Extracellular signal-regulated kinases mediate the enhancing effects of inflammatory mediators on resurgent currents in dorsal root ganglion neurons

Previously we reported that a group of inflammatory mediators significantly enhanced resurgent currents in dorsal root ganglion neurons. To understand the underlying intracellular signaling mechanism, we investigated the effects of inhibition of extracellular signal-regulated kinases and protein kinase C on the enhancing effects of inflammatory mediators on resurgent currents in rat dorsal root ganglion neurons. We found that the extracellular signal-regulated kinases inhibitor U0126 completely prevented the enhancing effects of the inflammatory mediators on both Tetrodotoxin-sensitive and Tetrodotoxin-resistant resurgent currents in both small and medium dorsal root ganglion neurons. U0126 substantially reduced repetitive firing in small dorsal root ganglion neurons exposed to inflammatory mediators, consistent with prevention of resurgent current amplitude increases. The protein kinase C inhibitor Bisindolylmaleimide I also showed attenuating effects on resurgent currents, although to a lesser extent compared to extracellular signal-regulated kinases inhibition. These results indicate a critical role of extracellular signal-regulated kinases signaling in modulating resurgent currents and membrane excitability in dorsal root ganglion neurons treated with inflammatory mediators. It is also suggested that targeting extracellular signal-regulated kinases-resurgent currents might be a useful strategy to reduce inflammatory pain

Strong quantum fluctuation of vortices in the new superconductor MgB2MgB_2

By using transport and magnetic measurement, the upper critical field $H_{c2}(T)$ and the irreversibility line $H_{irr}(T)$ has been determined. A big separation between $H_{c2}(0)$ and $H_{irr}(0)$ has been found showing the existence of a quantum vortex liquid state induced by quantum fluctuation of vortices in the new superconductor $MgB_2$. Further investigation on the magnetic relaxation shows that both the quantum tunneling and the thermally activated flux creep weakly depends on temperature. But when the melting field $H_{irr}$ is approached, a drastic rising of the relaxation rate is observed. This may imply that the melting of the vortex matter at a finite temperature is also induced by the quantum fluctuation of vortices.Comment: 4 pages, 4 figure

Protein kinase C enhances human sodium channel hNav1.7 resurgent currents via a serine residue in the domain III-IV linker

Resurgent sodium currents likely play a role in modulating neuronal excitability. Here we studied whether protein kinase C (PKC) activation can increase resurgent currents produced by the human sodium channel hNav1.7. We found that a PKC agonist significantly enhanced hNav1.7-mediated resurgent currents and this was prevented by PKC antagonists. The enhancing effects were replicated by two phosphorylation-mimicking mutations and were prevented by a phosphorylation-deficient mutation at a conserved PKC phosphorylation site (Serine 1479). Our results suggest that PKC can increase sodium resurgent currents through phosphorylation of a conserved Serine residue located in the domain III-IV linker of sodium channels

Evidence That the P\u3csub\u3ei\u3c/sub\u3e Release Event Is the Rate-Limiting Step in the Nitrogenase Catalytic Cycle

Nitrogenase reduction of dinitrogen (N2) to ammonia (NH3) involves a sequence of events that occur upon the transient association of the reduced Fe protein containing two ATP molecules with the MoFe protein that includes electron transfer, ATP hydrolysis, Pi release, and dissociation of the oxidized, ADP-containing Fe protein from the reduced MoFe protein. Numerous kinetic studies using the nonphysiological electron donor dithionite have suggested that the rate-limiting step in this reaction cycle is the dissociation of the Fe protein from the MoFe protein. Here, we have established the rate constants for each of the key steps in the catalytic cycle using the physiological reductant flavodoxin protein in its hydroquinone state. The findings indicate that with this reductant, the rate-limiting step in the reaction cycle is not protein–protein dissociation or reduction of the oxidized Fe protein, but rather events associated with the Pi release step. Further, it is demonstrated that (i) Fe protein transfers only one electron to MoFe protein in each Fe protein cycle coupled with hydrolysis of two ATP molecules, (ii) the oxidized Fe protein is not reduced when bound to MoFe protein, and (iii) the Fe protein interacts with flavodoxin using the same binding interface that is used with the MoFe protein. These findings allow a revision of the rate-limiting step in the nitrogenase Fe protein cycle

Tetrodotoxin-resistant sodium channels in sensory neurons generate slow resurgent currents that are enhanced by inflammatory mediators

Resurgent sodium currents contribute to the regeneration of action potentials and enhanced neuronal excitability. Tetrodotoxin-sensitive (TTX-S) resurgent currents have been described in many different neuron populations, including cerebellar and dorsal root ganglia (DRG) neurons. In most cases, sodium channel Nav1.6 is the major contributor to these TTX-S resurgent currents. Here we report a novel TTX-resistant (TTX-R) resurgent current recorded from rat DRG neurons. The TTX-R resurgent currents are similar to classic TTX-S resurgent currents in many respects, but not all. As with TTX-S resurgent currents, they are activated by membrane repolarization, inhibited by lidocaine, and enhanced by a peptide-mimetic of the β4 sodium channel subunit intracellular domain. However, the TTX-R resurgent currents exhibit much slower kinetics, occur at more depolarized voltages, and are sensitive to the Nav1.8 blocker A803467. Moreover, coimmunoprecipitation experiments from rat DRG lysates indicate the endogenous sodium channel β4 subunits associate with Nav1.8 in DRG neurons. These results suggest that slow TTX-R resurgent currents in DRG neurons are mediated by Nav1.8 and are generated by the same mechanism underlying TTX-S resurgent currents. We also show that both TTX-S and TTX-R resurgent currents in DRG neurons are enhanced by inflammatory mediators. Furthermore, the β4 peptide increased excitability of small DRG neurons in the presence of TTX. We propose that these slow TTX-R resurgent currents contribute to the membrane excitability of nociceptive DRG neurons under normal conditions and that enhancement of both types of resurgent currents by inflammatory mediators could contribute to sensory neuronal hyperexcitability associated with inflammatory pain

Sequential and Differential Interaction of Assembly Factors During Nitrogenase MoFe Protein Maturation

Nitrogenases reduce atmospheric nitrogen, yielding the basic inorganic molecule ammonia. The nitrogenase MoFe protein contains two cofactors, a [7Fe-9S-Mo-C-homocitrate] active-site species, designated FeMo-cofactor, and a [8Fe-7S] electron-transfer mediator called P-cluster. Both cofactors are essential for molybdenum-dependent nitrogenase catalysis in the nitrogen-fixing bacterium Azotobacter vinelandii. We show here that three proteins, NafH, NifW, and NifZ, copurify with MoFe protein produced by an A. vinelandii strain deficient in both FeMo-cofactor formation and P-cluster maturation. In contrast, two different proteins, NifY and NafY, copurified with MoFe protein deficient only in FeMo-cofactor formation. We refer to proteins associated with immature MoFe protein in the following as “assembly factors.” Copurifications of such assembly factors with MoFe protein produced in different genetic backgrounds revealed their sequential and differential interactions with MoFe protein during the maturation process. We found that these interactions occur in the order NafH, NifW, NifZ, and NafY/NifY. Interactions of NafH, NifW, and NifZ with immature forms of MoFe protein preceded completion of P-cluster maturation, whereas interaction of NafY/NifY preceded FeMo-cofactor insertion. Because each assembly factor could independently bind an immature form of MoFe protein, we propose that subpopulations of MoFe protein–assembly factor complexes represent MoFe protein captured at different stages of a sequential maturation process. This suggestion was supported by separate isolation of three such complexes, MoFe protein–NafY, MoFe protein–NifY, and MoFe protein–NifW. We conclude that factors involved in MoFe protein maturation sequentially bind and dissociate in a dynamic process involving several MoFe protein conformational states

A New Cationic Liposome DNA Complex Enhances the Efficiency of Arterial Gene Transfer In Vivo

Overview summary GAP-DLRIE/DOPE, a new cationic liposome preparation, is an efficient liposomal vector that increases gene expression in arteries compared to naked DNA or previously described cationic DNA–liposome complexes by more than 15-fold. Although less efficient than adenoviral gene transfer, these levels of gene expression represent a significant improvement in liposome transfection in vivo and approach levels observed with clinically acceptable doses of adenoviral vectors. The improvement in gene expression, together with the relative safety associated with liposomal gene transfer, suggests that such nonviral vectors may be appropriate for human gene therapy protocols which utilize catheter-based gene delivery.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/63105/1/hum.1996.7.15-1803.pd

Genomic diversity among Basmati rice (Oryza sativa L) mutants obtained through 60Co gamma radiations using AFLP markers

Mutation breeding can be considered successful in obtaining new cultivars and broadening the genetic base of rice crop. In order to obtain new varieties of rice with improved agronomic and grain characteristics, gamma radiation (60Co) has been used to generate novel mutants of the Basmati rice. In this study rice cultivars; Basmati-370 and Basmati-Pak, were exposed to different doses of gamma radiations and stable mutants along with parents were studied for genomic diversity on the basis of molecular marker (AFLP). Morphological data showed that mutants of Basmati-370 performed well for yield and yield components and grain physical parameters whereas, the mutant EL-30-2-1 has extra long rain trait as compared to the parent (Basmati-Pak). The genetic variations determined through AFLP revealed a total of 282 scorable bands, out of which 108 (37.81%) were polymorphic. The number of fragments produced by various primers combinations ranged from 11 - 26 with an average of 17.63fragments per primer combination. Maximum 26 bands were amplified with P-AAG/M-CAG primer combination and minimum one band was amplified with P-ATG/M-CTA primer combination. Two groups of genotypes were detected; group-A had DM-1-30-3-99, DM-1-30-34-99 and EF-1-20-52-04 mutants along with parent Basmati-370, whereas the group-B contained EL-30-2-1 and parent Basmati-Pak. The results of AFLP analysis indicated that the rate of polymorphism was 4.43% (DM-1-30-3-99), 4.25% (DM-1-30-34-99) and 6.38% (EF-1-20-52-04) among the genomes of mutants and parent Basmati-370, respectively, whereas polymorphism rate was 5.32% between genome of EL-30-2-1 and Basmati-Pak. The study further confirmed that the use of gamma radiations is an effective approach for creating new rice germplasm

Pair production of the T-odd leptons at the LHC

The T-odd leptons predicted by the littlest $Higgs$ model with T-parity can be pair produced via the subprocesses $gg\to \ell^{+}_{H}\ell^{-}_{H}$, $q\bar{q}\to \ell^{+}_{H}\ell^{-}_{H}$, $\gamma\gamma\to \ell^{+}_{H}\ell^{-}_{H}$ and $VV \to \ell^{+}_{H}\ell^{-}_{H}$ ($V$=$W$ or $Z$) at the $CERN$ Large Hadron Collider $(LHC)$. We estimate the hadronic production cross sections for all of these processes and give a simply phenomenology analysis. We find that the cross sections for most of the above processes are very small. However, the value of the cross section for the $Drell-Yan$ process $q\bar{q}\to \ell^{+}_{H}\ell^{-}_{H}$ can reach $270fb$.Comment: 12 pages, 2 figure