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Human Monocytes, Macrophages, And Dendritic Cells: Alcohol Treatment Methods, Gyongyi Szabo, Pranoti Mandrekar 2010 University of Massachusetts Medical School

Human Monocytes, Macrophages, And Dendritic Cells: Alcohol Treatment Methods, Gyongyi Szabo, Pranoti Mandrekar

Gyongyi Szabo

Both acute and chronic alcohol consumption have significant immunomodulatory effects of which alterations in innate immune functions contribute to impaired antimicrobial defense and inflammatory responses. Blood monocytes, macrophages, and dendritic cells play a central role in innate immune recognition as these cells recognize pathogens, respond with inflammatory cytokine production, and induce antigen-specific T-lymphocyte activation. All of these innate immune cell functions are affected in humans by alcohol intake. Here, we summarize the different effects of acute and chronic alcohol on monocyte, macrophage, and dendritic cell functions in humans and describe methods for separation and functional evaluation of these cell types.


Acetylation Of Wrn Protein Regulates Its Stability By Inhibiting Ubiquitination, Kai Li, Rui Wang, Enerlyn M. Lozada, Wei Fan, David K. Orren, Jianyuan Luo 2010 University of Massachusetts Medical School

Acetylation Of Wrn Protein Regulates Its Stability By Inhibiting Ubiquitination, Kai Li, Rui Wang, Enerlyn M. Lozada, Wei Fan, David K. Orren, Jianyuan Luo

Open Access Articles

BACKGROUND: WRN is a multi-functional protein involving DNA replication, recombination and repair. WRN acetylation has been demonstrated playing an important role in response to DNA damage. We previously found that WRN acetylation can regulate its enzymatic activities and nuclear distribution.

METHODOLOGY/PRINCIPAL FINDING: Here, we investigated the factors involved in WRN acetylation and found that CBP and p300 are the only major acetyltransferases for WRN acetylation. We further identified 6 lysine residues in WRN that are subject to acetylation. Interestingly, WRN acetylation can increase its protein stability. SIRT1-mediated deacetylation of WRN reverses this effect. CBP dramatically increases the half-life of ...


Determinants For Stop-Transfer And Post-Import Pathways For Protein Targeting To The Chloroplast Inner Envelope Membrane, Antonio A. B. Viana, Ming Li, Danny Schnell 2010 University of Massachusetts - Amherst

Determinants For Stop-Transfer And Post-Import Pathways For Protein Targeting To The Chloroplast Inner Envelope Membrane, Antonio A. B. Viana, Ming Li, Danny Schnell

Danny Schnell

he inner envelope membrane (IEM) of the chloroplast plays key roles in controlling metabolite transport between the organelle and cytoplasm and is a major site of lipid and membrane synthesis within the organelle. IEM biogenesis requires the import and integration of nucleus-encoded membrane proteins. Previous reports have led to the conclusion that membrane proteins are inserted into the IEM during protein import from the cytoplasm via a stop-transfer mechanism or are completely imported into the stroma and then inserted into the IEM in a post-import mechanism. In this study, we examined the determinants for each pathway by comparing the targeting ...


Oxidation Of Methane By A Biological Dicopper Centre, Ramakrishnan Balasubramanian, Stephen M. Smith, Swati Rawat, Liliya A. Yatsunyk, Timothy L. Stemmler, Amy C. Rosenzweig 2010 Northwestern University

Oxidation Of Methane By A Biological Dicopper Centre, Ramakrishnan Balasubramanian, Stephen M. Smith, Swati Rawat, Liliya A. Yatsunyk, Timothy L. Stemmler, Amy C. Rosenzweig

Biochemistry and Molecular Biology Faculty Publications

Vast world reserves of methane gas are underutilized as a feedstock for the production of liquid fuels and chemicals owing to the lack of economical and sustainable strategies for the selective oxidation of methane to methanol1. Current processes to activate the strong C–H bond (104 kcal mol−1) in methane require high temperatures, are costly and inefficient, and produce waste2. In nature, methanotrophic bacteria perform this reaction under ambient conditions using metalloenzymes called methane monooxygenases (MMOs). MMOs thus provide the optimal model for an efficient, environmentally sound catalyst3. There are two types of MMO. Soluble MMO (sMMO),expressed by ...


Partitioning Of Minimotifs Based On Function With Improved Prediction Accuracy, Sanguthevar Rajasekaran, Tian Mi, Jerlin Camilus Merlin, Aaron Oommen, Patrick R. Gradie, Martin R. Schiller 2010 University of Connecticut - Storrs

Partitioning Of Minimotifs Based On Function With Improved Prediction Accuracy, Sanguthevar Rajasekaran, Tian Mi, Jerlin Camilus Merlin, Aaron Oommen, Patrick R. Gradie, Martin R. Schiller

Life Sciences Faculty Publications

Background

Minimotifs are short contiguous peptide sequences in proteins that are known to have a function in at least one other protein. One of the principal limitations in minimotif prediction is that false positives limit the usefulness of this approach. As a step toward resolving this problem we have built, implemented, and tested a new data-driven algorithm that reduces false-positive predictions.

Methodology/Principal Findings

Certain domains and minimotifs are known to be strongly associated with a known cellular process or molecular function. Therefore, we hypothesized that by restricting minimotif predictions to those where the minimotif containing protein and target protein ...


Neuroprotection With Anesthetics In Two Models Of Cerebral Ischemia, Rafael Eduardo Chaparro-Buitrago 2010 University of South Florida

Neuroprotection With Anesthetics In Two Models Of Cerebral Ischemia, Rafael Eduardo Chaparro-Buitrago

Graduate Theses and Dissertations

Neuroprotection with anesthetics has been studied for many decades; important advances in this field have modified the way Anesthesiologists treat patients in the operating room. Animal models have played an important role in the study of ischemia in the operating room. Recent studies have demonstrated that the effect of anesthetics seems to be different in different animal models. We decided to evaluate anesthetics in a well-known model of cerebral ischemia and also in hypotensive models designed by us. We used a model of cerebral ischemia (MCAO) to test anesthetics neuroprotective effect in a two-week period. Then, we used a model ...


T10c12 Conjugated Linoleic Acid Causes Delipidation In 3t3-L1 Adipocytes And Mice, shan jiang 2010 University of Nebraska - Lincoln

T10c12 Conjugated Linoleic Acid Causes Delipidation In 3t3-L1 Adipocytes And Mice, Shan Jiang

Theses and Dissertations in Animal Science

T10c12 conjugated linoleic acid (t10c12 CLA) causes the reduction of triglyceride contents in adipocytes. T10c12 CLA’s delipidation effect is associated with decreased nutrient uptake, adipogenesis, lipogenesis, and increased energy expenditure, lipolysis, fatty acid oxidation in adipocytes. However, the molecular mechanisms of CLA’s delipidation effects are still unknown. AMP-activated protein kinase (AMPK), a central regulator of cellular energy levels, is activated by an increase in the cellular AMP:ATP ratio or various cellular stresses. We demonstrated that t10c12 CLA activated AMPK in 3T3-L1 adipocytes, leading to inhibition of anabolic biosynthesis and increase of energy expenditure. Strong activation of AMPK ...


Ribosomal Rna Mutations That Inhibit The Activity Of Transfer-Messenger Rna Of Stalled Ribosomes, Jacob N. Crandall 2010 Brigham Young University - Provo

Ribosomal Rna Mutations That Inhibit The Activity Of Transfer-Messenger Rna Of Stalled Ribosomes, Jacob N. Crandall

All Theses and Dissertations

In eubacteria, stalled ribosomes are rescued by a conserved quality-control mechanism involving transfer-messenger RNA (tmRNA) and its protein partner SmpB. Mimicking a tRNA, tmRNA enters stalled ribosomes, adds Ala to the nascent polypeptide, and serves as a template to encode a short peptide that tags the nascent protein for destruction. To further characterize the tagging process, we developed two genetic selections that link tmRNA activity to cell death. These negative selections can be used to identify inhibitors of tagging or to identify mutations in key residues essential for ribosome rescue. Little is known about which ribosomal elements are specifically required ...


Dynamics On Multiple Timescales In The Rna-Directed Rna Polymerase From The Cystovirus Φ6, Zhen Ren, Hsin Wang, Ranajeet Ghose 2010 CUNY City College

Dynamics On Multiple Timescales In The Rna-Directed Rna Polymerase From The Cystovirus Φ6, Zhen Ren, Hsin Wang, Ranajeet Ghose

Publications and Research

The de novoinitiating RNA-directed RNA polymerase (RdRP), P2, forms the central machinery in the infection cycle of the bacteriophage ϕ6 by performing the dual tasks of replication and transcription of the double-stranded RNA genome in the host cell. By measurement and quantitative analysis of multiple-quantum spin-relaxation data for the δ1 positions of Ile residues that are distributed over the 3D-fold of P2, we find that the enzyme is dynamic both on the fast (ps–ns) and slow (µs–ms) timescales. The characteristics of several motional modes including those that coincide with the catalytic timescale (500–800/s) are altered ...


Phosphorylation Of Su(Var)3–9 By The Chromosomal Kinase Jil-1, Joern Boeke, Catherine Regnard, Weili Cai, Jorgen Johansen, Kristen M. Johansen, Peter B. Becker, Axel Imhof 2010 Ludwig Maximilians University of Munich

Phosphorylation Of Su(Var)3–9 By The Chromosomal Kinase Jil-1, Joern Boeke, Catherine Regnard, Weili Cai, Jorgen Johansen, Kristen M. Johansen, Peter B. Becker, Axel Imhof

Biochemistry, Biophysics and Molecular Biology Publications

The histone methyltransferase SU(VAR)3–9 plays an important role in the formation of heterochromatin within the eukaryotic nucleus. Several studies have shown that the formation of condensed chromatin is highly regulated during development, suggesting that SU(VAR)3–9's activity is regulated as well. However, no mechanism by which this may be achieved has been reported so far. As we and others had shown previously that the N-terminus of SU(VAR)3–9 plays an important role for its activity, we purified interaction partners from Drosophila embryo nuclear extract using as bait a GST fusion protein containing ...


Inhibiting Mirna In Caenorhabditis Elegans Using A Potent And Selective Antisense Reagent, Genhua Zheng, Victor R. Ambros, Wen-hong Li 2010 University of Texas Southwestern Medical Center

Inhibiting Mirna In Caenorhabditis Elegans Using A Potent And Selective Antisense Reagent, Genhua Zheng, Victor R. Ambros, Wen-Hong Li

Program in Molecular Medicine Publications and Presentations

BACKGROUND: Antisense reagents can serve as efficient and versatile tools for studying gene function by inhibiting nucleic acids in vivo. Antisense reagents have particular utility for the experimental manipulation of the activity of microRNAs (miRNAs), which are involved in the regulation of diverse developmental and physiological pathways in animals. Even in traditional genetic systems, such as the nematode Caenorhabditis elegans, antisense reagents can provide experimental strategies complementary to mutational approaches. Presently no antisense reagents are available for inhibiting miRNAs in the nematode C. elegans.

RESULTS: We have developed a new class of fluorescently labelled antisense reagents to inhibit miRNAs in ...


Aspergillus Fumigatus Stimulates The Nlrp3 Inflammasome Through A Pathway Requiring Ros Production And The Syk Tyrosine Kinase, Najwane Saïd-Sadier, Eduardo V. Padilla, Gordon Langsley, David M. Ojcius 2010 University of California, Merced

Aspergillus Fumigatus Stimulates The Nlrp3 Inflammasome Through A Pathway Requiring Ros Production And The Syk Tyrosine Kinase, Najwane Saïd-Sadier, Eduardo V. Padilla, Gordon Langsley, David M. Ojcius

Dugoni School of Dentistry Faculty Articles

Invasive aspergillosis (IA) is a life-threatening disease that occurs in immunodepressed patients when infected with Aspergillus fumigatus. This fungus is the second most-common causative agent of fungal disease after Candida albicans. Nevertheless, much remains to be learned about the mechanisms by which A. fulmigatus activates the innate immune system. We investigated the inflammatory response to conidia and hyphae of A. fumigatus and specifically, their capacity to trigger activation of an inflammasome. Our results show that in contrast to conidia, hyphal fragments induce NLRP3 inflammasome assembly, caspase-1 activation and IL-1β release from a human monocyte cell line. The ability of Aspergillus ...


Computational Protein Design: Advances In The Design And Redesign Of Biomolecular Nanostructures, Jeffery G. Saven 2010 University of Pennsylvania

Computational Protein Design: Advances In The Design And Redesign Of Biomolecular Nanostructures, Jeffery G. Saven

Departmental Papers (Chemistry)

Computational protein design facilitates the continued development of methods for the design of biomolecular structure, sequence and function. Recent applications include the design of novel protein sequences and structures, proteins incorporating nonbiological components, protein assemblies, soluble variants of membrane proteins, and proteins that modulate membrane function.


Coupling Into Waveguide Evanescent Modes With Applications In Electron Paramagnetic Resonance, Jason Walter Sidabras 2010 Marquette University

Coupling Into Waveguide Evanescent Modes With Applications In Electron Paramagnetic Resonance, Jason Walter Sidabras

Master's Theses (2009 -)

The use of analytical and numerical techniques in solving the coupling of evanescent modes in a microwave waveguide through slots can be optimized to create a uniform magnetic field excitation on axis within a waveguide. This work has direct applications in Electron Paramagnetic Resonance (EPR) where a 100~kHz time-varying magnetic field is incident on a sample contained in a microwave cavity. Typical cavity designs do not take into consideration the uniformity of the 100~kHz field modulation and assume it to be uniform enough over the sample region from quasi-static principles. This work shows otherwise and uses Ansoft (Pittsburgh ...


Characterization Of Glycation Sites On Human Serum Albumin Using Mass Spectrometry, Omar S. Barnaby 2010 University of Nebraska-Lincoln

Characterization Of Glycation Sites On Human Serum Albumin Using Mass Spectrometry, Omar S. Barnaby

Student Research Projects, Dissertations, and Theses - Chemistry Department

The modification of proteins by reducing sugars is a process that occurs naturally in the body. This process, which is known as glycation, has been linked to many of the chronic complications encountered during diabetes. Glycation has also been linked to changes in the binding of human serum albumin (HSA) to several drugs and small solutes in the body. While these effects are known, there is little information that explains why these changes in binding occur. The goal of this project was to obtain qualitative and quantitative information about glycation that occurs on HSA. The first section of this dissertation ...


Synthesis Of Cifidabco And Mannose Co-Functionalized G(4)-Pamam Dendrimer For Use In Antibiotics, Harrison VanKoten 2010 Carroll College, Helena, MT

Synthesis Of Cifidabco And Mannose Co-Functionalized G(4)-Pamam Dendrimer For Use In Antibiotics, Harrison Vankoten

Chemistry and Physics Undergraduate Theses

Synthesis of C16DABCO and mannose co-functionalized G(4)-PAMAM dendrimer is expected to yield a useful antimicrobial that is effective in binding to mannose specific lectins on bacterial fimbriae. Thus far, mannose has been attached to the dendrimer with near complete functionalization. C16DABCO and mannose cofunctionalized G(4)-PAMAM dendrimer have been synthesized, but not in sufficient quantities to allow examination of its the antibacterial properties. Future work includes optimizing the final synthesis and performing toxicity studies.


The Effects Of Glycosylation On Prion Protein Conversion, Christopher Casebeer 2010 Carroll College, Helena, MT

The Effects Of Glycosylation On Prion Protein Conversion, Christopher Casebeer

Life and Environmental Sciences Undergraduate Theses

Glycosylation of the prion protein may be experimentally altered through expression in different insect cell lines. The presence or absence of several terminal sugars may or may not have an effect on the conversion efficiency of the prion protein into its infectious form. The main focus of this research was to characterize the prion protein as it is produced in two varying insect cell lines that are able to glycosylate proteins to different extents. Glycosidase treatment demonstrated that the expressed prion protein was glycosylated. Lectin blotting was used to assess the presence or absence of terminal sugar moieties existing on ...


The Development Of A Quantitative Assay To Measure The Effects Of Protein Disulfide Isomerase On Prion Protein Misfolding, Lauren Vogl 2010 Carroll College, Helena, MT

The Development Of A Quantitative Assay To Measure The Effects Of Protein Disulfide Isomerase On Prion Protein Misfolding, Lauren Vogl

Life and Environmental Sciences Undergraduate Theses

The objective ofthis project was to develop a quantitative assay to measure the misfolding ofprion proteins that can be used to assess the effects of Protein Disulfide Isomerase (PDI) on the misfolding process ofthe Chronic Wasting Disease (CWD) prion protein. CWD is a spongiform encephalopathy native to animals in the family Cervidae, which includes whitetail deer, mule deer, elk and moose. CWD is caused by the misfolding ofprion proteins. Qualitative observations from previous studies indicate that disulfide bond rearrangement might be a rate limiting step in the misfolding process. There is a need to be able to quantitate the amount ...


Characterization And Normalization Of Rat Salivary Cholinesterase, Elise Klohe 2010 Carroll College, Helena, MT

Characterization And Normalization Of Rat Salivary Cholinesterase, Elise Klohe

Life and Environmental Sciences Undergraduate Theses

Cholinesterase (ChE) is an enzyme which allows neurons to return to a resting state by hydrolyzing the neurotransmitter acetylcholine. Organophosphate pesticides, which are commonly used in agriculture, inhibit the activity of ChE. The inhibition of ChE activity in plasma and red blood cells is used as a biomarker for human pesticide exposure. Currently, salivary ChE activity is being examined as a non-invasive biomarker; however little is known about baseline ChE activity in saliva. In order to characterize baseline salivary ChE, saliva was collected from 5 adult male SpragueDawley rats, salivary ChE was measured using the Ellman assay, and total salivary ...


Effect Of Membrane Cholesterol Levels And Allelic Variation On Prion Conversion, Ross Hartman 2010 Carroll College, Helena, MT

Effect Of Membrane Cholesterol Levels And Allelic Variation On Prion Conversion, Ross Hartman

Life and Environmental Sciences Undergraduate Theses

Chronic Wasting Disease (CWD) is caused by an accumulation of misfolded prion proteins (PrPsc) and subsequent plaque formation in the central nervous system. CWD is horizontally transferable; misfolded prions from one animal can enter another and cause normal prion proteins (PrPc) to misfold. This misfolding process is termed prion conversion. In natural deer populations an allelic variation in the prion gene is thought to confer resistance to CWD. Wild type mule deer are serine (S) homozygotes at codon 225. Mule deer that are Serine/ Phenyalanine (S/F) heterozygotes exhibit resistance to prion infection. In this study the F encoding allele ...


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