We report here the expression and properties of the
intermediate-conductance Ca2+-activated K+ (IKCa)
channel in the GL-15 human glioblastoma cell line.
Macroscopic IKCa currents on GL-15 cells displayed
a mean amplitude of 7.2±0.8 pA/pF at 0 mV, at day 1
after plating. The current was inhibited by clotrimazole
(CTL, IC50=257 nM), TRAM-34 (IC50=55 nM), and
charybdotoxin (CTX, IC50=10.3 nM). RT-PCR analysis
demonstrated the expression of mRNA encoding
the IKCa channel in GL-15 cells. Unitary currents recorded
using the inside-out configuration had a conductance
of 25 pS, a KD for Ca2+ of 188 nM at -100
mV, and no voltage dependence. We tested whether
the IKCa channel expression in GL-15 cells could be
the result of an increased ERK activity. Inhibition of
the ERK pathway with the MEK antagonist PD98059
(25 µM, for 5 days) virtually suppressed the IKCa current
in GL-15 cells. PD98059 treatment also increased
the length of cellular processes and up-regulated the
astrocytic differentiative marker GFAP. A significant
reduction of the IKCa current amplitude was also observed
with time in culture, with mean currents of
7.17±0.75 pA/pF at 1-2 days, and 3.11±1.35 pA/pF
at 5-6 days after plating. This time-dependent downregulation
of the IKCa current was not accompanied
by changes in the ERK activity, as assessed by
immunoblot analysis. Semiquantitative RT-PCR analysis
demonstrated a ~35% reduction of the IKCa channel
mRNA resulting from ERK inhibition and a ~50%
reduction with time in culture.
It has been demonstrated that in hepatocyte nuclei the chromatin phospholipid fraction is localized near the RNA in decondensed chromatin. The aim of the present study was to see if there is any linkage between phospholipids and other nuclear components. Isolated hepatocyte nuclei and nuclear membranes were treated with deoxyribonuclease and ribonuclease. No loss of phospholipids was observed after DNA digestion, whereas 48% was lost following enzymatic RNA removal. This loss of phospholipids, localized either near the membrane or inside the nucleus, was not homogeneous for all phospholipids: phosphatidylserine and sphingomyelin being the most affected. It can be concluded that 48% of nuclear phospholipids, in particular sphingomyelin, is lost with RNA removal. This result is discussed in view of a possible role of phospholipids in DNA synthesis and RNA transcription.
model innovation, Diversity, Inter-organizational networks, ProactivenessThe relevance of business model innovation (BMI) for strategic renewal and competitive advantage has been increasingly recognized by both strategy scholars and managers. In this paper, we explore how inter-organizational networks and firms' proactiveness influence BMI. By examining Dutch design companies, we show that having a network of companies with different sizes has a positive effect on BMI. We also observe a positive effect of ties change. Finally, we show the importance of firms' proactiveness. Through the discussion of network elements and proactiveness as antecedents of BMI, we contribute to the business model and social network literature.
How can organizations capitalize on core stigma through spectacles? In this paper, we adopt a performativity perspective to address this question. Specifically, we analyze how an assemblage of different actors within and around an organization contributes to the spectacularization of stigma and thus brings a new reality into being. In this new reality, rather than stigma being concealed, it is normalized, and the organization capitalizes on it to enhance success. We focus on RuPaul's Drag Race as a stigmatized organization that has built its success through the active spectacularization of its core stigma. In our analyses, we identify three mechanisms (i.e., reiteration of transgressions, awakening of social consciousness, and language modelling) the organization strategically uses to spectacularize the transgressions associated with the social deviance of drag queens. Through these mechanisms, a new reality around the stigma of drag queens is constructed and stigma is normalized. Overall, we contribute to a better understanding of how organizations can capitalize on their core stigma through spectacularization; we also explore the role of audiences in co-creating organizational realities around stigma.
Myogenesis is mainly sustained by a subpopulation of myogenic cells known as satellite cells (SC). In this paper we studied alpha-smooth muscle (alphaSMA) and alpha-sarcomeric muscle (alphaSRA) actin isoform expression in cultures of human satellite cells (HSC) isolated from skeletal muscle biopsies from a 5-day-old newborn, a 34-year-old young adult and a 71-year-old donor. Myogenicity of cultures was assessed using immunocytochemical detection of desmin and myosin heavy chain. Time-course expression of alphaSRA and alphaSMA were studied with both immunocytochemistry and western blotting procedures. Although alphaSMA was never detected in whole skeletal muscle, both alphaSMA and alphaSRA were detected in proliferating and differentiating HSC derived from donors of all examined ages. The expression level experiments showed that alphaSRA was gradually up-regulated during HSC differentiation, but no significant differences were observed between newborn, young, and elderly HSC cultures. Our data demonstrated that HSC, isolated from subjects of different ages, re-expressed alphaSMA, but its levels and expression pattern varied considerably in the newborn with respect to the young adult and elderly donors. These results are discussed in relation to the myogenic differentiation capability of HSC during human muscle senescence.
Chloramphenicol resistance (Cmlr) of Streptomyces coelicolor A3(2) behaves like a transposon locus, not being localisable in any region of the map and yet being transferable in crosses at a rate comparable to that of chromosomal markers. It can also be transposed onto a plasmid (SCP1) and back to the chromosome. Some traits, such as arginino-succinate synthase production (ArgG), aerial mycelium formation (AmyA), resistance to tetracycline and to rifamycin C appear to be joined to Cml in three processes: co-mutation, i.e. simultaneous loss, post-mutation, i.e. spontaneous loss at high frequency in subclones from Cmls strains, co-transfer, i.e. joint transfer with the cml locus in crosses or during infection by the aggregate SCP1::SCTn1 plasmid. All these processes have been consistently observed with special attention to the argG locus.
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