919 research outputs found
Separation of v-Src-induced mitogenesis and morphological transformation by inhibition of AP-1
v-Src activity results in both morphological transformation and reentry of quiescent chick embryo fibroblasts (CEF) into cell cycle. We have previously used temperature-sensitive v-Src mutants to show that enhanced activity of cellular AP-1 in the first few hours after activation of v-Src invariably precedes the biological consequences. Here we have investigated whether the early activation of AP-1 is essential for any or all of the v-Src responses by using a mutant c-Fos that comprises the leucine zipper and a disrupted basic region. Expression of the c-Fos mutant partially reduced cellular AP-1 activity in exponentially growing cells. However, in CEF that had been made quiescent by serum deprivation, v-Src-induced stimulation of AP-1 DNA binding activity was substantially reduced. In addition, quiescent CEF stably transfected with this mutant show an impaired mitogenic response to v-Src, indicating that the AP-1 stimulation is a necessary prerequisite for cell-cycle reentry. The ability of v-Src to morphologically transform quiescent CEF was not impaired by the inhibition of AP-1 stimulation, indicating that the mitogenic and morphological consequences of v-Src have distinguishable biochemical mediators. Focal adhesion kinase, a recently identified determinant of cell morphology, undergoes a gel mobility shift, characteristic of its hyperphosphorylated state, in response to v-Src activation in cells expressing the inhibitory AP-1 protein. This provides further evidence that the pathways that regulate morphological transformation are independent of AP-1
Hydrodynamic interactions in colloidal ferrofluids: A lattice Boltzmann study
We use lattice Boltzmann simulations, in conjunction with Ewald summation
methods, to investigate the role of hydrodynamic interactions in colloidal
suspensions of dipolar particles, such as ferrofluids. Our work addresses
volume fractions of up to 0.20 and dimensionless dipolar interaction
parameters of up to 8. We compare quantitatively with Brownian
dynamics simulations, in which many-body hydrodynamic interactions are absent.
Monte Carlo data are also used to check the accuracy of static properties
measured with the lattice Boltzmann technique. At equilibrium, hydrodynamic
interactions slow down both the long-time and the short-time decays of the
intermediate scattering function , for wavevectors close to the peak of
the static structure factor , by a factor of roughly two. The long-time
slowing is diminished at high interaction strengths whereas the short-time
slowing (quantified via the hydrodynamic factor ) is less affected by the
dipolar interactions, despite their strong effect on the pair distribution
function arising from cluster formation. Cluster formation is also studied in
transient data following a quench from ; hydrodynamic interactions
slow the formation rate, again by a factor of roughly two
The amino acid sequence of Neurospora NADP-specific glutamate dehydrogenase. Peptic and chymotryptic peptides and the complete sequence
The amino acid sequence of Neurospora NADP-specific glutamate dehydrogenase. The tryptic peptides
One-Step Purification of Recombinant Human Amelogenin and Use of Amelogenin as a Fusion Partner
Amelogenin is an extracellular protein first identified as a matrix component important for formation of dental enamel during tooth development. Lately, amelogenin has also been found to have positive effects on clinical important areas, such as treatment of periodontal defects, wound healing, and bone regeneration. Here we present a simple method for purification of recombinant human amelogenin expressed in Escherichia coli, based on the solubility properties of amelogenin. The method combines cell lysis with recovery/purification of the protein and generates a >95% pure amelogenin in one step using intact harvested cells as starting material. By using amelogenin as a fusion partner we could further demonstrate that the same method also be can explored to purify other target proteins/peptides in an effective manner. For instance, a fusion between the clinically used protein PTH (parathyroid hormone) and amelogenin was successfully expressed and purified, and the amelogenin part could be removed from PTH by using a site-specific protease
The International Fertility Education Initiative: research and action to improve fertility awareness
Attachment, infidelity, and loneliness in college students involved in a romantic relationship: the role of relationship satisfaction, morbidity and prayer for partner
This study examined the mediating effects of relationship satisfaction, prayer
for a partner, and morbidity in the relationship between attachment and loneliness, infidelity
and loneliness, and psychological morbidity and loneliness, in college students
involved in a romantic relationship. Participants were students in an introductory course on
family development. This study examined only students (n = 345) who were involved in a
romantic relationship. The average age of participants was 19.46 (SD = 1.92) and 25 %
were males. Short-form UCLA Loneliness Scale (ULS-8), (Hays and DiMatteo in J Pers
Assess 51:69â81, doi:10.1207/s15327752jpa5101_6, 1987); Relationship Satisfaction
Scale (Funk and Rogge in J Fam Psychol 21:572â583, doi:10.1037/0893-3200.21.4.572,
2007); Rotterdam Symptom Checklist (De Haes et al. in Measuring the quality of life of
cancer patients with the Rotterdam Symptom Checklist (RSCL): a manual, Northern
Centre for Healthcare Research, Groningen, 1996); Prayer for Partner Scale, (Fincham
et al. in J Pers Soc Psychol 99:649â659, doi:10.1037/a0019628, 2010); Infidelity Scale,
(Drigotas et al. in J Pers Soc Psychol 77:509â524, doi:10.1037/0022-3514.77.3.509, 1999);
and the Experiences in Close Relationship Scale-short form (Wei et al. in J Couns Psychol
52(4):602â614, doi:10.1037/0022-0167.52.4.602, 2005). Results showed that relationship
satisfaction mediated the relationship between avoidance attachment and loneliness and
between infidelity and loneliness. Physical morbidity mediated the relationship between
anxious attachment and psychological morbidity. Psychological morbidity mediated the
relationship between anxious attachment and physical morbidity. The present results
expand the literature on attachment by presenting evidence that anxious and avoidant partners experience loneliness differently. Implications for coupleâs therapy are addressed.
Future research should replicate these results with older samples and married couples.Acknowledgments This research was supported by Grant Number 90FE0022 from the United States
Department of Health and Human Services awarded to the last author
Elevated c-Src is linked to altered cellâmatrix adhesion rather than proliferation in KM12C human colorectal cancer cells
Elevated expression and/or activity of c-Src, the prototype of the Src family of protein tyrosine kinases, is associated with the development of human colon cancer. However, despite the known pleiotropic effects of these kinases in promoting (a) cell growth downstream of growth factor receptors, and (b) the dynamic regulation of integrin adhesions in fibroblast model systems, their precise role in epithelial cancer cells is unknown. Here we addressed whether elevated expression and activity of cellular Src alters cell proliferation and/or cellâmatrix adhesion in cancer cells from the Fidler model of colorectal metastasis. Although elevated Src correlates with ability to metastasise to the liver after intrasplenic injection, we found that this was not linked to enhanced growth, either in vitro or in vivo as sub-cutaneous tumours. However, elevated Src was associated with enhanced attachment to extracellular matrix. In addition, adhesion to fibronectin, was suppressed by agents that inhibited Src activity, while enforced elevation of Src in non-metastatic cells was sufficient to stimulate adhesion to fibronectin and enhanced assembly of adhesion complexes, without influencing cell growth. Thus, we conclude that one role of elevated Src in human colon cancer cells is to modulate integrin-dependent cellâmatrix attachment and formation of adhesion structures, which may, in turn, influence cell motility and integrin-dependent cellular responses
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