Fluorescent Antibody Technique

Publication Title: 
In Vitro Cellular & Developmental Biology: Journal of the Tissue Culture Association

Human microvascular endothelial cells (HMVEC) from adult adipose tissue were cultured in MCDB 131 medium supplemented with 10% fetal bovine serum. Under these conditions, HMVEC from seven different donors had finite proliferative life spans ranging from 14.5 to 23.5 population doublings (PD), with a mean life span of 19 PD. Addition of 10% conditioned medium from activated human leukocyte cultures (BM Condimed) extended the life span of HMVEC to 31 to 41 PD, with a mean life span of 37 PD.

Author(s): 
Stein, G. H.
St Clair, J. A.
Publication Title: 
Experimental Cell Research

Lymphocytes have a finite and predictable proliferative life span in culture similar to that observed in fibroblasts. In general, the senescence of human fibroblasts is inevitable and irreversible, but their proliferative life span can be extended by certain DNA tumor virus oncogenes, such as the large T antigen of the SV40 virus. Here, we show that human T lymphocytes (HTL) can be stably transfected with SV40 large T and that expression of T antigen extended the life span of T cell cultures.

Author(s): 
Ryan, Q. C.
Goonewardene, I. M.
Murasko, D. M.
Publication Title: 
Aging

Excessive production of reactive oxygen species (ROS) contributes to progression of atherosclerosis, at least in part by causing endothelial dysfunction and inflammatory activation. The class III histone deacetylase SIRT1 has been implicated in extension of lifespan. In the vasculature,SIRT1 gain-of-function using SIRT1 overexpression or activation has been shown to improve endothelial function in mice and rats via stimulation of endothelial nitric oxide (NO) synthase (eNOS). However, the effects of SIRT1 loss-of-function on the endothelium in atherosclerosis remain to be characterized.

Author(s): 
Stein, Sokrates
Sch‰fer, Nicola
Breitenstein, Alexander
Besler, Christian
Winnik, Stephan
Lohmann, Christine
Heinrich, Kathrin
Brokopp, Chad E.
Handschin, Christoph
Landmesser, Ulf
Tanner, Felix C.
L¸scher, Thomas F.
Matter, Christian M.
Publication Title: 
Neurobiology of Aging

Epigenetic processes play a key role in the central nervous system and altered levels of 5-methylcytosine have been associated with a number of neurologic phenotypes, including Alzheimer's disease (AD). Recently, 3 additional cytosine modifications have been identified (5-hydroxymethylcytosine, 5-formylcytosine, and 5-carboxylcytosine), which are thought to be intermediate steps in the demethylation of 5-methylcytosine to unmodified cytosine. Little is known about the frequency of these modifications in the human brain during health or disease.

Author(s): 
Condliffe, Daniel
Wong, Andrew
Troakes, Claire
Proitsi, Petroula
Patel, Yogen
Chouliaras, Leonidas
Fernandes, Cathy
Cooper, Jonathan
Lovestone, Simon
Schalkwyk, Leonard
Mill, Jonathan
Lunnon, Katie
Publication Title: 
Antimicrobial Agents and Chemotherapy

Endocytosis is a fundamental process of eukaryotic cells and fulfills numerous functions, most notably, that of macromolecular nutrient uptake. Malaria parasites invade red blood cells and during their intracellular development endocytose large amounts of host cytoplasm for digestion in a specialized lysosomal compartment, the food vacuole. In the present study we have examined the effects of artemisinin and the quinoline drugs chloroquine and mefloquine on endocytosis in Plasmodium falciparum.

Author(s): 
Hoppe, Heinrich C.
van Schalkwyk, Donelly A.
Wiehart, Ursula I. M.
Meredith, Sandra A.
Egan, Joanne
Weber, Brandon W.
Publication Title: 
Cancer Research

Artesunate is a semisynthetic derivative from artemisinin, a natural product from the Chinese herb Artemisia annua L. It exerts antimalarial activity, and, additionally, artemisinin and its derivatives are active against cancer cells. The active moiety is an endoperoxide bridge. Its cleavage leads to the formation of reactive oxygen species and carbon-centered radicals. These highly reactive molecules target several proteins in Plasmodia, which is thought to result in killing of the microorganism. DNA damage induced by artemisinins has not yet been described.

Author(s): 
Li, Paul C. H.
Lam, Elena
Roos, Wynand P.
Zdzienicka, Malgorzata Z.
Kaina, Bernd
Efferth, Thomas
Publication Title: 
Asian Pacific Journal of Allergy and Immunology / Launched by the Allergy and Immunology Society of Thailand

Plasmodium falciparum, the protozoan parasite responsible for severe malaria infection, undergoes a complex life cycle. Infected red blood cells (iRBC) sequester in host cerebral microvessels, which underlies the pathology of cerebral malaria. Using immunohistochemistry on post mortem brain samples, we demonstrated positive staining for vascular endothelial growth factor (VEGF) on iRBC.

Author(s): 
Sachanonta, Navakanit
Medana, Isabelle M.
Roberts, Rachel
Jones, Margaret
Day, Nicholas P. J.
White, Nicholas J.
Ferguson, David J. P.
Turner, Gareth D. H.
Pongponratn, Emsri
Publication Title: 
Antimicrobial Agents and Chemotherapy

We previously reported that among a series of artemisinin-derived monomers and dimers, dimer diphenyl phosphate (838) was the most potent inhibitor of human cytomegalovirus (CMV) replication. Our continued investigation of a prototypic artemisinin monomer (artesunate [AS]) and dimer (838) now reveals that both compounds have specific activity against CMV but do not inhibit lytic replication of human herpesvirus 1 or 2 or Epstein-Barr virus.

Author(s): 
He, Ran
Park, Kyoungsook
Cai, Hongyi
Kapoor, Arun
Forman, Michael
Mott, Bryan
Posner, Gary H.
Arav-Boger, Ravit
Publication Title: 
Human Molecular Genetics

Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by the impairment of cognitive functions and by beta amyloid (Abeta) plaques in the cerebral cortex and the hippocampus. Our objective was to determine genes that are critical for cellular changes in AD progression, with particular emphasis on changes early in disease progression.

Author(s): 
Reddy, P. Hemachandra
McWeeney, Shannon
Park, Byung S.
Manczak, Maria
Gutala, Ramana V.
Partovi, Dara
Jung, Youngsin
Yau, Vincent
Searles, Robert
Mori, Motomi
Quinn, Joseph
Publication Title: 
Cellular Immunology

OBJECTIVE: Bromelain, a clinically used pineapple extract and natural product, has reported anti-inflammatory and immunomodulatory activities. The purpose of this study was to determine the effect of bromelain treatment in an ovalbumin (OVA)-induced murine model of allergic airway disease (AAD). METHODS: To establish AAD, mice were sensitized with intraperitoneal (i.p.) OVA/alum and challenged with daily OVA aerosols. Mice were treated i.p. with either saline, 2 or 6 mg/kg bromelain, twice daily for four consecutive days.

Author(s): 
Secor, Eric R.
Carson, William F.
Cloutier, Michelle M.
Guernsey, Linda A.
Schramm, Craig M.
Wu, Carol A.
Thrall, Roger S.

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