growth. The influence of medium composition on the toxicity of cadmium was also studied using serum-free and serum-containing media. The experiments using immortalised cell lines …

Las Vegas Wash Water Quality Monitoring Program: 1996 Report Of Findings, Richard A. Roline, James J. Sartoris, U.S. Bureau Of Reclamation, U.S. Geological Survey

Publications (WR)

Las Vegas Wash, a natural wash east of the city of Las Vegas, Nevada, carries stormwater, groundwater drainage, and sewage effluent from three sewage treatment plants to Lake Mead. The Wash provides nearly the only surface water outlet for the entire 2,193 mi² of Las Vegas Valley. A drainage area of 1,586 mi² contributes directly to the Wash through surface flow which is channeled to Las Vegas Bay of Lake Mead, while drainage of the remaining 607 mi² is presumably subsurface and may drain toward Las Vegas Wash.

In the 1930's and 1940's, sewage treatment plants were …

Synthesis And Characterization Of Modified Silica Gel As An Intermediate In The Generation Of Gaseous Standard Mixtures, M. Prokopowicz, K. Lewandowska, A. Skwierawska, Andrzej Przyjazny, Jan F. Biernat, J. Namiesnik

Chemistry & Biochemistry Publications

A possibility of extending analytical applications of chemically modified silica gels is described. This involves their utilization for the generation of gaseous standard mixtures consisting of methyl chloride as the analyte and nitrogen as a carrier gas to be used for the calibration of the GC-FID system. N-methylmorpholine was chemically bonded to the propylsilylated surface of silica gel forming chloride of an appropriate immobilized compound which, under certain conditions, undergoes thermal decomposition yielding a single, volatile component (methyl chloride). Such a method of generating specific amounts of a standard substance can be used both for a single point calibration and …

Endosome To Golgi Retrieval Of The Vacuolar Protein Sorting Receptor, Vps10p, Requires The Function Of The Vps29, Vps30, And Vps35 Gene Products, Matthew N. J. Seaman, Eric G. Marcusson, Joan Lin-Cereghino, Scott D. Emr

College of the Pacific Faculty Articles

Mutations in the S. cerevisiae VPS29 and VPS30 genes lead to a selective protein sorting defect in which the vacuolar protein carboxypeptidase Y (CPY) is missorted and secreted from the cell, while other soluble vacuolar hydrolases like proteinase A (PrA) are delivered to the vacuole. This phenotype is similar to that seen in cells with mutations in the previously characterized VPS10 and VPS35 genes. Vps10p is a late Golgi transmembrane protein that acts as the sorting receptor for soluble vacuolar hydrolases like CPY and PrA, while Vps35p is a peripheral membrane protein which cofractionates with membranes enriched in Vps10p. The …

Both An N-Terminal 65-Kda Domain And A C-Terminal 30-Kda Domain Of Seca Cycle Into The Membrane At Secyeg During Translocation, Jerry Eichler, William Wickner

Dartmouth Scholarship

SecA, a 102-kDa hydrophilic protein, couples the energy of ATP binding to the translocation of preprotein across the bacterial inner membrane. SecA function and topology were studied with metabolically labeled [35S]SecA and with inner membrane vesicles from cells that overex- pressed SecYEGDFyajC, the integral domain of preprotein translocase. During translocation in the presence of ATP and preprotein, a 65-kDa N-terminal domain of SecA is protected from proteolytic digestion through insertion into the mem- brane, as previously reported for a 30-kDa C-terminal domain [Economou, A. & Wickner, W. (1994) Cell 78, 835–843]. Insertion of both domains occurs at saturable SecYEGDFyajC sites …

I2b Is A Small Cytosolic Protein That Participates In Vacuole Fusion, Paul Slusarewicz, Zuoyu Xu, Kimberly Seefeld, Albert Haas, William T. Wickner

Dartmouth Scholarship

Saccharomyces cerevisiae vacuole inheritance requires two low molecular weight activities, LMA1 and LMA2. LMA1 is a heterodimer of thioredoxin and protease B inhibitor 2 (I^B2). Here we show that the second low molecular weight activity (LMA2) is monomeric I^B2. Though LMA2 / I^B2 was initially identified as a protease B inhibitor, this protease inhibitor activity is not related to its ability to promote vacuole fusion: ( i ) Low M r protease B inhibitors cannot substitute for LMA1 or LMA2, ( ii ) LMA1 and LMA2 promote the fusionof vacuoles from a strain that …

Characterization Of Recombinant Soybean Leghemoglobin A And Apolar Distal Histidine Mutants, Mark S. Hargrove, Jennifer K. Barry, Eric Allen Brucker, Michael B. Berry, George N. Phillips, Jr., John S. Olson, Raúl Arredondo-Peter, Jeanenne M. Dean, Robert V. Klucas, Gautam Sarath

Gautam Sarath Publications

The cDNA for soybean leghemoglobin a (Lba) was cloned from a root nodule cDNA library and expressed in Escherichia coli. The crystal structure of the ferric acetate complex of recombinant wild-type Lba was determined at a resolution of 2.2 Å. Rate constants for O₂, CO and NO binding to recombinant Lba are identical with those of native soybean Lba. Rate constants for hemin dissociation and auto-oxidation of wild-type Lba were compared with those of sperm whale myoglobin. At 37°C and pH 7, soybean Lba is much less stable than sperm whale myoglobin due both to a fourfold …

Ifn-Gamma, Igif, And Iddm., Nora Sarvetnick

Journal Articles: Regenerative Medicine

No abstract provided.

Docking Of Yeast Vacuoles Is Catalyzed By The Ras-Like Gtpase Ypt7p After Symmetric Priming By Sec18p (Nsf), Andreas Mayer, William Wickner

Dartmouth Scholarship

Vacuole inheritance in yeast involves the for- mation of tubular and vesicular “segregation struc- tures” which migrate into the bud and fuse there to es- tablish the daughter cell vacuole. Vacuole fusion has been reconstituted in vitro and may be used as a model for an NSF-dependent reaction of priming, docking, and fusion. We have developed biochemical and micro- scopic assays for the docking step of in vitro vacuole fusion and characterized its requirements. The vacu- oles must be primed for docking by the action of Sec17p ( a -SNAP) and Sec18p (NSF). Priming is neces- sary for both fusion …

A Heterodimer Of Thioredoxin And Ib2 Cooperates With Sec18p (Nsf) To Promote Yeast Vacuole Inheritance, Zuoyu Xu, Andreas Mayer, Eric Muller, William Wickner

Dartmouth Scholarship

Early in S phase, the vacuole (lysosome) of Saccharomyces cerevisiae projects a stream of vesicles and membranous tubules into the bud where they fuse and establish the daughter vacuole. This inheritance reaction can be studied in vitro with isolated vacuoles. Rapid and efficient homotypic fusion between saltwashed vacuoles requires the addition of only two purified soluble proteins, Sec18p (NSF) and LMA1, a novel heterodimer with a thioredoxin subunit. We now report the identity of the second subunit of LMA1 as IB2, a previously identified cytosolic inhibitor of vacuolar proteinase B. Both subunits are needed for efficient vacuole inheritance in vivo …

Molecular Engineering Of Side-Chain Liquid Crystalline Polymers By Living Polymerizations, Coleen Pugh, Alan L. Kiste

Chemistry and Biochemistry

“Living” anionic, cationic, metalloporphyrin and ring-opening metathesis polymerizations have been used to prepare well-defined side-chain liquid crystalline homopolymers, block and graft copolymers and statistical copolymers. This paper analyzes their successes and failures by reviewing the mechanistic aspects and experimental conditions of each type of polymerization, and identifies other classes of mesogenic monomers that could be polymerized in a controlled manner in the future. The emerging structure/property relationships are then identified using well-defined SCLCPs in which only one structural feature is varied while all others remain constant.

The thermal transitions of liquid crystalline polymethacrylates, polynorbomenes and poly(viny1 ether)s reach their limiting …

Genetic Analysis Of Chlorophyll Biosynthesis, David Bollivar

Scholarship

No abstract provided.

Identification Of The Adp-L-Glycero-D-Manno-Heptose-6-Epimerase (Rfad) And Heptosyltransferase Ii (Rfaf) Biosynthesis Genes From Nontypeable Haemophilus Influenzae 2019, W A. Nichols, B W. Gibson, William Melaugh, N G. Lee, M Sunshine, M A. Apicella

Chemistry Faculty Publications

Haemophilus influenzae is an important human pathogen. The lipooligosaccharide (LOS) of H. influenzae has been implicated as a virulence determinant. To better understand the assembly of LOS in nontypeable H. influenzae (NtHi), we have cloned and characterized the rfaD and rfaF genes of NtHi 2019, which encode the ADP-L-glycero-D-manno-heptose-6-epimerase and heptosyltransferase II enzymes, respectively. This cloning was accomplished by the complementation of Salmonella typhimurium lipopolysaccharide (LPS) biosynthesis gene mutants. These deep rough mutants are novobiocin susceptible until complemented with the appropriate gene. In this manner, we are able to use novobiocin resistance to select for specific NtHi LOS inner core …

Characterization Of A Transposon Tn916-Generated Mutant Of Haemophilus Ducreyi 35000 Defective In Lipooligosaccharide Biosynthesis, B W. Gibson, A A. Campagnari, William Melaugh, M A. Apicella, S Grass, Jing Wang, Katherine L. Palmer, R S. Munson

Chemistry Faculty Publications

To define the role of the surface lipooligosaccharide (LOS) of Haemophilus ducreyi in the pathogenesis of chancroid, Tn916 mutants of H. ducreyi 35000 defective in expression of the murine monoclonal antibody (MAb) 3F11 epitope on H. ducreyi LOS were identified by immunologic screening. One mutant, designated 1381, has an LOS which lacks the MAb 3F11 epitope and migrates with an increased mobility on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The gene disrupted by the Tn916 element in strain 1381 was identified by cloning the sequences flanking the Tn916 element. The sequences were then used to probe a lambda DASHII genomic library. …

Sodium Nitrite Alone Protects The Brain O _I_ Microsomal Ca -Atpase Against Potassium Cyanide-Induced Neurotoxicity In Rats, Odutayo O. Odunuge, G.. A. Adenuga

Faculty Publications

The effect of a short-term oral administration of potassium cyanide (KCN) (200 ppm in diet) with or without sodium nitrite (NaNO2) pretreatment on rat brain microsomal Ca2* ATPase was investigated. The specific activity value of the enzyme significantly decreased (p<0.05) by 50% compared with control and by 63% for KCN-treated rats compared with KCN-treated rats pretreated with NaNO;,. There was no significant difference at the h = 0.05 level between the values obtained for the control and KCN-treated rats pretreated with NaNO,. These results show both that feeding lowers brain microsomal Ca2f-ATPase activity and that NaNO, has a protective role (antidote function) in that respect.

Rna Base-Amino Acid Interaction Strengths Derived From Structures And Sequences, Brooke Lustig, S Arora, R L. Jernigan

Faculty Publications, Chemistry

We investigate RNA base-amino acid interactions by counting their contacts in structures and their implicit contacts in various functional sequences where the structures can be assumed to be preserved. These frequencies are cast into equations to extract relative interaction energetics. Previously we used this approach in considering the major groove interactions of DNA, and here we apply it to the more diverse interactions observed in RNA. Structures considered are the three different tRNA synthetase complexes, the U1A spliceosomal protein with an RNA hairpin and the BIV TARTat complex. We use binding data for the base frequencies for the seryl, aspartyl …

Exercise The Power Of The Purse With Hussein, Porcher L. Taylor Iii

School of Professional and Continuing Studies Faculty Publications

In the same shrewd diplomatic spirit displayed by the Pentagon in its recent purchase of a squadron of MIG-29 fighter jets from Moldova in order to preempt the sale of the planes to Iran, the U.S. should offer to buy Iraq's entire biochemical weapons arsenal and then destroy it in place. Let's call this emerging instrument of U.S. foreign policy "preemptive economic diplomacy."