bluuebunny/crossref_metadata_2025_split huggingface dataset:
Preoperative staging of sinonasal inverted papilloma by magnetic resonance imaging
Authors: Oikawa Keita , Furuta Yasushi , Oridate Nobuhiko , Nagahashi Tatsumi , Homma Akihiro , Ryu Takusyu , Fukuda Satoshi 
Published: November 2003

AbstractObjectives/Hypothesis: Sinonasal inverted papilloma is a rare but locally aggressive benign tumor. Inverted papilloma tends to recur after surgical resection and is occasionally associated with squamous cell carcinoma. Radical en bloc resection by lateral rhinotomy and medial maxillectomy has been recommended for initial management; however, endoscopic sinus surgery is effective in selected cases. To determine adequate surgical approaches, a staging system for inverted papilloma based on extent and location of the tumor has been advocated. The study investigated whether preoperative assessment by magnetic resonance imaging (MRI) accurately predicts the extent of inverted papilloma. Study Design: Magnetic resonance imaging was retrospectively reviewed in 21 cases of inverted papilloma without knowledge of the surgical and pathological findings. Methods: Patients were categorized into stages based on MRI findings, according to the staging system proposed by Krouse. The involvement of inverted papilloma in each sinus was also assessed. Results: Stages graded by MRI were coincident with postoperative staging verified by surgical and pathological findings in 18 of the 21 cases (86%), which included 1 case of stage T1, 3 cases of stage T2, and 14 cases of stage T3. Two cases of stage T2 were judged as T3, and one case of T3 was judged as T2 by MRI. Positive predictive value of MRI diagnosis for tumor involvement was 68% to 89% in each sinus, and negative predictive value was more than 93%. Conclusion: In most cases, MRI assessment of inverted papilloma can accurately predict the extent of tumor involvement. Preoperative staging of inverted papilloma by MRI may be useful for planning an appropriate surgical approach.
α‐Lactalbumin unfolding is not sufficient to cause apoptosis, but is required for the conversion to HAMLET (human α‐lactalbumin made lethal to tumor cells)
Authors: Svensson Malin , Fast Jonas , Mossberg Ann‐Kristin , Düringer Caroline , Gustafsson Lotta , Hallgren Oskar , Brooks Charles L. , Berliner Lawrence , Linse Sara , Svanborg Catharina 
Published: November 2003

AbstractHAMLET (human α‐lactalbumin made lethal to tumor cells) is a complex of human α‐lactalbumin and oleic acid (C18:1:9 cis) that kills tumor cells by an apoptosis‐like mechanism. Previous studies have shown that a conformational change is required to form HAMLET from α‐lactalbumin, and that a partially unfolded conformation is maintained in the HAMLET complex. This study examined if unfolding of α‐lactalbumin is sufficient to induce cell death. We used the bovine α‐lactalbumin Ca2+ site mutant D87A, which is unable to bind Ca2+, and thus remains partially unfolded regardless of solvent conditions. The D87A mutant protein was found to be inactive in the apoptosis assay, but could readily be converted to a HAMLET‐like complex in the presence of oleic acid. BAMLET (bovine α‐lactalbumin made lethal to tumor cells) and D87A‐BAMLET complexes were both able to kill tumor cells. This activity was independent of the Ca2+site, as HAMLET maintained a high affinity for Ca2+ but D87A‐BAMLET was active with no Ca2+ bound. We conclude that partial unfolding of α‐lactalbumin is necessary but not sufficient to trigger cell death, and that the activity of HAMLET is defined both by the protein and the lipid cofactor. Furthermore, a functional Ca2+‐binding site is not required for conversion of α‐lactalbumin to the active complex or to cause cell death. This suggests that the lipid cofactor stabilizes the altered fold without interfering with the Ca2+site.
Lipids as cofactors in protein folding: Stereo‐specific lipid–protein interactions are required to form HAMLET (human α‐lactalbumin made lethal to tumor cells)
Authors: Svensson Malin , Mossberg Ann‐Kristin , Pettersson Jenny , Linse Sara , Svanborg Catharina 
Published: November 2003

AbstractProteins can adjust their structure and function in response to shifting environments. Functional diversity is created not only by the sequence but by changes in tertiary structure. Here we present evidence that lipid cofactors may enable otherwise unstable protein folding variants to maintain their conformation and to form novel, biologically active complexes. We have identified unsaturated C18 fatty acids in the cis conformation as the cofactors that bind apo α‐lactalbumin and form HAMLET (human α‐lactalbumin made lethal to tumor cells). The complexes were formed on an ion exchange column, were stable in a molten globule‐like conformation, and had attained the novel biological activity. The protein–fatty acid interaction was specific, as saturated C18 fatty acids, or unsaturated C18:1trans conformers were unable to form complexes with apo α‐lactalbumin, as were fatty acids with shorter or longer carbon chains. Unsaturated cis fatty acids other than C18:1:9cis were able to form stable complexes, but these were not active in the apoptosis assay. The results demonstrate that stereo‐specific lipid–protein interactions can stabilize partially unfolded conformations and form molecular complexes with novel biological activity. The results offer a new mechanism for the functional diversity of proteins, by exploiting lipids as essential, tissue‐specific cofactors in this process.
PorphyromonasgingivalisLipopolysaccharide AntagonizesEscherichiacoliLipopolysaccharide at Toll-Like Receptor 4 in HumanEndothelialCells
Authors: Coats Stephen R. , Reife Robert A. , Bainbridge Brian W. , Pham Thu-Thao T. , Darveau Richard P. 
Published: November 2003

ABSTRACTE. colilipopolysaccharide (LPS) induces cytokine and adhesion molecule expression via the toll-like receptor 4 (TLR4) signaling complex in human endothelial cells. In the present study, we investigated the mechanism by whichPorphyromonas gingivalisLPS antagonizesE. coliLPS-dependent activation of human endothelial cells.P. gingivalisLPS at 1 μg/ml inhibited bothE. coliLPS (10 ng/ml) andMycobacterium tuberculosisheat shock protein (HSP) 60.1 (10 μg/ml) stimulation of E-selectin mRNA expression in human umbilical vein endothelial cells (HUVEC) without inhibiting interleukin-1 beta (IL-1β) stimulation.P. gingivalisLPS (1μ g/ml) also blocked bothE. coliLPS-dependent andM. tuberculosisHSP60.1-dependent but not IL-1β-dependent activation of NF-κB in human microvascular endothelial (HMEC-1) cells, consistent with antagonism occurring upstream from the TLR/IL-1 receptor adaptor protein, MyD88. Surprisingly,P. gingivalisLPS weakly but significantly activated NF-κB in HMEC-1 cells in the absence ofE. coliLPS, and theP. gingivalisLPS-dependent agonism was blocked by transient expression of a dominant negative murine TLR4. Pretreatment of HUVECs withP. gingivalisLPS did not influence the ability ofE. coliLPS to stimulate E-selectin mRNA expression. Taken together, these data provide the first evidence thatP. gingivalisLPS-dependent antagonism ofE. coliLPS in human endothelial cells likely involves the ability ofP. gingivalisLPS to directly compete withE. coliLPS at the TLR4 signaling complex.
Th1-Directing Adjuvants Increase the Immunogenicity of Oligosaccharide-Protein Conjugate Vaccines Related toStreptococcus pneumoniaeType 3
Authors: Lefeber Dirk J. , Benaissa-Trouw Barry , Vliegenthart Johannes F. G. , Kamerling Johannis P. , Jansen Wouter T. M. , Kraaijeveld Kees , Snippe Harm 
Published: November 2003

ABSTRACTOligosaccharide (OS)-protein conjugates are promising candidate vaccinesagainst encapsulated bacteria, such asHaemophilus influenzae,Neisseria meningitidis, andStreptococcus pneumoniae. Although the effects of several variables such as OS chain length and protein carrier have been studied, little is known about the influence of adjuvants on the immunogenicity of OS-protein conjugates. In this study, a minimal protective trisaccharide epitope ofStreptococcus pneumoniaetype 3 conjugated to the cross-reacting material of diphtheria toxin was used for immunization of BALB/c mice in the presence of different adjuvants. Subsequently, half of the mice received a booster immunization with conjugate alone. Independent of the use and type of adjuvant, all mice produced long-lasting anti-polysaccharide type 3 (PS3) antibody levels, which provided full protection against challenge with pneumococcal type 3 bacteria. All adjuvants tested increased the anti-PS3 antibody levels and opsonic capacities as measured by an enzyme-linked immunosorbent assay and an in vitro phagocytosis assay. The use of QuilA or a combination of the adjuvants CpG and dimethyl dioctadecyl ammonium bromide resulted in the highest phagocytic capacities and the highest levels of Th1-related immunoglobulin G (IgG) subclasses. Phagocytic capacity correlated strongly with Th1-associated IgG2a and IgG2b levels, to a lesser extent with Th2-associated IgG1 levels, and weakly with thiocyanate elution as a measure of avidity. Thus, the improved immunogenicity of OS-protein conjugates was most pronounced for Th1-directing adjuvants.
Mycobacterium tuberculosisGrowth at theCavity Surface: a Microenvironment with FailedImmunity
Authors: Kaplan Gilla , Post Frank A. , Moreira Andre L. , Wainwright Helen , Kreiswirth Barry N. , Tanverdi Melike , Mathema Barun , Ramaswamy Srinivas V. , Walther Gabi , Steyn Lafras M. , Barry Clifton E. , Bekker Linda-Gail 
Published: November 2003

ABSTRACTProtective immunity against pulmonary tuberculosis (TB) is characterized by the formation in the lungs of granulomas consisting of macrophages and activated T cells producing tumor necrosis factor alpha and gamma interferon, both required for the activation of the phagocytes. In 90% of immunocompetent humans, this response controls the infection. To understand why immunity fails in the other 10%, we studied the lungs of six patients who underwent surgery for incurable TB. Histologic examination of different lung lesions revealed heterogeneous morphology and distribution of acid-fast bacilli; only at the surface of cavities, i.e., in granulomas with a patent connection to the airways, were there numerous bacilli. The mutation profile of the isolates suggested that a single founder strain ofMycobacterium tuberculosismay undergo genetic changes during treatment, leading to acquisition of additional drug resistance independently in discrete physical locales. Additional drug resistance was preferentially observed at the cavity surface. Cytokine gene expression revealed that failure to control the bacilli was not associated with a generalized suppression of cellular immunity, since cytokine mRNA was up regulated in all lesions tested. Rather, a selective absence of CD4+and CD8+T cells was noted at the luminal surface of the cavity, preventing direct T-cell-macrophage interactions at this site, probably allowing luminal phagocytes to remain permissive for bacillary growth. In contrast, in the perinecrotic zone of the granulomas, the two cell types colocalized and bacillary numbers were substantially lower, suggesting that in this microenvironment an efficient bacteriostatic or bactericidal phagocyte population was generated.
GeneticAnalysis of the Capsule Locus of
Haemophilus influenzae
Serotypef
Authors: Satola Sarah W. , Schirmer Patricia L. , Farley Monica M. 
Published: November 2003

ABSTRACT

A
19-kb DNA region containing genes involved in the biosynthesis of the
capsule of
Haemophilus influenzae
serotype f (Hif) has been
cloned and characterized. The Hif
cap
locus organization is
typical of group II capsule biosynthetic loci found in other
H.
influenzae
serotype b bacteria and other gram-negative bacteria.
However, the Hif
cap
locus was not associated with an
IS
1016
element. Three new open reading frames, Fcs1, Fcs2, and
Fcs3, were identified in the Hif capsule-specific region II. The
chromosomal location of the Hif
cap
locus and the organization
of the flanking sequences differed significantly from previously
described division I
H. influenzae
serotypes.
VariedImmunity Generated in Mice by DNA Vaccines with Large and SmallHepatitis DeltaAntigens
Authors: Huang Yi-Hsiang , Wu Jaw-Ching , Hsu Sheng-Chieh , Syu Wan-Jr 
Published: November 2003

ABSTRACT

Whether
the hepatitis delta virus (HDV) DNA vaccine can induce anti-HDV
antibodies has been debatable. The role of the isoprenylated motif of
hepatitis delta antigens (HDAg) in the generation of immune responses
following DNA-based immunization has never been studied. Plasmids
p2577L, encoding large HDAg (L-HDAg), p2577S, expressing small HDAg
(S-HDAg), and p25L-211S, encoding a mutant form of L-HDAg with a
cysteine-to-serine mutation at codon 211, were constructed in this
study. Mice were intramuscularly injected with the plasmids. The
anti-HDV antibody titers, T-cell proliferation responses, T-helper
responses, and HDV-specific, gamma interferon (IFN-γ)-producing
CD8
+
T cells were analyzed. Animals immunized with
p2577S showed a strong anti-HDV antibody response. Conversely, only a
low titer of anti-HDV antibodies was detected in mice immunized with
p2577L. Epitope mapping revealed that the anti-HDV antibodies generated
by p2577L vaccination hardly reacted with epitope amino acids 174 to
194, located at the C terminus of S-HDAg. All of the HDAg-encoding
plasmids could induce significant T-cell proliferation responses and
generate Th1 responses and HDV-specific, IFN-γ-producing
CD8
+
T cells. In conclusion, HDAg-specific
antibodies definitely exist following DNA vaccination. The magnitudes
of the humoral immune responses generated by L-HDAg- and
S-HDAg-encoding DNA vaccines are different. The isoprenylated motif can
mask epitope amino acids 174 to 195 of HDAg but does not interfere with
cellular immunity following DNA-based immunization. These findings are
important for the choice of a candidate HDV DNA vaccine in the
future.
Functional Analysis of nsP3 Phosphoprotein Mutants ofSindbisVirus
Authors: Dé Indra , Fata-Hartley Cori , Sawicki Stanley G. , Sawicki Dorothea L. 
Published: November 2003

ABSTRACT

Alphavirus
nsP3 phosphoprotein is essential for virus replication and functions
initially within polyprotein P123 or P23 components of the short-lived
minus-strand replicase, and upon polyprotein cleavage, mature nsP3
likely functions also in plus-strand synthesis. We report the
identification of a second nsP3 mutant from among the A complementation
group of Sindbis virus (SIN) heat-resistant strain,
ts
RNA-negative mutants. The
ts
138 mutant possessed a change of
G4303 to C, predicting an Ala68-to-Gly alteration that altered a
conserved His-Ala-Val tripeptide in the ancient (pre-eukaryotic),“
X” or histone 2A phosphoesterase-like macrodomain that
in SIN encompasses nsP3 residues 1 to 161 and whose role is unknown. We
undertook comparative analysis of three nsP3 N-terminal
region mutants and observed (i) that nsP3 and nsP2
functioned initially as a single unit as deduced from complementation
analysis and in agreement with our previous studies, (ii) that the
degree of phosphorylation varied among the nsP3 mutants, and (iii) that
reduced phosphorylation of nsP3 correlated with reduced minus-strand
synthesis. The most striking phenotype was exhibited by
ts
4
(Ala268 to Val), which after shift to 40°C made significantly
underphosphorylated P23/nsP3 and lost selectively the ability to make
minus strands. After shift to 40°C, mutant
ts
7 (Phe312
to Ser) made phosphorylated P23/nsP3 and minus strands but failed to
increase plus-strand synthesis. Macrodomain mutant
ts
138 was
intermediate, making at 40°C partially phosphorylated P23/nsP3
and reduced amounts of minus strands. The mutants were able to assemble
their nsPs at 40°C into complexes that were membrane
associated. Our analyses argue that P23/P123 phosphorylation is
affected by macrodomain and Ala268 region sequences and in turn affects
the efficient transcription of the alphavirus
genome.
Intravenous Inoculation of Replication-DeficientRecombinant Vaccinia Virus DIs Expressing Simian ImmunodeficiencyVirus Gag Controls Highly Pathogenic Simian-HumanImmunodeficiency Virus inMonkeys
Authors: Izumi Yasuyuki , Ami Yasushi , Matsuo Kazuhiro , Someya Kenji , Sata Tetsutaro , Yamamoto Naoki , Honda Mitsuo 
Published: November 2003

ABSTRACTTo be effective, a vaccine against human immunodeficiency virus type 1 (HIV-1) must induce virus-specific T-cell responses and it must be safe for use in humans. To address these issues, we developed a recombinant vaccinia virus DIs vaccine (rDIsSIVGag), which is nonreplicative in mammalian cells and expresses the full-lengthgaggene of simian immunodeficiency virus (SIV). Intravenous inoculation of 106PFU of rDIsSIVGag in cynomologus macaques induced significant levels of gamma interferon (IFN-γ) spot-forming cells (SFC) specific for SIV Gag. Antigen-specific lymphocyte proliferative responses were also induced and were temporally associated with the peak of IFN-γ SFC activity in each macaque. In contrast, macaques immunized with a vector control (rDIsLacZ) showed no significant induction of antigen-specific immune responses. After challenge with a highly pathogenic simian-human immunodeficiency virus (SHIV), CD4+T lymphocytes were maintained in the peripheral blood and lymphoid tissues of the immunized macaques. The viral set point in plasma was also reduced in these animals, which may be related to the enhancement of virus-specific intracellular IFN-γ+CD8+cell numbers and increased antibody titers after SHIV challenge. These results demonstrate that recombinant DIs has potential for use as an HIV/AIDS vaccine.
Lytic Viral Replication as a Contributor to theDetection of Epstein-Barr Virus in BreastCancer
Authors: Huang J. , Chen H. , Hutt-Fletcher L. , Ambinder R. F. , Hayward S. D. 
Published: November 2003

ABSTRACT
Epstein-Barr
virus (EBV) has an accepted association with the epithelial malignancy
nasopharyngeal carcinoma and has also been reported in other more
controversial carcinoma settings. Evaluation of EBV association with
epithelial carcinomas such as breast cancer would benefit from a better
understanding of the outcome of EBV infection of these cells. Cell-free
preparations of a green fluorescent protein-expressing virus, BX1, were
used to infect breast cancer cell lines, which were then examined for
EBV gene expression and viral genome copy number. Reverse
transcription-PCR analyses revealed that the cells supported a mixture
of latency II and lytic EBV gene expression. Lytic Zta and BMRF1
protein expression was detected by immunohistochemistry, and DNA PCR
analyses estimated an EBV copy number of 300 to 600 genomes per
infected cell. Evidence for lytic EBV expression was also found in
breast tissue, where reverse transcription-PCR analyses detected lytic
Zta transcripts in 7 of 10 breast carcinoma tissues and 4 of 10 normal
tissues from the same patients. Scattered cells immunoreactive for Zta
protein were also detectable in breast carcinoma. Quantitative
real-time PCR analysis of EBV-positive breast carcinoma tissues
suggested that less than 0.1% of the cells contained viral
genomes. We suggest that sporadic lytic EBV infection may contribute to
PCR-based detection of EBV in traditionally nonvirally associated
epithelial
malignancies.
Neutralizationof Enteric Coronaviruses withEscherichia coliCellsExpressing Single-Chain Fv-AutotransporterFusions
Authors: Veiga Esteban , de Lorenzo Víctor , Fernández Luis Angel 
Published: November 2003

ABSTRACTWe report here that fusions of single-chain antibodies (scFvs) to the autotransporter β domain of the IgA protease ofNeisseria gonorrhoeaeare instrumental in locating virus-neutralizing activity on the cell surface ofEscherichia coli.E. colicells displaying scFvs against the transmissible gastroenteritis coronavirus on their surface blocked in vivo the access of the infectious agent to cultured epithelial cells. This result raises prospects for antiviral strategies aimed at hindering the entry into target cells by bacteria that naturally colonize the same intestinal niches.
Expressionof Unmodified Hepatitis C Virus Envelope Glycoprotein-Coding SequencesLeads to Cryptic Intron Excision and Cell Surface Expression of E1/E2Heterodimers Comprising Full-Length and Partially DeletedE1
Authors: Dumonceaux Julie , Cormier Emmanuel G. , Kajumo Francis , Donovan Gerald P. , Roy-Chowdhury Jayanta , Fox Ira J. , Gardner Jason P. , Dragic Tatjana 
Published: November 2003

ABSTRACT
Hepatitis
C virus (HCV) is a positive-strand RNA virus that replicates
exclusively in the cytoplasm of infected cells. The viral envelope
glycoproteins, E1 and E2, appear to be retained in the endoplasmic
reticulum, where viral budding is thought to occur. Surprisingly, we
found that the expression system used to generate HCV envelope
glycoproteins influences their subcellular localization and processing.
These findings have important implications for optimizing novel HCV
fusion and entry assays as well as for budding and virus particle
formation.
CpG-A and CpG-B oligonucleotides differentially enhance human peptide–specific primary and memory CD8+ T-cell responses in vitro
Authors: Rothenfusser Simon , Hornung Veit , Ayyoub Maha , Britsch Stefanie , Towarowski Andreas , Krug Anne , Sarris Anja , Lubenow Norbert , Speiser Daniel , Endres Stefan , Hartmann Gunther 
Published: November 2003

AbstractTwo distinct types of CpG oligodeoxynucleotide (ODN) have been identified that differ in their capacity to stimulate antigen-presenting cells: CpG-A induces high amounts of interferon-α (IFN-α) and IFN-β in plasmacytoid dendritic cells (PDCs), whereas CpG-B induces PDC maturation and is a potent activator of B cells but stimulates only small amounts of IFN-α and IFN-β. Here we examined the ability of these CpG ODNs to enhance peptide-specific CD8+ T-cell responses in human peripheral blood mononuclear cells (PBMCs). The frequency of influenza matrix–specific “memory” CD8+ T cells was increased by both types of CpG ODN, whereas the frequency of Melan-A specific “naive” CD8+ T cells increased on stimulation with CpG-B but not with CpG-A. The presence of PDCs in PBMCs was required for this CpG ODN-mediated effect. The expanded cells were cytotoxic and produced IFN-γ on peptide restimulation. Soluble factors induced by CpG-A but not CpG-B increased the granzyme-B content and cytotoxicity of established CD8+ T-cell clones, each of which was IFN-α/-β dependent. In conclusion, CpG-B seems to be superior for priming CD8+ T-cell responses, and CpG-A selectively enhances memory CD8+ T-cell responses and induces cytotoxicity. These results demonstrate distinct functional properties of CpG-A and CpG-B with regard to CD8 T cells.