bluuebunny/crossref_metadata_2025 huggingface dataset:
Medial and Lateral Orbital Wall Surgery for Balanced Decompression in Thyroid Eye Disease
Authors: Graham Scott M. , Brown Christopher L. , Carter Keith D. , Song Alice , Nerad Jeffrey A.
Published: November 2003

Objectives/Hypothesis Diplopia remains a major potential complication of orbital decompression performed for thyroid eye disease. We sought to examine the effect of medial and lateral wall surgery with sparing of the orbital floor on the incidence of postoperative diplopia.

Study Design Retrospective review.

Methods A retrospective review was made of 63 consecutive medial and lateral orbital wall surgeries (40 patients) performed between December 1996 and May 2002 for orbital decompression of thyroid eye disease.

Results The average patient age was 50.9 years of age (age range, 14–83 y). The studied group was predominantly female (sex ratio, 36:4). Indications were compressive optic neuropathy (34), exposure keratopathy (25), and aesthetic concerns (4). The mean time since surgery was 31.5 months (range, 3–69 mo). The medial wall was approached by a transcaruncular (59) or endoscopic (4) technique. Two patients had fat removal. The average improvement in exophthalmos was 4.1 mm (range, 0–10 mm). The average palpebral fissure improvement was 2.0 mm (range, 0–7 mm). Two patients had cerebrospinal fluid leaks during excavation of the sphenoid bone. These were diagnosed and repaired primarily without complication. New‐onset diplopia occurred in four patients. Two of these patients required strabismus surgery.

Conclusion A 10% new‐onset diplopia rate compares favorably with other surgical series. Medial and lateral wall surgery only, sparing the floor, may reduce diplopia after surgery for orbital decompression in thyroid eye disease.
Expiratory Pharyngeal Airway Obstruction During Sleep: A Multiple Element Model
Authors: Woodson B. Tucker
Published: November 2003

Objective/Hypothesis In patients with obstructive sleep apnea and snoring, airway obstruction during sleep is not limited to inspiration but may also occur with expiration. The aim of this study was to assess the segmental mechanics of expiratory obstruction.

Design Experimental study of a convenience sample of 20 patients with snoring and mild obstructive sleep apnea.

Methods During sedated sleep, airflow, airway pressure measurements (supraglottic, oropharyngeal, nasopharyngeal, and nasal mask), and either supraglottic/retroglossal or retropalatal areas were simultaneously measured. Nasal continuous positive airway pressures were experimentally adjusted during single breath tests (SBTs) to modify upper airway size. Airway mechanics were evaluated during pressure drops on expiration.

Results The predominant level of expiratory obstruction was supraglottic/retroglossal level alone (65%) or combined supraglottic/retroglossal and retropalatal (17.6%). In nonobstructed SBTs, compliance curves derived from supraglottic/retroglossal and retropalatal pressures were similar but diverged in obstructed breaths. Compliance during expiration was greater in the supraglottic/retroglossal segment compared to the retropalatal segment. Retropalatal cross‐sectional size was smaller during early and late expiration on obstructed than on nonobstructed breaths independent of airway pressure measures. The rate of expiratory collapse was increased at all time points measured (P < .005) in the retropalatal segment on obstructed as compared with nonobstructed breaths.

Conclusions During expiration, the supraglottic/retroglossal level is obstructed more frequently and has greater compliance than the retropalatal segment. Failure of upstream pressures to describe pharyngeal obstruction supports a multi‐element model of collapse. Segments interact during expiration, with increased retropalatal collapse on obstructed as compared with nonobstructed breaths. Increased collapse on expiration provides a mechanism for increased obstruction on subsequent inspiratory breaths.
Safe tracheostomy for patients with severe acute respiratory syndrome
Authors: Wei William I. , Tuen Henry H. , Ng Raymond W. M. , Lam Lai Kun
Published: November 2003

Objectives/Hypothesis: Severe acute respiratory syndrome (SARS) caused by coronavirus has become an epidemic affecting many regions worldwide. Fourteen percent to 20% of patients require endotracheal intubation and ventilator support. Some of these patients may require tracheostomy subsequently. This procedure, when performed without protection, may lead to infection of the medical and nursing staff taking care of the patient. Study Design: Based on clinical information of three patients. Methods: The authors carried out an emergency tracheostomy and changed the tracheostomy tube for one patient and performed elective tracheostomy in another two patients. Results: No medical or nursing staff member was infected after carrying out the procedure while taking all the precautions and wearing the appropriate protective apparel. Conclusion: The authors have prepared guidelines for performing a safe tracheostomy under both elective and emergency conditions. Surgeons who might be involved in performing the tracheostomy should become familiar with these guidelines and the appropriate protective apparel.
External auditory canal foreign body removal: Management practices and outcomes
Authors: Thompson Scott K. , Wein Richard O. , Dutcher Paul O.
Published: November 2003

Objectives/Hypothesis: The purpose of the study was to evaluate the effectiveness of external auditory canal foreign body removal attempts by health care practitioners and to explore outcomes of patients with unsuccessful initial removal attempts. Study Design: Retrospective case series. Methods: A case series of patients presenting with external auditory canal foreign body to the emergency department of the authors' institution (Strong Memorial Hospital, University of Rochester Medical Center) over a 3‐year period was studied retrospectively. Medical records were reviewed, and information including age at presentation, type of foreign body, side of presentation, length of time in place, signs and symptoms at presentation, management practices, and outcomes was recorded. Results: One hundred sixty‐two patients with a diagnosis of external auditory canal foreign body were identified. Emergency personnel successfully managed 67% of patients using direct visualization techniques, and 33% required otolaryngological consultation. Otolaryngologists used otomicroscopy with standard otological instruments as their mainstay of management. Analysis of successfully managed emergency department cases revealed that 82% of foreign bodies were irregularly shaped objects with soft, graspable parts. Conversely, 72% of foreign bodies in otolaryngology referrals were firm, rounded objects such as beads and beans. Analysis of patients referred to otolaryngologists revealed a requirement for removal under anesthesia of 19%. Patients with a history of one or more removal attempts before emergency department evaluation universally failed further direct visualization techniques. A disproportionate number of these patients eventually required operative intervention and/or had tympanic membrane perforation. Patients with a history of previous removal attempt(s) who were referred directly to otolaryngologists were more likely to be successfully managed without general anesthesia. Conclusion: Emergency room personnel successfully manage the majority of patients with foreign bodies of the external auditory canal. For patients with firm, rounded objects, direct otolaryngology consultation without further manipulation should be strongly considered. Patients who have had previous removal attempts should not undergo further manipulation in the emergency department but rather should be referred directly to an otolaryngologist.
The accuracy of the modified Evan's blue dye test in predicting aspiration
Authors: Belafsky Peter C. , Blumenfeld Liza , Lepage Amanda , Nahrstedt Kristen
Published: November 2003

Objectives/Hypothesis: The modified Evan's blue dye test (MEBDT) is a relatively simple, inexpensive bedside procedure for the assessment of aspiration in the tracheotomized patient. Recent investigations have questioned its diagnostic accuracy. The purpose of the study was to evaluate the accuracy of the MEBDT in predicting aspiration among tracheotomized patients. Study Design: Prospective observational study. Methods: In the setting of a long‐term acute care hospital, all persons with a tracheotomy tube undergoing a bedside swallowing evaluation between October 1, 2001, and March 31, 2002, were prospectively evaluated. All individuals underwent a MEBDT and a subsequent fiberoptic endoscopic evaluation of swallowing (FEES) using a standardized protocol. The sensitivity and specificity of the MEBDT in predicting aspiration were determined. Results: Thirty persons were evaluated. The mean age of the cohort was 65 years (SD ± 11 y). Sixty percent (18 of 30) were men. The sensitivity and specificity of the MEBDT for the entire cohort were 82% and 38%, respectively. The sensitivity of the MEBDT for patients receiving mechanical ventilation was 100% compared with 76% for individuals not receiving mechanical ventilation. The specificity of the MEBDT remained low, regardless of ventilator status (33%–40%). Conclusion: The sensitivity of the MEBDT in predicting aspiration among individuals in our cohort was 82%. The sensitivity was even higher (100%) when performed on persons receiving mechanical ventilation. These results support the use of the MEBDT as a screening tool for persons with a tracheotomy tube. The specific technique of performing the MEBDT is imperative, and the results of the study must be differentiated from other reports evaluating the MEBDT that use a different test protocol.
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

Objectives/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

HAMLET (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

Proteins 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

E. 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

Oligosaccharide (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

Protective 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

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

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

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

To 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

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

We 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

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.
Activation of Apoptosis Signal-Regulating Kinase 1 in Injured Artery and Its Critical Role in Neointimal Hyperplasia
Authors: Izumi Yasukatsu , Kim Shokei , Yoshiyama Minoru , Izumiya Yasuhiro , Yoshida Kaoru , Matsuzawa Atsushi , Koyama Hidenori , Nishizawa Yoshiki , Ichijo Hidenori , Yoshikawa Junichi , Iwao Hiroshi
Published: November 2003

Background—

Apoptosis signal-regulating kinase 1 (ASK1), recently identified as one of the mitogen-activated protein kinase kinase kinases, is activated by various extracellular stimuli and involved in a variety of cellular function. Therefore, we first examined the role of ASK1 in vascular remodeling.

Methods and Results—

We used rat balloon injury model and cultured vascular smooth muscle cells (VSMCs). Arterial ASK1 activity was rapidly and dramatically increased after balloon injury. To specifically inhibit endogenous ASK1 activation, dominant-negative mutant of ASK1 (DN-ASK1) was transfected into rat carotid artery before balloon injury. Gene transfer of DN-ASK1 significantly prevented neointimal formation at 14 days after injury. Bromodeoxyuridine labeling index at 7 days after injury showed that DN-ASK1 remarkably suppressed VSMC proliferation in both the intima and the media. We also examined the role of ASK1 in cultured rat VSMCs. Infection with DN-ASK1 significantly attenuated serum-induced VSMC proliferation and migration. We also compared neointimal formation after cuff placement around the femoral artery between mice deficient in ASK1 (ASK1
−/−
mice) and wild-type (WT) mice. Neointimal formation at 28 days after cuff injury in ASK1
−/−
mice was significantly attenuated compared with WT mice. Furthermore, we compared the proliferation and migration of VSMCs isolated from ASK1
−/−
mice with WT mice. Both proliferation and migration of VSMCs from ASK1
−/−
mice were significantly attenuated compared with VSMCs from WT mice.

Conclusions—

ASK1 activation plays the key role in vascular intimal hyperplasia. ASK1 may provide the basis for the development of new therapeutic strategy for vascular diseases.
Recurrent Thromboembolism in Infants and Children Suffering From Symptomatic Neonatal Arterial Stroke
Authors: Kurnik Karin , Kosch Andrea , Sträter Ronald , Schobess Rosemarie , Heller Christine , Nowak-Göttl Ulrike
Published: November 2003

Background and Purpose—

The present study was performed to evaluate the rate of recurrent symptomatic thromboembolism with respect to prothrombotic risk factors and underlying clinical conditions.

Methods—

In a series of 215 consecutively enrolled neonates with arterial ischemic stroke (AIS), the factor V G1691A mutation, factor II G20210A variant, methylenetetrahydrofolate reductase (MTHFR) T677T genotype, lipoprotein (Lp) (a), antithrombin, protein C, protein S, and anticardiolipin antibodies (ACA) were investigated. Patient median follow-up was 3.5 years (range, 1 to 8 years).

Results—

During follow-up, 7 infants and children (3.3%) showed recurrent symptomatic thromboembolism (AIS, n=4; venous sinus thrombosis, n=2; deep vein thrombosis of the leg, n=1). The factor V mutation, factor II variant, elevated Lp(a) >30 mg/dL, protein C deficiency, and protein S or antithrombin deficiency were associated with first stroke onset. In 5 of 7 cases (71.4%), prothrombotic risk factors [MTHFR T677T, elevated Lp(a), hyperhomocysteinemia, protein C deficiency] were involved at the time of recurrence. Furthermore, a second thromboembolic event was triggered additionally by underlying diseases (71%), eg, cardiac malformation and immobilization, diarrhea, mastoiditis, and moyamoya syndrome.

Conclusions—

Data shown here give evidence that symptomatic recurrent thromboembolism is not common in children with neonatal AIS. The risk of a second event, however, is increased when underlying diseases occur and prothrombotic risk factors are involved.
Thomas Wilfred and Intermedia: Seeking a Framework for Lumia
Authors: Eskilson Stephen
Published: November 2003

The most successful early-20th-century artist of colored light in the United States was undoubtedly Thomas Wilfred (1889–1968). In the 1920s, his “Lumia” compositions were praised by art critics and performed throughout the U.S. After initially embracing a musical analogy to explain Lumia, in the early 1930s he shifted to an analogy based on painting. In pursuit of this new context, Wilfred sought to legitimize Lumia through a relationship with the Museum of Modern Art in New York. His career is emblematic of the difficulties inherent in the creation of art using technology early in the 20th century, years before the postmodern embrace of pluralism.
Did Diplomatic Immunity Exist in the Ancient Near East?
Authors:   Elgavish
Published: November 2003

No Abstract
Humphrey and the High Commissioner
Authors:   Hobbins
Published: November 2003

No Abstract
Antony Carty and Richard Smith. Sir Gerald Fitzmaurice and the World Crisis
Authors:   Johnston
Published: November 2003

No Abstract
Legal Education in the People's Republic of China
Authors:   Mo,   Li
Published: November 2003

No Abstract
My Visits with Professor Wang Tieya
Authors:   van Dijk
Published: November 2003

No Abstract
King Magnus Eriksson's Law of the Realm (a Medieval Swedish code)
Authors:   Thomas
Published: November 2003

No Abstract
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

Two 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.
The FIP1L1-PDGFRα fusion tyrosine kinase in hypereosinophilic syndrome and chronic eosinophilic leukemia: implications for diagnosis, classification, and management
Authors: Gotlib Jason , Cools Jan , Malone James M. , Schrier Stanley L. , Gilliland D. Gary , Coutré Steven E.
Published: November 2003

Idiopathic hypereosinophilic syndrome (HES) and chronic eosinophilic leukemia (CEL) comprise a spectrum of indolent to aggressive diseases characterized by unexplained, persistent hypereosinophilia. These disorders have eluded a unique molecular explanation, and therapy has primarily been oriented toward palliation of symptoms related to organ involvement. Recent reports indicate that HES and CEL are imatinib-responsive malignancies, with rapid and complete hematologic remissions observed at lower doses than used in chronic myelogenous leukemia (CML). These BCR-ABL–negative cases lack activating mutations or abnormal fusions involving other known target genes of imatinib, implicating a novel tyrosine kinase in their pathogenesis. A bedside-to-benchtop translational research effort led to the identification of a constitutively activated fusion tyrosine kinase on chromosome 4q12, derived from an interstitial deletion, that fuses the platelet-derived growth factor receptor-α gene (PDGFRA) to an uncharacterized human gene FIP1-like-1 (FIP1L1). However, not all HES and CEL patients respond to imatinib, suggesting disease heterogeneity. Furthermore, approximately 40% of responding patients lack the FIP1L1-PDGFRA fusion, suggesting genetic heterogeneity. This review examines the current state of knowledge of HES and CEL and the implications of the FIP1L1-PDGFRA discovery on their diagnosis, classification, and management. (Blood. 2004;103:2879-2891)
Role of NKG2D signaling in the cytotoxicity of activated and expanded CD8+ T cells
Authors: Verneris Michael R. , Karami Mobin , Baker Jeanette , Jayaswal Anishka , Negrin Robert S.
Published: November 2003

Activating and expanding T cells using T-cell receptor (TCR) cross-linking antibodies and interleukin 2 (IL-2) results in potent cytotoxic effector cells capable of recognizing a broad range of malignant cell targets, including autologous leukemic cells. The mechanism of target cell recognition has previously been unknown. Recent studies show that ligation of NKG2D on natural killer (NK) cells directly induces cytotoxicity, whereas on T cells it costimulates TCR signaling. Here we demonstrate that NKG2D expression is up-regulated upon activation and expansion of human CD8+ T cells. Antibody blocking, redirected cytolysis, and small interfering RNA (siRNA) studies using purified CD8+ T cells demonstrate that cytotoxicity against malignant target cells occurs through NKG2D-mediated recognition and signaling and not through the TCR. Activated and expanded CD8+ T cells develop cytotoxicity after 10 to 14 days of culture, coincident with the expression of the adapter protein DAP10. T cells activated and expanded in low (30 U/mL) and high (300 U/mL) concentrations of IL-2 both up-regulated NKG2D expression equally, but only cells cultured in high-dose IL-2 expressed DAP10 and were cytotoxic. Collectively these results establish that NKG2D triggering accounts for the majority of major histocompatibility complex (MHC)–unrestricted cytotoxicity of activated and expanded CD8+ T cells, likely through DAP10-mediated signaling. (Blood. 2004;103: 3065-3072)
Effect of Mthfr genotype on diet-induced hyperhomocysteinemia and vascular function in mice
Authors: Devlin Angela M. , Arning Erland , Bottiglieri Teodoro , Faraci Frank M. , Rozen Rima , Lentz Steven R.
Published: November 2003

Deficiency of methylenetetrahydrofolate reductase (MTHFR) predisposes to hyperhomocysteinemia and vascular disease. We tested the hypothesis that heterozygous disruption of the Mthfr gene sensitizes mice to diet-induced hyperhomocysteinemia and endothelial dysfunction. Mthfr+/- and Mthfr+/+ mice were fed 1 of 4 diets: control, high methionine (HM), low folate (LF), or high methionine/low folate (HM/LF). Plasma total homocysteine (tHcy) was higher with the LF and HM/LF diets than the control (P < .01) or HM (P < .05) diets, and Mthfr+/- mice had higher tHcy than Mthfr+/+ mice (P < .05). With the control diet, the S-adenosylmethionine (SAM) to S-adenosylhomocysteine (SAH) ratio was lower in the liver and brain of Mthfr+/- mice than Mthfr+/+ mice (P < .05). SAM/SAH ratios decreased further in Mthfr+/+ or Mthfr+/- mice fed LF or LF/HM diets (P < .05). In cerebral arterioles, endothelium-dependent dilation to 1 or 10 μM acetylcholine was markedly and selectively impaired with the HM/LF diet compared with the control diet for both Mthfr+/+ (maximum dilation 5% ± 2% versus 21% ± 4%; P < .01) and Mthfr+/- (6% ± 2% versus 21% ± 3%; P < .01) mice. These findings demonstrate that the Mthfr+/- genotype sensitizes mice to diet-induced hyperhomocysteinemia and that hyperhomocysteinemia alters tissue methylation capacity and impairs endothelial function in cerebral microvessels.
Immunodeficiency virus uptake, turnover, and 2-phase transfer in human dendritic cells
Authors: Turville Stuart G. , Santos John J. , Frank Ines , Cameron Paul U. , Wilkinson John , Miranda-Saksena Monica , Dable Joanne , Stössel Hella , Romani Nikolaus , Piatak Michael , Lifson Jeffrey D. , Pope Melissa , Cunningham Anthony L.
Published: November 2003

HIV-1 subverts antigen processing in dendritic cells (DCs) resulting in viral uptake, infection, and transfer to T cells. Although DCs bound monomeric gp120 and HIV-1 similarly, virus rarely colocalized with endolysosomal markers, unlike gp120, suggesting HIV-1 alters endolysosomal trafficking. Virus within DC intracellular compartments rapidly moved to DC-CD4+ lymphocyte synapses when introduced to CD4+ lymphocyte cultures. Although viral harboring and transfer from nonlysosomal compartments was transient, given DC-associated virus protein, nucleic acids, and infectious HIV-1 transfer to CD4+, lymphocytes decayed within 24 hours. However a second long-term transfer phase was apparent in immature DCs after 48 hours as a zidovudine-sensitive rise in proviral DNA. Therefore, DCs transfer HIV-1 to CD4+ lymphocytes in 2 distinct phases. Immature and mature DCs first divert virus from the endolysosomal pathway to the DC–T-cell synapse. Secondly, the later transfer phase from immature DCs is through de novo HIV-1 production. Thus, the controversy of DCs being infected or not infected for the mechanics of viral transfer to CD4+ lymphocytes can be addressed as a function of time.
Monoclonal CD5+ and CD5- B-lymphocyte expansions are frequent in the peripheral blood of the elderly
Authors: Ghia Paolo , Prato Giuseppina , Scielzo Cristina , Stella Stefania , Geuna Massimo , Guida Giuseppe , Caligaris-Cappio Federico
Published: November 2003

The responsiveness and diversity of peripheral B-cell repertoire decreases with age, possibly because of B-cell clonal expansions, as suggested by the incidence of serum monoclonal immunoglobulins and of monoclonal chronic lymphocytic leukemia (CLL)–like B lymphocytes in clinically silent adults. We phenotyped peripheral blood cells from 500 healthy subjects older than 65 years with no history or suspicion of malignancies and no evidence of lymphocytosis. In 19 cases (3.8%) a κ/λ ratio of more than 3:1 or less than 1:3 was found: 9 were CD5+, CD19+, CD23+, CD20low, CD79blow, sIglow (classic CLL-like phenotype); 3 were CD5+, CD19+, CD23+, CD20high, CD79blow, sIglow (atypical CLL-like), and 7 were CD5-, CD19+, CD20high, CD23-, CD79bbright, FMC7+, sIgbright (non–CLL-like). In 2 subjects, 2 phenotypically distinct unrelated clones were concomitantly evident. No cases were CD10+. Polymerase chain reaction (PCR) analysis demonstrated a monoclonal rearrangement of IgH genes in 15 of 19 cases. No bcl-1 or bcl-2 rearrangements were detected. Using a gating strategy based on CD20/CD5/CD79 expression, 13 additional CLL-like B-cell clones were identified (cumulative frequency of classic CLL-like: 5.5%). Thus, phenotypically heterogeneous monoclonal B-lymphocyte expansions are common among healthy elderly individuals and are not limited to classic CLL-like clones but may have the phenotypic features of different chronic lymphoproliferative disorders, involving also CD5- B cells.
The bHLH genesGLABRA3 (GL3)andENHANCER OF GLABRA3(EGL3)specify epidermal cell fate in theArabidopsisroot
Authors: Bernhardt Christine , Lee Myeong Min , Gonzalez Antonio , Zhang Fan , Lloyd Alan , Schiefelbein John
Published: November 2003

The position-dependent specification of the hair and non-hair cell types in the Arabidopsis root epidermis provides a simple model for the study of cell fate determination in plants. Several putative transcriptional regulators are known to influence this cell fate decision. Indirect evidence from studies with the maize R gene has been used to suggest that a bHLH transcription factor also participates in this process. We show that two Arabidopsis genes encoding bHLH proteins, GLABRA3(GL3) and ENHANCER OF GLABRA3 (EGL3), act in a partially redundant manner to specify root epidermal cell fates. Plants homozygous for mutations in both genes fail to specify the non-hair cell type,whereas plants overexpressing either gene produce ectopic non-hair cells. We also find that these genes are required for appropriate transcription of the non-hair specification gene GL2 and the hair cell specification gene CPC, showing that GL3 and EGL3 influence both epidermal cell fates. Furthermore, we show that these bHLH proteins require a functional WER MYB protein for their action, and they physically interact with WER and CPC in the yeast two-hybrid assay. These results suggest a model in which GL3 and EGL3 act together with WER in the N cell position to promote the non-hair cell fate, whereas they interact with the incomplete MYB protein CPC in the H position, which blocks the non-hair pathway and leads to the hair cell fate.