621 research outputs found
Ellen Carolyn Larkins, circa 1950
No full textWritten on verso: Ellen Carolyn Larkins.The Atlanta University Center Robert W. Woodruff Library acknowledges the generosity of the Digital Public Library of America for supporting in part the digitization of this collection as part of the Black Women's Suffrage Digital Collection, a project made possible through funding from Pivotal Ventures, A Melinda Gates Company
Construction of a Retroviral Vector for Production of Transgenic Cells Expressing Feline Immunodeficiency Virus (FIV) Viral Protein
No full textProgram year: 1994/1995Digitized from print original stored in HDRRetroviral vectors are being used as vehicles for transporting foreign genes into mammalian cells for the purpose of human gene therapy to treat viral disease like human immunodeficiency virus (HIV). In efforts to develop a preventative vaccine against a lentiviral infection, research is centering on developing a protective vaccine that will identify antigens which induce protective cell mediated immunity, especially cytotoxic T lymphocytes (CTL). Retroviral vectors provide the required endogenous expression of a retrovirus antigen for identificatoin of CTL responses against that individual antigen. Feline immunodeficiency virus (FIV) is an animal lentivirus that shares common clinical properties with HIV. The commonalities shared between HIV and FIV allows the use of FIV as an animal model for developing a protective vaccine that will identify antigens which induce protective cell mediated immunity or CTL responses. The FIV gag nucleocapsid (p10) protein will be cloned into the retroviral vector pLXSN for production of transgenic cells expressing the p10 protein for study of specific T-cell responses to this FIV antigen.
PCR will be used to amplify the nucleocapsid from the pUC119 plasmid containing the FIV genome. The amplified p10 PCR product will be EcoRI and BamHI digested for directional insertion into the EcoRI and BamHI digested retroviral vector pLXSN. Transformation of competent E. coli cells will provide colonies containing the recombinant retroviral vector DNA. Analysis of the recombinant retroviral vector DNA will be carried out by PCR.
Amplification by PCR of the nucleocapsid protein provided the p10 DNA sequence. Subsequent EcoRI and BamHI digestion revealed a natural EcoRI site at base pair 1871, therefore cutting off 160 bp from the p10 sequence. The upstream primer was reconstructed with an XhoI cut site for reamplification of the p10 protein from the pUC119 plasmid and will be inserted into the retroviral vector by XhoI and BamHI digestion. The 204 bp sequence generated from the EcoRI and BamHI digestion was inserted into the retroviral vector and awaits analysis of the idetification of recombinant retroviral vector colonies
Looking Inside the Black Box of "Attendance at Services": New Measures for Exploring an Old Dimension in Religion and Health Research
No full textResearch in religion and health has spurred new interest in measuring religiousness. Measurement efforts have focused on subjective facets of religiousness such as spirituality and beliefs, and less attention has been paid to congregate aspects, beyond the single item measuring attendance at services. We evaluate some new measures for religious experiences occurring during congregational worship services. Respondents (N=576) were religiously-diverse community dwelling adults interviewed prior to cardiac surgery. Exploratory factor analysis of the new items with a pool of standard items yielded a readily interpretable solution, involving seven correlated but distinct factors and one index variable, with high levels of internal consistency. We describe religious affiliation and demographic differences in these measures. Attendance at religious services provides multifaceted physical, emotional, social, and spiritual experiences that may promote physical health through multiple pathways.This research was supported by grants from the National Institute on Aging (AG15160 and AG16750, Richard Contrada, PI).Published 2009 in International Journal for the Psychology of Religion at http://www.informaworld.com/smpp/content~db=all~content=a907482564~frm=titlelin
Molecular characterization of oct4-expressing yolk sac endoderm stem cell lines.
No full textThe extraembryonic endoderm (XEN) defines the yolk sac, a set of membranes that
provide essential support for mammalian embryos. Recently, the committed XENprecursor
was identified in the embryonic Inner Cell Mass (ICM) as a group of cells that
intermingles with the closely related, anatomically indistinguishable epiblast (EPI)-
precursor that gives rise to the fetus. In vitro, the EPI-precursor is represented by the
well-known embryonic stem (ES) cell lines, but cell lines representing the XENprecursor
are not known. Furthermore, since the XEN-precursor cells were discovered
only very recently, the unexpected fact that they express the key pluripotency marker
Oct4 has not been explored. Recently, however, our laboratory has isolated rat XEN cell
lines that express Oct4, leading to the following two questions: (i) Do these new XEN
cell lines represent XEN-precursor cells? (ii) Is their Oct4 expression regulated similarly
as previously known from ES cells? These two questions are addressed here by lineage
marker and reporter gene analyses. Whole culture analyses showed that rat XEN cell lines expressed markers of all
XEN stages including XEN-precursor, primitive endoderm (PrE) and/or visceral
endoderm (VE), and parietal endoderm (PE) but trophoectoderm and EPI-precursor
markers were missing. In line with this, immunocytochemistry demonstrated
heterogeneity and directly visualized the XEN-precursor, PrE/VE, and PE
subpopulations. Low-density plating and time-dependent immunocytochemistry on
resulting colonies strongly suggested that XEN-precursor cells generate the other XEN
stages. Moreover, by analyzing single-cell derived clones, it was shown that culture
heterogeneity results from the self-renewal and differentiation of a single cell. Reporter
gene analyses using the 5��� regulatory region of the mouse Oct4 gene revealed that a
DNA fragment containing the previously described distal enhancer drove reporter gene
expression only in ES cells whereas inclusion of an upstream fragment led to high
expression in both mouse ES and rat XEN cells.
In conclusion, our rat XEN cell lines contain XEN-precursor cells that differentiate
extensively, providing for the first time an in vitro model that mimics the natural process
of early XEN differentiation. In addition, they regulate Oct4 gene transcription
differently than ES cells suggesting heterogeneous Oct4 regulation within the
mammalian ICM
First Characterization of Avian Memory T Lymphocyte Responses to Avian Influenza Virus Proteins
No full textAlthough wild birds are natural hosts of avian influenza viruses (AIVs), these
viruses can be highly contagious to poultry and a zoonotic threat to humans. The
propensity of AIV for genetic variation through genetic shift and drift allows virus to
evade vaccine mediated humoral immunity. An alternative approach to current vaccine
development is induction of CD8+ T cells which responds to more conserved epitopes
than humoral immunity and targets a broader spectrum of viruses. Since the memory
CD8+ T lymphocyte responses in chickens to individual AIV proteins have not been
defined, the modulation of responses of the memory CD8+ T lymphocytes to H5N9 AIV
hemagglutinin (HA) and nucleocapsid (NP) proteins over a time course were evaluated.
CD8+ T lymphocyte responses induced by intramuscular inoculation of chickens with
AIV HA and NP expressing cDNA plasmids or a non-replicating human adenovirus
vector were identified through ex vivo stimulation with virus infected, major
histocompatibility complex (MHC) matched antigen presenting cells (APCs). The IFN?
production by activated lymphocytes was evaluated by macrophage production of nitric
oxide and ELISA. MHC-I restricted memory T lymphocyte responses were determined at 10 days and 3, 5, 7 and 9 weeks post-inoculation (p.i). The use of non-professional
APCs and APC driven proliferation of cells with CD8+ phenotype correlated with the
activation of CD8+ T lymphocytes. The responses specific to nucleocapsid protein (NP)
were consistently greater than those to the hemagglutinin (HA) at 5 weeks when the
CD8+ T cell responses were maximum. By 8 to 9 weeks p.i., responses to either protein
were undetectable. The T lymphocytes also responded to stimulation with a heterologous
H7N2 AIV infected APCs. Administration of booster dose induced secondary effector
cell mediated immune responses which had greater magnitudes than primary effector
responses at 10 days p.i. Flow cytometric analysis (FACS) of the T lymphocytes
demonstrated that memory CD8+ T lymphocytes of chickens can be distinguished from
naive lymphocytes by their higher expression of CD44 and CD45 surface antigens.
CD45 expression of memory lymphocytes further increases upon ex vivo stimulation
with APCs expressing AIV. This is the first characterization of avian memory responses
following both primary and secondary expression of any individual viral protein
Intracellular trafficking and plasma membrane microdomain distribution of the NSP4 enterotoxin during rotavirus infection in epithelial cells
No full textRotavirus (RV) nonstructural protein 4 (NSP4) is a multifunctional glycoprotein
that induces secretory diarrhea in mouse pups in the absence of other viral proteins. The
intracellular transport route(s) and functional mechanism(s) of NSP4 are poorly
understood; however, the recent association of the enterotoxin with cellular caveolin-1
may provide a link between NSP4 transport and function. To determine if NSP4 traffics
to a specific subset of lipid rafts at the plasma membrane (PM), we isolated caveolae
from a PM-enriched fraction with a new method that yielded endoplasmic reticulum
(ER)-free caveolae membranes with a unique membrane structure and composition.
Comparison of these caveolae with other detergent- and non-detergent-extracted
membranes revealed that each caveolae/raft fraction contained caveolae markers;
however, only our PM caveolae fraction mimicked the membrane structure and sterol
exchange dynamics of intact PM without ER or non-raft PM contaminants. When these
PM caveolae were isolated from RV-infected cells, full-length, high-mannose
glycosylated NSP4 was present. Confocal imaging showed association of NSP4 with
caveolin-1 moving from perinuclear and cytoplasmic sites toward the PM as the
infection progressed. Fluorescent imaging also indicated exposure of the NSP4 Cterminus
at the exofacial PM surface without transport of the enterotoxin through the
Golgi apparatus. Surface-specific biotinylation was used to confirm NSP4 exposure at
the surface of infected MDCK cells and to determine that the exposed protein was fulllength
and high-mannose glycosylated. This study presents an ER contaminant-free PM
caveolae isolation methodology, identifies the presence of full-length, high-mannose glycosylated NSP4 in both PM caveolae and exposed at the cell surface, and confirms
the Golgi-bypassing nature of NSP4 ER to PM transport in RV-infected MDCK cells
The effect of stress on the neuropathogenesis of Theiler's virus-induced demyelination as an animal model of multiple sclerosis
No full textStressful life events have been associated with the onset and/or exacerbation of
multiple sclerosis (MS). To investigate the effects of stress on the pathogenesis of MS,
we employed restraint stress (RST) in the Theiler��������s virus-induced demyelination
(TVID) model, an animal model for human MS. Intracerebral inoculation of
susceptible strain of mice with Theiler��������s murine encephalomyelitis virus (TMEV)
results in a biphasic disease �������� an acute encephalomyelitis and chronic demyelination.
The establishment of persistent viral infection is critical in inducing immune-mediated
demyelination during the chronic disease. The exposure of mice to RST prior to viral
infection produced a stress response as evidenced by elevated circulating corticosterone
(CORT). To further study the effect of stress on the immune response to TMEV
infection and demyelination, we first examined the cytokine and chemokine response
during the acute TMEV infection. We demonstrated that RST down-regulated the
virus-induced expression of chemokines, Ltn, IP-10, RANTES, and pro-inflammatory
cytokines, TNF, IFN and LT in both the brain and spleen during early infection.
Histologically, a decreased pattern of inflammation was observed in the brain of
restrained mice as compared to non-restrained mice. The increased viral titer was noted in the CNS of restrained mice and was correlated with the decreased production
of pro-inflammatory cytokine, suggesting an impaired immune response by RST.
Secondly, the duration of stress on the late demyelination was investigated. Repeated
and chronically stressed SJL/J mice developed an early onset of clinical signs and a
delayed onset was observed in acutely stressed mice. Both acute and chronic RST
suppressed the antibody response to TMEV and stressed displayed a higher incidence
of demyelination than non-restrained mice. Axonal loss was also noted in chronic
stressed mice. Additionally, RST caused an increased systemic viral infection in
extraneural organs during the acute infection and cardiotropic TMEV was isolated
from the heart of stressed mice. Taken together, stress resulted in profound
immunsuppression during acute infection, which may consequently increase the
incidence of demyelination. The present study may be generalized in human MS
which is potentially triggered by viral infection
Adair, Dr. Thelma, Liberia
No full textEllen Sandamini becomes a member of the board of directors at a school in Monrovia. She is honored in song by the children of the school. Thelma Adair also introduces Ellen Sandamini who gives an acceptance speech at the presidents mansion in Liberia. The audio also includes a sermon by Dr. Henry Mitchell delivered at the Bethany Baptist church (William Augustus Jones pastor) in Brooklyn, New York. In the sermon Dr. Mitchell uses the life of Dr. Martin Luther King Jr. and the relationship with his father Rev. Martin Luther King Sr. to discuss how children learn from the patterns of their parents.The Atlanta University Center Robert W. Woodruff Library acknowledges the generous support of the National Endowment for Humanities - Humanities Collections and Reference Resources Implementation Project Grant in supporting the processing and digitization of a number of its major archival collections as part of the project: Spreading the Word: Expanding Access to African American Religious Archival Collections at the Atlanta University Center Robert W. Woodruff Library.</em
Differential cytokine mRNA expression induced by binding of virulent and avirulent molecularly cloned equine infectious anemia viruses to equine macrophages
No full textEquine infectious anemia virus (EIAV) causes rapid development of acute disease followed by recurring episodes of fever, thrombocytopenia and viremia, termed chronic EIA. Most infected horses control the virus by immune mechanisms and become inapparent carriers. To further our understanding of the equine immune response to EIAV, a multi-probe ribonuclease protection assay (RPA) was developed to quantitate equine-specific cytokine mRNAs. Eleven template plasmids specific to ten equine cytokine genes and the ?-actin gene were generated, from which radiolabeled anti-sense RNA probes were produced. The RPA simultaneously quantitated mRNA levels of interleukin (IL)-1���, IL-1���, IL-6, IL-8, IL-10, IL-12 p35, IL-12 p40, interferon (IFN)-���, transforming growth factor (TGF)-1��� and tumor necrosis factor (TNF)-��� in equine peripheral blood mononuclear cells and equine monocyte-derived macrophages (EMDM). The assay detected as few as 5���105 RNA molecules and displayed coefficients of variation of 0.03-0.08 when normalized to ���-actin expression. Using this RPA, cytokine expression in EMDM infected with 2 molecularly cloned viruses (EIAV17 and EIAV19) was determined. EIAV17 varies from EIAV19 only in env, rev and LTR and causes fatal disease in Shetland ponies. When added to EMDM cultures, virulent EIAV17 stimulated expression of IL-1���, IL-1���, IL-6, IL-10 and TNF-���. These cytokine mRNAs were significantly elevated by 0.5 to 1 hr post infection (hpi) and returned to basal levels by 12 to 24 hpi, indicating modulation by early event(s), such as receptor binding. In contrast to EIAV17, EIAV19 is avirulent in vivo and failed to induce any of the tested cytokines in EMDM. These data show a direct correlation between the virulence of the EIAV clone and the induction of cytokines. The cytokines stimulated by EIAV17 may contribute to EIA-associated symptoms, enhance viral replication in the host, and regulate the host immune response. To determine whether cytokine induction requires EIAV17 replication, EMDM cultures were exposed to UV-inactivated EIAV17 and cytokine induction was monitored. UV-inactivation did not block cytokine induction by EIAV17, suggesting dispensability of viral replication. Given that EIAV17 induces cytokines in a rapid and replication-independent manner, the activation of cytokine expression is likely mediated by binding of EIAV17 to equine macrophage receptor(s)
Liftings for noncomplete probability spaces
No full textThe current state of knowledge concerning liftings for noncomplete probability spaces is discussed. This is a somewhat expanded version of the author's talk given at the 1991 Summer Conference on General Topology and Applications in Honor of Mary Ellen Rudin and Her Work.PT: S; CR: BURKE MR, IN PRESS P AM MATH S BURKE MR, 1991, ISRAEL J MATH, V73, P33 BURKE MR, 1992, ISRAEL J MATH, V79, P289 CARLSON T, THEOREM LIFTING CHRISTENSEN JPR, 1974, TOPOLOGY BOREL STRUC FREMLIN DH, 1989, HDB BOOLEAN ALGEBRAS, P877 INOESCUTULCEA A, 1966, 5TH P BERK S MATH ST, V2 IONESCUTULCEA A, 1967, CONTRIBUTIONS PROB 1, P63 IONESCUTULCEA A, 1969, TOPICS THEORY LIFTIN JECH TJ, 1978, SET THEORY JOHNSON RA, 1980, P AM MATH SOC, V80, P234 JUST W, IN PRESS T AM MATH S KUPKA J, 1983, INDIANA U MATH J, V32, P717 LOSERT V, 1983, LNM, V1080, P95 MAHARAM D, 1958, P AM MATH SOC, V9, P987 SHELAH S, 1983, ISRAEL J MATH, V45, P90 TALAGRAND M, 1982, P AM MATH SOC, V84, P379 VONNEUMANN J, 1931, CRELLES J MATH, V165, P109; NR: 18; TC: 0; J9: ANN N Y ACAD SCI; PG: 4; GA: BZ86BSource type: Electronic(1
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