bluuebunny/crossref_metadata_2025_split huggingface dataset:
Overexpression of the β‐catenin binding domain of cadherin selectively kills colorectal cancer cells
Authors: Pierce Michael , Wang Chunwei , Stump Mark , Kamb Alexander 
Published: August 2003

AbstractThe β‐catenin pathway is involved in growth, differentiation and tumor formation. Suppression of pathway activity by expressed inhibitors can cause growth arrest or apoptosis in certain colon carcinoma lines. We compare the effects of 2 pathway inhibitors, a VE‐cadherin cytoplasmic domain fragment (Cad5CD) and a truncated, dominant‐negative Tcf4 (TcfDN), using a microplate assay for cell death and microarray gene expression analysis. The cell‐lethal assay shows that Cad5CD, when expressed in HT29 human colon tumor cells and 3 non‐colon lines, selectively kills the HT29 cells. Cad5CD overexpression inhibits β‐catenin/Tcf4 transcriptional activity, as determined by results from microarray experiments. Our results support the view that β‐catenin is an attractive anti‐cancer target, especially if the Cad5CD binding site or Cad5CD itself can be exploited for drug development. In addition, therapeutically relevant phenotypes such as drug selectivity may be difficult to predict from gene expression analysis alone. Other more specialized phenotypic tests such as cell‐lethal assays may be required. © 2003 Wiley‐Liss, Inc.
ASC/TMS1, a caspase‐1 activating adaptor, is downregulated by aberrant metylation in human melanoma
Authors: Guan Xin , Sagara Junji , Yokoyama Taro , Koganehira Yoko , Oguchi Misae , Saida Toshiaki , Taniguchi Shun'ichiro 
Published: August 2003

AbstractASC/TMS1 is an adaptor protein activating caspase‐1 that stimulates processing of proIL‐1β and proIL‐18. ASC was reported to be aberrantly methylated and silenced in human breast cancers. In our present study, ASC expression was examined in 12 melanoma cell lines by Western blot analysis and in 18 benign melanocytic nevi and 32 melanoma tissues by immunohistochemical staining. ASC expression was absent or reduced in 7 of 12 (58.3%) cell lines and in 20 of 32 (62.5%) melanoma tissues, whereas all 18 benign melanocytic nevi showed intensive ASC expression. To investigate whether ASC silencing in melanoma is involved in aberrant methylation, methylation specific PCR was carried out. Five of ten (50%) melanoma tissues exhibited methylation in CpG island of ASC companied with reduced ASC expression. Six of twelve (50%) melanoma cell lines showed aberrant methylation in the ASC gene, and 4 of the 6 (66.7%) methylation positive cell lines exhibited reduced ASC expression. We characterized methylation patterns in melanoma cell lines by using bisulfite genomic sequencing, and found that the degree of aberrant methylation correlated with the level of reductive ASC expression. Treatment with demethylating agent 5‐aza‐2′‐deoxycytidine resulted in both demethylation of the ASC gene and the upregulation of ASC expression in the methylation positive melanoma cell lines. Our study shows that ASC is downregulated in melanoma, and that its suppression is partially mediated by aberrant methylation. © 2003 Wiley‐Liss, Inc.
CYP17 polymorphisms in relation to risks of prostate cancer and benign prostatic hyperplasia: A population‐based study in China
Authors: Madigan M. Patricia , Gao Yu‐Tang , Deng Jie , Pfeiffer Ruth M. , Chang Bao‐Li , Zheng Siqun , Meyers Deborah A. , Stanczyk Frank Z. , Xu Jianfeng , Hsing Ann W. 
Published: August 2003

AbstractBecause androgens likely play a key role in prostate growth and prostate cancer development, variants of genes involved in androgen biosynthesis may be related to prostate cancer risk. The enzyme P450c17α, encoded by the CYP17 gene, catalyzes the conversion of progesterone and pregnenolone into precursors of potent androgens. In the 5′ promoter region of the CYP17 gene, a T (A1 allele) to C substitution (A2 allele) has been hypothesized to increase CYP17 gene expression, resulting in higher levels of androgens. To investigate a possible role of CYP17 in prostate diseases, we evaluated the risk of prostate cancer and benign prostatic hyperplasia (BPH) in relation to variation in CYP17 genotype in a population‐based case‐control study conducted in Shanghai, China. The study included 174 prostate cancer cases, 182 BPH cases and 274 population controls. We observed no statistically significant overall associations of CYP17 genotypes with prostate cancer risk, although associations of the A1/A1 (odds ratio (OR) =1.42, 95% confidence interval (CI) 0.83–2.48) and A1/A2 (OR 1.41, 95% CI 0.91–2.17) genotypes with prostate cancer were suggested. A similar association of the A1/A1 genotype with BPH was suggested. We found no associations of CYP17 genotypes with serum sex hormone levels or other biomarkers after correction for multiple comparisons. Large population‐based studies are needed to clarify whether CYP17 plays a role in prostate cancer risk and whether genotype effects vary in different racial/ethnic and other subgroups. © 2003 Wiley‐Liss, Inc.
Granulocyte/macrophage‐colony stimulating factor and interleukin‐4 expand and activate type‐1 dendritic cells (DC1) when administered in vivo to cancer patients
Authors: Kiertscher Sylvia M. , Gitlitz Barbara J. , Figlin Robert A. , Roth Michael D. 
Published: August 2003

AbstractTwo rare populations of cells with the features of dendritic cell precursors (preDC) can be identified in human peripheral blood. PreDC1 are HLA‐DR+/CD11c+ cells which mature into DC1 capable of stimulating Th1 responses. In contrast, preDC2 are HLA‐DR+/CD11c−/CD123+ cells that promote Th2 responses when matured into DC2. We hypothesized that administration of GM‐CSF and IL‐4, growth factors for DC1, would specifically augment the number and function of circulating DC1 in vivo. Patients with advanced metastatic cancer were treated with GM‐CSF (2.5 μg/kg/day) and IL‐4 (4 or 6 μg/kg/day) for 7 days. Cytokine administration at the highest IL‐4 dose produced an average 2.3‐fold increase in preDC2 number, but a 6.5‐fold increase in preDC1, resulting in an increased ratio of circulating preDC1:preDC2 from 1.4:1 pre‐treatment to 4.3:1 after cytokine therapy. DC1 precursors identified after in vivo therapy were larger, more complex and expressed higher levels of HLA‐DR, CD11c and CD80 than pre‐treatment cells. DC1 isolated from the peripheral blood of patients receiving GM‐CSF/IL‐4 therapy demonstrated MLR activity comparable to that of monocyte‐derived DC generated in vitro from the patients' pre‐treatment blood using GM‐CSF and IL‐4. We conclude that systemic administration of GM‐CSF and IL‐4 preferentially expands and matures the preDC1 population in vivo. These effects correlate with antigen‐presenting activity, providing a mechanism by which systemic GM‐CSF and IL‐4 might stimulate anti‐tumor immunity in vivo. © 2003 Wiley‐Liss, Inc.
Fas ligand expressed in colon cancer is not associated with increased apoptosis of tumor cells in vivo
Authors: Houston Aileen , Waldron‐Lynch Frank D. , Bennett Michael W. , Roche Desmond , O'Sullivan Gerald C. , Shanahan Fergus , O'Connell Joe 
Published: August 2003

AbstractExpression of Fas ligand (FasL/CD95L) may help to maintain colon cancers in a state of immune privilege by inducing apoptosis of antitumor immune effector cells. Colon tumor‐derived cell lines appear to be relatively insensitive to apoptosis mediated by their own or exogenous FasL in vitro, despite expression of cell surface Fas. In our present study, we sought to investigate if FasL upregulated in human colon cancers leads to any increase in apoptosis of the tumor cells in vivo. FasL and Fas receptor (APO‐1/CD95) expression by tumor cells were detected immunohistochemically. Apoptotic tumor cell death was detected by immunohistochemistry for caspase‐cleaved cytokeratin‐18. FasL expression did not correlate with the extent of apoptosis of tumor cells. There was no significant local difference in the frequency of apoptosis of tumor cells between tumor nests that expressed FasL (mean = 2.4%) relative to those that did not (mean = 2.6%) (p = 0.625, n = 10; Wilcoxon signed rank). FasL expressed by the tumor cells appeared to be functional, since FasL expression in tumor nests correlated with diminished infiltration of tumor‐infiltrating lymphocytes (TILs). TILs were detected using immunohistochemistry for CD45. Expression of FasL by tumor nests was associated with a mean 4‐fold fewer TILs relative to FasL‐negative nests (range 2.4–33‐fold, n = 10, p < 0.003). Together, our results indicate that colon tumors are insensitive to FasL‐mediated apoptosis in vivo. © 2003 Wiley‐Liss, Inc.
Dual variant of Epstein‐Barr virus in Hodgkin/Reed‐Sternberg cells: Single‐cell PCR study on latent membrane protein‐1 gene
Authors: Kim Lian‐hua , Peh Suat‐cheng , Poppema Sibrand 
Published: August 2003

AbstractIsolation of single cells permits analysis of DNA or RNA from individual cells among heterogeneous populations. This technique is particularly useful in the study of classical Hodgkin's lymphoma (cHL) due to the scarcity of H/RS tumor cells among large numbers of reactive leukocytes. In a previous study, we found a high frequency of dual LMP‐1 variant (concurrent presence of deleted and nondeleted variants) in cHL from whole‐tissue sections. For the present study, we applied a single‐cell isolation technique to determine the LMP‐1 oncogene variant in EBV‐associated H/RS cells. Five cases of EBV‐infected cHL, containing nondeleted (n = 1), deleted (n = 1) and dual infection (n =3) based on whole‐tissue section analysis, were selected for study. Paraffin‐embedded tissue sections were stained with antibody to LMP‐1 and positively stained H/RS cells isolated using a semiautomated micromanipulator. Each isolated single cell was subjected to PCR for amplification of the LMP‐1 gene flanking the 30 bp deletion region and Xho1 restriction site. Cases with either nondeleted variant or the deleted variant showed similar LMP‐1 variant expression in isolated single H/RS cells. However, 1 of the 3 cases with dual variants showed only the deleted variant in H/RS cells. The other 2 cases showed mixed patterns of deleted, nondeleted and dual LMP‐1 variants in isolated single H/RS cells. All cases showed loss of the Xho1 restriction site, with the exception of the case with nondeleted LMP‐1. Results of single‐H/RS cell analysis of the Xho1 restriction site concur with those of whole‐tissue section amplification. A mixed pattern of LMP‐1 variants was observed in isolated H/RS cells, and it is speculated that this is due to the accumulation of mutation and deletion events. © 2003 Wiley‐Liss, Inc.
Adaptation to the land: The skin of reptiles in comparison to that of amphibians and endotherm amniotes
Authors: Alibardi Lorenzo 
Published: August 2003

AbstractThe adaptation to land from amphibians to amniotes was accompanied by drastic changes of the integument, some of which might be reconstructed by studying the formation of the stratum corneum during embryogenesis. As the first amniotes were reptiles, the present review focuses on past and recent information on the evolution of reptilian epidermis and the stratum corneum. We aim to generalize the discussion on the evolution of the skin in amniotes. Corneous cell envelopes were absent in fish, and first appeared in adult amphibian epidermis. Stem reptiles evolved a multilayered stratum corneum based on a programmed cell death, intensified the production of matrix proteins (e.g., HRPs), corneous cell envelope proteins (e.g., loricrine‐like, sciellin‐like, and transglutaminase), and complex lipids to limit water loss. Other proteins were later produced in association to the soft or hairy epidermis in therapsids (e.g., involucrin, profilaggrin‐filaggrin, trichohyalin, trichocytic keratins), or to the hard keratin of hairs, quills, horns, claws (e.g., tyrosine‐rich, glycine‐rich, sulphur‐rich matrix proteins). In sauropsids special proteins associated to hard keratinization in scales (e.g., scale beta‐keratins, cytokeratin associated proteins) or feathers (feather beta‐keratins and HRPs) were originated. The temporal deposition of beta‐keratin in lepidosaurian reptiles originated a vertical stratified epidermis and an intraepidermal shedding layer. The evolutions of the horny layer in Therapsids (mammals) and Saurospids (reptiles and birds) are discussed. The study of the molecules involved in the dermo‐epidermal interactions in reptilian skin and the molecular biology of epidermal proteins are among the most urgent future areas of research in the biology of reptilian skin. J. Exp. Zool. (Mol. Dev. Evol.) 298B:12–41, 2003. © 2003 Wiley‐Liss, Inc.
Avian skin development and the evolutionary origin of feathers
Authors: Sawyer Roger H. , Knapp Loren W. 
Published: August 2003

AbstractThe discovery of several dinosaurs with filamentous integumentary appendages of different morphologies has stimulated models for the evolutionary origin of feathers. In order to understand these models, knowledge of the development of the avian integument must be put into an evolutionary context. Thus, we present a review of avian scale and feather development, which summarizes the morphogenetic events involved, as well as the expression of the beta (β) keratin multigene family that characterizes the epidermal appendages of reptiles and birds. First we review information on the evolution of the ectodermal epidermis and its beta (β) keratins. Then we examine the morphogenesis of scutate scales and feathers including studies in which the extraembryonic ectoderm of the chorion is used to examine dermal induction. We also present studies on the scaleless (sc) mutant, and, because of the recent discovery of “four‐winged” dinosaurs, we review earlier studies of a chicken strain, Silkie, that expresses ptilopody (pti), “feathered feet.” We conclude that the ability of the ectodermal epidermis to generate discrete cell populations capable of forming functional structural elements consisting of specific members of the β keratin multigene family was a plesiomorphic feature of the archosaurian ancestor of crocodilians and birds. Evidence suggests that the discrete epidermal lineages that make up the embryonic feather filament of extant birds are homologous with similar embryonic lineages of the developing scutate scales of birds and the scales of alligators. We believe that the early expression of conserved signaling modules in the embryonic skin of the avian ancestor led to the early morphogenesis of the embryonic feather filament, with its periderm, sheath, and barb ridge lineages forming the first protofeather. Invagination of the epidermis of the protofeather led to formation of the follicle providing for feather renewal and diversification. The observations that scale formation in birds involves an inhibition of feather formation coupled with observations on the feathered feet of the scaleless (High‐line) and Silkie strains support the view that the ancestor of modern birds may have had feathered hind limbs similar to those recently discovered in nonavian dromaeosaurids. And finally, our recent observation on the bristles of the wild turkey beard raises the possibility that similar integumentary appendages may have adorned nonavian dinosaurs, and thus all filamentous integumentary appendages may not be homologous to modern feathers. J. Exp. Zool. (Mol. Dev. Evol.) 298B:57–72, 2003. © 2003 Wiley‐Liss, Inc.
Molecular biology of feather morphogenesis: A testable model for evo‐devo research
Authors: Widelitz Randall B. , Jiang Ting Xin , Yu Mingke , Shen Ted , Shen Jen‐Yee , Wu Ping , Yu Zhicao , Chuong Cheng‐Ming 
Published: August 2003

AbstractDarwin's theory describes the principles that are responsible for evolutionary change of organisms and their attributes. The actual mechanisms, however, need to be studied for each species and each organ separately. Here we have investigated the mechanisms underlying these principles in the avian feather. Feathers comprise one of the most complex and diverse epidermal organs as demonstrated by their shape, size, patterned arrangement and pigmentation. Variations can occur at several steps along each level of organization, leading to highly diverse forms and functions. Feathers develop gradually during ontogeny through a series of steps that may correspond to the evolutionary steps that were taken during the phylogeny from a reptilian ancestor to birds. These developmental steps include 1) the formation of feather tract fields on the skin surfaces; 2) periodic patterning of the individual feather primordia within the feather tract fields; 3) feather bud morphogenesis establishing anterio ‐ posterior (along the cranio ‐ caudal axis) and proximo ‐ distal axes; 4) branching morphogenesis to create the rachis, barbs and barbules within a feather bud; and 5) gradual modulations of these basic morphological parameters within a single feather or across a feather tract. Thus, possibilities for variation in form and function of feathers occur at every developmental step. In this paper, principles guiding feather tract formation, distributions of individual feathers within the tracts and variations in feather forms are discussed at a cellular and molecular level. J. Exp. Zool. (Mol. Dev. Evol.) 298B:109–122, 2003. © 2003 Wiley‐Liss, Inc.
Horse hooves and bird feathers: Two model systems for studying the structure and development of highly adapted integumentary accessory organs—the role of the dermo‐epidermal interface for the micro‐architecture of complex epidermal structures
Authors: Bragulla Hermann , Hirschberg Ruth M. 
Published: August 2003

AbstractAccessory organs of the integument are locally modified parts of the potentially feather‐bearing skin in birds (e.g., the rhamphotheca, claws, or scales), and of the potentially hairy skin in mammals (e.g., the rhinarium, nails, claws, or hooves). These special parts of the integument are characterised by a modified structure of their epidermal, dermal and subcutaneous layers. The developmental processes of these various integumentary structures in birds and mammals show both similarities and differences. For example, the development of the specialised epidermal structures of both feathers and the hoof capsule is influenced by the local three‐dimensional configuration of the dermis. However, in feathers, in contrast to hooves, the arrangement of the corneous cells is only partially a direct result of the particular arrangement and shape of the dermal surface of the papillary body. Whereas the diameter of the feather papilla, as well as the number, length, and width of dermal ridges on the surface of the feather papilla influence the three‐dimensional architecture of the feather rami, there is no apparent direct correlation between the dermo‐epidermal interface and the development of the highly ordered architecture of the radii and hamuli in the feather vane. In order to elucidate this morphogenic problem and the problem of locally different processes of keratinisation and cornification, the structure and development of feathers in birds are compared to those of the hoof capsule in horses. The equine hoof is the most complex mammalian integumentary structure, which is determined directly by the dermal surface of the papillary body. Perspectives for further research on the development of modified integumentary structures, such as the role of the dermal microangioarchitecture and the selective adhesion and various differentiation pathways of epidermal cells, are discussed. J. Exp. Zool. (Mol. Dev. Evol.) 298B:140–151, 2003. © 2003 Wiley‐Liss, Inc.
Long‐term evaluation of retinal function in Prph2Rd2/Rd2 mice following AAV‐mediated gene replacement therapy
Authors: Schlichtenbrede Frank C. , da Cruz Lyndon , Stephens Clare , Smith Alexander J. , Georgiadis Anastasios , Thrasher Adrian J. , Bainbridge James W. B. , Seeliger Mathias W. , Ali Robin R. 
Published: August 2003

AbstractBackgroundPrph2Rd2/Rd2 mice have a retinal degeneration due to a null mutation for the Prph2 gene which encodes a photoreceptor‐specific glycoprotein, peripherin2, essential for outer‐segment formation. We have previously shown improvement of photoreceptor function at a single time point following AAV‐mediated gene replacement therapy. Here we quantify the functional rescue over a 15‐week time course and present a detailed analysis of the improvement in retinal function.MethodsAn AAV2 vector, AAV.rho.rds, carrying a peripherin2 c‐DNA, was in injected subretinally into 10‐day‐old Prph2Rd2/Rd2 mice. One group was injected at a single time point while in a second group the injections were repeated after 5 days. The effect of treatment was analysed histologically using electron microscopy and electroretinography (ERG) was used to assess functional changes. Treated mice were recorded at regular intervals over 15 weeks. Untreated contralateral eyes served as internal control.ResultsA significant increase in b‐wave amplitude was first noted 3 weeks after treatment of 10‐day‐old Prph2Rd2/Rd2 mice and persisted for up to 14 weeks. An increase in the area of retina exposed to vector resulted in a significant increase in both b‐wave amplitude and persistence.ConclusionsIn this study AAV‐mediated gene replacement in Prph2Rd2/Rd2 mice resulted in a significant functional improvement over a period of 14 weeks. These results support the utility of gene therapy approaches as treatment for photoreceptor dystrophies. Copyright © 2003 John Wiley & Sons, Ltd.
Failure to generate atheroprotective apolipoprotein AI phenotypes using synthetic RNA/DNA oligonucleotides (chimeraplasts)
Authors: Manzano Anna , Mohri Zahra , Sperber Galia , Ogris Manfred , Graham Ian , Dickson George , Owen James S. 
Published: August 2003

AbstractBackgroundElevated plasma high‐density lipoprotein (HDL), and its major constituent apolipoprotein AI (apoAI), are cardioprotective. Paradoxically, two natural variants of apoAI, termed apoAIMilano and apoAIParis, are associated with low HDL, but nevertheless provide remarkable protection against heart disease for heterozygous carriers and may even lead to longevity. Both variants arise from point mutations and have Arg173 and Arg151 to Cys substitutions, respectively, which allow disulphide‐linked dimers to form. Potentially, synthetic RNA/DNA oligonucleotides (chimeraplasts) can permanently correct single point mutations in genomic DNA. Here, we use a variation of such targeted gene repair technology, ‘gain‐of‐function chimeraplasty’, and attempt to enhance the biological activity of apoAI by altering a single genomic base to generate the atheroprotective phenotypes, apoAIMilano and apoAIParis.MethodsWe targeted two cultured cell lines that secrete human apoAI, hepatoblastoma HepG2 cells and recombinant CHO‐AI cells, using standard 68‐mer chimeraplasts with polyethyleneimine (PEI) as carrier and then systematically varied several experimental conditions. As a positive control we targeted the dysfunctional APOE2 gene, which we have previously converted to wild‐type APOE3.ResultsConversion of wild‐type apoAI to apoAIMilano proved refractory, with limited correction in CHO‐AI cells only. However, a successful conversion to apoAIParis was achieved, as demonstrated by polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) analysis and direct genomic sequencing. Unexpectedly, attempts with a new batch of 68‐mer chimeraplast to enhance conversion, by using different delivery vehicles, including chemically modified PEI, failed to show a base change; nor could conversion be detected with an 80‐mer or a 52–76‐mer series. In contrast, when a co‐culture of CHO‐E2 and CHO‐AI cells was co‐targeted, a clear conversion of apoE2 to apoE3 was seen, whereas no apoAIParis could be detected. When the individual chimeraplasts were analysed by denaturing electrophoresis only the active apoE2‐to‐E3 chimeraplast gave a sharp band.ConclusionsOur findings suggest that different batches of chimeraplasts have variable characteristics and that their quality may be a key factor for efficient targeting and/or base conversion. We conclude that, although an evolving technology with enormous potential, chimeraplast‐directed gene repair remains problematical. Copyright © 2003 John Wiley & Sons, Ltd.