The roles of various plasma agents in the inactivation of bacteria have recently been investigated. However, up to now, the effect of the charged particles on the inactivation of bacteria is not well understood. In this paper, an atmospheric pressure plasma jet device, which generates a cold plasma plume carrying a peak current of 300 mA, is used to investigate the role of the charged particles in the inactivation process. It is found that the charged particles play a minor role in the inactivation process when He/N2(3%) is used as working gas. On the other hand, when He/O2(3%) is used, the charged particles are expected to play an important role in the inactivation of bacteria. Further analysis shows that the negative ions O2− might be the charged particles that are playing the role. Besides, it is found that the active species, including O, O3, and metastable state O2∗, can play a crucial role in the inactivation of the bacteria. However, the excited He∗, N2 C Π3u, and N2+ B Σ2u+ have no significant direct effect on the inactivation of bacteria. It is also concluded that heat and UV play no or minor role in the inactivation process.

RNA helicase A (RHA) belongs to the DEAH family of proteins that are capable of unwinding double-stranded RNA structure. In addition to its involvement in the metabolism of cellular RNA, RHA has been shown to stimulate RNA transcription from the long terminal repeat promoter of human immunodeficiency virus type 1 (HIV-1) as well as to enhance Rev/Rev response element-mediated gene expression. In this study, we provide evidence that RHA associates with HIV-1 Gag in an RNA-dependent manner. This interaction results in specific incorporation of RHA into HIV-1 particles. Knockdown of endogenous RHA in virus producer cells leads to generation of HIV-1 particles that are less infectious in part as a result of restricted reverse transcription. Therefore, RHA represents the first example of cellular RNA helicases that participate in HIV-1 particle production and promote viral reverse transcription. RNA helicase A (RHA)2 is a member of the DEXH-box (where X can be any amino acid) family of proteins and is also termed DHX9 (1, 2). The DEXH-box proteins, together with the DEAD-box and the Ski2 family members, are referred to as RNA helicases that are able to rearrange the structures of RNA molecules (3). RHA contains a helicase core domain consisting of seven motifs that are conserved for all RNA helicases. Within the N-terminal region of RHA there are two copies of type A double-stranded RNA binding domains. Together with an RGG-box domain located at the C terminus, double-stranded RNA binding domains regulate RNA binding as well as helicase activities of RHA (4). RHA is a nuclear protein and shuttles between the nucleus and the cytoplasm with the assistance of a bidirectional nuclear transport domain consisting of 110 amino acids at the C terminus (5). This function of the RHA nuclear transport domain is subject to regulation of arginine methylation catalyzed by PRMT1 (protein-arginine methyltransferase 1) (6). RHA is able to unwind double-stranded RNA or DNA with the energy derived from hydrolysis of NTPs by virtue of its NTPase activity (1). This property enables RHA to participate in multiple cellular processes from RNA transcription to RNA processing to RNA nuclear export (7). These multiple functions underlie the vital role of RHA in the germ line proliferation and development of Caenorhabditis elegans (8) and also account for the early embryonic lethality observed with RHA knock-out mice (9).The regulation activity of RHA in RNA transcription is implicated by its presence within the RNA polymerase II holoenzyme complex. For example, RHA has been shown to bridge the interactions between RNA polymerase II and transcription co-activators such as CREB-binding protein and BRAC1 (breast cancer-specific tumor suppressor protein 1) (10, 11). RHA also directly interacts with the p65 subunit of NF-B and stimulates NF-B-mediated reporter gene expression (12). Involvement of RHA in transcription is further indicated by the function of its homologue in Drosophila, named the maleless (MLE) gene, that increases gene expression fr...

A 14-amino-acid spacer peptide termed SP1 that separates the capsid (CA) and nucleocapsid (NC) sequences plays an active role in the assembly of human immunodeficiency virus type 1. This activity of SP1 involves its amino-terminal residues that, together with adjacent CA residues, constitute a putative ␣-helical structure spanning Gag residues from positions 359 to 371. In this study, we have determined that the virus assembly determinants within this putative ␣-helix were residues H359, K360, A361, L364, A367, and M368, of which K360 and A367 contribute to virus production to lesser extents. Notably, changes of the two basic amino acids H359 and K360 to arginine (R) impaired virus production, whereas mutations L364I and M368I, in contrast to L364A and M368A, generated near-wild-type levels of virus particles. This suggests that within Gag complexes, amino acids H359 and K360 are involved in stricter steric interactions than L364 and M368. Since L364 and M368 are separated by four residues and thus presumably located on the same side of the helical surface, they may initiate synergistic hydrophobic interactions to stabilize Gag association. Further analysis in the context of the protease-negative mutation D185H confirmed the key roles of amino acids H359, A361, L364, and M368 in virus assembly. Importantly, when transfected cells were subjected to Dounce homogenization and the cell lysates were treated by ultracentrifugation at 100,000 ؋ g, Gag molecules containing each of the H359A, A361V, L364A, and M368A mutations were found mainly in the supernatant fraction (S100), whereas approximately 80% of wild-type Gag proteins were found in the pellet. Therefore, these four mutations must have prevented Gag from generating large complexes.

Crystal structures of human immunodeficiency virus type 1 (HIV-1) capsid protein (CA) reveal that the last 11 C-terminal amino acids are disordered. This disordered region contains a glycine-rich sequence 353-GVGGP-357 (numbering refers to the initiation methionine of Gag) that is highly conserved within the Gag proteins of HIV-1, HIV-2, and simian immunodeficiency virus, which suggests the importance of this sequence in virus replication. In the present study, we demonstrate that changing any individual residue within this short region in the context of the full-length HIV-1 genome virtually abolishes production of extracellular virus particles, in either the presence or absence of viral protease activity. This severe defect in virus particle production results from impaired Gag multimerization, as well as from decreased Gag association with the cellular membranes, as demonstrated by the results of gradient sedimentation and membrane flotation centrifugation assays. These findings are further supported by the diffuse distribution pattern of the mutant Gag within the cytoplasm, as opposed to the punctate distribution of the wild-type Gag on the plasma membrane. On the basis of these results, we propose that the disordered feature of amino acid stretch 353-GVGGP-357 in the CA crystal forms may have allowed Gag to adopt multiple conformations and that such structural flexibility is needed by Gag in order to construct geometrically complex particles.

The propagation behavior of cold atmospheric pressure plasma jets has recently attracted lots of attention. In this paper, a cold He plasma jet generated by a single plasma electrode jet device is studied. The spatial-temporal resolved optical emission spectroscopy measurements are presented. It is found that the emission intensity of the He 706.5 nm line of the plasma behaves similarly both inside the syringe and in the surrounding air (plasma plume). It decreases monotonously, which is different from the emission lines, such as N2 337.1 nm line, N2+ 391.4 nm line, and O 777.3 nm line. For the discharge inside the syringe, the emission intensity of the He 706.5 nm line decays more rapidly than that of the other three spectral lines mentioned above. The N2 337.1 nm line behaves a similar time evolution with the discharge current. For the N2+ 391.4 nm line and the atomic O 777.3 nm line, both of them decay slower than that of the He 706.5 nm and the N2 337.1 nm. When the plasma plume propagates further away from the nozzle, the temporal behaviors of the emission intensities of the four lines tend to be similar gradually. Besides, it is found that, when the size of the plasma bullet appears biggest, the propagation velocity of the bullet achieves its highest value while the emission intensity of the N2+ 391.4 nm line reaches its maximum. Detailed analysis shows that the Penning effect between the metastable state Hem and the air molecules may play a significant role in the propagation of the plasma bullet in the open air.

The effects of various discharge parameters and ambient gas on the length of He atmospheric plasma jet plumes expanding into the open air are studied. It is found that the voltage and width of the discharge-sustaining pulses exert significantly stronger effects on the plume length than the pulse frequency, gas flow rate, and nozzle diameter. This result is explained through detailed analysis of the I-V characteristics of the primary and secondary discharges which reveals the major role of the integrated total charges of the primary discharge in the plasma dynamics. The length of the jet plume can be significantly increased by guiding the propagating plume into a glass tube attached to the nozzle. This increase is attributed to elimination of the diffusion of surrounding air into the plasma plume, an absence which facilitates the propagation of the ionization front. These results are important for establishing a good level of understanding of the expansion dynamics and for enabling a high degree of control of atmospheric pressure plasmas in biomedical, materials synthesis and processing, environmental and other existing and emerging industrial applications.

The Gag protein of human immunodeficiency virus type 1 contains a 14-amino-acid region, termed SP1, between the capsid and downstream nucleocapsid sequences. Although SP1 is known to be indispensable for virus production, the mechanisms involved are mostly unclear. In this study, we demonstrate that an M368A mutation within SP1 severely diminished the ability of Gag to associate with cellular membranes. Although wild-type levels of membrane binding were restored to the M368A Gag by a second-site L20K mutation within matrix, the resultant Gag mutant L20K-M368A remained defective in virus production. This latter deficit was partially consequent to the binding of L20K-M368A Gag to nonraft membranes as opposed to raft association seen for wild-type Gag. Further analysis revealed that the majority of membrane-bound M368A Gag proteins were small oligomers, indicating a multimerization defect. In support of this observation, purified recombinant Gag derivatives containing the M368A mutation formed much lower amounts of high-molecular-weight complexes that were pelletable at 21,000 ؋ g than did wild-type Gag. Based on the myristyl switch model, we propose that the M368A mutation inhibits Gag multimerization and, as a result, restricts the binding of Gag to cellular membranes.

The “plasma bullet” behavior of atmospheric pressure plasma plumes has recently attracted significant interest. In this paper, a specially designed plasma jet device is used to study this phenomenon. It is found that a helium primary plasma can propagate through the wall of a dielectric tube and keep propagating inside the dielectric tube (secondary plasma). High-speed photographs show that the primary plasma disappears before the secondary plasma starts to propagate. Both plumes propagate at a hypersonic speed. Detailed studies on the dynamics of the plasma plumes show that the local electric field induced by the charges on the surface of the dielectric tube plays an important role in the ignition of the secondary plasma. This indicates that the propagation of the plasma plumes may be attributed to the local electric field induced by the charges in the bulletlike plasma volume.