bluuebunny/crossref_metadata_2025 huggingface dataset:
Parental oral health care of mentally retarded children in Ilala municipality, Dar es Salaam, Tanzania
Authors: Kahabuka FK , Ndalahwa M
Published: July 2006

Objective: The aim of this study was to investigate oral hygiene practices and oral health care given by parents to mentally retarded children in Dar es Salaam, Tanzania. Methods: A total of 140 questionnaires were distributed; only 100 questionnaires were completed and thus were included in the analysis, a 71.4% response rate. Questionnaires inquired on oral hygiene practices, supervision or assistance during tooth brushing, examination of the oral cavity, and measures taken in an event of oral health problems. Results: Sixty‐five per cent of the children are able to brush their teeth, whereas 35% are brushed by a family member. About 74% of the children who brush themselves brush occasionally or once per day while 26% brush more than once per day. More than 58% of the children who brush themselves brush under supervision. A statistically significant difference was observed in respect of bleeding on brushing which was among 61% of the children who brush occasionally or once per day and among 29% of the children who brush more than once per day (Pearson χ2 = 5.13, P = 0.024). Sixty‐four per cent of parents occasionally examine their children's oral cavity. Few parents take their children to a dentist in an event of toothache/cavity on a tooth (26%) or bleeding on brushing (37%). Conclusion: Most children are able to brush their teeth themselves. Majority of those who cannot brush are assisted. Bleeding on brushing and tooth cavities are common problems but few children are taken to a dentist for care.
Special features of topical and systemic dermatologic therapy in children
Authors: Lukas Anne , Wolf Gerd , Fölster‐Holst Regina
Published: July 2006

Since children differ greatly from adults in terms of physiology and pathophysiology, which in turn influences pharmacokinetics and pharmacodynamics, pediatric clinical trials on the effectiveness and safety of drugs must be supported. Such studies are still lacking. To avoid excluding pediatric patients from the benefit of optimal treatment, treatment strategies for children are often derived from trials with adult patients. Pediatric prescribing is also assisted by years of experience in the use of drugs and empirical data found in the literature.

This article discusses typical side effects that can occur with pediatric topical and systemic drug use. Salicylic acid and lindane intoxication are discussed as are Reye syndrome after taking acetylsalicylic acid to treat viral infection. Additional examples and tables listing drugs often used in pediatric dermatology as well as general information on prescribing off‐label pharmaceuticals should help children with skin disorders benefit from effective and tolerable treatment.
Acne infantum as presenting symptom of congenital adrenal hyperplasia due to 11‐beta‐hydroxylase deficiency
Authors: Harde Viola , Müller Marina , Sippell Wolfgang G. , Schwarz Thomas , Fölster‐Holst Regina
Published: July 2006

Infantile acne is a rare condition which usually begins after the third month of life and appears mainly on the cheeks. Spontaneous healing typically occurs within 2 years. A 2‐year‐old boy developed acne infantum during the first days after birth; it failed to respond to topical treatment. Detailed physical examination and endocrinologic evaluation confirmed the presumed diagnosis of congenital adrenal hyperplasia (CAH), and showed it was caused by 11‐beta‐hydroxylase deficiency.

CAH comprises a group of autosomal‐recessively inherited disorders. These hereditary enzyme defects in steroid biosynthesis cause glucocorticoid deficiency and an overproduction of biosynthetic precursor steroids. Adrenal androgen biosynthesis is not impaired but shows a massive reactive overproduction due to the increased ACTH secretion within the up‐regulated hypothalamo‐pituitary‐adrenal system. The characteristic features of CAH in male infants are demonstrated on the basis of this case report.
Trichophyton eboreum– A recently described dermatophyte
Authors: Brasch Jochen , Gräser Yvonne
Published: July 2006

Background: The distribution of distinct species of dermatophytes is continuously changing and different species of dermatophytes can cause the same type of tinea. Therefore, it is necessary to always identify the pathogenic agents in dermatophytoses.

Patients and methods: An unusual fungus was isolated from moccasin type tinea pedis in a HIV‐positive immunocompromised patient from the Ivory Coast and characterized by morphological, physiological and genetic methods.

Results: The strain under study showed typical characteristics of dermatophytes but considering all its morphologic, physiologic and genetic features it unambiguously differed from all previously described species.

Conclusion: The strain was recently described as a new dermatophyte species (Trichophyton eboreum). Distribution and pathogenicity of this species are as yet unknown.
A chromosomal event resulting in possession and expression of multiple HLA‐B and Cw genes in a renal patient
Authors: Poole K. , Montague B. , Roberts R. , Stoves J. , Bendukidze N. , Clark B.
Published: July 2006

Routine HLA typing of a renal patient for purposes of registration for transplantation revealed an unusual human leucocyte antigen (HLA)‐B and Cw genotype, with three specificities detected. Results were confirmed in a second sample, and in a second laboratory. The possibility of these results reflecting a chimaeric state was rejected following short tandem repeat (STR) analysis. Although cytogenetic analysis has failed to detect a chromosomal abnormality, these findings support the view that the aberrant expression of HLA in this patient resulted from an unequal crossover event, occurring during meiosis in a previous generation.
DRB3.2 gene polymorphism and its association with pashmina production in Changthangi goat
Authors: Sheikh F. D. , Bhattacharya T. K. , Kumar P. , Sharma A.
Published: July 2006

PstI and TaqI restriction fragment length polymorphism (RFLP) of caprine DRB3.2 revealed three genotypes under each RFLP in Changthangi goat. The genotypic and allelic frequency in PstI RFLP was calculated as 0.07, 0.72 and 0.20 for PP, Pp and pp genotypes and 0.43 and 0.57 for P and p alleles. In TaqI RFLP the frequencies were observed as 0.11, 0.61 and 0.27 for TT, Tt and tt genotypes and 0.41, 0.59 for T and t alleles. Alignment study indicates variability of nucleotides and amino acids between alleles and different breeds of goats. The nucleotide sequences reported in this paper have been submitted to the European Molecular Biology Laboratory GenBank and the accession numbers were AY496935, AY496061 and AY496062. A significant affect (P ≤ 0.05) of MHC DRB3.2 PstI polymorphism was found on the first and second year of pashmina production.
SiPep: a system for the prediction of tissue‐specific minor histocompatibility antigens
Authors: Halling‐Brown M. , Quartey‐Papafio R. , Travers P. J. , Moss D. S.
Published: July 2006

Approximately 50 years ago it was found that inbred strains of mice were able to reject tumours and skin grafts from major histocompatibility complex (MHC) identical donors. They proposed that additional transplantation antigens must exist outside the MHC. These were described as minor histocompatibility antigens (mHAgs). Since then, related studies in humans have identified 16 human mHAgs. The aim of this work is to increase the number of known mHAgs by prediction of candidate minor histocompatibility loci by identifying coding single nucleotide polymorphisms (SNPs) where the amino acid variation lies within an MHC‐binding peptide and alters the ability of that peptide to bind.

We have developed an algorithm called SiPep which uses peptide sequences derived from the flanking regions of known non‐synonymous SNPs, various MHC‐binding and proteolytic cleavage evaluation methods and protein expression data to predict mHAgs. We have processed 45094 SNPs using the SiPep algorithm and have stored the results in a database called SNPBinder. The facilities to process submitted proteins through the SiPep algorithm as well as the SNPBinder database are available to the public. A set of peptides that are predicted as possible mHAgs by the SiPep algorithm have been tested using refolding assays and gel filtration and the results are presented in this paper.

The SiPep tools and SNPBinder database are available free of charge via the internet. An HTML interface providing search facilities can be found at the following address: http://www.sipep.org/.
A functional variant in the CARD4 gene and risk of premature coronary heart disease
Authors: El Mokhtari N. E. , Ott S. J. , Nebel A. , Simon R. , Schreiber S.
Published: July 2006

Infection and innate immunity have been suggested as playing an important role in the pathogenesis of atherosclerosis. The recently discovered pattern‐recognition receptor (PRR) proteins initiate signalling after host–pathogen interactions and several PRRs, especially the Toll‐like receptor 4 (TLR4), have been shown to be involved in the development and progression of atherosclerosis. A new addition to the PRRs is CARD4, a gene that encodes the protein nucleotide‐binding oligomerization domain 1 (NOD1) and that seems to be associated with barrier function in chronic inflammatory disorders. Recently, a functional variant in the CARD4 gene, the insertion–deletion polymorphism ND1+32656, has been associated with inflammatory barrier diseases (inflammatory bowel diseases and asthma). We analysed the frequencies of this known functional mutation in the CARD4 gene and of the two adjacent variants, rs2075822 and rs2907748, in a German sample of 1440 unrelated early onset coronary heart disease (CHD) patients and healthy controls. Genotype and haplotype data showed no evidence for a significant association of these CARD4 variants with CHD. Our results suggest that the analysed CARD4 mutations do not play a major role in the aetiology of CHD.
Effect of copper on the development, protein and esterase isozymes ofDrosophila melanogaster
Authors: DING Ling , WANG Yufeng
Published: July 2006

Copper is an important ingredient in fungicides, which are used for many economic plants. However, there is concern about the side‐effects of copper‐based fungicides due to their potential to affect beneficial mites. The purpose of the present paper was to investigate the effects of copper on the development of the fruit fly,Drosophila melanogaster.It was found that higher doses of copper significantly prolonged the developmental time of the fruit flies, especially during the larval stages. When 320 mg L‐1copper‐contaminated food was used, most of the larvae died when they were small and before the pupal stage. The protein contents and esterase isozymes extracted from the larvae changed according to the doses of copper. The small‐molecular‐weight protein bands gradually became weaker or were lost as the copper levels increased. However, low doses of copper stimulated a stronger expression of a few proteins. These results indicate that low doses of copper generally have no lethal effects onD. melanogasterbecause a specific group of genes, which encode specific proteins, are probably activated in order to withstand the onslaught of stressful conditions. At high doses of copper in food (e.g. 320 mg L‐1), fly development and viability are significantly affected.
Online Zoological Collections of Australian Museums (OZCAM): a national approach to making zoological data available on the web
Authors: WALLIS Elycia J.
Published: July 2006

Museums increasingly seek to publish data on the internet. However, individual records from a natural sciences database are not meaningful. The combined results of a number of datasets can be made much more meaningful by using a graphical display. In this paper, a project undertaken by a group of Australian natural sciences collecting institutions is described. Issues about data quality and other aspects of data management are discussed.
Muscle protein expression of pikeperches (Stizostedion lucioperca and S. volgense)
Authors: ISLAM Asiful
Published: July 2006

The purpose of the present study was to investigate the muscle protein expression in two pikeperches (Stizostedion lucioperca and S. volgense) through intra‐ and intermyomeric composition of white muscles. Using denaturing 10% sodium dodecylsulfate‐polyacrylamide gel electrophoresis, muscle protein expression was studied in relation to within‐ and between‐species morphological development, sex, maturity and age of pikeperches. Myosin, actin and troponin have a distinct role in the contraction and length tension of muscle fibers of these species. No obvious intramyomeric differences were found in the myosin heavy chain of both species. Myosin light chains (15–38 kDa) have different expression in different age groups. The muscle protein of the fingerling and adult S. lucioperca had high molecular weight (50 kDa) myosin in contrast to the other Percid species. The molecular weight of actins increased comparatively in low‐age‐group fish. ATP is stored in myosin and released to cause contraction when myosin comes in contact with actin of the experimental fish. Troponin regulates increasing concentration of light‐chain myosin in mature fish. Because troponin T has been implicated in the regulation of skeletal muscle kinetics, muscle contraction kinetics was predicted in different age groups. The muscle proteins of both sexes of these species have polymorphism in various age groups but have no difference in similar aged fish. No muscle protein dimorphism was found in these Percid species. The white muscle protein composition and contractile properties affect power production during fast, unsteady movement and swimming.
The effectiveness of graded activity for low back pain in occupational healthcare
Authors: Steenstra I A , Anema J R , Bongers P M , de Vet H C W , Knol D L , van Mechelen W
Published: July 2006

Background:Low back pain is a common medical and social problem associated with disability and absence from work. Knowledge on effective return to work (RTW) interventions is scarce.

Objective:To determine the effectiveness of graded activity as part of a multistage RTW programme.

Design:Randomised controlled trial.

Setting:Occupational healthcare.

Subjects:112 workers absent from work for more than eight weeks due to low back pain were randomised to either graded activity (n = 55) or usual care (n = 57).

Intervention:Graded activity, a physical exercise programme aimed at RTW based on operant-conditioning behavioural principles.

Main outcome measures:The number of days off work until first RTW for more then 28 days, total number of days on sick leave during follow up, functional status, and severity of pain. Follow up was 26 weeks.

Results:Graded activity prolonged RTW. Median time until RTW was equal to the total number of days on sick leave and was 139 (IQR = 69) days in the graded activity group and 111 (IQR = 76) days in the usual care group (hazard ratio = 0.52, 95% CI 0.32 to 0.86). An interaction between a prior workplace intervention and graded activity, together with a delay in the start of the graded activity intervention, explained most of the delay in RTW (hazard ratio = 0.86, 95% CI 0.40 to 1.84 without prior intervention and 0.39, 95% CI 0.19 to 0.81 with prior intervention). Graded activity did not improve pain or functional status clinically significantly.

Conclusions:Graded activity was not effective for any of the outcome measures. Different interventions combined can lead to a delay in RTW. Delay in referral to graded activity delays RTW. In implementing graded activity special attention should be paid to the structure and process of care.
Cancer incidence among semiconductor and electronic storage device workers
Authors: Bender T J , Beall C , Cheng H , Herrick R F , Kahn A R , Matthews R , Sathiakumar N , Schymura M J , Stewart J H , Delzell E
Published: July 2006

Aims:To evaluate cancer incidence among workers at two facilities in the USA that made semiconductors and electronic storage devices.

Methods:89 054 men and women employed by International Business Machines (IBM) were included in the study. We compared employees’ incidence rates with general population rates and examined incidence patterns by facility, duration of employment, time since first employment, manufacturing era, potential for exposure to workplace environments other than offices and work activity.

Results:For employees at the semiconductor manufacturing facility, the standardised incidence ratio (SIR) for all cancers combined was 81 (1541 observed cases, 95% confidence interval (CI) 77 to 85) and for those at the storage device manufacturing facility the SIR was 87 (1319 observed cases, 95% CI 82 to 92). The subgroups of employees with ≥15 years since hiring and ≥5 years worked had 6–16% fewer total incidents than expected. SIRs were increased for several cancers in certain employee subgroups, but analyses of incidence patterns by potential exposure and by years spent and time since starting in specific work activities did not clearly indicate that the excesses were due to occupational exposure.

Conclusions:This study did not provide strong or consistent evidence of causal associations with employment factors. Data on employees with long potential induction time and many years worked were limited. Further follow-up will allow a more informative analysis of cancer incidence that might be plausibly related to workplace exposures in the cohort.
The risk of lung cancer with increasing time since ceasing exposure to asbestos and quitting smoking
Authors: Reid A , de Klerk N H , Ambrosini G L , Berry G , Musk A W
Published: July 2006

Objectives:To examine if the risk of lung cancer declines with increasing time since ceasing exposure to asbestos and quitting smoking, and to determine the relative asbestos effect between non-smokers and current smokers.

Methods:A cohort study of 2935 former workers of the crocidolite mine and mill at Wittenoom, who responded to a questionnaire on smoking first issued in 1979 and on whom quantitative estimates of asbestos exposure are known. Conditional logistic regression was used to relate asbestos exposure, smoking category, and risk of lung cancer.

Results:Eighteen per cent of the cohort reported never smoking; 66% of cases and 50% of non-cases were current smokers. Past smokers who ceased smoking within six years of the survey (OR = 22.1, 95% CI 5.6 to 87.0), those who ceased smoking 20 or more years before the survey (OR = 1.9, 95% CI 0.50 to 7.2), and current smokers (<20 cigarettes per day (OR = 6.8, 95% CI 2.0 to 22.7) or >20 cigarettes per day (OR = 13.2, 95% CI 4.1 to 42.5)) had higher risks of lung cancer compared to never smokers after adjusting for asbestos exposure and age. The asbestos effect between non-smokers and current smokers was 1.23 (95% CI 0.35 to 4.32).

Conclusion:Persons exposed to asbestos and tobacco but who subsequently quit, remain at an increased risk for lung cancer up to 20 years after smoking cessation, compared to never smokers. Although the relative risk of lung cancer appears higher in never and ex-smokers than in current smokers, those who both smoke and have been exposed to asbestos have the highest risk; this study emphasises the importance of smoking prevention and smoking cessation programmes within this high risk cohort.
Continuous cardiac output monitoring in humans by invasive and noninvasive peripheral blood pressure waveform analysis
Authors: Lu Zhenwei , Mukkamala Ramakrishna
Published: July 2006

We present an evaluation of a novel technique for continuous (i.e., automatic) monitoring of relative cardiac output (CO) changes by long time interval analysis of a peripheral arterial blood pressure (ABP) waveform in humans. We specifically tested the mathematical analysis technique based on existing invasive and noninvasive hemodynamic data sets. With the former data set, we compared the application of the technique to peripheral ABP waveforms obtained via radial artery catheterization with simultaneous thermodilution CO measurements in 15 intensive care unit patients in which CO was changing because of disease progression and therapy. With the latter data set, we compared the application of the technique to noninvasive peripheral ABP waveforms obtained via a finger-cuff photoplethysmography system with simultaneous Doppler ultrasound CO measurements made by an expert in 10 healthy subjects during pharmacological and postural interventions. We report an overall CO root-mean-squared normalized error of 15.3% with respect to the invasive hemodynamic data set and 15.1% with respect to the noninvasive hemodynamic data set. Moreover, the CO errors from the invasive and noninvasive hemodynamic data sets were only mildly correlated with mean ABP (ρ = 0.41, 0.37) and even less correlated with CO (ρ = −0.14, −0.17), heart rate (ρ = 0.04, 0.19), total peripheral resistance (ρ = 0.38, 0.10), CO changes (ρ = −0.26, −0.20), and absolute CO changes (ρ = 0.03, 0.38). With further development and successful prospective testing, the technique may potentially be employed for continuous hemodynamic monitoring in the acute setting such as critical care and emergency care.
Combination Therapy with Anti–CTL Antigen-4 and Anti-4-1BB Antibodies Enhances Cancer Immunity and Reduces Autoimmunity
Authors: Kocak Ergun , Lute Kenneth , Chang Xing , May Kenneth F. , Exten Katie R. , Zhang Huiming , Abdessalam Shahab F. , Lehman Amy M. , Jarjoura David , Zheng Pan , Liu Yang
Published: July 2006

The majority of cancer antigens identified thus far have limited expression in normal tissues. It has been suggested that autoimmune disease is a necessary price for cancer immunity. This notion is supported by a recent clinical trial involving an anti–CTL antigen-4 (CTLA-4) antibody that showed significant clinical responses but severe autoimmune diseases in melanoma patients. To selectively modulate cancer immunity and autoimmunity, we used anti-CTLA-4 and anti-4-1BB antibodies to treat mice with a preexisting cancer, MC38. The combination of the two antibodies led to CD8 T-cell-mediated rejection of large established MC38 tumors and long-lasting immunity to the same tumor cells, although the same regimen was not effective for B16 melanoma. More importantly, whereas individual antibodies induced inflammation and autoimmune manifestations, combination therapy increased cancer immunity while reducing autoimmunity. The reduction of autoimmune effects correlates with an increased function of regulatory T cells. Our results suggest a novel approach to simultaneously enhance cancer immunity and reduce autoimmunity. (Cancer Res 2006; 66(14): 7276-84)
Oncolytic Capacity of Attenuated Replicative Semliki Forest Virus in Human Melanoma Xenografts in Severe Combined Immunodeficient Mice
Authors: Vähä-Koskela Markus J.V. , Kallio Janne P. , Jansson Linda C. , Heikkilä Jari E. , Zakhartchenko Vasily A. , Kallajoki Markku A. , Kähäri Veli-Matti , Hinkkanen Ari E.
Published: July 2006

Oncolytic viruses have gained attention as a novel form of cancer treatment. Many viral vectors in use today have been rendered safe by deletion of genes encoding viral structural proteins, thus making them unable to spread beyond the first infected cells. Hence, such replication-deficient constructs may lack efficacy. Here, we analyzed the oncolytic potential of the replication-competent vector VA7-EGFP, based on the avirulent Semliki Forest virus (SFV) strain A7(74), to kill cancer cells in culture as well as to target s.c. human melanoma xenografts in severe combined immunodeficient (SCID) mice. VA7-EGFP was able to infect most cancer cell lines studied, leading to complete lysis of the cells within 72 hours after infection. In SCID mice grafted with A2058 human melanoma, marked regression of the xenografts was observed following a single injection of 106 plaque-forming units of virus given either i.p., i.v., or intratumorally. Histologic analysis revealed the presence of virus not only in all treated tumors but also in the brains of the treated mice, causing progressing neuropathology beginning at day 16 after infection. Following initial oncolysis, clusters of viable tumor cells were observed embedded in connective tissue, and at later stages, encapsulated tumor nodules had formed. Infection of melanoma cells from explant cultures of these nodules revealed that a portion of the cells were resistant to virus. To be eligible for use in virotherapy, the ability of avirulent SFV to spread within tumor tissue may have to be improved and the biological safety of the virus may have to be addressed thoroughly in higher animals. (Cancer Res 2006; 66(14): 7185-94)
Cell Adhesion Regulates CDC25A Expression and Proliferation in Acute Myeloid Leukemia
Authors: Fernandez-Vidal Anne , Ysebaert Loïc , Didier Christine , Betous Remy , De Toni Fabienne , Prade-Houdellier Naïs , Demur Cécile , Contour-Galcéra Marie-Odile , Prévost Grégoire P. , Ducommun Bernard , Payrastre Bernard , Racaud-Sultan Claire , Manenti Stéphane
Published: July 2006

The effects of cell adhesion on leukemia cell proliferation remain poorly documented and somehow controversial. In this work, we investigated the effect of adhesion to fibronectin on the proliferation of acute myeloid leukemia (AML) cell lines (U937 and KG1a) and CD34+ normal or leukemic primary cells. We observed an increased rate of proliferation of AML cells when adhered to fibronectin, concomitant with accelerated S-phase entry and accumulation of CDC25A. Conversely, normal CD34+ cell proliferation was decreased by adhesion to fibronectin with a concomitant drop in CDC25A expression. Importantly, we showed that both small interfering RNA (siRNA)–mediated CDC25A down-regulation and a recently developed CDC25 pharmacologic inhibitor impaired this adhesion-dependent proliferation, establishing a functional link between CDC25A accumulation and adhesion-dependent proliferation in leukemic cells. CDC25A accumulation was found only slightly dependent on transcriptional regulation and essentially due to modifications of the proteasomal degradation of the protein as shown using proteasome inhibitors and reverse transcription-PCR. Interestingly, CDC25A regulation was Chk1 dependent in these cells as suggested by siRNA-mediated down-regulation of this protein. Finally, we identified activation of the phosphatidylinositol 3-kinase/Akt pathway as an adhesion-dependent regulation mechanism of CDC25A protein expression. Altogether, our data show that in leukemic cells adhesion to fibronectin increases CDC25A expression through proteasome- and Chk1-dependent mechanisms, resulting in enhanced proliferation. They also suggest that these adhesion-dependent proliferation properties of hematopoietic cells may be modified during leukemogenesis. (Cancer Res 2006; 66(14): 7128-35)
AIMP3 Haploinsufficiency Disrupts Oncogene-Induced p53 Activation and Genomic Stability
Authors: Park Bum-Joon , Oh Young Sun , Park Seung Yong , Choi So Jung , Rudolph Cornelia , Schlegelberger Brigitte , Kim Sunghoon
Published: July 2006

AIMP3 (previously known as p18) was shown to up-regulate p53 in response to DNA damage. Here, we show that AIMP3 couples oncogenic stresses to p53 activation to prevent cell transformation. Growth factor– or Ras-dependent induction of p53 was blocked by single allelic loss of AIMP3 as well as by suppression of AIMP3. AIMP3 heterozygous cells became susceptible to cell transformation induced by oncogenes such as Ras or Myc alone. The transformed AIMP3+/− cells showed severe abnormality in cell division and chromosomal structure. Thus, AIMP3 plays crucial roles in p53-mediated tumor-suppressive response against oncogenic stresses via differential activation of ATM and ATR, and in the maintenance of genomic stability. (Cancer Res 2006; 66(14): 6913-8)
Expression of the FUS-CHOP Fusion Protein in Primary Mesenchymal Progenitor Cells Gives Rise to a Model of Myxoid Liposarcoma
Authors: Riggi Nicolò , Cironi Luisa , Provero Paolo , Suvà Mario-Luca , Stehle Jean-Christophe , Baumer Karine , Guillou Louis , Stamenkovic Ivan
Published: July 2006

A subset of sarcomas is associated with specific chromosomal translocations that give rise to fusion genes believed to participate in transformation and oncogenesis. Identification of the primary cell environment that provides permissiveness for the oncogenic potential of these fusion genes is essential to understand sarcoma pathogenesis. We have recently shown that expression of the EWS-FLI-1 fusion protein in primary mesenchymal progenitor cells (MPCs) suffices to develop Ewing's sarcoma-like tumors in mice. Because most sarcomas bearing unique chromosomal translocations are believed to originate from common progenitor cells, and because MPCs populate most organs, we expressed the sarcoma-associated fusion proteins FUS/TLS-CHOP, EWS-ATF1, and SYT-SSX1 in MPCs and tested the tumorigenic potential of these cells in vivo. Whereas expression of EWS-ATF1 and SYT-SSX1 failed to transform MPCs, FUS-CHOP–expressing cells formed tumors resembling human myxoid liposarcoma. Transcription profile analysis of these tumors revealed induction of transcripts known to be associated with myxoid liposarcoma and novel candidate genes, including PDGFA, whose expression was confirmed in human tumor samples. MPCFUS-CHOP and the previously described MPCEWS-FLI-1 tumors displayed distinct transcription profiles, consistent with the different target gene repertoires of their respective fusion proteins. Unexpectedly, a set of genes implicated in cell survival and adhesion displayed similar behavior in the two tumors, suggesting events that may be common to primary MPC transformation. Taken together, our observations suggest that expression of FUS-CHOP may be the initiating event in myxoid liposarcoma pathogenesis, and that MPCs may constitute one cell type from which these tumors originate. (Cancer Res 2006; 66(14): 7016-23)
Regulation of Human Nitric Oxide Synthase 2 Expression by Wnt β-Catenin Signaling
Authors: Du Qiang , Park Kyung Soo , Guo Zhong , He Peijun , Nagashima Makoto , Shao Lifang , Sahai Rohit , Geller David A. , Hussain S. Perwez
Published: July 2006

Nitric oxide (NO·), an important mediator of inflammation, and β-catenin, a component of the Wnt–adenomatous polyposis coli signaling pathway, contribute to the development of cancer. We have identified two T-cell factor 4 (Tcf-4)-binding elements (TBE1 and TBE2) in the promoter of human inducible NO synthase 2 (NOS2). We tested the hypothesis that β-catenin regulates human NOS2 gene. Mutation in either of the two TBE sites decreased the basal and cytokine-induced NOS2 promoter activity in different cell lines. The promoter activity was significantly reduced when both TBE1 and TBE2 sites were mutated (P < 0.01). Nuclear extract from HCT116, HepG2, or DLD1 cells bound to NOS2 TBE1 or TBE2 oligonucleotides in electrophoretic mobility shift assays and the specific protein-DNA complexes were supershifted with anti-β-catenin or anti-Tcf-4 antibody. Overexpression of β-catenin and Tcf-4 significantly increased both basal and cytokine-induced NOS2 promoter activity (P < 0.01), and the induction was dependent on intact TBE sites. Overexpression of β-catenin or Tcf-4 increased NOS2 mRNA and protein expression in HCT116 cells. Lithium chloride (LiCl), an inhibitor of glycogen synthase kinase-3β, increased cytosolic and nuclear β-catenin level, NOS2 expression, and NO· production in primary human and rat hepatocytes and cancer cell lines. Treatment with Wnt-3A-conditioned medium increased β-catenin and NOS2 expression in fetal human hepatocytes. When administered in vivo, LiCl increased hepatic β-catenin level in a dose-dependent manner with simultaneous increase in NOS2 expression. These data are consistent with the hypothesis that β-catenin up-regulates NOS2 and suggest a novel mechanism by which the Wnt/β-catenin signaling pathway may contribute to cancer by increasing NO· production. (Cancer Res 2006; 66(14): 7024-31)
Differential Effect of Bryostatin 1 and Phorbol 12-Myristate 13-Acetate on HOP-92 Cell Proliferation Is Mediated by Down-regulation of Protein Kinase Cδ
Authors: Choi Sung Hee , Hyman Tehila , Blumberg Peter M.
Published: July 2006

Bryostatin 1 is currently in clinical trials as a cancer chemotherapeutic agent. Although bryostatin 1, like phorbol 12-myristate 13-acetate (PMA), is a potent activator of protein kinase C (PKC), it induces only a subset of those responses induced by PMA and antagonizes others. We report that, in the HOP-92 non–small cell lung cancer line, bryostatin 1 induced a biphasic proliferative response, with maximal proliferation at 1 to 10 nmol/L. This biphasic response mirrored a biphasic suppression of the level of PKCδ protein, with maximal suppression likewise at 1 to 10 nmol/L bryostatin 1. The typical phorbol ester PMA, in contrast to bryostatin 1, had no effect on the level of PKCδ and modest suppression of cell proliferation, particularly evident at later treatment times. Flow cytometric analysis revealed changes in the fraction of cells in the G0-G1 and S phases corresponding to the effects on proliferation. Cells overexpressing PKCδ exhibited a lower rate of cell proliferation compared with control untreated cells and showed neither a proliferative response nor a loss of PKCδ in response to bryostatin 1. Conversely, treatment with PKCδ small interfering RNA significantly increased the cellular growth compared with controls. We conclude that the differential effect on cellular proliferation induced by bryostatin 1 compared with PMA reflects the differential suppression of PKCδ. (Cancer Res 2006; 66(14): 7261-9)
Gene Expression Preferentially Regulated by Tamoxifen in Breast Cancer Cells and Correlations with Clinical Outcome
Authors: Frasor Jonna , Chang Edmund C. , Komm Barry , Lin Chin-Yo , Vega Vinsensius B. , Liu Edison T. , Miller Lance D. , Smeds Johanna , Bergh Jonas , Katzenellenbogen Benita S.
Published: July 2006

The beneficial effect of the selective estrogen receptor (ER) modulator tamoxifen in the treatment and prevention of breast cancer is assumed to be through its ability to antagonize the stimulatory actions of estrogen, although tamoxifen can also have some estrogen-like agonist effects. Here, we report that, in addition to these mixed agonist/antagonist actions, tamoxifen can also selectively regulate a unique set of >60 genes, which are minimally regulated by estradiol (E2) or raloxifene in ERα-positive MCF-7 human breast cancer cells. This gene regulation by tamoxifen is mediated by ERα and reversed by E2 or ICI 182,780. Introduction of ERβ into MCF-7 cells reverses tamoxifen action on ∼75% of these genes. To examine whether these genes might serve as markers of tamoxifen sensitivity and/or the development of resistance, their expression level was examined in breast cancers of women who had received adjuvant therapy with tamoxifen. High expression of two of the tamoxifen-stimulated genes, YWHAZ/14-3-3z and LOC441453, was found to correlate significantly with disease recurrence following tamoxifen treatment in women with ER-positive cancers and hence seem to be markers of a poor prognosis. Our data indicate a new dimension in tamoxifen action, involving gene expression regulation that is tamoxifen preferential, and identify genes that might serve as markers of tumor responsiveness or resistance to tamoxifen therapy. This may have a potential effect on the choice of tamoxifen versus aromatase inhibitors as adjuvant endocrine therapy. (Cancer Res 2006; 66(14): 7334-40)
Differential Binding of Plasminogen, Plasmin, and Angiostatin4.5 to Cell Surface β-Actin: Implications for Cancer-Mediated Angiogenesis
Authors: Wang Hao , Doll Jennifer A. , Jiang Keyi , Cundiff Deborah L. , Czarnecki Jarema S. , Wilson Mindy , Ridge Karen M. , Soff Gerald A.
Published: July 2006

Angiostatin4.5 (AS4.5) is the product of plasmin autoproteolysis and consists of kringles 1 to 4 and ∼85% of kringle 5. In culture, cancer cell surface globular β-actin mediates plasmin autoproteolysis to AS4.5. We now show that plasminogen binds to prostate cancer cells and that the binding colocalizes with surface β-actin, but AS4.5 does not bind to the cell surface. Plasminogen and plasmin bind to immobilized β-actin similarly, with a Kd of ∼140 nmol/L. The binding is inhibited by ε-aminocaproic acid (εACA), indicating the requirement for a lysine-kringle domain interaction. Using a series of peptides derived from β-actin in competitive binding studies, we show that the domain necessary for plasminogen binding is within amino acids 55 to 69 (GDEAQSKRGILTLKY). Substitution of Lys61 or Lys68 with arginine results in the loss of the ability of the peptide to block plasminogen binding, indicating that Lys61 and Lys68 are essential for plasminogen binding. Other actin peptides, including peptides with lysine, did not inhibit the plasminogen-actin interaction. AS4.5 did not bind actin at concentrations up to 40 μmol/L. Plasminogen, plasmin, and AS4.5 all contain kringles 1 to 4; however, kringle 5 is truncated in AS4.5. Isolated kringle 5 binds to actin, suggesting intact kringle 5 is necessary for plasminogen and plasmin to bind to cell surface β-actin, and the truncated kringle 5 in AS4.5 results in its release from β-actin. These data may explain the mechanism by which AS4.5 is formed locally on cancer cell surfaces and yet acts on distant sites. (Cancer Res 2006; 66(14): 7211-5)
Prx1 Suppresses Radiation-Induced c-Jun NH2-Terminal Kinase Signaling in Lung Cancer Cells through Interaction with the Glutathione S-Transferase Pi/c-Jun NH2-Terminal Kinase Complex
Authors: Kim Yun-Jeong , Lee Weon-Sup , Ip Clement , Chae Ho-Zoon , Park Eun-Mi , Park Young-Mee
Published: July 2006

Radiotherapy is one of the major treatment modalities for lung cancer. Cell killing by ionizing radiation is mediated primarily through the reactive oxygen species (ROS) and ROS-driven oxidative stress. Prx1, a peroxiredoxin family member, was shown to be frequently elevated in lung cancer cells and tissues. Although the antioxidant function of Prx1 is expected to affect the radiotherapy response of lung cancer, the physiologic significance of its peroxidase activity in irradiated cells is unclear because the catalytic Cys52 is easily inactivated by ROS due to its overoxidation to sulfinic or sulfonic acid. In this study, we investigated the role of Prx1 in radiation sensitivity of human lung cancer cells, with special emphasis on the redox status of the catalytic Cys52. We found that overexpression of Prx1 enhances the clonogenic survival of irradiated cells and suppresses ionizing radiation–induced c-Jun NH2-terminal kinase (JNK) activation and apoptosis. The peroxidase activity of Prx1, however, is not essential for inhibiting JNK activation. The latter effect is mediated through its association with the glutathione S-transferase pi (GSTpi)-JNK complex, thereby preventing JNK release from the complex. Reduced JNK activation is observed when the peroxidase activity of Prx1 is compromised by Cys52 overoxidation or in the presence of the Cys52 to Ser52 mutant (Prx1C52S) lacking peroxidase activity. We show that both Prx1 and Prx1C52S interact with the GSTpi-JNK complex and suppress the release of JNK from the complex. Our study provides new insight into the antiapoptotic function of Prx1 in modulating radiosensitivity and provides the impetus to monitor the influence of Prx1 levels in the management of lung cancer. (Cancer Res 2006; 66(14): 7136-42)
Periostin Promotes Invasion and Anchorage-Independent Growth in the Metastatic Process of Head and Neck Cancer
Authors: Kudo Yasusei , Ogawa Ikuko , Kitajima Shojiro , Kitagawa Masae , Kawai Hidehiko , Gaffney Patrick M. , Miyauchi Mutsumi , Takata Takashi
Published: July 2006

Head and neck squamous cell carcinoma (HNSCC) is one of the most common types of human cancer. Typically, HNSCC cells show persistent invasion that frequently leads to local recurrence and distant lymphatic metastasis. However, molecular mechanisms associated with the invasion and metastasis of HNSCC remain poorly understood. Here, we identified periostin as an invasion-promoting factor in HNSCC by comparing the gene expression profiles between parent HNSCC cells and a highly invasive clone. Indeed, periostin overexpression promoted invasion and anchorage-independent growth both in vitro and in vivo in HNSCC cells. Moreover, periostin-overexpressing cells spontaneously metastasized to cervical lymph nodes and to the lung through their aggressive invasiveness in an orthotopic mouse model of HNSCC. Interestingly, periostin was highly expressed in HNSCCs in comparison with normal tissues, and the level of periostin expression was well correlated with the invasiveness of HNSCC cases. In summary, these findings suggest that periostin plays an important role in the invasion and anchorage-independent growth of HNSCC. (Cancer Res 2006; 66(14): 6928-35)
Identification and Clinical Significance of Circulating Endothelial Progenitor Cells in Human Non–Small Cell Lung Cancer
Authors: Dome Balazs , Timar Jozsef , Dobos Judit , Meszaros Livia , Raso Erzsebet , Paku Sandor , Kenessey Istvan , Ostoros Gyula , Magyar Melinda , Ladanyi Andrea , Bogos Krisztina , Tovari Jozsef
Published: July 2006

Until recently, it was generally accepted that vascularization of tumors arises exclusively from endothelial sprouting. Whether circulating bone marrow–derived endothelial progenitor cells (EPC) participate in the progression of non–small cell lung cancer (NSCLC) has not yet been evaluated. EPCs labeled with CD34, CD133, and vascular endothelial growth factor receptor-2 (VEGFR2) antibodies were counted by flow cytometry in the peripheral blood of 53 NSCLC patients. Furthermore, by means of a quantitative reverse transcription-PCR approach, we measured VEGFR2, CD133, CD34, and VE-cadherin mRNA in the peripheral blood samples of the same patient population. EPCs in tumor samples were identified by confocal microscopy using CD31, CD34, CD133, and VEGFR2 antibodies. Although immunofluorescent labeling of microvessels made clear that incorporation of EPCs is a rare phenomenon in NSCLC tissue (9 of 22 cases), circulating EPC levels before therapeutic intervention were increased in NSCLC patients (P < 0.002, versus healthy controls), and high pretreatment circulating EPC numbers correlated with poor overall survival (P < 0.001). Furthermore, in the subgroup of responders to treatment, the posttreatment EPC numbers in the peripheral blood were significantly lower compared with nonresponding patients. Interestingly, pretreatment mRNA levels of CD133, VE-cadherin, and CD34 were not significantly increased in NSCLC patients, whereas VEGFR2 expression was increased by 80-fold. Moreover, posttreatment VEGFR2 mRNA level in the peripheral blood was significantly higher in the subgroup of nonresponding patients when compared with posttreatment level of patients responding to antitumor therapy. Circulating levels of bone marrow–derived EPCs are significantly increased in NSCLC patients and correlate with clinical behavior. (Cancer Res 2006; 66(14): 7341-7)
Synergy between In situ Cryoablation and TLR9 Stimulation Results in a Highly Effective In vivo Dendritic Cell Vaccine
Authors: den Brok Martijn H.M.G.M. , Sutmuller Roger P.M. , Nierkens Stefan , Bennink Erik J. , Toonen Liza W.J. , Figdor Carl G. , Ruers Theo J.M. , Adema Gosse J.
Published: July 2006

Dendritic cells (DC) are professional antigen-presenting cells that play a pivotal role in the induction of immunity. Ex vivo–generated, tumor antigen–loaded mature DC are currently exploited as cancer vaccines in clinical studies. However, antigen loading and maturation of DC directly in vivo would greatly facilitate the application of DC-based vaccines. We have previously shown that in situ tumor destruction by ablative treatments efficiently delivers antigens for the in vivo induction of antitumor immunity. In this article, we show that although 20% of the draining lymph node DCs acquire intratumorally injected model antigens after in situ cryoablation, only partial protection against a subsequent tumor rechallenge is observed. However, we also show that a combination treatment of cryoablation plus TLR9 stimulation via CpG-oligodeoxynucleotides is far more effective in the eradication of local and systemic tumors than either treatment modality alone. Analysis of the underlying mechanism revealed that in situ tumor ablation synergizes with TLR9 stimulation to induce DC maturation and efficient cross-presentation in tumor-bearing mice, leading to superior DC function in vivo. Therefore, in situ tumor destruction in combination with CpG-oligodeoxynucleotide administration creates a unique “in situ DC vaccine” that is readily applicable in the clinic. (Cancer Res 2006; 66(14): 7285-92)
Polo-Like Kinase 1: Target and Regulator of Anaphase-Promoting Complex/Cyclosome–Dependent Proteolysis
Authors: Eckerdt Frank , Strebhardt Klaus
Published: July 2006

Polo-like kinase 1 (Plk1) is a key regulator of progression through mitosis. Although Plk1 seems to be dispensable for entry into mitosis, its role in spindle formation and exit from mitosis is crucial. Recent evidence suggests that a major role of Plk1 in exit from mitosis is the regulation of inhibitors of the anaphase-promoting complex/cyclosome (APC/C), such as the early mitotic inhibitor 1 (Emi1) and spindle checkpoint proteins. Thus, Plk1 and the APC/C control mitotic regulators by both phosphorylation and targeted ubiquitylation to ensure the fidelity of chromosome separation at the metaphase to anaphase transition. The mechanisms underlying the control of genomic stability by Plk1 are discussed in this review. (Cancer Res 2006; 66(14): 6895-8)
Systemic Induction of the Angiogenesis Switch by the Tetraspanin D6.1A/CO-029
Authors: Gesierich Sabine , Berezovskiy Igor , Ryschich Eduard , Zöller Margot
Published: July 2006

Expression of the tetraspanin CO-029 is associated with poor prognosis in patients with gastrointestinal cancer. In a pancreatic tumor line, overexpression of the rat homologue, D6.1A, induces lethally disseminated intravascular coagulation, suggesting D6.1A engagement in angiogenesis. D6.1A-overexpressing tumor cells induce the greatest amount of angiogenesis in vivo, and tumor cells as well as exosomes derived thereof strikingly increase endothelial cell branching in vitro. Tumor cell–derived D6.1A stimulates angiogenic factor transcription, which includes increased matrix metalloproteinase and urokinase-type plasminogen activator secretion, pronounced vascular endothelial growth factor expression in fibroblasts, vascular endothelial growth factor receptor expression, and strong D6.1A up-regulation in sprouting endothelium. Thus, D6.1A initiates an angiogenic loop that, probably due to the abundance of D6.1A in tumor-derived exosomes, reaches organs distant from the tumor. Most importantly, because of the strong D6.1A up-regulation on sprouting capillaries, angiogenesis could be completely inhibited by a D6.1A-specific antibody, irrespective of whether or not the tumor expresses D6.1A. Tetraspanins have been suggested to be involved in morphogenesis. This is the first report that a tetraspanin, CO-029/D6.1A, promotes tumor growth by its capacity to induce systemic angiogenesis that can effectively, and with high selectivity for sprouting endothelium, be blocked by a D6.1A-specific antibody. (Cancer Res 2006; 66(14): 7083-94)