bluuebunny/crossref_metadata_2025 huggingface dataset:
Performance of the PHQ-9 as a Screening Tool for Depression After Stroke
Authors: Williams Linda S. , Brizendine Edward J. , Plue Laurie , Bakas Tamilyn , Tu Wanzhu , Hendrie Hugh , Kroenke Kurt
Published: January 2005

Background and Purpose—

The purpose of this study was to examine the performance of the Patient Health Questionnaire (PHQ)-9, a 9-item depression scale, as a screening and diagnostic instrument for assessing depression in stroke survivors.

Methods—

As part of a randomized treatment trial for poststroke depression (PSD), subjects with and without PSD completed the PHQ-9, a 9-item summed scale, with scores ranging from 0 (no depressive symptoms) to 27 (all symptoms occurring daily). Subjects endorsing 2 or more symptoms of depression were administered the criterion standard Structured Clinical Interview for Depression (SCID). Receiver operating characteristic analysis was used to examine the sensitivity and specificity of the PHQ-9

Results—

Of 316 subjects enrolled, 145 met SCID criteria for major depression or other depressive disorder, and 171 were not depressed. PHQ-9 scores discriminated well between subjects with any versus no depressive disorder, with an area under the curve (AUC) of 0.96, as well as between subjects with and without major depression (AUC=0.96). The AUC was similar regardless of patient age, gender, or ethnicity. A PHQ-9 score ≥10 had 91% sensitivity and 89% specificity for major depression, and 78% sensitivity and 96% specificity for any depression diagnosis.

Conclusions—

The PHQ-9 performs well as a brief screener for PSD with operating characteristics similar or superior to other depression measures and similar to its characteristics in a primary care population. Moreover, PHQ-9 scores discriminate equally well between those with and without PSD regardless of age, gender, or ethnicity.
Are We Saving Enough? The Macroeconomics of the Savings Gap
Authors: Pomerantz Olga , Weale Martin
Published: January 2005

We assess the adequacy of saving in the United Kingdom from a macroeconomic perspective. We present conditions needed for wealth to grow in line with income and discuss the circumstances in which capital gains can represent a substitute for saving. We show that reference to balance sheets can be misleading because they fail to identify the cost of future consumption. A savings gap of between £16.5 bn and £66 bn is identified before taking account of the effects of demographic change. The outcome depends on whether one regards rising house prices as a consequence of fixed supplies of land or as the outcome of failing to invest enough in housing with the upper figure arising in the latter case. Policy measures to reduce the savings gap are discussed. These include appropriate use of fiscal policy and taxation of credit.
Policy Design and Macroeconomic Stability in Europe
Authors: Barrell Ray , Davis E. Philip
Published: January 2005

Macroeconomic policy in Europe is now oriented to creating a stable environment in which the scope for output growth is enhanced. However, we maintain that not all dimensions of a stability-oriented policy framework appear to be in place. Fiscal policy rules and arrangements have been much discussed, but their design is not yet settled. The Single Market Programme has transformed competition in Europe, but its full implications for macroeconomic stability, especially its implications for financial market stability in combination with the Single Currency, have not yet been fully appreciated by policymakers. Future pension issues in the context of population ageing will pose a major challenge. We discuss the design of fiscal policy in (a Single Market) Europe, looking at fiscal pacts and the need for Europe-wide financial regulation in an integrated financial market as well as pension reform aspects.
Breastfeeding the Premature Infant: Experience of a Baby-Friendly Hospital in Brazil
Authors: Reinert do Nascimento Maria Beatriz , Issler Hugo
Published: January 2005

Breastfeeding is the natural way of feeding infants, providing nutritional, immunological, psychological, and economic advantages. These qualities are especially important in premature neonates because of their vulnerability. Nevertheless, a lower incidence of successful breastfeeding has been observed in this population. This prospective study investigated the frequency of mother’s milk use on discharge of premature neonates from an at-risk neonatal unit. It involved 244 premature neonates and took place at a Brazilian Baby-Friendly Hospital. The observed frequency of mother’s milk use at neonatal unit discharge was 94.6%. The finding that most premature neonates were receiving mother’s milk reflects a well-structured program for promoting breastfeeding of premature neonates at this institution, which relies on an interdisciplinary team to provide adequate support to the mothers.
Probiotic Potential of 3 Lactobacilli Strains Isolated From Breast Milk
Authors: Martín Rocío , Olivares Mónica , Marín María L. , Fernández Leonides , Xaus Jordi , Rodríguez Juan M.
Published: January 2005

Breast milk is an important factor in the initiation, development, and composition of the neonatal gut microbiota. In a previous study, the authors isolated lactic acid bacteria from milk of healthy mothers. Since some of the identified isolates belonged to the genus Lactobacillus, the objective of this work was to evaluate the probiotic potential of 2 Lactobacillus gasseri and 1 Lactobacillus fermentu strains. Different assays, including survival to conditions simulating those existing in the gastrointestinal tract, production of antimicrobial compounds, adherence to intestinal cells, production of biogenic amines, degradation of mucin, enzymatic profile, and pattern of antibiotic resistance, were performed. Globally, the results showed that the probiotic potential of lactobacilli isolated from milk of healthy mothers is, at least, similar to that of the strains commonly used in commercial probiotic products. This fact, together with the presence of prebiotic substances, indicates that breast milk is a natural synbiotic food. J Hum Lact. 21(1):8-17.
Comparison of Milk Output Between Mothers of Preterm and Term Infants: The First 6 Weeks After Birth
Authors: Hill Pamela D. , Aldag Jean C. , Chatterton Robert T. , Zinaman Michael
Published: January 2005

This study reports the temporal pattern of milk output in 2 groups of lactating mothers during the first 6 weeks postpartum. A study aim was to examine if the average milk output for postpartum days 6 and 7 (baseline) predicts milk adequacy at week 6 postpartum. Mothers of preterm (= 31 weeks) infants (n = 95) used mechanical expression to initiate and maintain their milk supply; mothers of a singleton healthy term infant (n = 98) fed their infant at the breast. Baseline milk output was predictive of milk adequacy, defined as = 500 mL/d at week 6 (P = .000). Preterm mothers were 2.81 times more at risk of not producing adequately than term mothers were. Study findings suggest that interventions that promote the initiation and maintenance of an adequate milk supply during the first week postbirth are critical.
Safety and Efficacy of Testosterone Gel 1% Augmentation in Depressed Men With Partial Response to Antidepressant Therapy
Authors: Orengo Claudia A. , Fullerton Laura , Kunik Mark E.
Published: January 2005

The current study evaluates the efficacy and safety of testosterone (T) gel 1% augmentation on depressive symptoms and quality of life in treatment-resistant, depressed, hypogonadal men older than 50 years of age who are receiving antidepressants. The authors hypothesized that T augmentation would improve depressive symptoms and quality of life. Eighteen hypogonadal men entered the study who had had an adequate trial of antidepressant therapy and had significant depressive symptoms. Participants were continued on their antidepressant and were randomized to receive either placebo or active T gel (5 g) to be applied once a day. Participants were tested on 6 occasions: screening visit, an initial session (pretreatment), at 6 and 12 weeks during the first treatment condition, and at 18 and 24 weeks during the crossover condition. The authors found a significant improvement in depressive symptoms from baseline to 12 weeks of testosterone treatment. However, a statistical difference between placebo and testosterone treatment phases was not demonstrated. The limitations of the study, including the chronicity and severity of patients’ depression, variability in T levels, and a small sample size, probably influenced the ability to detect a discernable difference. Nevertheless, the study shows that T gel augmentation may be helpful in hypogonadal males with depression. ( J Geriatr Psychiatry Neurol 2005; 18:20-24)
Regulation of endothelial thrombomodulin expression by inflammatory cytokines is mediated by activation of nuclear factor-kappa B
Authors: Sohn Richard H. , Deming Clayton B. , Johns David C. , Champion Hunter C. , Bian Ce , Gardner Kevin , Rade Jeffrey J.
Published: January 2005

Inflammation and thrombosis are increasingly recognized as interrelated biologic processes. Endothelial cell expression of thrombomodulin (TM), a key component of the anticoagulant protein C pathway, is potently inhibited by inflammatory cytokines. Because the mechanism underlying this effect is largely unknown, we investigated a potential role for the inflammatory transcription factor nuclear factor-kappa B (NF-κB). Blocking NF-κB activation effectively prevented cytokine-induced down-regulation of TM, both in vitro and in a mouse model of tumor necrosis factor-α (TNF-α)–mediated lung injury. Although the TM promoter lacks a classic NF-κB consensus site, it does contain tandem Ets transcription factor binding sites previously shown to be important for both constitutive TM gene expression and cytokine-induced repression. Using electrophoretic mobility shift assay and chromatin immunoprecipitation, we found that multiple Ets species bind to the TNF-α response element within the TM promoter. Although cytokine exposure did not alter Ets factor binding, it did reduce binding of p300, a coactivator required by Ets for full transcriptional activity. Overexpression of p300 also prevented TM repression by cytokines. We conclude that NF-κB is a critical mediator of TM repression by cytokines. Further evidence suggests a mechanism involving competition by NF-κB for limited pools of the transcriptional coactivator p300 necessary for TM gene expression.
Induction of polyploidization in leukemic cell lines and primary bone marrow by Src kinase inhibitor SU6656
Authors: Lannutti Brian J. , Blake Noel , Gandhi Manish J. , Reems Jo Anna , Drachman Jonathan G.
Published: January 2005

Megakaryocytes (MKs) undergo successive rounds of endomitosis during differentiation, resulting in polyploidy (typically, 16-64N). Previous studies have demonstrated that this occurs through an interruption of normal cell cycle progression during anaphase. However, the molecular mechanism(s) controlling this unique process is undefined. In the present report, we examine the effect of an Src kinase inhibitor, SU6656, on thrombopoietin (TPO)-induced growth and differentiation. Remarkably, when SU6656 (2.5 μM) was added to a megakaryocytic cell line, UT-7/TPO, the cells ceased cell division but continued to accumulate DNA by endomitosis. During this interval, CD41 and CD61 expression on the cell surface increased. Similar effects on polyploidization and MK differentiation were seen with expanded primary MKs, bone marrow from 2 patients with myelodysplastic syndrome, and other cell lines with MK potential. Our data suggest that SU6656 might be useful as a differentiation-inducing agent for MKs and is an important tool for understanding the molecular basis of MK endomitosis.
Autocrine- and paracrine-activated receptor tyrosine kinases in classic Hodgkin lymphoma
Authors: Renné Christoph , Willenbrock Klaus , Küppers Ralf , Hansmann Martin-Leo , Bräuninger Andreas
Published: January 2005

The pathogenesis of Hodgkin lymphoma (HL) is still largely unknown. Based on a search for footprints of pathogenetic mechanisms in global RNA expression data of Hodgkin/Reed-Sternberg (HRS) cell lines, we analyzed the expression and activation of 6 receptor tyrosine kinases (RTKs) in classic HL. Immunohistochemistry revealed that the RTKs platelet-derived growth factor receptor A (PDGFRA), DDR2, EPHB1, RON, TRKB, and TRKA were each expressed in HRS cells in 30% to 75% of patients. These RTKs were not expressed in normal B cells, the origin of HRS cells, or in most B-cell non-Hodgkin lymphoma (NHL). In the majority of patients at least one RTK was expressed, and in most patients several RTKs were coexpressed, most prominently in Hodgkin lymphoma of the nodular sclerosis subtype. Phosphotyrosine-specific antibodies revealed exemplarily the activation of PDGFRA and TRKA/B and an elevation of cellular phosphotyrosine content. Immunohistochemistry for RTK ligands indicated that DDR2 and TRKA are likely activated in a paracrine fashion, whereas PDGFRA and EPHB1 seem to be activated by autocrine loops. Activating mutations were not detected in cDNA encoding the RTKs in HRS cell lines. These findings show the unprecedented coexpression of multiple RTKs in a tumor and indicate that aberrant RTK signaling is an important factor in HL pathogenesis and that it may be a novel therapeutic target.
Characterization of gana-1, a Caenorhabditis elegans gene encoding a single ortholog of vertebrate α-galactosidase and α-N-acetylgalactosaminidase
Authors: Hujová Jana , Sikora Jakub , Dobrovolný Robert , Poupětová Helena , Ledvinová Jana , Kostrouchová Marta , Hřebíček Martin
Published: January 2005

Background

Human α-galactosidase A (α-GAL) and α-N-acetylgalactosaminidase (α-NAGA) are presumed to share a common ancestor. Deficiencies of these enzymes cause two well-characterized human lysosomal storage disorders (LSD) – Fabry (α-GAL deficiency) and Schindler (α-NAGA deficiency) diseases. Caenorhabditis elegans was previously shown to be a relevant model organism for several late endosomal/lysosomal membrane proteins associated with LSDs. The aim of this study was to identify and characterize C. elegans orthologs to both human lysosomal luminal proteins α-GAL and α-NAGA.

Results

BlastP searches for orthologs of human α-GAL and α-NAGA revealed a single C. elegans gene (R07B7.11) with homology to both human genes (α-ga lactosidase and α-N-a cetylgalactosaminidase) – gana-1. We cloned and sequenced the complete gana-1 cDNA and elucidated the gene organization.

Phylogenetic analyses and homology modeling of GANA-1 based on the 3D structure of chicken α-NAGA, rice α-GAL and human α-GAL suggest a close evolutionary relationship of GANA-1 to both human α-GAL and α-NAGA.

Both α-GAL and α-NAGA enzymatic activities were detected in C. elegans mixed culture homogenates. However, α-GAL activity on an artificial substrate was completely inhibited by the α-NAGA inhibitor, N-acetyl-D-galactosamine.

A GANA-1:: GFP fusion protein expressed from a transgene, containing the complete gana-1 coding region and 3 kb of its hypothetical promoter, was not detectable under the standard laboratory conditions. The GFP signal was observed solely in a vesicular compartment of coelomocytes of the animals treated with Concanamycin A (CON A) or NH4Cl, agents that increase the pH of the cellular acidic compartment.

Immunofluorescence detection of the fusion protein using polyclonal anti-GFP antibody showed a broader and coarsely granular cytoplasmic expression pattern in body wall muscle cells, intestinal cells, and a vesicular compartment of coelomocytes.

Inhibition of gana-1 by RNA interference resulted in a decrease of both α-GAL and α-NAGA activities measured in mixed stage culture homogenates but did not cause any obvious phenotype.

Conclusions

GANA-1 is a single C. elegans ortholog of both human α-GAL and α-NAGA proteins. Phylogenetic, homology modeling, biochemical and GFP expression analyses support the hypothesis that GANA-1 has dual enzymatic activity and is localized in an acidic cellular compartment.
CASCAD: a database of annotated candidate single nucleotide polymorphisms associated with expressed sequences
Authors: Guryev Victor , Berezikov Eugene , Cuppen Edwin
Published: January 2005

Background

With the recent progress made in large-scale genome sequencing projects a vast amount of novel data is becoming available. A comparative sequence analysis, exploiting sequence information from various resources, can be used to uncover hidden information, such as genetic variation. Although there are enormous amounts of SNPs for a wide variety of organisms submitted to NCBI dbSNP and annotated in most genome assembly viewers like Ensembl and the UCSC Genome Browser, these platforms do not easily allow for extensive annotation and incorporation of experimental data supporting the polymorphism. However, such information is very important for selecting the most promising and useful candidate polymorphisms for use in experimental setups.

Description

The CASCAD database is designed for presentation and query of candidate SNPs that are retrieved by in silico mining of high-throughput sequencing data. Currently, the database provides collections of laboratory rat (Rattus norvegicus) and zebrafish (Danio rerio) candidate SNPs. The database stores detailed information about raw data supporting the candidate, extensive annotation and links to external databases (e.g. GenBank, Ensembl, UniGene, and LocusLink), verification information, and predictions of a potential effect for non-synonymous polymorphisms in coding regions. The CASCAD website allows search based on an arbitrary combination of 27 different parameters related to characteristics like candidate SNP quality, genomic localization, and sequence data source or strain. In addition, the database can be queried with any custom nucleotide sequences of interest. The interface is crosslinked to other public databases and tightly coupled with primer design and local genome assembly interfaces in order to facilitate experimental verification of candidates.

Conclusions

The CASCAD database discloses detailed information on rat and zebrafish candidate SNPs, including the raw data underlying its discovery. An advanced web-based search interface http://cascad.niob.knaw.nl allows universal access to the database content and allows various queries supporting many types of research utilizing single nucleotide polymorphisms.
FAM20: an evolutionarily conserved family of secreted proteins expressed in hematopoietic cells
Authors: Nalbant Demet , Youn Hyewon , Nalbant S Isil , Sharma Savitha , Cobos Everardo , Beale Elmus G , Du Yang , Williams Simon C
Published: January 2005

Background

Hematopoiesis is a complex developmental process controlled by a large number of factors that regulate stem cell renewal, lineage commitment and differentiation. Secreted proteins, including the hematopoietic growth factors, play critical roles in these processes and have important biological and clinical significance. We have employed representational difference analysis to identify genes that are differentially expressed during experimentally induced myeloid differentiation in the murine EML hematopoietic stem cell line.

Results

One identified clone encoded a previously unidentified protein of 541 amino acids that contains an amino terminal signal sequence but no other characterized domains. This protein is a member of family of related proteins that has been named family with sequence similarity 20 (FAM20) with three members (FAM20A, FAM20B and FAM20C) in mammals. Evolutionary comparisons revealed the existence of a single FAM20 gene in the simple vertebrate Ciona intestinalis and the invertebrate worm Caenorhabditis elegans and two genes in two insect species, Drosophila melanogaster and Anopheles gambiae. Six FAM20 family members were identified in the genome of the pufferfish, Fugu rubripes and five members in the zebrafish, Danio rerio. The mouse Fam20a protein was ectopically expressed in a mammalian cell line and found to be a bona fide secreted protein and efficient secretion was dependent on the integrity of the signal sequence. Expression analysis revealed that the Fam20a gene was indeed differentially expressed during hematopoietic differentiation and that the other two family members (Fam20b and Fam20c) were also expressed during hematcpoiesis but that their mRNA levels did not vary significantly. Likewise FAM20A was expressed in more limited set of human tissues than the other two family members.

Conclusions

The FAM20 family represents a new family of secreted proteins with potential functions in regulating differentiation and function of hematopoietic and other tissues. The Fam20a mRNA was only expressed during early stages of hematopoietic development and may play a role in lineage commitment or proliferation. The expansion in gene number in different species suggests that the family has evolved as a result of several gene duplication events that have occurred in both vertebrates and invertebrates.
Half versus full vacuum suction drainage after modified radical mastectomy for breast cancer- a prospective randomized clinical trial[ISRCTN24484328]
Authors: Chintamani  , Singhal Vinay , Singh JP , Bansal Anju , Saxena Sunita
Published: January 2005

Background

Suction drains are routinely used after modified radical mastectomy and are an important factor contributing to increased hospital stay as the patients are often discharged only after their removal. Amongst various factors that influence the amount of postoperative drainage, the negative suction pressure applied to the drain has been reported to be of great significance. While a high negative suction pressure is expected to drain the collection and reduce the dead space promptly, it may also prevent the leaking lymphatics from closing and lead to increased drainage from the wound. Against this background a prospective randomized clinical study was conducted to compare the amount and duration of drainage between a half negative suction and full vacuum suction drainage in patients following modified radical mastectomy. The associated postoperative morbidity was also compared between the two groups.

Methods

85 FNAC (fine needle aspiration cytology) proven cases of locally advanced breast cancer were randomized. (Using randomly ordered sealed envelops, which were opened immediately before the closure of the wound) in to 50 patients with full vacuum suction (pressure = 700 g/m2) and 35 cases in to half vacuum suction drainage (pressure = 350 g/m2) groups. The two groups were comparable in respect of age, weight, and technique of operation and extent of axillary dissection. Surgery was performed by the same surgical team comprising of five surgeons (two senior and three resident surgeons) using a standardized technique with electrocautery. External compression dressing was provided over the axilla for first 48 hrs and following that patients were encouraged to do active and passive shoulder exercises. The outcomes measured were postoperative morbidity and the length of hospital stay.

Statistical methods used: Descriptive studies were performed with SPSS version 10 and group characteristics were compared using student t-test.

Results

Half vacuum suction drains were removed earlier than the full suction vacuum suction drains. There was no significant difference in the incidence of seroma formation in the two groups and there was a significant reduction in the total hospital stay in patients with half vacuum suction drainage systems as compared to the full suction drainage group (p < 0.001) without any added morbidity.

Conclusions

Half negative suction drains provide an effective compromise between no suction and full or high suction drainage after modified radical mastectomy by reducing the hospital stay and the post operative morbidity including post operative seromas.
The importance of socio-economic context for social marketing models for improving reproductive health: Evidence from 555 years of program experience
Authors: Meekers Dominique , Rahaim Stephen
Published: January 2005

Background

Over the past two decades, social marketing programs have become an important element of the national family planning and HIV prevention strategy in several developing countries. As yet, there has not been any comprehensive empirical assessment to determine which of several social marketing models is most effective for a given socio-economic context. Such an assessment is urgently needed to inform the design of future social marketing programs, and to avoid that programs are designed using an ineffective model.

Methods

This study addresses this issue using a database of annual statistics about reproductive health oriented social marketing programs in over 70 countries. In total, the database covers 555 years of program experience with social marketing programs that distribute and promote the use of oral contraceptives and condoms. Specifically, our analysis assesses to what extent the model used by different reproductive health social marketing programs has varied across different socio-economic contexts. We then use random effects regression to test in which socio-economic context each of the models is most successful at increasing use of socially marketed oral contraceptives and condoms.

Results

The results show that there has been a tendency to design reproductive health social marketing program with a management structure that matches the local context. However, the evidence also shows that this has not always been the case. While socio-economic context clearly influences the effectiveness of some of the social marketing models, program maturity and the size of the target population appear equally important.

Conclusions

To maximize the effectiveness of future social marketing programs, it is essential that more effort is devoted to ensuring that such programs are designed using the model or approach that is most suitable for the local context.
Use of email for patient communication in student health care: a cross-sectional study
Authors: Castrén Johanna , Niemi Marja , Virjo Irma
Published: January 2005

Background

Citizens increasingly use email in personal communication. It is not however clear to what extent physicians utilize it for patient communication. Our study was designed to examine physicians' activity in using email and to estimate the proportion of email messages missing from documentation in electronic patient records (EPR).

Methods

All physicians (n = 76; 48 general practitioners and 28 specialists) at the Finnish Student Health Service received a questionnaire by email, and were asked to print it and keep a daily tally of visits, phone calls and email messages over the study period of one working week (5.5. – 9.5.2003). The response rate was 70%. The data originating from the questionnaire were compared with statistical data from the EPR during the study period.

Results

The majority (79%, 41/52) of doctors reported using email with patients, averaging 8.6 (range: 0–96) email contacts and a percentage rate of "email / visit" 20% (range: 0–185%) in one working week. Doctors in the capital city region and those doctors who had a positive attitude toward email for patient communication were most active in email use. Up to 73% of email contacts were not documented in the EPR.

Conclusion

The activity in using email with patients verified among Finnish physicians is compatible with recent study results elsewhere. The notable proportion of un-recorded email messages establishes the need for an electric communication system built into the EPR to improve the quality of patient care and to limit medico-legal risks.
Clinical response in Japanese metastatic melanoma patients treated with peptide cocktail-pulsed dendritic cells
Authors: Akiyama Yasuto , Tanosaki Ryuji , Inoue Naoki , Shimada Makiko , Hotate Yukie , Yamamoto Akifumi , Yamazaki Naoya , Kawashima Ichiro , Nukaya Ikuei , Takesako Kazutoh , Maruyama Kouji , Takaue Yoichi , Yamaguchi Ken
Published: January 2005

Background

Metastatic, chemotherapy-resistant melanoma is an intractable cancer with a very poor prognosis. As to immunotherapy targeting metastatic melanoma, HLA-A2+ patients were mainly enrolled in the study in Western countries. However, HLA-A24+ melanoma patients-oriented immunotherapy has not been fully investigated. In the present study, we investigated the effect of dendritic cell (DC)-based immunotherapy on metastatic melanoma patients with HLA-A2 or A24 genotype.

Methods

Nine cases of metastatic melanoma were enrolled into a phase I study of monocyte-derived dendritic cell (DC)-based immunotherapy. HLA-genotype analysis revealed 4 cases of HLA-A*0201, 1 of A*0206 and 4 of A*2402. Enriched monocytes were obtained using OptiPrep™ from leukapheresis products, and then incubated with GM-CSF and IL-4 in a closed serum-free system. After pulsing with a cocktail of 5 melanoma-associated synthetic peptides (gp100, tyrosinase, MAGE-2, MAGE-3 and MART-1 or MAGE-1) restricted to HLA-A2 or A24 and KLH, cells were cryopreserved until used. Finally, thawed DCs were washed and injected subcutaneously (s.c.) into the inguinal region in a dose-escalation manner.

Results

The mean percentage of DCs rated as lin-HLA-DR+ in melanoma patients was 46.4 ± 15.6 %. Most of DCs expressed high level of co-stimulatory molecules and type1 phenotype (CD11c+HLA-DR+), while a moderate number of mature DCs with CD83 and CCR7 positive were contained in DC products. DC injections were well tolerated except for transient liver dysfunction (elevation of transaminases, Grade I-II). All 6 evaluable cases except for early PD showed positive immunological responses to more than 2 melanoma peptides in an ELISPOT assay. Two representative responders demonstrated strong HLA-class I protein expression in the tumor and very high scores of ELISPOT that might correlate to the regression of metastatic tumors. Clinical response through DC injections was as follows : 1CR, 1 PR, 1SD and 6 PD. All 59 DC injections in the phase I study were tolerable in terms of safety, however, the maximal tolerable dose of DCs was not determined.

Conclusions

These results suggested that peptide cocktail-treated DC-based immunotherapy had the potential for utilizing as one of therapeutic tools against metastatic melanoma in Japan.
Oral Tolerance: Overview and Historical Perspectives
Authors: MOWAT ALLAN McI. , PARKER LUCY A. , BEACOCK‐SHARP HELEN , Millington Owain R. , CHIRDO FERNANDO
Published: January 2005

Abstract: Oral tolerance was first detailed almost 100 years ago, and since then, it has been shown repeatedly that feeding a wide variety of nonpathogenic antigens can inhibit subsequent systemic immune responses. All systemic immune responses are susceptible, but the degree and scope of the suppression depends on the nature and dose of the fed antigen. Oral tolerance has been described in most mammals, including humans, and it may be the homeostatic mechanism that prevents hypersensitivity to food antigens, as is found in celiac disease. A similar process may prevent the aberrant immune responses to commensal bacteria that occur in inflammatory bowel disease. The ability of oral tolerance to modulate experimental models of autoimmune and inflammatory disease has led to clinical trials in such diseases as rheumatoid arthritis, multiple sclerosis, and type I diabetes, with only variable success. Despite intense research, the exact mechanisms responsible for the systemic tolerance and the reasons why tolerance is the default response to many fed antigens remain controversial. Early studies suggested that CD8+“suppressor” T cells were important, but it is now accepted that it may involve either anergy/deletion of CD4+ T cells, or the induction of regulatory CD4+ T cells that produce IL‐10 and/or TGFβ. There may also be a role for CD4+ CD25+ Treg, but how and when all these different mechanisms operate is still unclear. The ability of fed antigens to induce tolerance probably reflects their uptake by “quiescent” antigen‐presenting cells in the intestine, with presentation to specific CD4+ T cells in the absence of costimulation, or with the involvement of inhibitory costimulatory molecules. Dendritic cells in the Peyer's patches or mucosal lamina propria are the most likely APCs involved, but it remains to be determined exactly where these interactions occur and what the precise nature of the relevant dendritic cells is.
Activation of a Unique Population of CD8+ T Cells by Intestinal Epithelial Cells
Authors: ALLEZ MATTHIEU , BRIMNES JENS , SHAO LING , DOTAN IRIS , NAKAZAWA ATSUSHI , MAYER LLOYD
Published: January 2005

Abstract: Intestinal epithelial cells may play a role in the regulation of immune responses toward luminal antigens. We show that a subset of CD8+ T cells undergoes oligoclonal expansion in the intestinal mucosa, probably through interaction with a unique complex expressed on epithelial cells, formed by a CEA subfamily member (gp180) and CD1d. This subset, which is regulatory in vitro, may play a role in the control of intestinal immune responses toward luminal antigens. A lack of expansion of these CD8+ regulatory T cells, probably related to the defective expression of the gp180/CD1d complex, is observed in inflammatory bowel disease.
Involvement of Dendritic Cell Subsets in the Induction of Oral Tolerance and Immunity
Authors: FLEETON MARINA , CONTRACTOR NIKHAT , LEON FRANCISCO , HE JIANPING , WETZEL DENISE , DERMODY TERENCE , IWASAKI AKIKO , KELSALL BRIAN
Published: January 2005

Abstract: Dendritic cells (DCs) play a central role in the generation of immune responses in the intestine. DCs induce differentiation and tolerance of T cells, and may have a direct role in B cell switching to IgA. Four distinct subsets of CD11c+ DCs are present in murine Peyer's patches, which represent primary sites for the induction of mucosal T and B cell responses. Studies suggest that CD11b+ DCs or plasmacytoid DCs may be specialized for the induction of regulatory T cells, and CD8α+ DCs for the induction of clonal deletion in response to soluble oral antigen, while all DC subsets (including CD8α−/CD11b− DCs) may be involved in responses to pathogens. We are currently using reovirus type‐1 Lang (TIL) to explore the role of DC populations in mucosal immunity in vivo, as oral administration of live T1L to mice induces strong mucosal and systemic antiviral immune responses, whereas oral administration of inactivated T1L results in tolerance to viral proteins. We found that primary infection with T1L occurs in epithelial cells of the PP follicle‐associated epithelium, but that CD8α−/CD11b− DCs in the subepithelial dome region (SED) are loaded with T1L antigens in the absence of active DC infection. At least a portion of this antigen is associated with cell fragments from apoptotic epithelial cells, demonstrating that SED DCs cross‐present antigens from apoptotic epithelial cells. In vitro, in contrast to exposure to several TLR‐ligands or anti‐CD40, exposure to T1L does not activate DCs to mature or to produce cytokines, despite clear loading of the DCs with viral antigens. These data suggest that T1L is taken up by a “silent” receptor on DCs, and that the induction of immunity to T1L is dependent on signals from non‐DCs following active viral infection that induce DC maturation. Thus, the decision between tolerance and immunity to inactive and live virus, respectively, likely depends on whether there is active infection of epithelial cells by T1L, which results in the elaboration of molecules, such as cytokines, that induce DC maturation.
Intestinal Epithelial Cells Control Dendritic Cell Function
Authors: RIMOLDI MONICA , CHIEPPA MARCELLO , VULCANO MARISA , ALLAVENA PAOLA , RESCIGNO MARIA
Published: January 2005

Abstract: Dendritic cells (DCs) comprise a family of cells specializing in antigen capture and presentation to T cells. We have recently shown that DC play an active role in bacterial uptake across mucosal surfaces. Indeed, DC are able to open tight junctions and to sample antigens directly across epithelia, both in vitro and in vivo. Because DC express tight junction proteins, the integrity of the epithelial barrier is preserved. In this study we have analyzed the possible involvement of epithelial cells in controlling DC function. We developed an in vitro model in our laboratory consisting of a three‐player system of dendritic cells, epithelial cell monolayers, and bacteria. The crosstalk between epithelial cells and dendritic cells was analyzed, and epithelial cells were tested for their capacity to release cytokines and chemokines that induce the migration and activation of DC. We show that the capacity of epithelial cells to produce cytokines and activate DC is dependent on the invasiveness of the bacteria tested. In particular, invasive bacteria stimulate epithelial cells to release proinflammatory cytokines and to induce the maturation state of DC. By contrast, noninvasive bacteria are unable to stimulate epithelial cells, but can activate DC directly when DC translocate to the apical side. In conclusion, epithelial cells are not simply a barrier to bacteria entering via the oral route, but actively influence the activating properties of bystander DC.
In Vivo Enhancement of Dendritic Cell Function
Authors: BILSBOROUGH JANINE , VINEY JOANNE L.
Published: January 2005

Abstract: Despite the apparent positive recognition of antigen by mucosal T cells after ingesting food, the default functional response in the gut is tolerance. Although dendritic cells (DCs) are classically defined as potent stimulatory antigen‐presenting cells, we have previously shown that tolerance is enhanced in vivo in the presence of elevated numbers of DCs. In order to more closely investigate the mechanistic basis of tolerance induction, we have focused our subsequent studies on identifying features peculiar to mucosal dendritic cells and the functional involvement of mucosal DCs in driving the early T cell response to fed antigen. These studies have revealed a population of DCs in the mucosae that exhibit the plasmacytoid phenotype and secrete IFN‐α following stimulation with CpG, and can drive differentiation of naive T cells into cells that exhibit regulatory properties. The activity of these DCs also failed to sustain robust T cell proliferation and, rather, functioned to enhance the suppressive efficacy of CD4+CD25+ T regulatory cells. Given their significant presence in mucosal tissue, these DCs likely provide a mechanistic basis for the homeostatic regulation prominent in the gut, presumably by eliciting regulatory cell suppressor function and poorly supporting T helper cell proliferation. These studies further underscore the critical role for intestinal DCs in promoting tolerogenic antigen presentation at a site of high antigenic stimulation.
Study of Oral Tolerance and Its Indirect Effects in Adoptive Cell Transfer Experiments
Authors: da CUNHA ANDRÉ PIRES , VAZ NELSON MONTEIRO , CARVALHO CLAUDIA ROCHA
Published: January 2005

Abstract: Parenteral exposure to antigens to which oral tolerance had been previously induced results in the inhibition of immune responses to other unrelated antigens. Herein we tested whether indirect effects of oral tolerance could be adoptively transferred. Anti‐Ova‐ and antihemoglobin‐specific responsiveness as well as oral tolernace to Ova were transferred to irradiated, but not to normal, nonirradiated recepients. Irradiation, thus, facilitated adoptive transfer of oral tolerance. However, the inhibitory (indirect) effects upon the unrelated immunogen were not adoptively transferred, even to irradiated recepients. In addition, we studied adoptively transferred CFSE‐labeled spleen cells by flow cytometry in recipient spleen, inguinal lymph nodes, and bone marrow, both in irradiated and nonirradiated recipients, 1, 3, or 5 days after cell transfer. Comparing the percent and absolute number of CFSE‐labeled cells in each organ displayed significant differences in the dynamics of decay of adoptively transferred cells from tolerant or immune donors.
Peyer's Patch Dendritic Cells Capturing Oral Antigen Interact with Antigen‐Specific T Cells and Induce Gut‐Homing CD4+CD25+ Regulatory T Cells in Peyer's Patches
Authors: NAGATANI KATSUYA , SAGAWA KAYO , KOMAGATA YOSHINORI , YAMAMOTO KAZUHIKO
Published: January 2005

Abstract: Antigen‐specific naive T cells accumulated in Peyer's patches only after the feeding of antigen. DCs that captured oral antigen interacted with these T cells in the IFR of PP. Some of these T cells acquired a similar phenotype to CD4+ CD25+ regulatory T cells and CCR9+ gut‐homing T cells.
Commonly Used Drugs Impair Oral Tolerance in Mice
Authors: PECQUET SOPHIE , PRIOULT GUÉNOLÉE , CAMPBELL JOHN , GERMAN BRUCE , TURINI MARCO
Published: January 2005

Abstract: Ibuprofen and antibiotics are commonly prescribed during early childhood. When given to mice at the time at which oral tolerance is induced, both treatments affect either the induction or the maintenance of oral tolerance. These results suggest that the coadministration of these and similarly acting drugs should be considered cautiously for infants at risk of allergy.
Selective Generation of Gut‐Tropic T Cells in Gut‐Associated Lymphoid Tissues: Requirement for GALT Dendritic Cells and Adjuvant
Authors: SVENSSON MARCUS , JOHANSSON‐LINDBOM BENGT , WURBEL MARC‐ANDRÉ , MALISSEN BERNARD , MÁRQUEZ GABRIEL , AGACE WILLIAM
Published: January 2005

Abstract: To gain entry into peripheral tissues, effector T lymphocytes express specific combinations of adhesion molecules and chemokine receptors. Expression of these are induced during activation in regional secondary lymphoid organs. Herein, we describe a role for GALT DCs in the generation of CCR9+α4β7+ gut tropic lymphocytes.
Insights into the Mechanism of Oral Tolerance Derived from the Study of Models of Mucosal Inflammation
Authors: STROBER WARREN , FUSS IVAN , BOIRIVANT MONICA , KITANI ATSUSHI
Published: January 2005

Abstract: Murine models of mucosal inflammation are frequently due to the inability of the mouse to mount a regulatory T cell response. To the extent that such responses arise from oral tolerance mechanisms, these models provide a unique way of studying oral tolerance. In this paper we focus on the regulatory cells generated in two of the most well‐studied of such models, the cell‐transfer model and the TNBS‐colitis model. Our analysis leads to the view that regulatory cells generated by the oral tolerance seen in mucosal inflammation are, at least in part, cells that recognize self‐antigens or antigens in the mucosal microflora whose effector function relies on the expression of TGF‐β.
Regulation of Autoreactive T Cell Function by Oral Tolerance to Self‐Antigens
Authors: WHITACRE CAROLINE C. , SONG FEI , WARDROP RICHARD M. , CAMPBELL KIM , McCLAIN MELANIE , BENSON JACQUELINE , GUAN ZHEN , GIENAPP INGRID
Published: January 2005

Abstract: The oral administration of neuroantigens can suppress as well as treat autoimmune disease. Using EAE as a model system, we examined the antigen‐presenting cell in oral tolerance. Expansion of dendritic cells (DCs) prior to or after disease is established facilitated oral tolerance. Transfer of oral antigen‐loaded DCs resulted in protection from EAE by induction of IL‐4 and IL‐5 in recipient animals. LPS treatment of donors abrogated the ability of DCs to transfer protection from EAE, emphasizing the importance of the DC activation state. T cells exposed to orally administered antigen were monitored in TCR transgenic mice and found to undergo activation followed by deletion. The thymus plays a critical role in oral tolerance since thymectomized mice could not be tolerized. The thymus is postulated to be a site for deletion of autoreactive T cells or a site for generation of regulatory T cells.
Differential Immune Induction with Subcutaneous versus Oral Administration of a Diabetogenic Insulin Peptide in the NOD Mouse
Authors: DEVENDRA DEVASENAN , PARONEN JOHANNA , LIU EDWIN , MORIYAMA HIROAKI , MIAO DONGMEI , YU LIPING , EISENBARTH GEORGE S.
Published: January 2005

Abstract: The B chain insulin peptide 9 to 23 (B:9–23) is a dominant T cell epitope of the NOD mouse. Given in oral form with multiple different vehicles, it did not alter expression of insulin autoantibodies in contrast to subcutaneous administration.
CD45 Knockout Mice, Diet, and Colitis: In the Absence of CD45, Colitis Follows Dietary Changes
Authors: LÓPEZA MARÍA C. , HOLMES NICK
Published: January 2005

Abstract: Healthy CD45 KO mice show a Th2‐type cytokine secretion profile in their small intestines. Dietary supplementation with human infant formula induced colitis and a shift toward a Th1‐type profile, suggesting that diet can alter cytokine secretion.
The Thymus Plays a Role in Oral Tolerance Induction in Experimental Autoimmune Encephalomyelitis
Authors: SONG FEI , GIENAPP INGRID E. , SHAWLER TODD , GUAN ZHEN , WHITACRE CAROLINE C.
Published: January 2005

Abstract: Mice are protected from experimental autoimmune encephalomyelitis (EAE) when fed myelin basic protein (MBP). Thymectomized mice do not exhibit oral tolerance. We found evidence for two mechanisms to explain the role of the thymus in oral tolerance: a site for deletion of autoreactive T cells and a source of regulatory T cells.
IL‐18 and Antigen‐Specific CD4+ Regulatory T Cells in Peyer's Patches
Authors: TSUJI NORIKO M. , NOWAK BERNADETA
Published: January 2005

Abstract: A study using gene‐deficient mice revealed that IL‐18 is essential for inducing antigen‐specific regulatory T cells and oral tolerance. It is also suggested that IL‐18 is a functional downregulator for immunogenic dendritic cells, contributing to the maintenance of homeostasis for intestinal immunity.
Oral Tolerance in Humans: Failure to Suppress an Existing Immune Response by Oral Antigen Administration
Authors: MOLDOVEANU ZINA , OLIVER FRED , MESTECKY JIRI , ELSON CHARLES O.
Published: January 2005

Abstract: Orally administered antigens can induce systemic tolerance. In animal models, oral tolerance can both prevent and treat experimental autoimmune diseases. The induction of oral tolerance to human autoantigens has been envisioned as a potential treatment for human autoimmune conditions. The results of previous human studies from our laboratory have provided evidence that oral administration of a model antigen, keyhole limpet hemocyanin (KLH), prior to systemic immunization, can decrease the magnitude of subsequent T cell proliferative and skin test responses to KLH. The present study was designed to test the hypothesis that orally administered KLH could attenuate a preexisting immune response to KLH. Thus, human subjects (n= 8) were primed subcutaneously with KLH without adjuvant prior to a 42‐day feeding regimen of 100 mg of KLH per day. At the end of the feeding regimen, the subjects were boosted with KLH, again without adjuvant. Eight control subjects were immunized as above with KLH, but fed ovalbumin. The measurement of antigen‐driven T cell proliferation, serum and salivary antibody, and dermal delayed hypersensitivity responses to KLH failed to reveal significant differences between subjects fed KLH and those fed ovalbumin. These results indicate that the KLH dose and feeding regimen used in this study failed to attenuate the primary response or to prevent the secondary response to KLH. Therefore, some form of immunomodulation greater than that provided by oral administration of antigen alone is required in humans for suppression of an existing immune response.