bluuebunny/crossref_metadata_2025_split huggingface dataset:
BAD Ser128 Is Not Phosphorylated by c-Jun NH2-Terminal Kinase for Promoting Apoptosis
Authors: Zhang Jiyan , Liu Jing , Yu Chenfei , Lin Anning 
Published: September 2005

Abstract
The phosphorylation and regulation of the proapoptotic Bcl-2 family protein BAD by c-Jun NH2-terminal kinase (JNK) is controversial. JNK can suppress interleukin-3 withdrawal-induced apoptosis via phosphorylation of BAD at Thr201. However, it has also been reported that JNK promotes apoptosis through phosphorylation of BAD at Ser128. Here, we report that JNK is not a BAD Ser128 kinase. JNK phosphorylates murine BAD (mBAD), but not human BAD (hBAD), in which Ser91 is equivalent to Ser128 in mBAD. In contrast, Cdc2, which phosphorylates Ser128, phosphorylates both mBAD and hBAD. Replacement of Ser128 by alanine has no effects on BAD phosphorylation by JNK in vitro and in vivo. Two-dimensional phosphopeptide mapping in combination with phosphoamino acid analysis reveals that JNK does not phosphorylate BAD at Ser128. Elimination of Ser128 phosphorylation has no effects on the proapoptotic activity of BAD in apoptosis induced by UV via JNK or growth factor withdrawal. Thus, our results show that Ser128 is not phosphorylated by JNK for promoting cell death.
Side Population Is Enriched in Tumorigenic, Stem-Like Cancer Cells, whereas ABCG2+ and ABCG2− Cancer Cells Are Similarly Tumorigenic
Authors: Patrawala Lubna , Calhoun Tammy , Schneider-Broussard Robin , Zhou Jianjun , Claypool Kent , Tang Dean G. 
Published: September 2005

Abstract
Recently, several human cancers including leukemia and breast and brain tumors were found to contain stem-like cancer cells called cancer stem cells (CSC). Most of these CSCs were identified using markers that identify putative normal stem cells. In some cases, stem-like cancer cells were identified using the flow cytometry-based side population technique. In this study, we first show that ∼30% of cultured human cancer cells and xenograft tumors examined (∼30 in total) possess a detectable side population. Purified side population cells from two cell lines (U373 glioma and MCF7 breast cancer) and a xenograft prostate tumor (LAPC-9) are more tumorigenic than the corresponding non–side population cells. These side population cells also possess some intrinsic stem cell properties as they generate non–side population cells in vivo, can be further transplanted, and preferentially express some “stemness” genes, including Notch-1 and β-catenin. Because the side population phenotype is mainly mediated by ABCG2, an ATP-binding cassette half-transporter associated with multidrug resistance, we subsequently studied ABCG2+ and ABCG2− cancer cells with respect to their tumorigenicity in vivo. Although side population cells show increased ABCG2 mRNA expression relative to the non–side population cells and all cancer cells and xenograft tumors examined express ABCG2 in a small fraction (0.5-3%) of the cells, highly purified ABCG2+ cancer cells, surprisingly, have very similar tumorigenicity to the ABCG2− cancer cells. Mechanistic studies indicate that ABCG2 expression is associated with proliferation and ABCG2+ cancer cells can generate ABCG2− cells. However, ABCG2− cancer cells can also generate ABCG2+ cells. Furthermore, the ABCG2− cancer cells form more and larger clones in the long-term clonal analyses and the ABCG2− population preferentially expresses several “stemness” genes. Taken together, our results suggest that (a) the side population is enriched with tumorigenic stem-like cancer cells, (b) ABCG2 expression identifies mainly fast-cycling tumor progenitors, and (c) the ABCG2− population contains primitive stem-like cancer cells.
Concurrent Induction of Antitumor Immunity and Autoimmune Thyroiditis in CD4+CD25+ Regulatory T Cell–Depleted Mice
Authors: Wei Wei-Zen , Jacob Jennifer B. , Zielinski John F. , Flynn Jeffrey C. , Shim K. David , Alsharabi Ghazwan , Giraldo Alvaro A. , Kong Yi-chi M. 
Published: September 2005

AbstractWhen CD4+CD25+ regulatory T cells are depleted or inactivated for the purpose of enhancing antitumor immunity, the risk of autoimmune disease may be significantly elevated because these regulatory T cells control both antitumor immunity and autoimmunity. To evaluate the relative benefit and risk of modulating CD4+CD25+ regulatory T cells, we established a new test system to measure simultaneously the immune reactivity to a tumor-associated antigen, neu, and an unrelated self-antigen, thyroglobulin. BALB/c mice were inoculated with TUBO cells expressing an activated rat neu and treated with anti-CD25 monoclonal antibody to deplete CD25+ cells. The tumors grew, then regressed, and neu-specific antibodies and IFN-γ–secreting T cells were induced. The same mice were also exposed to mouse thyroglobulin by chronic i.v. injections. These mice produced thyroglobulin-specific antibody and IFN-γ–secreting T cells with inflammatory infiltration in the thyroids of some mice. The immune responses to neu or thyroglobulin were greater in mice undergoing TUBO tumor rejection and thyroglobulin injection than in those experiencing either alone. To the best of our knowledge, this is the first experimental system to assess the concurrent induction and possible synergy of immune reactivity to defined tumor and self-antigens following reduction of regulatory T cells. These results illustrate the importance of monitoring immune reactivity to self-antigens during cancer immunotherapy that involves immunomodulating agents, and the pressing need for novel strategies to induce antitumor immunity while minimizing autoimmunity.
Blockade of Epidermal Growth Factor Receptors Chemosensitizes Breast Cancer Cells through Up-Regulation of Bnip3L
Authors: Real Pedro J. , Benito Adalberto , Cuevas Jorge , Berciano Maria T. , de Juan Ana , Coffer Paul , Gomez-Roman Javier , Lafarga Miguel , Lopez-Vega Jose M. , Fernandez-Luna Jose L. 
Published: September 2005

Abstract
Epidermal growth factor receptor-1 (EGFR) and EGFR-2 (HER2) have become major targets for cancer treatment. Blocking antibodies and small-molecule inhibitors are being used to silence the activity of these receptors in different tumors with varying efficacy. Thus, a better knowledge on the signaling pathways activated by EGFR and HER2 may help unravel novel therapeutic targets and molecular markers of response. Here, we show that treatment of breast cancer cell lines with blocking antibodies against EGFR (cetuximab) or HER2 (trastuzumab) promotes the specific induction of proapoptotic Bnip3L and chemosensitization. Moreover, we found that the Bnip3L gene is transcriptionally activated by FoxO3a. Trastuzumab-mediated induction of Bnip3L and nuclear translocation of FoxO3a was also shown in pleural effusion cells from a breast cancer patient. Transfection of breast cancer cells with constitutively active FoxO3a or with Bnip3L promotes sensitization to chemotherapy-induced apoptosis. On the contrary, blockade of Bnip3L expression by a small interfering RNA strategy significantly diminished the chemosensitizing effect of cetuximab. We found also an inverse correlation between EGFR and Bnip3L expression in surgical specimens from patients with breast cancer. Therefore, blockading EGFR or HER2 specifically up-regulates Bnip3L, which is required for chemosensitization of breast cancer cells. This novel pathway provides also the rationale for therapeutic strategies aimed to induce the expression of Bnip3L.
Suppression of Mammary Carcinoma Growth by Retinoic Acid: Proapoptotic Genes Are Targets for Retinoic Acid Receptor and Cellular Retinoic Acid–Binding Protein II Signaling
Authors: Donato Leslie J. , Noy Noa 
Published: September 2005

Abstract
Retinoic acid (RA) displays pronounced anticarcinogenic activities in several types of cancer. Whereas the mechanisms that underlie this activity remain incompletely understood, tumor suppression by RA is believed to emanate primarily from its ability to regulate transcription of multiple target genes. Here, we investigated molecular events through which RA inhibits the growth of MCF-7 mammary carcinoma cells, focusing on the involvement of the two proteins that mediate transcriptional activation by RA, the nuclear hormone receptor retinoic acid receptor (RAR) and the cellular retinoic acid-binding protein (CRABP) II, in this process. RA treatment of MCF-7 cells did not affect cell cycle distribution but triggered pronounced apoptosis. Accordingly, expression array analyses revealed that RA induces the expression of several proapoptotic genes, including caspase 7 and caspase 9. Whereas caspase 7 is an indirect responder to RA signaling, caspase 9 is a novel direct target for RAR, and it harbors a functional retinoic acid response element in its second intron. In agreement with the known role of CRABP-II in enhancing the transcriptional activity of RAR, the binding protein augmented RA-induced up-regulation of caspase 9, cooperated with RA in activating both caspase 7 and 9, and amplified the ability of RA to trigger apoptosis. Surprisingly, the data indicate that CRABP-II also displays proapoptotic activities on its own. Specifically, overexpression of CRABP-II, in the absence of RA, up-regulated the expression of Apaf1 and triggered caspase 7 and caspase 9 cleavage. These observations suggest that, in addition to its known role in direct delivery of RA to RAR, CRABP-II may have an additional, RA-independent, function.
Distinct and Overlapping Roles of Interleukin-10 and CD25+ Regulatory T Cells in the Inhibition of Antitumor CD8 T-Cell Responses
Authors: Dercamp Christophe , Chemin Karine , Caux Christophe , Trinchieri Giorgio , Vicari Alain P. 
Published: September 2005

AbstractLack of antitumor immunity is often related to impaired CD8 T-cell responses that could result from a poor priming capacity by tumor-infiltrating dendritic cells (TIDC) and/or further inhibition by regulatory T cells (Treg). Interleukin-10 (IL-10) has been implicated in the inhibition of TIDC as well as in the generation and functions of Treg. Here, we address some of the respective and possibly overlapping roles of IL-10 and CD25+ Treg in CD8 antitumor immunity. Whereas tumor antigen–specific CD8 T cells proliferated in vivo in the presence of IL-10 or Treg, optimal effector functions were observed in mice lacking both IL-10 and Treg. Indeed, tumors grown in normal but not in IL-10–deficient or CD25-depleted mice induced tumor antigen–specific CD8 suppressor T cells. Suppression involved transforming growth factor-β. Similarly, both IL-10 and Treg were responsible for impaired CD8 T cell priming by TIDCs, but IL-12 production by TIDCs was prevented only by Treg-independent IL-10. Subsequently, IL-10 defect and Treg depletion were required to achieve optimal induction of CD8 T-cell effectors by TIDC following CpG activation. Our results point out major redundant and nonredundant roles for IL-10 and Treg in the inhibition of TIDC-mediated generation of antitumor CD8 T-cell response.
Darpp-32: a Novel Antiapoptotic Gene in Upper Gastrointestinal Carcinomas
Authors: Belkhiri Abbes , Zaika Alexander , Pidkovka Nataliya , Knuutila Sakari , Moskaluk Christopher , El-Rifai Wa'el 
Published: September 2005

Abstract
We show the molecular mechanisms involved in Darpp-32 overexpression and its biological role in upper gastrointestinal adenocarcinomas (UGC). A tumor tissue array of 377 samples was developed and used to detect DARPP-32 DNA amplification and protein overexpression, which occurred in 32% and 60% of UGCs, respectively. Concomitant overexpression of mRNA for Darpp-32 and its truncated isoform t-Darpp was observed in 68% of tumors (P &lt; 0.001). When Darpp-32 and t-Darpp were overexpressed in AGS and RKO gastrointestinal cells, up to a 4-fold reduction in the apoptosis rate was observed (terminal deoxynucleotidyl transferase–mediated nick-end labeling and Annexin V assays) in response to camptothecin, sodium butyrate, and ceramide. However, the introduction of mutations in phosphorylation sites abrogated this effect. Expression of Darpp-32 and t-Darpp preserved the mitochondrial transmembrane potential and was associated with increased levels of Bcl2 protein. A reversal of Bcl2 protein level was obtained using small interfering RNAs for Darpp-32 and t-Darpp. Luciferase assays using the p53 and p21 reporter plasmids and probing of immunoblots with antibodies specific for p53 transcriptional targets, such as Hdm2 and p21, indicated that neither Darpp-32 nor t-Darpp interfere with p53 function. Altogether, we show more frequent mRNA and protein overexpression of Darpp-32 than DNA amplification, suggesting that, in addition to amplification, transcriptional or posttranscriptional mechanisms may play an important role. The expression of Darpp-32 and t-Darpp is associated with a potent antiapoptotic advantage for cancer cells through a p53-independent mechanism that involves preservation of mitochondrial potential and increased Bcl2 levels.
Two Distinct Activities Contribute to Human Papillomavirus 16 E6's Oncogenic Potential
Authors: Simonson Sara J.S. , Difilippantonio Michael J. , Lambert Paul F. 
Published: September 2005

Abstract
High-risk human papillomaviruses, such as HPV16, cause cervical cancers, other anogenital cancers, and a subset of head and neck cancers. E6 and E7, two viral oncogenes expressed in these cancers, encode multifunctional proteins best known for their ability to bind and inactivate the tumor suppressors p53 and pRb, respectively. In skin carcinogenesis experiments using E6 transgenic (K14E6WT) mice, HPV16 E6 was found to contribute to two distinct stages in skin carcinogenesis: promotion, a step involved in the formation of benign papillomas, and progression, the step involved in the malignant conversion of benign tumors to frank cancer. In this study, we compared the tumorigenic properties of K14E6WT mice with those of K14E6Δ146-151 mice, which express a mutant form of E6 that cannot bind a family of cellular proteins known as PDZ domain proteins but retains the ability to inactivate p53. In skin carcinogenesis experiments, the K14E6Δ146-151 transgene failed to contribute to the promotion stage of skin carcinogenesis but retained the ability to contribute to the progression stage. Cytogenetic analysis indicated that, although gains of chromosome 6 are consistently seen in tumors arising on K14E6WT mice, they are infrequently seen in tumors arising on K14E6Δ146-151 mice. This observation supports the premise that the nature of cancer development in these two mouse strains is distinct. Based on these studies, we conclude that E6 contributes to cancer through its disruption of multiple cellular pathways, one of which is mediated through its interaction with PDZ domain partners and the other through E6's inactivation of p53.
The APC Tumor Suppressor Promotes Transcription-Independent Apoptosis In vitro
Authors: Steigerwald Kira , Behbehani Gregory K. , Combs Kelly A. , Barton Michelle Craig , Groden Joanna 
Published: September 2005

Abstract
The APC tumor suppressor is found in nonproliferating epithelial cells of the colonic crypts and is mutated in most colorectal tumors. To understand the function of APC in normal epithelium and how its loss leads to tumor formation, we tested whether APC is a mediator of apoptosis using an in vitro assay that monitors caspase-3-mediated cleavage of lamin B protein or a colorimetric substrate in a cell-free Xenopus egg extract. Recombinant APC protein accelerates apoptosis-associated caspase activity independently of ongoing transcription and protein synthesis. Conversely, the addition of mutant APC and immunodepletion of Xenopus APC decelerates apoptosis-associated caspase activity. Acceleration of apoptosis by APC is abolished by the caspase-8 inhibitor Z-IETD-FMK, demonstrating that caspase-8 is an essential component of APC-mediated apoptosis. These results suggest that the induction of apoptosis may be one role of APC in tumor suppression and that this mechanism is independent of β-catenin-mediated effects on transcription.
Methylation Silencing of the Apaf-1 Gene in Acute Leukemia
Authors: Furukawa Yutaka , Sutheesophon Krittaya , Wada Taeko , Nishimura Miki , Saito Yasushi , Ishii Hideshi , Furukawa Yusuke 
Published: September 2005

Abstract
Apaf-1 is important for tumor suppression and drug resistance because it plays a central role in DNA damage–induced apoptosis. Inactivation of the Apaf-1 gene is implicated in disease progression and chemoresistance of some malignancies. In this study, we attempted to clarify the role of Apaf-1 in leukemogenesis. Apaf-1 mRNA levels were below the detection limit or very low in 5 of 20 human leukemia cell lines (25%) and 5 of 12 primary acute myeloblastic leukemia cells (42%). There were no gross structural abnormalities in the Apaf-1 gene in these samples. Expression of factors regulating Apaf-1 transcription, such as E2F-1, p53, and Sp-1, did not differ between Apaf-1-positive and Apaf-1-negative cells. Methylation of CpG in the region between +87 and +128 of the Apaf-1 gene was almost exclusively observed in Apaf-1-defective cell lines. Treatment of these cells with 5-aza-2′-deoxycytidine, a specific inhibitor of DNA methylation, restored the expression of Apaf-1. Furthermore, we showed that the region between +87 and +128 could act as a repressor element by recruiting corepressors such as methylated DNA-binding domain 2 and histone deacetylase 1 upon methylation. Overexpression of Dnmt1, a mammalian maintenance DNA methyltransferase, was associated with Apaf-1 gene methylation. DNAs from Dnmt1-overexpressing cells were more resistant to digestion with methylation-sensitive enzyme HpaII than those from cells with low Dnmt1 expression, suggesting that Dnmt1 mediates aberrant methylation of multiple genes. In conclusion, methylation silencing is a mechanism of the inactivation of Apaf-1 in acute leukemia, and Dnmt1 overexpression may underlie hypermethylation of the Apaf-1 gene.
A Nonclassic CCAAT Enhancer Element Binding Protein Binding Site Contributes to α-Methylacyl-CoA Racemase Expression in Prostate Cancer
Authors: Zha Shan , Isaacs William B. 
Published: September 2005

Abstract
α-Methylacyl-CoA racemase (AMACR), an enzyme involved in branched-chain fatty acid β-oxidation that is normally expressed at high levels in human liver, is specifically and consistently overexpressed at both mRNA and protein levels in human prostate cancer and potentially other cancer types. To characterize the mechanisms underlying transcriptional regulation of AMACR at the genetic and epigenetic levels, we performed a series of methylation and reporter assays in prostate cancer tissues and cell lines. The results ruled out altered methylation patterns as the cause of overexpression in prostate cancer cells. However, promoter deletion analysis identified an 8-bp nonclassic CCAAT enhancer element located ∼80 bp upstream of the transcriptional initiation site that is responsible for AMACR expression in both prostate cancer cell lines and cell lines of liver origin. Deletion or mutation of this element completely abolished AMACR promoter activity. Ectopic expression of CCAAT/enhancer binding protein β increased luciferase activity driven by a wild-type AMACR promoter sequence but not by the sequence in which the putative CCAAT/enhancer binding protein binding element had been mutated. These results implicate a nonclassic CCAAT enhancer element in the AMACR gene promoter as playing a critical role in the regulation of AMACR gene expression.
Comparison of prevalence and severity of asthma among adolescents in the Caribbean islands of Trinidad and Tobago: results of a nationwide cross-sectional survey
Authors: Monteil Michele A , Joseph Gina , Changkit Catherine , Wheeler Gillian , Antoine Robin M 
Published: September 2005

Abstract

Background
Asthma is a growing problem in the Caribbean but the prevalence in most islands is unknown and possible inter-island variation in prevalence has not been determined. A nationwide cross-sectional survey was conducted to compare the prevalence of asthma symptoms among high school students in the two islands of the Republic of Trinidad and Tobago.

Methods
Questionnaire and video instruments based on those developed by the International Study of Asthma & Allergy in Childhood (ISAAC) were used to assess asthma prevalence among 6394 children (age range, 11–19 years; mean age, 14.08 yrs) in the second and third years of 35 randomly selected high schools in Trinidad and Tobago. This cross sectional survey was conducted between September and December 2002.

Results
A total of 4988 questionnaires were available for analysis (3519 in Trinidad and 1469 in Tobago). Among respondents from the two islands, there were no significant differences in the prevalence of ever wheezing (24.1% and 24.3% for Trinidad and Tobago, respectively, RR 0.99, 95% CI, 0.90–1.08); wheezing in the previous 12 months (13.1% & 13.4%, RR 0.98, 95% CI 0.84–1.15); a previous or current diagnosis of asthma (12.8% & 13.5%, RR 0.95, 95% CI 0.82–1.12) and night cough in the past 12 months (35.4% & 38.3%, RR0.93, 95% CI 0.86–1.00). However, symptoms of severe asthma were significantly more common among students from Tobago and included having had more than one acute attack in the past year (13.4% & 15.8%, RR 0.85, 95% CI 0.73–1.00, p = 0.0004), night waking as a result of wheeze (7.4% & 10.9%, RR 0.68, 95% CI 0.56–0.83, p < 0.0001) and speech limitation in the past year (5.2% & 8.7%, RR 0.59, 95% CI 0.47–0.74, p < 0.001) Exercise-associated wheezing was also more frequent among Tobagonian adolescents (17.5% & 20.2%, RR 0.87, 95% CI 0.76 – 0.98, p = 0.04).

Conclusion
Self-reported wheeze is common among adolescents in Trinidad and Tobago. Variation in symptoms was found between the two territories; high school students from Tobago, the less industrialized of the two islands, reported more symptoms of severe asthma and exercise-induced wheeze. Difference in the ethnic composition rather than socio-economic factors may be contributing to the observed differences in symptom prevalence.
Lessons learned in developing family medicine residency training programs in Japan
Authors: Murai Mitsuya , Kitamura Kazuya , Fetters Michael D 
Published: September 2005

Abstract

Background
While family medicine is not well established as a discipline in Japan, a growing number of Japanese medical schools and training hospitals have recently started sougoushinryoubu (general medicine departments). Some of these departments are incorporating a family medicine approach to residency training. We sought to learn from family medicine pioneers of these programs lessons for developing residency training.

Methods
This qualitative project utilized a long interview research design. Questions focused on four topics: 1) circumstances when becoming chair/faculty member; 2) approach to starting the program; 3) how Western ideas of family medicine were incorporated; and 4) future directions. We analyzed the data using immersion/crystallization to identify recurring themes. From the transcribed data, we selected representative quotations to illustrate them. We verified the findings by emailing the participants and obtaining feedback.

Results
Participants included: five chairpersons, two program directors, and three faculty members. We identified five lessons: 1) few people understand the basic concepts of family medicine; 2) developing a core curriculum is difficult; 3) start with undergraduates; 4) emphasize clinical skills; and 5) train in the community.

Conclusion
While organizational change is difficult, the identified lessons suggest issues that merit consideration when developing a family medicine training program. Lessons from complexity science could inform application of these insights in other countries and settings newly developing residency training.