Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 62
Filter
Add more filters










Publication year range
1.
Phys Rev Lett ; 130(15): 156901, 2023 Apr 14.
Article in English | MEDLINE | ID: mdl-37115858

ABSTRACT

The determination of depth profiles across interfaces is of primary importance in many scientific and technological areas. Photoemission spectroscopy is in principle well suited for this purpose, yet a quantitative implementation for investigations of liquid-vapor interfaces is hindered by the lack of understanding of electron-scattering processes in liquids. Previous studies have shown, however, that core-level photoelectron angular distributions (PADs) are altered by depth-dependent elastic electron scattering and can, thus, reveal information on the depth distribution of species across the interface. Here, we explore this concept further and show that the experimental anisotropy parameter characterizing the PAD scales linearly with the average distance of atoms along the surface normal obtained by molecular dynamics simulations. This behavior can be accounted for in the low-collision-number regime. We also show that results for different atomic species can be compared on the same length scale. We demonstrate that atoms separated by about 1 Å along the surface normal can be clearly distinguished with this method, achieving excellent depth resolution.

2.
Phys Rev Lett ; 112(24): 240503, 2014 Jun 20.
Article in English | MEDLINE | ID: mdl-24996074

ABSTRACT

Optimal quantum control theory carries a huge promise for quantum technology. Its experimental application, however, is often hindered by imprecise knowledge of the input variables, the quantum system's parameters. We show how to overcome this by adaptive hybrid optimal control, using a protocol named Ad-HOC. This protocol combines open- and closed-loop optimal control by first performing a gradient search towards a near-optimal control pulse and then an experimental fidelity estimation with a gradient-free method. For typical settings in solid-state quantum information processing, adaptive hybrid optimal control enhances gate fidelities by an order of magnitude, making optimal control theory applicable and useful.

3.
Phys Rev Lett ; 111(16): 163601, 2013 Oct 18.
Article in English | MEDLINE | ID: mdl-24182265

ABSTRACT

Quantum transmission lines are central to superconducting and hybrid quantum computing. In this work we show how coupling them to a left-handed transmission line allows circuit QED to reach a new regime: multimode ultrastrong coupling. Out of the many potential applications of this novel device, we discuss the preparation of multipartite entangled states and the simulation of the spin-boson model where a quantum phase transition is reached up to finite size effects.

5.
J Virol ; 75(20): 9808-18, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11559814

ABSTRACT

Poliovirus (PV) replicates its genome in association with membranous vesicles in the cytoplasm of infected cells. To elucidate the origin and mode of formation of PV vesicles, immunofluorescence labeling with antibodies against the viral vesicle marker proteins 2B and 2BC, as well as cellular markers of the endoplasmic reticulum (ER), anterograde transport vesicles, and the Golgi complex, was performed in BT7-H cells. Optical sections obtained by confocal laser scanning microscopy were subjected to a deconvolution process to enhance resolution and signal-to-noise ratio and to allow for a three-dimensional representation of labeled membrane structures. The mode of formation of the PV vesicles was, on morphological grounds, similar to the formation of anterograde membrane traffic vesicles in uninfected cells. ER-resident membrane markers were excluded from both types of vesicles, and the COPII components Sec13 and Sec31 were both found to be colocalized on the vesicular surface, indicating the presence of a functional COPII coat. PV vesicle formation during early time points of infection did not involve the Golgi complex. The expression of PV protein 2BC or the entire P2 and P3 genomic region led to the production of vesicles carrying a COPII coat and showing the same mode of formation as vesicles produced after PV infection. These results indicate that PV vesicles are formed at the ER by the cellular COPII budding mechanism and thus are homologous to the vesicles of the anterograde membrane transport pathway.


Subject(s)
COP-Coated Vesicles/ultrastructure , Carrier Proteins/physiology , Phosphoproteins/physiology , Poliovirus/physiology , Saccharomyces cerevisiae Proteins , Virus Replication , Animals , COP-Coated Vesicles/virology , Carrier Proteins/metabolism , Carrier Proteins/ultrastructure , Cell Membrane/virology , Cells, Cultured , Endoplasmic Reticulum/virology , Haplorhini , Microscopy, Confocal , Phosphoproteins/ultrastructure , Poliovirus/ultrastructure , Vesicular Transport Proteins , Viral Nonstructural Proteins/metabolism
6.
J Virol ; 75(8): 3841-50, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11264373

ABSTRACT

HeLa cells were transfected with several plasmids that encoded all poliovirus (PV) nonstructural proteins. Viral RNAs were transcribed by T7 RNA polymerase expressed from recombinant vaccinia virus. All plasmids produced similar amounts of viral proteins that were processed identically; however, RNAs were designed either to serve as templates for replication or to contain mutations predicted to prevent RNA replication. The mutations included substitution of the entire PV 5' noncoding region (NCR) with the encephalomyocarditis virus (EMCV) internal ribosomal entry site, thereby deleting the 5'-terminal cloverleaf-like structure, or insertion of three nucleotides in the 3Dpol coding sequence. Production of viral proteins was sufficient to induce the characteristic reorganization of intracellular membranes into heterogeneous-sized vesicles, independent of RNA replication. The vesicles were stably associated with viral RNA only when RNA replication could occur. Nonreplicating RNAs localized to distinct, nonoverlapping regions in the cell, excluded from the viral protein-membrane complexes. The absence of accumulation of positive-strand RNA from both mutated RNAs in transfected cells was documented. In addition, no minus-strand RNA was produced from the EMCV chimeric template RNA in vitro. These data show that the 5'-terminal sequences of PV RNA are essential for initiation of minus-strand RNA synthesis at its 3' end.


Subject(s)
Poliovirus/genetics , Poliovirus/physiology , RNA, Viral/biosynthesis , Virus Replication , Cytoplasmic Vesicles/metabolism , Cytoplasmic Vesicles/ultrastructure , Cytoplasmic Vesicles/virology , Fluorescent Antibody Technique , Genome, Viral , HeLa Cells , Humans , In Situ Hybridization, Fluorescence , Microscopy, Electron , Plasmids/genetics , Protein Processing, Post-Translational , RNA Stability , RNA, Viral/genetics , Regulatory Sequences, Nucleic Acid/genetics , Transcription, Genetic , Transfection , Viral Nonstructural Proteins/genetics , Viral Nonstructural Proteins/metabolism
8.
Anal Chem ; 73(23): 5743-51, 2001 Dec 01.
Article in English | MEDLINE | ID: mdl-11774916

ABSTRACT

The physical and chemical properties of PPI dendrimers' interior were investigated using the fluorescent, solvatochromic probe phenol blue. In aqueous solutions of each generation studied, two discrete dye populations were clearly observed. PPI dendrimers were shown to form a tight, nonpolar association with the vast majority of available dye, within the dendrimer interior, near the core. In the steady-state fluorescence emission spectra, a microenvironment of decreasing polarity in increasingly larger-generation PPI dendrimers (up to G3) was seen for the associated probe. Each of the remaining larger-generation dendrimers provided a microenvironment of essentially equal polarity. Fluorescence anisotropy values for phenol blue in the PPI dendrimers demonstrated the dye's sensitivity to the changing molecular volumes of the dendrimer generations. Model compounds that mimicked PPI's surface groups and branching moieties were used to better define the associated dye's location. The mimics further confirmed that phenol blue was associated inside the dendrimer, where it did not interact with the dendrimer surface groups. The comparison of amine-terminated PPI and PAMAM dendrimers clearly demonstrated the effects of their structural differences and the ability of phenol blue to have sensed those differences, including the initiator core length, branching unit length, and branching unit chemical composition.

9.
J Virol ; 74(14): 6570-80, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10864671

ABSTRACT

Poliovirus (PV) infection induces the rearrangement of intracellular membranes into characteristic vesicles which assemble into an RNA replication complex. To investigate this transformation, endoplasmic reticulum (ER) membranes in HeLa cells were modified by the expression of different cellular or viral membrane-binding proteins. The membrane-binding proteins induced two types of membrane alterations, i.e., extended membrane sheets and vesicles similar to those found during a PV infection. Cells expressing membrane-binding proteins were superinfected with PV and then analyzed for virus replication, location of membranes, viral protein, and RNA by immunofluorescence and fluorescent in situ hybridization. Cultures expressing cellular or viral membrane-binding proteins, but not those expressing soluble proteins, showed a markedly reduced ability to support PV replication as a consequence of the modification of ER membranes. The altered membranes, regardless of their morphology, were not used for the formation of viral replication complexes during a subsequent PV infection. Specifically, membrane sheets were not substrates for PV-induced vesicle formation, and, surprisingly, vesicles induced by and carrying one or all of the PV replication proteins did not contribute to replication complexes formed by the superinfecting PV. The formation of replication complexes required active viral RNA replication. The extensive alterations induced by membrane-binding proteins in the ER resulted in reduced viral protein synthesis, thus affecting the number of cells supporting PV multiplication. Our data suggest that a functional replication complex is formed in cis, in a coupled process involving viral translation, membrane modification and vesicle budding, and viral RNA synthesis.


Subject(s)
Endoplasmic Reticulum/virology , Poliovirus/physiology , RNA, Viral/biosynthesis , Viral Proteins/biosynthesis , Virus Replication/physiology , Coated Vesicles/ultrastructure , Coated Vesicles/virology , Endoplasmic Reticulum/metabolism , Endoplasmic Reticulum/ultrastructure , Fluorescent Antibody Technique , HeLa Cells , Humans , In Situ Hybridization, Fluorescence , Intracellular Membranes/metabolism , Intracellular Membranes/ultrastructure , Intracellular Membranes/virology , Poliovirus/metabolism , Poliovirus/ultrastructure , Protein Binding , Protein Biosynthesis , Viral Proteins/metabolism
10.
J Virol ; 74(12): 5534-41, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10823859

ABSTRACT

Productive poliovirus infection of HeLa cells leads to the canonical cytopathic effect (CPE), whereas certain types of abortive infection result in apoptosis. To define the time course of commitment to the different types of poliovirus-induced death, inhibitors of viral replication (guanidine HCl) or translation (cycloheximide) were added at different times postinfection (p.i.). Early in the infection (during the first approximately 2 h p.i.), predominantly proapoptotic viral function was expressed, rendering the cells committed to apoptosis, which developed several hours after viral expression was arrested. In the middle of infection, concomitantly with the onset of fast generation of viral progeny, the implementation of the viral apoptotic program was abruptly interrupted. In particular, activation of an Asp-Glu-Val-Asp (DEVD)-specific caspase(s) occurring in the apoptosis-committed cells was prevented by the ongoing productive infection. Simultaneously, the cells retaining normal or nearly normal morphology became committed to CPE, which eventually developed regardless of whether or not further viral expression was allowed to proceed. The implementation of the poliovirus-induced apoptotic program was suppressed in HeLa cells overexpressing the Bcl-2 protein, indicating that the fate of poliovirus-infected cells depends on the balance of host and viral pro- and antiapoptotic factors.


Subject(s)
Apoptosis , Cytopathogenic Effect, Viral , Poliovirus/physiology , Apoptosis/drug effects , Caspase Inhibitors , Caspases/metabolism , Cell Size/drug effects , Cycloheximide/pharmacology , Cytopathogenic Effect, Viral/drug effects , DNA Fragmentation/drug effects , Enzyme Activation/drug effects , Gene Expression , Genes, bcl-2/genetics , Genes, bcl-2/physiology , Guanidine/pharmacology , HeLa Cells , Humans , Microscopy, Electron , Poliovirus/drug effects , Poliovirus/genetics , Poliovirus/pathogenicity , Protein Biosynthesis/drug effects , Time Factors , Transfection , Virus Replication/drug effects
11.
Virology ; 266(1): 157-69, 2000 Jan 05.
Article in English | MEDLINE | ID: mdl-10612670

ABSTRACT

Aside from a common gene organization shared with other picornaviruses, hepatitis A virus (HAV) is characterized by its slow-growth phenotype, the inability to shut off host macromolecular synthesis, and, in general, lack of cytopathic (cp) effects in permissive cell cultures. Nevertheless, several cp HAV strains have been isolated during the past decade. In FRhK-4 cells infected with HM175/24a, a fast-growing cp strain, increasing amounts of viral RNA, detected by fluorescence in situ hybridization, indicated viral RNA replication. An ultrastructural analysis of the infected cells revealed a tubular-vesicular network in close proximity to the rough endoplasmic reticulum. Infection of the same cell type with a cell culture adapted (cc) strain, HM175/P35, divulged membrane alterations indistinguishable from the network induced by the cp strain. The overall appearance of the tubular-vesicular network resembles membrane alterations induced by other picornaviruses. However, the shape of the vesicle-like structures is rather oblong and tubular and, thus, seems to be specific for HAV. By electron microscopic immunocytochemistry (IEM), proteins 2B and 2C were found exclusively on the membranes of the network. Proteins expressed from the open reading frame of the cc HAV variant or 2B proteins originating from HM175 cp, cc, or the wt strain expressed in the absence of other HAV proteins induced membrane alterations resembling those seen in HAV-infected cells. The induction of similar structures suggests that protein 2B is involved in the rearrangement of cellular membranes. In all cases, IEM demonstrated that the 2B protein was closely associated with altered membranes. The extent of membrane changes did not seem to increase for both the cp strain and the cc strain during the infectious cycle. Late in the infection and shortly before the culture died off, a large number of cells infected with HM175/24a showed typical signs of apoptosis, whereas the cc strain did not induce cell killing in the same type of cells. Therefore, we conclude that cell death in HM175/24a-infected cells is induced by apoptosis rather than by cytopathology.


Subject(s)
Apoptosis , Cytopathogenic Effect, Viral , Hepatovirus/physiology , Hepatovirus/pathogenicity , Intracellular Membranes/ultrastructure , Cell Line , HeLa Cells , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Intracellular Membranes/metabolism , Microscopy, Electron , Microscopy, Fluorescence , Viral Nonstructural Proteins/metabolism
12.
Histochem Cell Biol ; 111(4): 319-24, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10219632

ABSTRACT

Detection of target RNA by in situ hybridization (ISH) in the classic and confocal fluorescence microscope was performed using strand-specific single-stranded RNA probes labeled directly with the fluorochromes fluorescein isothiocyanate or Texas red. The probes, produced by in vitro transcription from PCR-generated templates with T7 RNA polymerase and fluorochromized UTP, gave ISH signals directly visible by fluorescence microscopy without the use of any immunological detection step. In avoiding antibodies, it was possible to strongly increase the sensitivity of the ISH since antibodies may contain RNase which can reduce hybridization signals considerably, even beyond the detection limit. Fluorescent RNA probes thus allowed for the detection of low numbers of target molecules per cell, such as minus strand intermediates in picornavirus RNA replication. Using appropriate denaturing conditions, the targets could be visualized in a double-stranded configuration as well as in the presence of a 100-fold excess of complementary RNA. Furthermore, double ISH for the simultaneous detection of two different RNA species, such as plus and minus strand RNA of poliovirus, or of different regions of the viral genomic RNA was possible with appropriate fluorescent strand-specific probes labeled with different fluorochromes. Combination of ISH and immunofluorescence was found feasible if RNA was present in relatively large amounts. In addition to the investigation of virus replication, possible applications of fluorochromized RNA probes might include antisense RNA detection as well as plant virus resistance and gene silencing.


Subject(s)
Fluorescein-5-isothiocyanate , Fluorescent Dyes , RNA Probes , Xanthenes , Humans , In Situ Hybridization, Fluorescence/methods , Tumor Cells, Cultured
13.
Gene ; 228(1-2): 85-91, 1999 Mar 04.
Article in English | MEDLINE | ID: mdl-10072761

ABSTRACT

AU-rich elements function as instability elements which direct rapid mRNA degradation. AUH protein exhibits an AU-specific RNA-binding property and an intrinsic enoyl-CoA hydratase activity and may therefore function to link mRNA decay to metabolic processes (. Proc. Natl. Acad. Sci. USA 92, 2051-2055). The sequence encoding the murine protein, muAUH, was established by cloning, and the corresponding polypeptide predicted to have a molecular mass of 37kDa. As shown for the human protein, muAUH is expressed in a 32kDa form and there is 94% homology between the two species. Recombinant muAUH was shown to be an RNA-binding enoyl-CoA hydratase. All murine cells studied contained a single AUH transcript of approx. 1.7kb and an investigation of tissue-specific expression revealed highest levels in kidney, skeletal muscle, heart, liver and spleen. It was further determined, using immunoelectron microscopy, that AUH is located in the mitochondria of mouse cells.


Subject(s)
Enoyl-CoA Hydratase/genetics , Mitochondria/enzymology , RNA-Binding Proteins/genetics , Amino Acid Sequence , Animals , Blotting, Northern , Blotting, Western , DNA, Complementary/chemistry , DNA, Complementary/genetics , DNA, Complementary/isolation & purification , Enoyl-CoA Hydratase/analysis , Fluorescent Antibody Technique , Gene Expression Regulation, Enzymologic , Humans , Kidney/enzymology , Male , Mast Cells/cytology , Mast Cells/enzymology , Mast Cells/ultrastructure , Mice , Mice, Inbred BALB C , Mitochondria/ultrastructure , Molecular Sequence Data , RNA, Messenger/genetics , RNA, Messenger/metabolism , RNA-Binding Proteins/analysis , Sequence Analysis, DNA , Sequence Homology, Amino Acid , Tissue Distribution
14.
J Virol ; 72(11): 8578-85, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9765396

ABSTRACT

The time courses of poliovirus plus- and minus-strand RNA synthesis in infected HEp-2 cells were monitored separately, using a quantitative RNase assay. In parallel, viral RNA and proteins were located in situ by confocal microscopy within cells fixed by a protocol determined to retain their native size and shape. Plus- and minus-strand RNAs were visualized by fluorescent in situ hybridization (FISH) with strand-specific riboprobes. The probes were labelled with different fluorochromes to allow for the simultaneous detection of plus- and minus-strand RNA. The FISH experiments showed minus-strand RNA to be present in distinct, regularly sized, round structures throughout the viral replication cycle. Plus-strand RNA was found in the same structures and also in smaller clusters of vesicles. Association of viral RNA with membranes was demonstrated by combining FISH with immunofluorescence (IF) detection of the viral 2B- and 2C-containing P2 proteins, which are known to be markers for virus-induced membranes. At early times postinfection, the virus-induced membranous structures were distributed through most of the cytoplasm, whereas around peak RNA synthesis, both RNA-associated membranous structures migrated to the center of the cell. During this process, the plus- and minus-strand-containing larger structures stayed as recognizable entities, whereas the plus-strand-containing granules coalesced into a juxtanuclear area of membranous vesicles. An involvement of Golgi-derived membranes in the formation of virus-induced vesicles and RNA synthesis early in infection was investigated by IF with 2C- and Golgi-specific antibodies.


Subject(s)
Poliovirus/genetics , Poliovirus/metabolism , RNA, Viral/genetics , RNA, Viral/metabolism , Cell Line , Golgi Apparatus/virology , Humans , In Situ Hybridization, Fluorescence , Kinetics , Microscopy, Confocal , Microscopy, Electron , Poliovirus/physiology , RNA, Viral/biosynthesis , Viral Proteins/metabolism , Virus Replication
17.
Biochem Pharmacol ; 55(4): 447-53, 1998 Feb 15.
Article in English | MEDLINE | ID: mdl-9514079

ABSTRACT

Bleomycin and asparaginase are widely used antineoplastic agents which may induce allergic or inflammatory side-effects. Mast cells are implicated as effector cells in allergic and inflammatory responses. The aim of this study was to establish whether bleomycin or asparaginase modulate leukotriene production in vitro and in vivo. Leukotriene C4 (LTC4) production by murine bone marrow-derived mast cells (BMMC) was determined by radioimmunoassay (RIA). Leukotriene production in patients was assessed by determining leukotriene E4 and N-acetyl-leukotriene E4 in urine by means of combined HPLC and RIA. Bleomycin induced an up to 2.1-fold increase in LTC4 production both in unstimulated and in calcium ionophore-stimulated mast cells. In 3 of 7 patients treated with bleomycin, a greater than 2-fold increase in endogenous leukotriene production was observed. This effect was associated with febrile responses and was most pronounced in a patient who developed an Adult Respiratory Distress Syndrome (ARDS). Asparaginase increased leukotriene production up to 10-fold in stimulated but not in unstimulated BMMC. In a patient who developed an anaphylactic reaction after treatment with asparaginase, a pronounced increase in urinary leukotriene concentration was observed. In contrast to bleomycin or asparaginase, a number of other cytostatic agents did not significantly change leukotriene production by BMMC. Our data indicate that some of the inflammatory and allergic side-effects of bleomycin and asparaginase could be mediated by leukotrienes, a possible source of which may be mast cells.


Subject(s)
Antineoplastic Agents/pharmacology , Asparaginase/pharmacology , Bleomycin/pharmacology , Leukotrienes/biosynthesis , Mast Cells/drug effects , Mast Cells/metabolism , Adult , Anaphylaxis/chemically induced , Anaphylaxis/metabolism , Animals , Calcimycin/pharmacology , Drug Hypersensitivity/etiology , Humans , In Vitro Techniques , Inflammation/chemically induced , Ionophores/pharmacology , Leukotriene C4/biosynthesis , Leukotriene E4/analogs & derivatives , Leukotriene E4/urine , Lymphoma, Non-Hodgkin/drug therapy , Lymphoma, Non-Hodgkin/metabolism , Mast Cells/immunology , Mice , Mice, Inbred BALB C , Respiratory Distress Syndrome/chemically induced , Respiratory Distress Syndrome/metabolism
18.
Virology ; 252(2): 343-53, 1998 Dec 20.
Article in English | MEDLINE | ID: mdl-9878613

ABSTRACT

The death of poliovirus-infected cells may occur in two forms: canonical cytopathic effect (CPE) (on productive infections) or apoptosis (when the viral reproduction is hindered by certain drugs or some other restrictive conditions). Morphological manifestations of the CPE and apoptosis, being distinct, share some traits (e.g., chromatin condensation and nuclear deformation). It was shown here that a permeable caspase inhibitor, benzyloxycarbonyl-Val-Ala-Asp-(OMe) fluoromethyl ketone (zVAD.fmk), prevented the development of the poliovirus-induced apoptosis on abortive infection. The apoptotic pathway could be dissected by an inhibitor of chymotrypsin-like serine proteases, N-tosyl-l-phenylalanine chloromethyl ketone (TPCK), which prevented the cleavage of DNA to oligonucleosome-sized pieces and nuclear fragmentation but did not suppress cellular shrinkage, cytoplasmic blebbing, and partial chromatin condensation. These results demonstrate that caspase activation is involved in the execution phase of the viral apoptosis and suggest that a nuclear subset of the apoptotic program is under a separate control, involving a TPCK-sensitive event. Neither zVAD.fmk nor TPCK, at the concentrations affecting the apoptotic response, exerted appreciable influence on the virus growth or cellular pathological changes on productive infection, indicating that the pathways leading to the poliovirus-evoked CPE and apoptosis are different.


Subject(s)
Apoptosis/physiology , Caspases/metabolism , Cell Death/physiology , Poliovirus/physiology , Amino Acid Chloromethyl Ketones/pharmacology , Apoptosis/drug effects , Caspase Inhibitors , Cell Nucleus/drug effects , Cell Nucleus/ultrastructure , Cell Nucleus/virology , Cysteine Proteinase Inhibitors/pharmacology , Guanidine/pharmacology , HeLa Cells , Humans , Poliovirus/pathogenicity , Tosylphenylalanyl Chloromethyl Ketone/pharmacology
19.
Br J Haematol ; 103(4): 1181-3, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9886339

ABSTRACT

It has previously been shown that a combination of macrophage inflammatory protein-1alpha (MIP-1alpha) and interleukin (IL)-3 maintained human bone marrow (BM)-derived long-term culture-initiating cells (LTC-IC) for at least 8 weeks in vitro. We investigated colony- and cobblestone area-formation potential of peripheral blood progenitor cells (PBPC) at week 6 of long-term culture (LTC) in the absence of exogenous MIP-1alpha. but using cells which had been pre-incubated in the presence of MIP-1alpha for 40 h in liquid culture. The recovery of colony-forming cells (CFC) and cobblestone-area-forming cells (CAFC) after pre-incubation of PBPC with MIP-1alpha was up to threefold higher (P<0.05, n=5) than that of untreated controls. These results demonstrate that short-term pre-treatment of PBPC with MIP-1alpha induces long-lasting survival effects on early PBPC-derived progenitors in vitro.


Subject(s)
Hematopoietic Stem Cells/drug effects , Macrophage Inflammatory Proteins/pharmacology , Cell Survival/drug effects , Cells, Cultured , Chemokine CCL3 , Chemokine CCL4 , Humans
20.
Virology ; 237(1): 66-77, 1997 Oct 13.
Article in English | MEDLINE | ID: mdl-9344908

ABSTRACT

Hepatitis A virus (HAV) is distinguished from other picornaviruses by its slow and relatively poor, noncytopathic growth in cultures of mammalian cells. The 2C and 2BC proteins of HAV have been implicated in the determination of virus growth in cultured cells. The homologous proteins from other picornaviruses, such as poliovirus, have been demonstrated to exhibit multiple activities, such as RNA binding, nucleotide binding and NTPase, and membrane binding and reorganization. At least some of these activities are required for viral RNA replication. We report here that HAV 2C and 2BC proteins, like their poliovirus counterparts, can induce rearrangement of intracellular membranes and directly or indirectly interact with membranes. Therefore, the inefficient replication properties of HAV are not consequences of the inherent ability of 2C (2BC) to interact with membranes. The effect of 2C (2BC) protein sequences derived from a cell culture-adapted (cc) strain of HAV was compared with that of corresponding protein sequences from either a wild-type (wt) strain of HAV or a faster replicating cytopathic (cp) strain. The analysis demonstrated that mutations acquired in wt virus during adaptation to cell culture do not change dramatically either the ability of these proteins to associate with membranes and induce membrane alterations or the specific architecture of the induced membrane structures. On the other hand, 2C, but not 2BC, protein from the cp strain of HAV induced different membrane structures.


Subject(s)
Gene Expression Regulation , Hepatovirus/metabolism , Intracellular Membranes/ultrastructure , Viral Proteins/physiology , HeLa Cells , Humans , Intracellular Membranes/virology , Microscopy, Electron , Plasmids
SELECTION OF CITATIONS
SEARCH DETAIL
...