bluuebunny/crossref_metadata_2025_split huggingface dataset:
Zeolin. A New Recombinant Storage Protein Constructed Using Maize γ-Zein and Bean Phaseolin
Authors: Mainieri Davide , Rossi Marika , Archinti Marco , Bellucci Michele , De Marchis Francesca , Vavassori Stefano , Pompa Andrea , Arcioni Sergio , Vitale Alessandro 
Published: November 2004

Abstract
The major seed storage proteins of maize (Zea mays) and bean (Phaseolus vulgaris), zein and phaseolin, accumulate in the endoplasmic reticulum (ER) and in storage vacuoles, respectively. We show here that a chimeric protein composed of phaseolin and 89 amino acids of γ-zein, including the repeated and the Pro-rich domains, maintains the main characteristics of wild-type γ-zein: It is insoluble unless its disulfide bonds are reduced and forms ER-located protein bodies. Unlike wild-type phaseolin, the protein, which we called zeolin, accumulates to very high amounts in leaves of transgenic tobacco (Nicotiana tabacum). A relevant proportion of the ER chaperone BiP is associated with zeolin protein bodies in an ATP-sensitive fashion. Pulse-chase labeling confirms the high affinity of BiP to insoluble zeolin but indicates that, unlike structurally defective proteins that also extensively interact with BiP, zeolin is highly stable. We conclude that the γ-zein portion is sufficient to induce the formation of protein bodies also when fused to another protein. Because the storage proteins of cereals and legumes nutritionally complement each other, zeolin can be used as a starting point to produce nutritionally balanced and highly stable chimeric storage proteins.
Conventional Protein Kinase C Inhibition Prevents Alpha Interferon-Mediated Hepatitis C Virus Replicon Clearance by Impairing STAT Activation
Authors: Fimia Gian Maria , Evangelisti Cristina , Alonzi Tonino , Romani Marta , Fratini Federica , Paonessa Giacomo , Ippolito Giuseppe , Tripodi Marco , Piacentini Mauro 
Published: November 2004

ABSTRACT
Hepatitis C virus (HCV) has evolved complex strategies to evade host immune responses and establish chronic infection. The only treatment available for HCV infections, alpha interferon (IFN-α), is effective in a limited percentage of patients. The mechanisms by which IFN-α interferes with the HCV life cycle and the reasons for limited effectiveness of IFN-α therapy have not yet been fully elucidated. Using a cell-based HCV replication system and specific kinase inhibitors, we examined the role played by various signaling pathways in the IFN-α-mediated HCV clearance. We reported that conventional protein kinase C (cPKC) activity is important for the effectiveness of IFN-α treatment. In cells treated with a cPKC-specific inhibitor, IFN-α failed to induce an efficient HCV RNA degradation. The lack of cPKC activity leads to a broad reduction of IFN-α-stimulated gene expression due to a significant impairment of STAT1 and STAT3 tyrosine phosphorylation. Thus, modulation of cPKC function by either host or viral factors could influence the positive outcome of IFN-α-mediated antiviral therapies.
Chimpanzee Fab Fragments and a Derived Humanized Immunoglobulin G1 Antibody That Efficiently Cross-Neutralize Dengue Type 1 and Type 2 Viruses
Authors: Goncalvez Ana P. , Men Ruhe , Wernly Claire , Purcell Robert H. , Lai Ching-Juh 
Published: November 2004

ABSTRACTPassive immunization with monoclonal antibodies from humans or nonhuman primates represents an attractive alternative to vaccines for prevention of illness caused by dengue viruses (DENV) and other flaviviruses, including the West Nile virus. In a previous study, repertoire cloning to recover Fab fragments from bone marrow mRNA of chimpanzees infected with all four DENV serotypes (dengue virus serotype 1 [DENV-1] to DENV-4) was described. In that study, a humanized immunoglobulin G1 (IgG1) antibody that efficiently neutralized DENV-4 was recovered and characterized. In this study, the phage library constructed from the chimpanzees was used to recover Fab antibodies against the other three DENV serotypes. Serotype-specific neutralizing Fabs were not identified. Instead, we recovered DENV-neutralizing Fabs that specifically precipitated the envelope protein and were cross-reactive with all four DENV serotypes. Three of the Fabs competed with each other for binding to DENV-1 and DENV-2, although each of these Fabs contained a distinct complementarity determining region 3 (CDR3)-H sequence. Fabs that shared an identical or nearly identical CDR3-H sequences cross-neutralized DENV-1 and DENV-2 at a similar high 50% plaque reduction neutralization test (PRNT50) titer, ranging from 0.26 to 1.33 μg/ml, and neutralized DENV-3 and DENV-4 but at a titer 10- to 20-fold lower. One of these Fabs, 1A5, also neutralized the West Nile virus most efficiently among other flaviviruses tested. Fab 1A5 was converted to a full-length antibody in combination with human sequences for production in mammalian CHO cells. Humanized IgG1 1A5 proved to be as efficient as Fab 1A5 for cross-neutralization of DENV-1 and DENV-2 at a titer of 0.48 and 0.95 μg/ml, respectively. IgG1 1A5 also neutralized DENV-3, DENV-4, and the West Nile virus at a PRNT50titer of approximately 3.2 to 4.2 μg/ml. This humanized antibody represents an attractive candidate for further development of immunoprophylaxis against DENV and perhaps other flavivirus-associated diseases.
The DNA Architectural Protein HMGB1 Facilitates RTA-Mediated Viral Gene Expression in Gamma-2 Herpesviruses
Authors: Jung Song Moon , Hwang Seungmin , Wong Wendy , Round June , Martinez-Guzman DeeAnn , Turpaz Yaron , Liang Jie , Wong Ben , Johnson Reid C. , Carey Michael , Sun Ren 
Published: November 2004

ABSTRACT

Replication and transcription activator (RTA), an immediate-early gene product of gamma-2 herpesviruses including Kaposi's sarcoma-associated herpesvirus (KSHV) and murine gamma herpesvirus 68 (MHV-68), plays a critical role in controlling the viral life cycle. RTA acts as a strong transcription activator for several downstream genes of KSHV and MHV-68 through direct DNA binding, as well as via indirect mechanisms. HMGB1 (also called HMG-1) protein is a highly conserved nonhistone chromatin protein with the ability to bind and bend DNA. HMGB1 protein promoted RTA binding to different RTA target sites in vitro, with greater enhancement to low-affinity sites than to high-affinity sites. Box A or box B and homologues of HMGB1 also enhanced RTA binding to DNA. Transient transfection of HMGB1 stimulated RTA transactivation of RTA-responsive promoters from KSHV and MHV-68. Furthermore, MHV-68 viral gene expression, as well as viral replication, was significantly reduced in
HMGB1
-deficient cells than in the wild type. This abated viral gene expression was partially restored by HMGB1 transfection into

HMGB1
−/−

cells. These results suggest an important function of the DNA architectural protein, HMGB1, in RTA-mediated gene expression, as well as viral replication in gamma-2 herpesviruses.
Amino Acid Substitutions in the F-Specific Domain in the Stalk of the Newcastle Disease Virus HN Protein Modulate Fusion and Interfere with Its Interaction with the F Protein
Authors: Melanson Vanessa R. , Iorio Ronald M. 
Published: November 2004

ABSTRACT
The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus mediates attachment to sialic acid receptors, as well as cleavage of the same moiety. HN also interacts with the other viral glycoprotein, the fusion (F) protein, to promote membrane fusion. The ectodomain of the HN spike consists of a stalk and a terminal globular head. The most conserved part of the stalk consists of two heptad repeats separated by a nonhelical intervening region (residues 89 to 95). Several amino acid substitutions for a completely conserved proline residue in this region not only impair fusion and the HN-F interaction but also decrease neuraminidase activity in the globular domain, suggesting that the substitutions may alter HN structure. Substitutions for L94 also interfere with fusion and the HN-F interaction but have no significant effect on any other HN function. Amino acid substitutions at other positions in the intervening region also modulate only fusion. In all cases, diminished fusion correlates with a decreased ability of the mutated HN protein to interact with F at the cell surface. These findings indicate that the intervening region is critical to the role of HN in the promotion of fusion and may be directly involved in its interaction with the homologous F protein.
Spread and Replication of and Immune Response to γ
1
34.5-Negative Herpes Simplex Virus Type 1 Vectors in BALB/c Mice
Authors: Broberg Eeva K. , Peltoniemi Jutta , Nygårdas Michaela , Vahlberg Tero , Röyttä Matias , Hukkanen Veijo 
Published: November 2004

ABSTRACT

We have previously shown that intracranial infection of herpes simplex virus type 1 (HSV-1) vector R8306 expressing interleukin-4 (IL-4) can abolish symptoms of experimental autoimmune encephalomyelitis, which is used as a model for human multiple sclerosis (Broberg et al., Gene Ther.
8:
769-777, 2001). The aim of the current study was to search for means other than intracranial injection to deliver HSV-derived vectors to the central nervous system of mice. We also aimed to study the replication efficiency of these vectors in nervous system tissues and to elucidate the effects of the viruses on the immune response. We studied the spread and replication of the following viruses with deletions in neurovirulence gene γ
1
34.5: R3616, R849 (
lacZ
transgene), R3659 (alpha-
tk
), R8306 (murine IL-4 transgene), and R8308 (murine IL-10 transgene). The samples were taken from trigeminal ganglia and brains of BALB/c mice after corneal, intralabial, and intranasal infection, and the viral load was examined by viral culture, HSV DNA PCR, and VP16 reverse transcription (RT)-PCR. The results show that (i) intranasal infection was the most efficient means of spread to the central nervous system (CNS) besides intracranial injection; (ii) the viruses did not grow in the culture from the brain samples, but the viral DNA persisted even until day 21 postinfection; (iii) viral replication, as observed by VP16 mRNA RT-PCR, occurred mainly on days 4 and 7 postinfection in trigeminal ganglia and to a low extent in brain; (iv) R3659, R8306, and R8308 showed reactivation from the trigeminal ganglia in explant cultures; (v) in the brain, the vectors spread to the midbrain more efficiently than to other brain areas; and (vi) the deletions in the R3659 genome significantly limited the ability of this virus to replicate in the nervous system. The immunological studies show that (i) the only recombinant to induce IL-4 mRNA expression in the brain was R8306, the gamma interferon response was very low in the brain for R3659 and R8306, and the IL-23p19 response to R8306 decreased by day 21 postinfection, unlike for the other viruses; (ii) Δγ
1
34.5 HSV vectors modulated the subsets of the splenocytes differently depending on the transgene; (iii) R3659 infection of the nervous system induces expression and production of cytokines from the stimulated splenocytes; and (iv) HSV vectors expressing IL-4 or IL-10 induce expression and production of both of the Th2-type cytokines from splenocytes. We conclude that the intranasal route of infection is a possible means of delivery of Δγ
1
34.5 HSV vectors to the CNS in addition to intracranial infection, although replication in the CNS remains minimal. The DNA of the HSV vectors is able to reside in the brain for at least 3 weeks. The features of the immune response to the vectors must be considered and may be exploited in gene therapy experiments with these vectors.
Sequence Variation within the Dominant Amino Terminus Epitope Affects Antibody Binding and Neutralization of Human Immunodeficiency Virus Type 1 Tat Protein
Authors: Ruckwardt Tracy J. , Tikhonov Ilia , Berg Shannon , Hatfield Glen S. , Chandra Angelika , Chandra Prakash , Gilliam Bruce , Redfield Robert R. , Gallo Robert C. , Pauza C. David 
Published: November 2004

ABSTRACTTat is among the required regulatory genes of human immunodeficiency virus type 1 (HIV-1). Tat functions both within infected cells as a transcription factor and as an extracellular factor that binds and alters bystander cells. Some functions of extracellular Tat can be neutralized by immune serum or monoclonal antibodies. In order to understand the antibody response to Tat, we are defining antibody epitopes and the effects of natural Tat sequence variation on antibody recognition. The dominant Tat epitope in macaque sera is within the first 15 amino acids of the protein amino terminus. Together with a subdominant response to amino acids 57 to 60, these two regions account for most of the macaque response to linear Tat epitopes and both regions are also sites for the binding of neutralizing antibodies. However, the dominant and subdominant epitope sequences differ among virus strains, and this natural variation can preclude antibody binding and Tat neutralization. We also examined serum samples from 31 HIV-positive individuals that contained Tat binding antibodies; 23 of the 31 sera recognized the amino terminus peptide. Similar to binding in macaques, human antibody binding to the amino terminus was affected by variations at positions 7 and 12, sequences that are distinct for clade B compared to other viral clades. Tat-neutralizing antibodies to the dominant amino terminus epitope are affected by HIV clade variation.
Herpes Simplex Virus Type 1 Glycoprotein K and the UL20 Protein Are Interdependent for Intracellular Trafficking andtrans-Golgi Network Localization
Authors: Foster Timothy P. , Melancon Jeffrey M. , Olivier Trisha L. , Kousoulas Konstantin G. 
Published: November 2004

ABSTRACTFinal envelopment of the cytoplasmic herpes simplex virus type 1 (HSV-1) nucleocapsid is thought to occur by budding intotrans-Golgi network (TGN)-derived membranes. The highly membrane-associated proteins UL20p and glycoprotein K (gK) are required for cytoplasmic envelopment at the TGN and virion transport from the TGN to extracellular spaces. Furthermore, the UL20 protein is required for intracellular transport and cell surface expression of gK. Independently expressed gK or UL20p via transient expression in Vero cells failed to be transported from the endoplasmic reticulum (ER). Similarly, infection of Vero cells with either gK-null or UL20-null viruses resulted in ER entrapment of UL20p or gK, respectively. In HSV-1 wild-type virus infections and to a lesser extent in transient gK and UL20p coexpression experiments, both gK and UL20p localized to the Golgi apparatus. In wild-type, but not UL20-null, viral infections, gK was readily detected on cell surfaces. In contrast, transiently coexpressed gK and UL20p predominantly localized to the TGN and were not readily detected on cell surfaces. However, TGN-localized gK and UL20p originated from endocytosed gK and UL20p expressed at cell surfaces. Retention of UL20p to the ER through the addition of an ER retention motif forced total ER retention of gK, indicating that transport of gK is absolutely dependent on UL20p transport. In all experiments, gK and UL20p colocalized at intracellular sites, including the ER, Golgi, and TGN. These results are consistent with the hypothesis that gK and UL20p directly interact and that this interaction facilitates their TGN localization, an important prerequisite for cytoplasmic virion envelopment and egress.
Transmission of Prions from Mule Deer and Elk with Chronic Wasting Disease to Transgenic Mice Expressing Cervid PrP
Authors: Browning Shawn R. , Mason Gary L. , Seward Tanya , Green Mike , Eliason Gwyneth A. J. , Mathiason Candace , Miller Michael W. , Williams Elizabeth S. , Hoover Ed , Telling Glenn C. 
Published: November 2004

ABSTRACT
We generated mice expressing cervid prion protein to produce a transgenic system simulating chronic wasting disease (CWD) in deer and elk. While normal mice were resistant to CWD, these transgenic mice uniformly developed signs of neurological dysfunction ∼230 days following intracerebral inoculation with four CWD isolates. Inoculated transgenic mice homozygous for the transgene array developed disease after ∼160 days. The brains of sick transgenic mice exhibited widespread spongiform degeneration and contained abnormal prion protein and abundant amyloid plaques, many of which were florid plaques. Transmission studies indicated that the same prion strain caused CWD in the analyzed mule deer and elk. These mice provide a new and reliable tool for detecting CWD prions.
Infection of Macrophage Primary Cultures by Persistent and Nonpersistent Strains of Theiler's Virus: Role of Capsid and Noncapsid Viral Determinants
Authors: Mena Ignacio , Roussarie Jean-Pierre , Brahic Michel 
Published: November 2004

ABSTRACT
We compared the infection of bone marrow macrophages by the DA and GDVII strains of Theiler's virus and by two viruses constructed by exchanging the DA and GDVII capsids. The replication of the GDVII strain and of both chimeric viruses was restricted in macrophages. Therefore, the infection of macrophages requires both capsid and noncapsid viral determinants.
OPTIMUM SCHEDULE PROBLEMS IN STORE AND FORWARD NETWORKS
Authors: CLEMENTI ANDREA , IANNI MIRIAM DI 
Published: November 2004

We investigate the problem of devising an optimal time strategy to send messages from one site to another in a Store and Forward network. We consider the problem both in the cases of messages transmitted through fixed paths and through dynamically variable paths, respectively. In particular, we prove that both of versions are NP-complete even when very restrictive hypotheses are imposed on the network topology and on the required amount of time. We thus turn our attention towards the study of the approximability of this scheduling problem. In particular, we prove the following negative results: (i) In the case of fixed paths, the problem is not approximable for any constant approximation ratio unless P is equal to NP. (ii) In the case of arbitrary paths, the problem is not approximable for any constant approximation ratio (unless P is equal to NP) when an arbitrary priority function is defined on the set of messages. Finally, we present a particular network topology in which the problem can be solved in deterministic polynomial time.
CHANGES OF BLOOD AND URINE PARAMETERS AND MICROFLORA IN THE GASTROINTESTINAL TRACT OF CARP EXPOSED TO DILUTED SEWAGE FOR A SHORT PERIOD
Authors: KAKUTA I. , TAKASAKI M. , SATO T. , ISHII K. 
Published: November 2004

Changes in blood and urine parameters of carp exposed to diluted (20%) raw sewage for a short period were investigated. Carp exposed to the sewage lost their balance within 30 minutes. When carp lost their balance, haematocrite(Ht)and plasma non-nitrogenous compounds(NPN), ammonia. Ca2+,Mg2+, Sand catecholamines (CAs) increased significantly, whereas no remarkable changes were observed in urine parameters. Four hours after the exposure, marked changes were found in blood and urine parameters as follows; much higher values for Ht, plasma NPN, ammonia, glucose, lactate. Mg2+, S, Cl, Br and CAs, and for urine ammonia, S, Cl, K, Ca, Zn and Br were observed. After 12 hours, some slight aftereffects were still found on blood and urine parameters. After 36 hours, both blood and urine parameters recovered to the control levels. Effects of the sewage exposure on microflora in the gastrointestinal tract of carp were also studied. Microflora were widely different between upper and lower parts of the gastrointestinal tract in the control. Remarkable bacteriological changes in the tract, particularly in the lower part, were found 12 hours after the exposure. This study suggests that (i) even a short-term exposure to sewage causes considerable stress responses on carp, (ii) urinalysis and examination of gut microbes as well as blood test are effective for the diagnosis of physiological dysfunction in carp, and (iii) microflora in the gastrointestinal tract of carp are strikingly affected by the physiological conditions of fish and by microflora in environmental water.
SMART SENSORY INFORMATION WITH FUZZY LOGIC FOR AN INTELLIGENT AUTOMOTIVE GEAR CHANGING CONTROL SYSTEM
Authors: NWAGBOSO CHRISTOPHER 
Published: November 2004

The automotive gear changing process can be a mechanically and physically demanding operation. Although self-changing gearboxes have reduced the skills demanded of a driver, the physical performance requirements of the gear changing skill remains the same. The basic requirement for safety, reliability and efficiency of the gear changing process as an integral part of the power train system demands that continued development is made towards the application of intelligent control methods. When the systems development is coupled with the benefits of smart sensors, the control technique employed can be made more intelligent. In this paper, aspect of intelligent gear changing process together with sensory information factors which form part of the gear changing process is presented. The application of fuzzy logic with the multi-sensory systems for the purpose of reliably selecting appropriate gear ratios under changing road conditions demanding changing vehicle performance is analysed.
Multidataset Study of Optimal Parameter and Uncertainty Estimation of a Land Surface Model with Bayesian Stochastic Inversion and Multicriteria Method
Authors: Xia Youlong , Sen Mrinal K. , Jackson Charles S. , Stoffa Paul L. 
Published: November 2004

Abstract
This study evaluates the ability of Bayesian stochastic inversion (BSI) and multicriteria (MC) methods to search for the optimal parameter sets of the Chameleon Surface Model (CHASM) using prescribed forcing to simulate observed sensible and latent heat fluxes from seven measurement sites representative of six biomes including temperate coniferous forests, tropical forests, temperate and tropical grasslands, temperate crops, and semiarid grasslands. Calibration results with the BSI and MC show that estimated optimal values are very similar for the important parameters that are specific to the CHASM model. The model simulations based on estimated optimal parameter sets perform much better than the default parameter sets. Cross-validations for two tropical forest sites show that the calibrated parameters for one site can be transferred to another site within the same biome. The uncertainties of optimal parameters are obtained through BSI, which estimates a multidimensional posterior probability density function (PPD). Marginal PPD analyses show that nonoptimal choices of stomatal resistance would contribute most to model simulation errors at all sites, followed by ground and vegetation roughness length at six of seven sites. The impact of initial root-zone soil moisture and nonmosaic approach on estimation of optimal parameters and their uncertainties is discussed.