761 research outputs found
Intestinal Subepithelial Myofibroblasts Support in vitro and in vivo Growth of Human Small Intestinal Epithelium
The intestinal crypt-niche interaction is thought to be essential to the function, maintenance, and proliferation of progenitor stem cells found at the bases of intestinal crypts. These stem cells are constantly renewing the intestinal epithelium by sending differentiated cells from the base of the crypts of Lieberkühn to the villus tips where they slough off into the intestinal lumen. The intestinal niche consists of various cell types, extracellular matrix, and growth factors and surrounds the intestinal progenitor cells. There have recently been advances in the understanding of the interactions that regulate the behavior of the intestinal epithelium and there is great interest in methods for isolating and expanding viable intestinal epithelium. However, there is no method to maintain primary human small intestinal epithelium in culture over a prolonged period of time. Similarly no method has been published that describes isolation and support of human intestinal epithelium in an in vivo model. We describe a technique to isolate and maintain human small intestinal epithelium in vitro from surgical specimens. We also describe a novel method to maintain human intestinal epithelium subcutaneously in a mouse model for a prolonged period of time. Our methods require various growth factors and the intimate interaction between intestinal sub-epithelial myofibroblasts (ISEMFs) and the intestinal epithelial cells to support the epithelial in vitro and in vivo growth. Absence of these myofibroblasts precluded successful maintenance of epithelial cell formation and proliferation beyond just a few days, even in the presence of supportive growth factors. We believe that the methods described here can be used to explore the molecular basis of human intestinal stem cell support, maintenance, and growth
Neutron Stars and Black Holes as MACHOs
We consider the contribution of neutron stars and black holes to the
dynamical mass of galactic halos. In particular, we show that if these compact
objects were produced by an early generation of stars with initial metallicity
less than about , they can contribute at most 30--40% of the
Galactic halo mass without creating supersolar levels of enrichment. We show
that the case for halo neutron stars and black holes cannot be rejected on
metal overproduction arguments alone, due to the critical factor of the choice
of progenitor metallicity in determining the yields. We show that this scenario
satisfies observational constraints, similar to but no more severe than those
faced by halo white dwarfs. We also discuss the recent results on halo
microlensing, the presence of enriched hot gas in clusters and groups of
galaxies, and other observations. If there are halo neutron stars and black
holes, they will be detected by microlensing experiments in the future as
longer-timescale events.Comment: 23 pages (LaTex), including 2 figures; final version, to appear in
The Astrophysical Journal, May 10, 1999 issu
Metabolic model of necrotizing enterocolitis in the premature newborn gut resulting from enteric dysbiosis
Necrotizing enterocolitis (NEC) is a leading cause of premature newborn morbidity and mortality. The clinical features of NEC consistently include prematurity, gut dysbiosis and enteral inflammation, yet the pathogenesis remains obscure. Herein we combine metagenomics and targeted metabolomics, with functional in vivo and in vitro assessment, to define a novel molecular mechanism of NEC. One thousand six hundred and forty seven publicly available metagenomics datasets were analyzed (NEC = 245; healthy = 1,402) using artificial intelligence methodologies. Targeted metabolomic profiling was used to quantify the concentration of specified fecal metabolites at NEC onset (n = 8), during recovery (n = 6), and in age matched controls (n = 10). Toxicity assays of discovered metabolites were performed in vivo in mice and in vitro using human intestinal epithelial cells. Metagenomic and targeted metabolomic analyses revealed significant differences in pyruvate fermentation pathways and associated intermediates. Notably, the short chain fatty acid formate was elevated in the stool of NEC patients at disease onset (P = 0.005) dissipated during recovery (P = 0.02) and positively correlated with degree of intestinal injury (r2 = 0.86). In vitro, formate caused enterocyte cytotoxicity in human cells through necroptosis (P \u3c 0.01). In vivo, luminal formate caused significant dose and development dependent NEC-like injury in newborn mice. Enterobacter cloacae and Klebsiella pneumoniae were the most discriminatory taxa related to NEC dysbiosis and increased formate production. Together, these data suggest a novel biochemical mechanism of NEC through the microbial production of formate. Clinical efforts to prevent NEC should focus on reducing the functional consequences of newborn gut dysbiosis associated metabolic pathways
Quantum inference of states and processes
The maximum-likelihood principle unifies inference of quantum states and
processes from experimental noisy data. Particularly, a generic quantum process
may be estimated simultaneously with unknown quantum probe states provided that
measurements on probe and transformed probe states are available. Drawbacks of
various approximate treatments are considered.Comment: 7 pages, 4 figure
Propulsion in a viscoelastic fluid
Flagella beating in complex fluids are significantly influenced by
viscoelastic stresses. Relevant examples include the ciliary transport of
respiratory airway mucus and the motion of spermatozoa in the mucus-filled
female reproductive tract. We consider the simplest model of such propulsion
and transport in a complex fluid, a waving sheet of small amplitude free to
move in a polymeric fluid with a single relaxation time. We show that, compared
to self-propulsion in a Newtonian fluid occurring at a velocity U_N, the sheet
swims (or transports fluid) with velocity U / U_N = [1+De^2 (eta_s)/(eta)
]/[1+De^2], where eta_s is the viscosity of the Newtonian solvent, eta is the
zero-shear-rate viscosity of the polymeric fluid, and De is the Deborah number
for the wave motion, product of the wave frequency by the fluid relaxation
time. Similar expressions are derived for the rate of work of the sheet and the
mechanical efficiency of the motion. These results are shown to be independent
of the particular nonlinear constitutive equations chosen for the fluid, and
are valid for both waves of tangential and normal motion. The generalization to
more than one relaxation time is also provided. In stark contrast with the
Newtonian case, these calculations suggest that transport and locomotion in a
non-Newtonian fluid can be conveniently tuned without having to modify the
waving gait of the sheet but instead by passively modulating the material
properties of the liquid.Comment: 21 pages, 1 figur
Two novel human cytomegalovirus NK cell evasion functions target MICA for lysosomal degradation
NKG2D plays a major role in controlling immune responses through the regulation of natural killer (NK) cells, αβ and γδ T-cell function. This activating receptor recognizes eight distinct ligands (the MHC Class I polypeptide-related sequences (MIC) A andB, and UL16-binding proteins (ULBP)1–6) induced by cellular stress to promote recognition cells perturbed by malignant transformation or microbial infection. Studies into human cytomegalovirus (HCMV) have aided both the identification and characterization of NKG2D ligands (NKG2DLs). HCMV immediate early (IE) gene up regulates NKGDLs, and we now describe the differential activation of ULBP2 and MICA/B by IE1 and IE2 respectively. Despite activation by IE functions, HCMV effectively suppressed cell surface expression of NKGDLs through both the early and late phases of infection. The immune evasion functions UL16, UL142, and microRNA(miR)-UL112 are known to target NKG2DLs. While infection with a UL16 deletion mutant caused the expected increase in MICB and ULBP2 cell surface expression, deletion of UL142 did not have a similar impact on its target, MICA. We therefore performed a systematic screen of the viral genome to search of addition functions that targeted MICA. US18 and US20 were identified as novel NK cell evasion functions capable of acting independently to promote MICA degradation by lysosomal degradation. The most dramatic effect on MICA expression was achieved when US18 and US20 acted in concert. US18 and US20 are the first members of the US12 gene family to have been assigned a function. The US12 family has 10 members encoded sequentially through US12–US21; a genetic arrangement, which is suggestive of an ‘accordion’ expansion of an ancestral gene in response to a selective pressure. This expansion must have be an ancient event as the whole family is conserved across simian cytomegaloviruses from old world monkeys. The evolutionary benefit bestowed by the combinatorial effect of US18 and US20 on MICA may have contributed to sustaining the US12 gene family
2016 AAPP Monograph Series: African American Professors Program
The African American Professors Program (AAPP) at the University of South Carolina is honored to publish this fifteenth edition of its annual monograph series. AAPP recognizes the significance of offering scholars a venue through which to engage actively in research and to publish their refereed papers. Parallel with the publication of their manuscripts is the opportunity to gain visibility among colleagues throughout postsecondary institutions at national and international levels.
Scholars who have contributed papers for this monograph are acknowledged for embracing the value of including this responsibility within their academic milieu. Writing across disciplines adds to the intellectual diversity of these manuscripts. From neophytes to quite experienced individuals, the chapters have been researched and written in depth.
Founded in 1997 through the Department of Educational Leadership and Policies in the College of Education, AAPP was designed originally to address the under-representation of African American professors on college and university campuses. Its mission is to expand the pool of these professors in critical academic and research areas. Sponsored historically by the University of South Carolina, the W.K. Kellogg Foundation, and the South Carolina General Assembly, the program recruits doctoral students for disciplines in which African Americans currently are underrepresented among faculty in higher education.
The continuation of this monograph series is seen as responding to a window of opportunity to be sensitive to academic expectation of graduates as they pursue career placement and, at the same time, to allow for the dissemination of products of scholarship to a broader community. The importance of this series has been voiced by one of our 2002 AAPP graduates, Dr. Shundelle LaTjuan Dogan, formerly an Administrative Fellow at Harvard University, a Program Officer for the Southern Education Foundation, and a Program Officer for the Arthur M. Blank Foundation in Atlanta, Georgia. She is currently a Corporate Citizenship and Corporate Affairs Manager for IBM-International Business Machines in Atlanta, Georgia and has written the Foreword for the 2014 monograph.
Dr. Dogan wrote: One thing in particular that I want to thank you for is having the African American Professors Program scholars publish articles for the monograph. I have to admit that writing the articles seemed like extra work at the time. However, in my recent interview process, organizations have asked me for samples of my writing. Including an article from a published monograph helped to make my portfolio much more impressive. You were \u27right on target\u27 in having us do the monograph series. (AAPP 2003 Monograph, p. xi)
The African American Professors Program continues the tradition as a promoter of international scholarship in higher education evidenced through the inspiration from this group of interdisciplinary manuscripts. I hope that you will envision these published papers to serve as an invaluable contribution to your own professional development and career enhancement.
John McFadden, PhD
The Benjamin Elijah Mays Distinguished Professor Emeritus
Director, African American Professors Program
University of South Carolinahttps://scholarcommons.sc.edu/mcfadden_monographs/1003/thumbnail.jp
Anti-cancer effects and mechanism of actions of aspirin analogues in the treatment of glioma cancer
INTRODUCTION: In the past 25 years only modest advancements in glioma treatment have been made, with patient prognosis and median survival time following diagnosis only increasing from 3 to 7 months. A substantial body of clinical and preclinical evidence has suggested a role for aspirin in the treatment of cancer with multiple mechanisms of action proposed including COX 2 inhibition, down regulation of EGFR expression, and NF-κB signaling affecting Bcl-2 expression. However, with serious side effects such as stroke and gastrointestinal bleeding, aspirin analogues with improved potency and side effect profiles are being developed. METHOD: Effects on cell viability following 24 hr incubation of four aspirin derivatives (PN508, 517, 526 and 529) were compared to cisplatin, aspirin and di-aspirin in four glioma cell lines (U87 MG, SVG P12, GOS – 3, and 1321N1), using the PrestoBlue assay, establishing IC50 and examining the time course of drug effects. RESULTS: All compounds were found to decrease cell viability in a concentration and time dependant manner. Significantly, the analogue PN517 (IC50 2mM) showed approximately a twofold increase in potency when compared to aspirin (3.7mM) and cisplatin (4.3mM) in U87 cells, with similar increased potency in SVG P12 cells. Other analogues demonstrated similar potency to aspirin and cisplatin. CONCLUSION: These results support the further development and characterization of novel NSAID derivatives for the treatment of glioma
Direct Observation of the Myosin Va Recovery Stroke That Contributes to Unidirectional Stepping along Actin
Myosins are ATP-driven linear molecular motors that work as cellular force
generators, transporters, and force sensors. These functions are driven by
large-scale nucleotide-dependent conformational changes, termed
“strokes”; the “power stroke” is the force-generating
swinging of the myosin light chain–binding “neck” domain
relative to the motor domain “head” while bound to actin; the
“recovery stroke” is the necessary initial motion that primes, or
“cocks,” myosin while detached from actin. Myosin Va is a processive
dimer that steps unidirectionally along actin following a “hand over
hand” mechanism in which the trailing head detaches and steps forward
∼72 nm. Despite large rotational Brownian motion of the detached head about
a free joint adjoining the two necks, unidirectional stepping is achieved, in
part by the power stroke of the attached head that moves the joint forward.
However, the power stroke alone cannot fully account for preferential forward
site binding since the orientation and angle stability of the detached head,
which is determined by the properties of the recovery stroke, dictate actin
binding site accessibility. Here, we directly observe the recovery stroke
dynamics and fluctuations of myosin Va using a novel, transient caged
ATP-controlling system that maintains constant ATP levels through stepwise
UV-pulse sequences of varying intensity. We immobilized the neck of monomeric
myosin Va on a surface and observed real time motions of bead(s) attached
site-specifically to the head. ATP induces a transient swing of the neck to the
post-recovery stroke conformation, where it remains for ∼40 s, until ATP
hydrolysis products are released. Angle distributions indicate that the
post-recovery stroke conformation is stabilized by ≥5
kBT of energy. The high kinetic
and energetic stability of the post-recovery stroke conformation favors
preferential binding of the detached head to a forward site 72 nm away. Thus,
the recovery stroke contributes to unidirectional stepping of myosin Va
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