Cell and Developmental Biology Commons^™

16s Ribosomal Dna Sequencing Confirms The Synonymy Of Vibrio Harveyi And V. Carchariae, Eric J. Gauger, Marta Gomez-Chiarri

Marta Gomez-Chiarri

Seventeen bacterial strains previously identified as Vibrio harveyi (Baumann et al. 1981) or V. carchariae (Grimes et al. 1984) and the type strains of V. harveyi, V. carchariae and V. campbellii were analyzed by 16S ribosomal DNA (rDNA) sequencing. Four clusters were identified in a phylogenetic analysis performed by comparing a 746 base pair fragment of the 16S rDNA and previously published sequences of other closely related Vibrio species. The type strains of V. harveyi and V. carchariae and about half of the strains identified as V. harveyi or V. carchariae formed a single, wellsupported cluster designed as ‘bona fide’ …

Novel Developmental Biology-Based Protocol Of Embryonic Stem Cell Differentiation To Morphologically Sound And Functional Yet Immature Hepatocytes, Terence Bukong, Tracie Lo, Gyongyi Szabo, Angela Dolganiuc

Gyongyi Szabo

BACKGROUND/AIMS: Liver diseases are common in the United States and often require liver transplantation; however, donated organs are limited and thus alternative sources for liver cells are in high demand. Embryonic stem cells (ESC) can provide a continuous and readily available source of liver cells. ESC differentiation to liver cells is yet to be fully understood and comprehensive differentiation protocols are yet to be defined. Here, we aimed to achieve human (h)ESC differentiation into mature hepatocytes using defined recombinant differentiation factors and metabolites. METHODS: Embryonic stem cell H1 line was sub-cultured on feeder layer. We induced hESCs into endodermal differentiation …

Regulation Of Atg4b Stability By Rnf5 Limits Basal Levels Of Autophagy And Influences Susceptibility To Bacterial Infection, E. Kuang, Cheryl Okumura, S. Sheffy-Levin, T. Varsano, V. Shu, J. Qi, I. Niesman, H. Yang, C. LóPez-OtíN, W. Yang, J. Reed, L. Broday, V. Nizet, Z. Ronai

Cheryl Okumura

Autophagy is the mechanism by which cytoplasmic components and organelles are degraded by the lysosomal machinery in response to diverse stimuli including nutrient deprivation, intracellular pathogens, and multiple forms of cellular stress. Here, we show that the membrane-associated E3 ligase RNF5 regulates basal levels of autophagy by controlling the stability of a select pool of the cysteine protease ATG4B. RNF5 controls the membranal fraction of ATG4B and limits LC3 (ATG8) processing, which is required for phagophore and autophagosome formation. The association of ATG4B with—and regulation of its ubiquitination and stability by—RNF5 is seen primarily under normal growth conditions. Processing of …

Gonadotropin Concentrations, Follicular Development, And Luteal Function In Pituitary Stalk-Transfected Ewes Treated With Bovine Follicular Fluid, G. Larson, David Mallory, R. Dailey, P. Lewis

David S. Mallory

Two experiments, each arranged as a 2 x 2 factorial, were conducted in ewes to examine direct effects of bovine follicular fluid (bFF) on follicular development and luteal function and to further characterize follicular development and luteal function after pituitary stalk transection (SS). In Exp. 1, ewes were sham-operated or SS on d 6 of an estrous cycle and received 5 ml of saline or bFF three times daily on d 5 through 11 of the same cycle. In Exp. 2, all ewes were SS on d 6 of an estrous cycle and treated with saline or bFF three times …

Mecp2 Binds To Nucleosome Free (Linker Dna) Regions And To H3k9/H3k27 Methylated Nucleosomes In The Brain, Anita Thambirajah, Marlee Ng, Lindsay Frehlick, Andra Li, Jason Serpa, Evgeniy Petrotchenko, Begonia Silva-Moreno, Kristal Missiaen, Christoph Borchers, J. Hall, Ryan Mackie, Frank Lutz, Brent Gowen, Michael Hendzel, Philippe Georgel, Juan Ausio

Philippe T. Georgel

Methyl-CpG-binding protein 2 (MeCP2) is a chromatin-binding protein that mediates transcriptional regulation, and is highly abundant in brain. The nature of its binding to reconstituted templates has been well characterized in vitro. However, its interactions with native chromatin are less understood. Here we show that MeCP2 displays a distinct distribution within fractionated chromatin from various tissues and cell types. Artificially induced global changes in DNA methylation by 3-aminobenzamide or 5-aza-2′-deoxycytidine, do not significantly affect the distribution or amount of MeCP2 in HeLa S3 or 3T3 cells. Most MeCP2 in brain is chromatin-bound and localized within highly nuclease-accessible regions. We also …

A New Pharmacological Agent (Akb-4924) Stabilizes Hypoxia Inducible Factor (Hif) And Increases Skin Innate Defenses Against Bacterial Infection., Cheryl Okumura, A. Hollands, D. Tran, J. Olson, S. Dahesh, M. Von KöCkritz-Blickwede, W. Thienphrapa, C. Corle, S. Jeung, A. Kotsakis, R. Shalwitz, R. Johnson, V. Nizet

Cheryl Okumura

Hypoxia inducible factor-1 (HIF-1) is a transcription factor that is a major regulator of energy homeostasis and cellular adaptation to low oxygen stress. HIF-1 is also activated in response to bacterial pathogens and supports the innate immune response of both phagocytes and keratinocytes. In this work, we show that a new pharmacological compound AKB-4924 (Akebia Therapeutics) increases HIF-1α levels and enhances the antibacterial activity of phagocytes and keratinocytes against both methicillin-sensitive and -resistant strains of Staphylococcus aureus in vitro. AKB-4924 is also effective in stimulating the killing capacity of keratinocytes against the important opportunistic skin pathogens Pseudomonas aeruginosa and Acinitobacter …

Library Impact Statement For Bps 455 Protein Molecular Modeling, Michael Vocino

michael c vocino

Library Impact Statement for BPS 455 Protein Molecular Modeling. No new resources were required to support this course.

Library Impact Statement For Bps 555 Protein Molecular Modeling, Michael Vocino

michael c vocino

Library Impact Statement for BPS 555 Protein Molecular Modeling. No new resources were required to support this course.

Novel Role Of The Transcription Factor Hif-1Α In The Formation Of Mast Cell Extracellular Traps., K. Branitzki-Heinemann, Cheryl Okumura, L. VöLlger, Y. Kawakami, T. Kawakami, H. Naim, V. Nizet, M. Von KöCkritz-Blickwede

Cheryl Okumura

MCs (mast cells) are critical components of the host innate immune defence against bacterial pathogens, providing a variety of intra- and extra-cellular antimicrobial functions. In the present study we show, for the first time, that the transcriptional regulator HIF-1α (hypoxia-inducible factor-1α) mediates the extracellular antimicrobial activity of human and murine MCs by increasing the formation of MCETs (MC extracellular traps).

Migration Of Tumor Cells In 3d Matrices Is Governed By Matrix Stiffness Along With Cell-Matrix Adhesion And Proteolysis, Alisha Sarang-Sieminski, Muhammad Zaman, Linda Trapani, Drew Mackellar, Haiyan Gong, Roger Kamm, Alan Wells, Douglas Lauffenburger, Paul Matsudaira

Alisha L. Sarang-Sieminski

Cell migration on 2D surfaces is governed by a balance between counteracting tractile and adhesion forces. Although biochemical factors such as adhesion receptor and ligand concentration and binding, signaling through cell adhesion complexes, and cytoskeletal structure assembly/disassembly have been studied in detail in a 2D context, the critical biochemical and biophysical parameters that affect cell migration in 3D matrices have not been quantitatively investigated. We demonstrate that, in addition to adhesion and tractile forces, matrix stiffness is a key factor that influences cell movement in 3D. Cell migration assays in which Matrigel density, fibronectin concentration, and β1 integrin binding are …

Functional Characterization Of An Epidermal Growth Factor Receptor/Ret Chimera, Helen Donis-Keller, Sunil Pandit, Timothy O'Hare, Linda Pike

Helen Donis-Keller

The RET (combined in ransfection) gene encodes a receptor tyrosine kinase homolog involved in innervation of the gut and renal development. A chimeric epidermal growth factor receptor (EGFR)/RET receptor was constructed which contained the extracellular and transmembrane domains of the EGF receptor fused to the intracellular domain of RET. This construct was expressed in NIH 3T3 cells, and the functional properties of the receptor were characterized and compared with those of the wild type EGF receptor. Whereas the EGF receptor exhibited both high and low affinity binding sites for ¹²⁵I-EGF, the EGFR/RET chimera exhibited only low affinity binding …

Forisome Performance In Artificial Sieve Tubes, Michael Knoblauch, Mike Stubenrauch, Aart J.E. Van Bel, Winfried S. Peters

Winfried S. Peters

Multifetal Pregnancy May Increase The Risk For Severe Maternal And Neonatal Vitamin D Deficiency, Benjamin U. Nwosu, Leslie A. Soyka, Amanda Angelescu, Olga T. Hardy, Mary M. Lee

Benjamin U. Nwosu

Vitamin D deficiency is prevalent in pregnant women. This deficiency could be exaggerated in multifetal pregnancies by the increased demand on maternal stores of vitamin D. We present 2 cases in which hypocalcemia and secondary hyperparathyroidism occurred in 2 sets of twins from mothers with vitamin D deficiency. The first mother had gastric bypass surgery and Crohn disease. The second mother had no apparent cause of vitamin D deficiency. Both women had iron deficiency anemia and lived in Northeastern United States. We speculate that in twins, the demand for 25-hydroxyvitamin D by 2 fetuses could deplete the 25-hydroxyvitamin D stores …

Guanosine Diphosphatase Is Required For Protein And Sphingolipid Glycosylation In The Golgi Lumen Of Saccharomyces Cerevisiae, Claudia Abeijon, Ken Yanagisawa, Elisabet Mandon, Alex Hausler, Kelley Moremen, Carlos Hirschberg, Phillips Robbins

Elisabet Mandon

Current models for nucleotide sugar use in the Golgi apparatus predict a critical role for the lumenal nucleoside diphosphatase. After transfer of sugars to endogenous macromolecular acceptors, the enzyme converts nucleoside diphosphates to nucleoside monophosphates which in turn exit the Golgi lumen in a coupled antiporter reaction, allowing entry of additional nucleotide sugar from the cytosol. To test this model, we cloned the gene for the S. cerevisiae guanosine diphosphatase and constructed a null mutation. This mutation should reduce the concentrations of GDP-mannose and GMP and increase the concentration of GDP in the Golgi lumen. The alterations should in turn …

Viability Of Lactic Acid Bacteria And Sensory Evaluation In Cinnamomum Verum And Allium Sativum-Bio-Yogurts Made From Camel And Cow Milk, Ahmad Salihin Hj Baba, Shori A. B.

Ahmad Salihin Hj Baba

The present study investigate the effect of herbal water extract prepared from Allium sativum and Cinnamomum verum on the viability of lactic acid bacteria (Lactobacillus spp and Streptococcus thermophilus) in cow- and camel-milk yogurts during 21 day refrigerated storage. The organoleptic properties of fresh-yogurts were evaluated. Lactobacillus spp count for fresh cow milk-yogurts (0 day) in both present and absent of C. verum and A. sativum was ranged from 1.4×0 6 to 2.1×10 6 cfu/mL. These values were not significantly changed throughout the 21 days of refrigerated storage. Lactobacillusspp count in fresh plain camel milk- yogurt was 13.2×10 6 cfu/mL …

Novel Phenol Soluble Modulin Derivatives In Community-Associated Methicillin- Resistant Staphylococcus Aureus Identified Through Imaging Mass Spectrometry., D. Gonzalez, Cheryl Okumura, A. Hollands, R. Kersten, K. Akong-Moore, M. Pence, C. Malone, J. Derieux, B. Moore, A. Horswill, J. Dixon, P. Dorrestein, V. Nizet

Cheryl Okumura

Staphylococcus aureus causes a wide range of human disease ranging from localized skin and soft tissue infections to potentially lethal systemic infections. S. aureus has the biosynthetic ability to generate numerous virulence factors that assist in circumventing the innate immune system during disease pathogenesis. Recent studies have uncovered a set of extracellular peptides produced by community-associated methicillin-resistant S. aureus (CA-MRSA) with homology to the phenol-soluble modulins (PSMs) from Staphylococcus epidermidis. CA-MRSA PSMs contribute to skin infection and recruit and lyse neutrophils, and truncated versions of these peptides possess antimicrobial activity. In this study, novel CA-MRSA PSM derivatives were discovered by …

Symbiosis (Addiction: Biology, Psychology And Society), Ahmed Mustafa, Sharon Morgillo, Shree Dhawale

Ahmed Mustafa Dr.

No abstract provided.

Do Larval Traits Re-Evolve? Evidence From The Embryogenesis Of A Direct-Developing Salamander, Plethodon Cinereus, Ryan Kerney, David Blackburn, Hendrik Muller, James Hanken

Ryan Kerney

Recent molecular phylogenies suggest the surprising reacquisition of posthatching metamorphosis within an otherwise directdeveloping clade of lungless salamanders (family Plethodontidae). Metamorphosis was long regarded as plesiomorphic for plethodontids, yet the genus Desmognathus, which primarily includes metamorphosing species, is now nested within a much larger clade of direct-developing species. The extent to which the putative reacquisition of metamorphosis in Desmognathus represents a true evolutionary reversal is contingent upon the extent to which both larva-specific features and metamorphosis were actually lost during the evolution of direct development. In this study we analyze development of the hyobranchial skeleton, which is dramatically remodeled during …

Glioblastoma Derived Exosomes Induce Apoptosis In Cytotoxic T Cells Through A Fas Ligand Mediated Mechanism, Keith Sabin, Richard Rovin, Johnathan Lawrence, Robert Belton, Robert Winn

Johnathan Lawrence

INTRODUCTION: Glioblastoma multiforme deploy s a number of weapons to thwart the immune system. Within the tumor microenvironment, cytotoxic T cells fall victim to Fas ligand (FasL) induced apoptosis. In prostate and colorectal cancer, exosomes can mediate this FasL induced T cell apoptosis. Exosomes are tiny, membrane bound vesicles that are released from a cell. They contain functional mRNA and protein and have cell surface molecules representative of their parent cell. It is not known if GBM derived exosomes can also mediate FasL triggered apoptosis. In this study, the role of tumor derived exosomes as the delivery vehicle for FasL …

Pharmacological Inhibition Of The Clpxp Protease Increases Bacterial Susceptibility To Host Cathelicidin Antimicrobial Peptides And Cell-Envelope Active Antibiotics., S. Mcgillivray, D. Tran, N. Ramadoss, J. Alumasa, Cheryl Okumura, G. Sakoulas, M. Vaughn, D. Zhang, K. Keiler, V. Nizet

Cheryl Okumura

The ClpXP protease is a critical bacterial intracellular protease that regulates protein turnover in many bacterial species. Here we identified a pharmacological inhibitor of the ClpXP protease, F2, and evaluated its action in Bacillus anthracis and Staphylococcus aureus. We found that F2 exhibited synergistic antimicrobial activity with cathelicidin antimicrobial peptides and antibiotics that target the cell well and/or cell membrane such as penicillin and daptomycin in B. anthracis and drug resistant strains of S. aureus. ClpXP inhibition represents a novel therapeutic strategy to simultaneously sensitize pathogenic bacteria to host defenses and pharmaceutical antibiotics.

Leptin Promotes Glioblastoma, Johnathan Lawrence, Nicholas Cook, Richard Rovin, Robert Winn

Johnathan Lawrence

The hormone leptin has a variety of functions. Originally known for its role in satiety and weight loss, leptin more recently has been shown to augment tumor growth in a variety of cancers. Within gliomas, there is a correlation between tumor grade and tumor expression of leptin and its receptor. This suggests that autocrine signaling within the tumor microenvironment may promote the growth of high-grade gliomas. Leptin does this through stimulation of cellular pathways that are also advantageous for tumor growth and recurrence: antiapoptosis, proliferation, angiogenesis, and migration. Conversely, a loss of leptin expression attenuates tumor growth. In animal models …

Effects Of Ss3-Adrenergic Receptor Agonist On Gene Expression Of Leptin In Glioblastoma, Johnathan E. Lawrence, Nicholas J. Cook, Richard A. Rovin, Robert J. Belton, Robert J. Winn

Johnathan Lawrence

In the 25 years since temozolomide entered phase I clinical trials, few new primary or adjuvant therapies have been developed for the treatment of glioblastoma multiforme (GBM) tumors. Our laboratory has been exploring novel methods for the treatment of GBMs. Recent studies indicate that the expression of the hormone leptin and its receptor (OBR) increases in gliomas and positively correlates with the malignancy of the tumor. Interestingly, ß3-adrenergic receptor agonists are known to decrease leptin expression in adipocytes but have not been examined in GBM cells. We hypothesized that b3-adrenergic agonists downregulate the expression of leptin and its receptor. In …

7 Alpha-Hydroxy-Beta-Sitosterol From Chisocheton Tomentosus Induces Apoptosis Via Dysregulation Of Cellular Bax/Bcl-2 Ratio And Cell Cycle Arrest By Downregulating Erk1/2 Activation, Noor Hasima Nagoor

Noor Hasima Nagoor

In continuation of our interest towards the elucidation of apoptotic pathways of cytotoxic phytocompounds, we have embarked upon a study on the anticancer effects of 7 alpha-hydroxy-beta-sitosterol (CT1), a rare natural phytosterol oxide isolated from Chisocheton tomentosus. CT1 was found to be cytotoxic on three different human tumor cell lines with minimal effects on normal cell controls, where cell viability levels were maintained >= 80% upon treatment. Our results showed that cell death in MCF-7 breast tumor cells was achieved through the induction of apoptosis via downregulation of the ERK1/2 signaling pathway. CT1 was also found to increase proapoptotic Bax …