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Full-Text Articles in Biochemistry, Biophysics, and Structural Biology

Computational Prediction Of The Agregated Structure Of Denatured Lysozyme, Pongsathorn Chotikasemsri Dec 2009

Computational Prediction Of The Agregated Structure Of Denatured Lysozyme, Pongsathorn Chotikasemsri

Masters Theses & Specialist Projects

Mis-folded proteins and their associated aggregates are a contributing factor in some human diseases. In this study we used the protein lysozyme as a model to define aggregation structures under denaturing conditions. Sasahara et al. (2007), Frare et al. (2009, 2006), and Rubin et al. (2008) observed conditions where heat denatured lysozyme formed fibril structures that were observed to be 8-17 nanometers in diameter under the electron microscope. Even though the crystal structure of lysozyme is known, the denatured form of this protein is still unknown. Therefore, we used Rosetta++ protein folding and blind docking software to create in silico ...


Confocal Laser Scanning Microscopy: A Flexible Tool For Polarized Light And Three-Dimensional Fluorescence Imaging Of Compact Bone, Corey Maggiano, Tosha Dupras, Michael Schultz, John Biggerstaff Sep 2009

Confocal Laser Scanning Microscopy: A Flexible Tool For Polarized Light And Three-Dimensional Fluorescence Imaging Of Compact Bone, Corey Maggiano, Tosha Dupras, Michael Schultz, John Biggerstaff

John Biggerstaff

Wide-field polarized light and epifluorescence microscopy have been used to enhance analysis of archaeological bone tissue, providing information on bone formation, modeling, pathology, preservation, age estimation, and biomechanics. Though valuable, these techniques are limited by their inability to remove out-of-focus light and view multiple levels of a sample, restricting our understanding of the three-dimensional (3-D) microarchitecture of compact bone. Modern technological advances, such as microscopic computerized tomography, allow increasing resolution in 3-D bone imaging, but do not allow fluorescence labeling or polarized-light analysis. Confocal laser scanning microscopy (CLSM) is a valuable tool for 3-D histology. However, its application to the ...


Hdl-Cholesterol-Raising Effect Of Orange Juice In Subjects With Hypercholesterolemia, Elzbieta Kurowska, J. Spence, John Jordan, Stephen Wetmore, David Freeman, Leonard Piché, Paula Serratore Aug 2009

Hdl-Cholesterol-Raising Effect Of Orange Juice In Subjects With Hypercholesterolemia, Elzbieta Kurowska, J. Spence, John Jordan, Stephen Wetmore, David Freeman, Leonard Piché, Paula Serratore

Leonard Piché

Background: Orange juice-a rich source of vitamin C, folate, and flavonoids such as hesperidin-induces hypocholesterolemic responses in animals. Objective: We determined whether orange juice beneficially altered blood lipids in subjects with moderate hypercholesterolemia. Design: The sample consisted of 16 healthy men and 9 healthy women with elevated plasma total and LDL-cholesterol and normal plasma triacylglycerol concentrations. Participants incorporated 1, 2, or 3 cups (250 mL each) of orange juice sequentially into their diets, each dose over a period of 4 wk. This was followed by a 5-wk washout period. Plasma lipid, folate, homocyst(e)ine, and vitamin C (a compliance ...


Identification Of Light-Independent Inhibition Of Human Immunodeficiency Virus-1 Infection Through Bioguided Fractionation Of Hypericum Perforatum, Wendy Maury, Jason P. Price, Melinda A. Brindley, Choonseok Oh, Jeffrey D. Neighbors, David F. Wiemer, Nickolas Wills, Susan L. Carpenter, Catherine C. Hauck, Patricia A. Murphy, Mark P. Widrlechner, Kathleen Delate, Ganesh Kumar, George A. Kraus, Ludmila Rizshsky, Basil J. Nikolau Jul 2009

Identification Of Light-Independent Inhibition Of Human Immunodeficiency Virus-1 Infection Through Bioguided Fractionation Of Hypericum Perforatum, Wendy Maury, Jason P. Price, Melinda A. Brindley, Choonseok Oh, Jeffrey D. Neighbors, David F. Wiemer, Nickolas Wills, Susan L. Carpenter, Catherine C. Hauck, Patricia A. Murphy, Mark P. Widrlechner, Kathleen Delate, Ganesh Kumar, George A. Kraus, Ludmila Rizshsky, Basil J. Nikolau

NCRPIS Publications and Papers

Background
Light-dependent activities against enveloped viruses in St. John's Wort (Hypericum perforatum) extracts have been extensively studied. In contrast, light-independent antiviral activity from this species has not been investigated.

Results
Here, we identify the light-independent inhibition of human immunodeficiency virus-1 (HIV-1) by highly purified fractions of chloroform extracts of H. perforatum. Both cytotoxicity and antiviral activity were evident in initial chloroform extracts, but bioassay-guided fractionation produced fractions that inhibited HIV-1 with little to no cytotoxicity. Separation of these two biological activities has not been reported for constituents responsible for the light-dependent antiviral activities. Antiviral activity was associated with more ...


Association Of Oxidative Stress, Iron, And Centralized Fat Mass In Healthy Postmenopausal Women, Betsy L. Crist, D. Lee Alekel, Laura M. Ritland, Laura N. Hanson, Ulrike Genschel, Manju B. Reddy Jun 2009

Association Of Oxidative Stress, Iron, And Centralized Fat Mass In Healthy Postmenopausal Women, Betsy L. Crist, D. Lee Alekel, Laura M. Ritland, Laura N. Hanson, Ulrike Genschel, Manju B. Reddy

Food Science and Human Nutrition Publications

Objective: Centralized adiposity, insulin resistance, excess iron, and elevated oxidative stress place postmenopausal women at risk for atherosclerotic cardiovascular disease (CVD). The objective of this study was to determine the relationship among excess iron, oxidative stress, and centralized fat mass in healthy postmenopausal women. Methods: The parent project recruited healthy women for a randomized, double-blind, clinical trial designed to examine the effect of soy isoflavones on bone. At baseline ( n = 122), we measured three antioxidant enzymes, iron status indices (serum ferritin among others), oxidative stress indices (oxidized low-density lipoprotein [oxLDL], urinary isoprostanes [PGF2α], protein carbonyls, DNA damage), and waist, hip ...


Quantification Of Ferritin From Staple Food Crops, Rebbecca J. Lukac, Maneesha R. Aluru, Manju B. Reddy Mar 2009

Quantification Of Ferritin From Staple Food Crops, Rebbecca J. Lukac, Maneesha R. Aluru, Manju B. Reddy

Food Science and Human Nutrition Publications

Ferritin-iron has been shown to be as bioavailable as ferrous sulfate in humans. Thus, biofortification to breed crops with high ferritin content is a promising strategy to alleviate the global iron deficiency problem. Although ferritin is present in all food crops, its concentration varies between species and varieties. Therefore, a successful ferritin biofortification strategy requires a method to rapidly measure ferritin concentrations in food crops. The objective of this study was to develop a simple and reliable ELISA using an anti-ferritin polyclonal antibody to detect ferritin in various crops. Crude seed extracts were found to have 10.2 +/- 1.0 ...


Inhibition Of Lentivirus Replication By Aqueous Extracts Of Prunella Vulgaris, Melinda A. Brindley, Mark P. Widrlechner, Joe-Ann Mccoy, Patricia A. Murphy, Catherine C. Hauck, Ludmila Rizshsky, Basil J. Nikolau, Wendy Maury Jan 2009

Inhibition Of Lentivirus Replication By Aqueous Extracts Of Prunella Vulgaris, Melinda A. Brindley, Mark P. Widrlechner, Joe-Ann Mccoy, Patricia A. Murphy, Catherine C. Hauck, Ludmila Rizshsky, Basil J. Nikolau, Wendy Maury

NCRPIS Publications and Papers

Background
Various members of the mint family have been used historically in Chinese and Native American medicine. Many of these same family members, including Prunella vulgaris, have been reported to have anti-viral activities. To further characterize the anti-lentiviral activities of P. vulgaris, water and ethanol extractions were tested for their ability to inhibit equine infectious anemia virus (EIAV) replication.

Results
Aqueous extracts contained more anti-viral activity than did ethanol extracts, displaying potent anti-lentiviral activity against virus in cell lines as well as in primary cell cultures with little to no cellular cytotoxicity. Time-of-addition studies demonstrated that the extracts were effective ...


Rosmarinic Acid In Prunella Vulgaris Ethanol Extract Inhibits Lipopolysaccharide-Induced Prostaglandin E2 And Nitric Oxide In Raw 264.7 Mouse Macrophages, Nan Huang, Cathy Hauck, Man-Yu Yum, Ludmila Rizshsky, Mark P. Widrlechner, Joe-Ann Mccoy, Patricia A. Murphy, Philip M. Dixon, Basil J. Nikolau, Diane F. Birt Jan 2009

Rosmarinic Acid In Prunella Vulgaris Ethanol Extract Inhibits Lipopolysaccharide-Induced Prostaglandin E2 And Nitric Oxide In Raw 264.7 Mouse Macrophages, Nan Huang, Cathy Hauck, Man-Yu Yum, Ludmila Rizshsky, Mark P. Widrlechner, Joe-Ann Mccoy, Patricia A. Murphy, Philip M. Dixon, Basil J. Nikolau, Diane F. Birt

Food Science and Human Nutrition Publications

Prunella vulgaris has been used therapeutically for inflammation-related conditions for centuries, but systematic studies of its anti-inflammatory activity are lacking and no specific active components have been identified. In this study, water and ethanol extracts of four P. vulgaris accessions were applied to RAW 264.7 mouse macrophages, and the ethanol extracts significantly inhibited lipopolysaccharide (LPS)-stimulated prostaglandin E2 (PGE2) and nitric oxide (NO) production at 30 μg/mL without affecting cell viability. Extracts from different accessions of P. vulgaris were screened for anti-inflammatory activity to identify accessions with the greatest activity. The inhibition of PGE2 and NO production by ...


Endogenous Levels Of Echinacea Alkylamides And Ketones Are Important Contributors To The Inhibition Of Prostaglandin E2 And Nitric Oxide Production In Cultured Macrophages, Carlie A. Lalone, Ludmila Rizshsky, Kimberly D.P. Hammer, Lankun Wu, Avery K.S. Solco, Man-Yu Yum, Basil J. Nikolau, Eve S. Wurtele, Patricia A. Murphy, Meehye Kim, Diane F. Birt Jan 2009

Endogenous Levels Of Echinacea Alkylamides And Ketones Are Important Contributors To The Inhibition Of Prostaglandin E2 And Nitric Oxide Production In Cultured Macrophages, Carlie A. Lalone, Ludmila Rizshsky, Kimberly D.P. Hammer, Lankun Wu, Avery K.S. Solco, Man-Yu Yum, Basil J. Nikolau, Eve S. Wurtele, Patricia A. Murphy, Meehye Kim, Diane F. Birt

Food Science and Human Nutrition Publications

Because of the popularity of Echinacea as a dietary supplement, researchers have been actively investigating which Echinacea constituent or groups of constituents are necessary for immunemodulating bioactivities. Our prior studies indicate that alkylamides may play an important role in the inhibition of prostaglandin E2 (PGE2) production. High-performance liquid chromatography fractionation, employed to elucidate interacting anti-inflammatory constituents from ethanol extracts of Echinacea purpurea, Echinacea angustifolia, Echinacea pallida, and Echinacea tennesseensis, identified fractions containing alkylamides and ketones as key anti-inflammatory contributors using lipopolysaccharideinduced PGE2 production in RAW264.7 mouse macrophage cells. Nitric oxide (NO) production and parallel cytotoxicity screens were also employed ...


Signal Pathway Of 17Β-Estradiol–Induced Muc5b Expression In Human Airway Epithelial Cells, H Choi, Y Chung, H Kim, U Moon, Y Choi, I Van Seuringen, Seung Baek, H Yoon, J Yoon Dec 2008

Signal Pathway Of 17Β-Estradiol–Induced Muc5b Expression In Human Airway Epithelial Cells, H Choi, Y Chung, H Kim, U Moon, Y Choi, I Van Seuringen, Seung Baek, H Yoon, J Yoon

Seung J Baek

MUC5B is a major mucin of the human respiratory tract, and it is not clear how MUC5B expression is regulated in various airway diseases. The goal of this study was to determine the mechanisms by which 17β-estradiol induces MUC5B gene expression in airway epithelial cells. It was found that E2, a sex hormone, stimulates MUC5B gene overexpression by interaction with estrogen receptor {alpha} (ER{alpha}) and by acting through extracellular signal–regulated kinase 1/2 (ERK1/2)-mitogen-activated protein kinase (MAPK). Pretreatment with ER antagonist ICI 182,780 blocked both E2-induced ERK1/2-MAPK activation and MUC5B gene expression. It was ...


Nonsteroidal Anti-Inflammatory Drug-Activated Gene-1 Expression Inhibits Urethane-Induced Pulmonary Tumorigenesis In Transgenic Mice, M Cekanova, S Lee, M Sukhthankar, R Donnell, T Eling, S Fischer, Seung Baek Dec 2008

Nonsteroidal Anti-Inflammatory Drug-Activated Gene-1 Expression Inhibits Urethane-Induced Pulmonary Tumorigenesis In Transgenic Mice, M Cekanova, S Lee, M Sukhthankar, R Donnell, T Eling, S Fischer, Seung Baek

Seung J Baek

The expression of nonsteroidal anti-inflammatory drug-activated gene-1 (NAG-1) inhibits gastrointestinal tumorigenesis in NAG-1 transgenic mice (C57/BL6 background). In the present study, we investigated whether the NAG-1 protein would alter urethane-induced pulmonary lesions in NAG-1 transgenic mice on an FVB background (NAG-1(Tg+/FVB)). NAG-1(Tg+/FVB) mice had both decreased number and size of urethane-induced tumors, compared with control littermates (NAG-1(Tg+/FVB) = 16 +/- 4 per mouse versus control = 20 +/- 7 per mouse, P < 0.05). Urethane-induced pulmonary adenomas and adenocarcinomas were observed in control mice; however, only pulmonary adenomas were observed in NAG-1(Tg+/FVB) mice. Urethane-induced tumors from control littermates and NAG-1(Tg+/FVB) mice highly expressed proteins in the arachidonic acid pathway (cyclooxygenases 1/2, prostaglandin E synthase, and prostaglandin E(2) receptor) and highly activated several kinases (phospho-Raf-1 and phosphorylated extracellular signal-regulated kinase 1/2). However, only urethane-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation was decreased in NAG-1(Tg+/FVB) mice. Furthermore, significantly increased apoptosis in tumors of NAG-1(Tg+/FVB) mice compared with control mice was observed as assessed by caspase-3/7 activity. In addition, fewer inflammatory cells were observed in the lung tissue isolated from urethane-treated NAG-1(Tg+/FVB) mice compared with control mice. These results paralleled in vitro assays using human A549 pulmonary carcinoma cells. Less phosphorylated p38 MAPK was observed in cells overexpressing NAG-1 compared with control cells. Overall, our study revealed for the first time that the NAG-1 protein inhibits urethane-induced tumor formation, probably mediated by the p38 MAPK pathway, and is a possible new target for lung cancer chemoprevention


Changes In Hepatic Gene Expression In Dogs With Experimentally Induced Nutritional Iron Deficiency, M Fry, C Kirk, J Liggett, G Daniel, Seung Baek, J Gouffon, P Chimakurthy, B Rekapalli Dec 2008

Changes In Hepatic Gene Expression In Dogs With Experimentally Induced Nutritional Iron Deficiency, M Fry, C Kirk, J Liggett, G Daniel, Seung Baek, J Gouffon, P Chimakurthy, B Rekapalli

Seung J Baek

BACKGROUND AND OBJECTIVE: We investigated hepatic gene expression in dogs with experimentally induced nutritional iron deficiency (ID). Our hypothesis was that ID would result in decreased hepcidin gene expression, and possibly in altered expression of other genes associated with iron metabolism. METHODS: Liver biopsies were collected from each of 3 dogs before induction of ID, at the point of maximal ID, and after resolution of ID. Using Affymetrix microarray technology and analytical tools specifically designed for microarray data, we identified genes that had at least a 2-fold change in expression in response to ID. Four genes were selected for further ...


Pparγ Agonists Down-Regulate The Expression Of Atp10c Mrna During Adipogenesis, A Peretich, M Cekanova, S Hurst, Seung Baek, M Dhar Dec 2008

Pparγ Agonists Down-Regulate The Expression Of Atp10c Mrna During Adipogenesis, A Peretich, M Cekanova, S Hurst, Seung Baek, M Dhar

Seung J Baek

We have shown that Atp10c, a type 4 P-type ATPase, is a strong candidate affecting glucose and lipid metabolism in humans and mice. Atp10c is a putative phospholipid translocase associated with cell signaling and intracellular protein trafficking. In order to examine the biological role of Atp10c, semiquantitative reverse transcriptase – polymerase chain reaction was carried out. Atp10c mRNA is expressed in 3T3-L1 cells and in primary preadipocytes and adipocytes generated from mice. Atp10c mRNA is regulated during fat cell differentiation and modulated by PPARγ agonists and antagonists as well as by hormonal factors (insulin and dexamethasone). Atp10c expression is regulated by ...


The Cyclooxygenase Inhibitor Sulindac Sulfide Inhibits Ep4 Expression And Suppresses The Growth Of Glioblastoma Cells, A Kambe, H Yoshioka, H Kamitani, T Watanabe, Seung Baek, T Eling Dec 2008

The Cyclooxygenase Inhibitor Sulindac Sulfide Inhibits Ep4 Expression And Suppresses The Growth Of Glioblastoma Cells, A Kambe, H Yoshioka, H Kamitani, T Watanabe, Seung Baek, T Eling

Seung J Baek

EP4 expression in human glioblastoma cells correlates with growth on soft agar. The cyclooxygenase inhibitor sulindac sulfide first altered specificity protein-1 (Sp-1) and early growth response gene-1 expression, then increased the expression of nonsteroidal anti-inflammatory drug-activated gene 1 and activating transcription factor 3, and then decreased EP4 expression. EP4 suppression was dependent on blocking the Sp-1 binding sites in the human EP4 promoter. Mutation in the Sp-1 sites in EP4 altered the promoter activity and abolished sulindac sulfide effects. The inhibitory effect of sulindac sulfide on EP4 expression was reversed by PD98059, a mitogen-activated protein/extracellular signal-regulated kinase kinase-1/extracellular ...


A Reciprocal Relationship Exists Between Non-Steroidal Anti-Inflammatory Drug-Activated Gene-1 (Nag-1) And Cyclooxygenase-2, G Iguchi, K Chrysovergis, S Lee, Seung Baek, R Langenbach, T Eling Dec 2008

A Reciprocal Relationship Exists Between Non-Steroidal Anti-Inflammatory Drug-Activated Gene-1 (Nag-1) And Cyclooxygenase-2, G Iguchi, K Chrysovergis, S Lee, Seung Baek, R Langenbach, T Eling

Seung J Baek

Non-steroidal anti-inflammatory drug (NSAID)-activated gene-1 (NAG-1) and COX-2 are involved in cellular processes such as inflammation, apoptosis, and tumorigenesis. To address the relationship between COX-2 and NAG-1 expression, we investigated the expression of NAG-1 and COX-2 in normal and tumor tissue from human patients, Apc(Min/+) mice, and COX-2(-/-) mice. While COX-2 expression is highly induced in tumor tissue, NAG-1 expression is reduced. Furthermore, PGE(2) reduces NAG-1 while celebrex induces NAG-1 expression. The results suggest that a possible inverse relationship exists between the expression of NAG-1 and COX-2 in tumor formation of colon tissue.