bluuebunny/crossref_metadata_2025_split huggingface dataset:
Magnitude and polarization of P53‐specific T‐helper immunity in connection to leukocyte infiltration of colorectal tumors
Authors: van der Burg Sjoerd H. , Menon Anand G. , Redeker Anke , Franken Kees L.M.C. , Drijfhout Jan W. , Tollenaar Rob A.E.M. , Hartgrink Henk H. , van de Velde Cornelis J.H. , Kuppen Peter J.K. , Melief Cornelis J.M. , Offringa Rienk 
Published: September 2003

AbstractThe tumor antigen p53 is mutated frequently and overexpressed in colorectal cancer. As a result, patients with this type of cancer commonly display p53‐specific T‐helper (Th) immunity. Examination of the cytokines produced by these Th‐cells showed that a majority of the proliferative p53‐specific T cell cultures produced none of the key cytokines (IFNγ, TNFα, IL‐4, IL‐5 or IL‐10), indicating that these p53‐specific Th‐responses are not polarized. In patients who exhibited p53‐specific reactivity against multiple p53‐epitopes, non‐polarized responses could be found side by side with polarized Th‐responses that produced INFγ or other cytokines such as IL‐10. Patients who exhibited p53‐specific IFNγ‐producing Th cell‐immunity before surgical excision of the tumor displayed higher numbers of tumor infiltrating intraepithelial leukocytes (p = 0.04) than patients lacking such responses, suggesting that the systemic presence of p53‐specific Th‐cells positively affects local tumor‐immunity. Our data concerning the polarization‐state of p53‐specific Th immunity in colorectal cancer patients support the use of vaccine formulations that induce strong Th1‐polarized p53‐specific immunity to ensure proper (re‐)programming of the anti‐tumor response. © 2003 Wiley‐Liss, Inc.
Mapping of candidate region for chordoma development to 1p36.13 by LOH analysis
Authors: Riva Paola , Crosti Francesca , Orzan Francesca , Dalprà Leda , Mortini Pietro , Parafioriti Antonina , Pollo Bianca , Fuhrman Conti Anna Maria , Miozzo Monica , Larizza Lidia 
Published: September 2003

AbstractVarious cytogenetic and molecular findings indicate 1p36 loss as a consistent change in sporadic and inherited chordoma, a rare embryogenetic neoplasm arising from notochord remnants. We studied 27 sporadic chordomas by means of loss of heterozygosity (LOH) of 31 microsatellites localized to the 1p36.32–36.11 region, and restricted the minimal LOH interval shared by 85% of the tumours to 1p36.13. We also used RT‐PCR analysis to investigate the role of the candidate genes CASP9, EPH2A, PAX7, DAN and DVL1, which were selected on the basis of the physical mapping of the LOH region and their plausible oncosuppressor function. RT‐PCR analysis showed the presence of DAN and PAX7 transcript fragments of the expected size in all of 8 chordoma samples, whereas the CASP9‐specific fragment was observed in only 3 and EPH2A was absent in one. Smaller than expected DVL1 transcripts were found in 4 tumours as well as in their normal counterpart (nucleus pulposus), which also showed a typically sized transcript. Sequencing revealed the skipping of 3 exons in the smallest DVL1 fragment, thus leading to a frameshift and predicting a truncated DVL1 gene product. Our study of the largest cohort of chordoma patients recruited so far indicates a common molecular lesion at 1p36.13, and suggests that the CASP9, EPH2A and DVL1 genes may play an onco‐suppressing role and be involved in the development of chordoma. © 2003 Wiley‐Liss, Inc.
Versican in epithelial ovarian cancer: Relation to hyaluronan, clinicopathologic factors and prognosis
Authors: Voutilainen Kirsi , Anttila Maarit , Sillanpää Sari , Tammi Raija , Tammi Markku , Saarikoski Seppo , Kosma Veli‐Matti 
Published: September 2003

AbstractVersican, a proteoglycan previously reported to increase in other malignant tumours, was studied immunohistochemically in 299 primary epithelial ovarian cancers, their 43 metastases and 6 normal ovaries to evaluate its prognostic value and relation to hyaluronan, another extracellular matrix molecule increased in cancer and a binding partner of versican. The stainings were scored according to the area percentage of strong versican signal of total peri‐ and intratumoural stroma as low (<15%) or high (≥15%). Epithelial staining of the tumours was scored as positive or negative. Low and high area percentage of strong stromal versican staining were observed in 133 and 166 carcinomas, respectively. A low area percentage of strong stromal versican staining correlated with mucinous histology (p= 0.019) and early International Federation of Gynecologists and Obstetritians (FIGO) stage (p< 0.0005), whereas a high percentage was associated with reduced 5‐year survival rate of the patients (44%vs.32%;p= 0.032). Versican was associated with the cancer cells in 151 tumours and correlated with clear cell histology (p< 0.0005), early FIGO stage (p= 0.049) and increased recurrence‐free survival (63%vs.47%;p= 0.032). However, in Cox's multivariate analyses with the conventional prognostic factors included, neither stromal nor cancer cell‐associated versican reached a significant prognostic value. Versican is thus enriched in the malignant stroma surrounding and promoting the growth of ovarian cancer, probably acting with hyaluronan, and associates with unfavourable prognosis but does not constitute an independent indicator of patient survival. © 2003 Wiley‐Liss, Inc.
Derivation and characterization of a Wilms' tumour cell line, WiT 49
Authors: Alami Jennifer , Williams Bryan R. , Yeger Herman 
Published: September 2003

AbstractWilms' tumour is a pediatric neoplasm exhibiting histologic features of developing kidney. Although the majority of Wilms' tumour patients are treated effectively, approximately 15% develop metastases and of these, 30% succumb to their disease. The biologic factors governing Wilms' tumour metastasis are largely unknown. Attempts at deriving representative Wilms' tumour cell lines, which could facilitate functional studies, have only been partially successful thus far. We now report on derivation and characterization of a Wilms' tumour cell line, WiT 49, from a first‐generation xenograft of a human Wilms' tumour lung metastasis. WiT 49 recapitulates the phenotype of the parent tumours (primary and lung metastasis) and expresses normal WT1, overexpresses IGFII and carries a frequently identified p53 mutation. We recently reported overexpression of hepatocyte growth factor(HGF) and its receptor met in a series of Wilms' tumours with higher levels in homotypic metastatic cases. We therefore examined WiT 49 for expression of HGF/met and for met signaling targets associated with cell adhesion and cytoplasmic mediators of transcription using Western blot, co‐immunoprecipitation, immunofluorescence labeling and zymography. Our results show co‐expression of HGF and met protein, absence of E‐cadherin, high levels of beta‐catenin co‐immunolocalized to met at the cell membrane and moderate levels of gamma‐catenin and ezrin protein expression. After cell fractionation, beta‐catenin was detected in the cytoplasm and nuclei of WiT 49 with relatively higher levels in the cytoplasm as compared to nuclei. Examination of MMP expression in WiT 49 showed constitutive activation of MMP 9 and latent MMP 2 supporting possible beta‐catenin‐mediated transcriptional activation. The WiT 49 cell line responded to recombinant human HGF by an increase in the expression of the met receptor, recruitment of the Gab‐1 adapter protein to met and release of bound beta‐catenin from met. Our studies therefore establish WiT 49 as a representative Wilms' tumour cell line derived from a lung metastasis that co‐expresses HGF/met and shows absence of the cadherin‐catenin complex supporting a role for these factors in regulation of the invasive and metastatic phenotype in Wilms' tumour. © 2003 Wiley‐Liss, Inc.
Association between NAD(P)H: Quinone oxidoreductase 1 (NQ01) inactivating C609T polymorphism and adenocarcinoma of the upper gastrointestinal tract
Authors: Sarbia Mario , Bitzer Monika , Siegel David , Ross David , Schulz Wolfgang A. , Zotz Rainer B. , Kiel Sybille , Geddert Helene , Kandemir Yasemin , Walter Alexandra , Willers Reinhart , Gabbert Helmut E. 
Published: September 2003

AbstractNQO1 is an antioxidant enzyme, important in the detoxification of environmental carcinogens. A single nucleotide polymorphism (C→T) at position 609 of the NQO1 cDNA has been associated with susceptibility to tumours induced by chemical carcinogens. In our case‐control study, we determined the prevalence of the C609T NQO1 polymorphism by PCR‐RFLP analysis in Caucasian patients with oesophageal adenocarcinoma (OAC; n=61), cardiac adenocarcinoma (CAC; n=120) or gastric adenocarcinoma (GAC; n=203) vs. a control group that consisted of 252 healthy blood donors. Additionally, NQO1 mRNA expression and NQO1 protein expression were determined by RT‐PCR and immunohistochemistry in a subset of cases. The NQO1 C609T genotype distribution was significantly different among controls (C/C, 73.4%; C/T, 25.0%; T/T, 1.6%) as compared to OAC patients (C/C, 49.2%; C/T, 47.5%; T/T, 3.3%; p=0.0004), CAC patients (C/C, 55.8%; C/T, 40.0%; T/T, 4.2%; p=0.0005) and with GAC patients (C/C, 65.5%; C/T; 30.6%, T/T; 3.9%; p=0.0377). The 609T allele overall frequency was 0.141 in controls, 0.270 in OAC patients, 0.241 in CAC patients and 0.192 in GAC patients. Individuals carrying 1 or 2 609T alleles had a 2.85‐fold higher risk (95% CI: 1.61–5.07; p=0.0003) for the development of OAC and a 2.18‐fold higher risk (95% CI: 1.38–3.44; p=0.0007) for the development of CAC than wild‐type gene homozygotes. Immunohistochemical analysis showed NQO1 protein expression in 133 carcinomas, whereas 17 carcinomas were negative. Negativity for NQO1 protein expression correlated strongly with the NQO1 genotype being present in 3.9% of cases with C/C, 13.9% of cases with C/T and 62.5% of cases with T/T genotype (p<0.001). In contrast, NQO1 mRNA expression was detectable irrespective of underlying genotype. In conclusion, determination of the NQO1 genotype may gain importance as a stratification marker in future prevention trials for adenocarcinoma of upper gastrointestinal tract. © 2003 Wiley‐Liss, Inc.
Human papillomavirus type 16 and 18 L1 protein peptide binding to VERO and HeLa cells inhibits their VLPs binding
Authors: Vera‐Bravo Ricardo , Ocampo Marisol , Urquiza Mauricio , García Javier E. , Rodríguez Luis E. , Puentes Alvaro , López Ramses , Curtidor Hernando , Suárez Jorge E. , Torres Elizabeth , Guzmán Fanny , Díaz Diana , Cortes Jimena , Bravo María M. , Cómbita Alba L. , Orozco Oscar , Patarroyo Manuel E. 
Published: September 2003

AbstractHuman papillomaviruses (HPVs) are the cause of epithelial lesions, HPV type 16 and type 18 being associated with the development of anogenital cancer. The L1 Major Capsid Protein (L1) represents about 90% of total HPV protein and is involved in virus‐host cell interaction, but little is known about this binding process. L1 sequences from HPV types 16 and 18 were synthesized in 56 20‐mer peptides, covering the entire protein, HPLC‐purified,125I‐radiolabeled and tested in VERO and HeLa cell‐binding assays to identify those peptides with high specific binding activity. Peptides 18283 (residues 54–77) and 18294 (274–308) from HPV16 L1, as well as 18312 (59–78) and 18322 (259–278) from HPV18 L1, presented high specific target cell binding activity. Peptide 18283 and 18294 affinity constants were 300 and 600 nM, respectively. Enzyme cell treatment before binding assay indicated that VERO and HeLa cell peptide receptor is a surface‐exposed protein. There was a 60% reduction in peptide 18283 binding to heparin lyase‐treated cells. Cross‐linking assays showed that these proteins molecular weights were around 69 and 54 kDa. Peptides 18283 and 18294 specifically inhibited HPV‐16 VLP binding to HeLa cells. According to the L1‐ and VLP‐reported structure, both peptides are close on the VLP‐surface, belonging to the outer surface broad pockets suggested as being potential receptor sites. Furthermore, it has been reported that a conserved motif from peptide 18294 is the target for neutralizing antibodies. These results suggest that such binding sequences are used by the virus as cell‐binding regions. © 2003 Wiley‐Liss, Inc.
Cyclopalladated compounds as chemotherapeutic agents: Antitumor activity against a murine melanoma cell line
Authors: Rodrigues Elaine G. , Silva Luiz S. , Fausto Daniela M. , Hayashi Marina S. , Dreher Simone , Santos Edson L. , Pesquero João B. , Travassos Luiz R. , Caires Antonio C.F. 
Published: September 2003

AbstractPalladacycle compounds obtained from N, N‐dimethyl‐1‐phenethylamine (dmpa), phenyl‐2‐pyridinyl‐acetylene and 1‐phenyl‐3‐N, N‐dimethylamine‐propine, respectively, were complexed to 1, 2 ethanebis (diphenylphosphine) (dppe) ligand to synthesize antitumor cyclopalladated complexes that were tested in vitro and in vivo against syngeneic B16F10‐Nex2 murine melanoma cells of low immunogenicity implanted subcutaneously in mice. Complexes were not toxic to mice injected 3 times i.p. with as much as 60 μM/animal/week. Of 3 cyclopalladated complexes that were inhibitory in vitro at low concentrations (<1.25 μM), complex 7a was the most active in vivo, delaying tumor growth and prolonging animal survival. In vitro, binucleate complex 7a caused a collapse of respiratory activity with an abrupt decrease of extracellular acidification on short incubation (up to 100 min), followed by DNA degradation after 24 hr. The apoptosis‐like reaction to this Pd‐complex was not accompanied by increased levels of caspases 1 and 3. Complex 7a bound to a bacterial plasmid DNA, causing late conformational changes after 24 hr. Two other complexes with different C, N‐cycles were also apoptotic and 2 binucleated ones were inactive. These results introduce the palladacycle‐dppe complexes as promising antitumor drugs with exquisite structural specificities and for action in vivo and in vitro. © 2003 Wiley‐Liss, Inc.
Hepatic stellate cell activation in vitro: Cell cycle arrest at G2/M and modification of cell motility
Authors: da Silva Fabiana M. , Guimarães Eduardo L.M. , Grivicich Ivana , Trindade Vera Maria T. , Guaragna Regina Maria , Borojevic Radovan , Guma Fátima C.R. 
Published: September 2003

AbstractHepatic fibrosis is a common response to chronic liver injury and is characterized by increased production of extracellular matrix components, whose major part is produced by hepatic stellate cells activated by inflammatory mediators to proliferate and migrate into the injured regions. GRX cells are a model of hepatic stellate cells characterized as myofibroblasts by morphological and biochemical criteria. We have recently shown that they respond to inflammatory mediators and cytokines present in the concanavalin A‐activated spleen cell supernatant (SCS) by quantitative changes in the expression of intermediate filaments. The present study investigated the effects of SCS and TNF‐α on the GRX cell proliferation and on the organization of the actin cytoskeleton. SCS and TNF‐α diminished the culture cell density, with an increase of cell [3H]thymidine incorporation and of cellular protein content, indicating an arrest in the G2/M phase of the cell cycle, which was reversible 48 h after removal of SCS. This effect was abrogated by dibutiryl‐cAMP. Actin cytoskeleton reorganization was observed after 24 h treatment, indicating increased cell motility. Our results suggest that inflammation‐dependent activation of stellate cells occurs in ordered interaction and coordination of proinflammatory agents. The increase of cAMP levels activates the conversion of lipocytes into myofibroblasts and increases the number of cells that can participate in repair. Since cAMP retains cells in the G1 phase, cytokines of the TNF‐α group are required for cell proliferation inducing the entry into the S phase. The progression through the G2/M checkpoint is mediated again by increased cAMP levels. J. Cell. Biochem. 90: 387–396, 2003. © 2003 Wiley‐Liss, Inc.
Method of platinum‐carbon coating of ultrathin sections for transmission and scanning electron microscopy: An application for study of biological composites
Authors: Schwarz H. , Gorb S. 
Published: September 2003

AbstractMany biological materials are composites containing two or more components with different mechanical properties. This study is concerned with the application of a method of platinum‐carbon coating (Pt/C) of ultrathin sections for TEM and SEM studies of the design of natural composite materials. The changes in profile of the ultrathin resin‐embedded sections during different stages of the preparation reflect the material properties of the various components: stiffer regions deform less than softer ones. Such changes in the section profile can be visualized by the Pt/C method and used as evidence of specific material properties in particular regions of composite materials. The method increases the relief contrast, improves the 3D‐view of structures, and in combination with standard TEM and SEM procedures can provide clear demonstrations of material design. The distribution of chitin crystallites in the insect cuticle and the ultrastructure of the pore canal system specialized for the transport of epidermal secretions to the cuticle surface were studied here as examples. Microsc. Res. Tech. 62:218–224, 2003. © 2003 Wiley‐Liss, Inc.
Technique to study three‐dimensional spatial arrangement of synaptic vesicles using data from single sections
Authors: Nikonenko Alexander G. 
Published: September 2003

AbstractSynaptic vesicles are membrane‐bound organelles storing neurotransmitters in presynaptic terminals and releasing them into the synaptic cleft. Coordinated movements of synaptic vesicles relate to synaptic function and their spatial arrangement can provide useful information about the activity of a synapse. This article presents a technique to extract quantitative information about three‐dimensional (3D) spatial arrangement of synaptic vesicles from measurements performed on single ultrathin random sections of a presynaptic terminal. The technique presumes quantification of a 2D density as well as 2D spatial pattern formed by vesicle profiles using a minimum spanning tree (MST) algorithm, in digitized micrographs of a presynaptic terminal. Further, original software was used to simulate a 3D spatial arrangement of synaptic vesicles and their random sectioning. A 3D density and pattern of synaptic vesicles were used as basic input parameters of the model, while a 2D density and MST quantities for vesicle profiles served as output, model‐derived parameters allowing one to compare and fit simulated distributions to experimental ones. Pilot simulations performed to check the validity of the technique have shown that a 2D density and MST quantities of vesicle profiles closely relate to a 3D density and spatial pattern of vesicles. The technique was demonstrated in the analysis of spatial distribution of synaptic vesicles in axonal terminals forming asymmetric synaptic densities in the stratum radiatum of the CA1 subfield of the murine hippocampus. Microsc. Res. Tech. 62:201–210, 2003. © 2003 Wiley‐Liss, Inc.
In vitro and in situ experimental model for X‐ray microanalysis of intestinal epithelium
Authors: Vanthanouvong Viengphet , Högman Marieann , Roomans Godfried M. 
Published: September 2003

AbstractIntestinal chloride (Cl) transport is disturbed in a number of diseases. X‐ray microanalysis can be used to study the distribution of Cl and other ions in intestinal epithelial cells. In this study it was attempted to establish an experimental system that retains the in vivo elemental composition of intestinal epithelial cells. An in vitro system was set up in which a segment of rat intestine was mounted in a bath and perfused with different fluids. The chloride in the bath or in the perfusion fluid could be exchanged for gluconate or bromide to determine the direction of chloride fluxes. An in situ system was set up in which the animal was anesthetized and a segment of the intestine was perfused with different solutions. In the in vitro experiments the concentration of Na and Cl in the epithelial cells increased and that of K decreased. These changes occurred within the first 30 minutes of incubation. Uptake of chloride occurred mainly from the bath, as seen in experiments where bromide was used as a chloride analog. The concentration gradient between bath and tissue determined the extent of chloride uptake. Addition of glucose to the perfusion fluid and bath did not improve the results. In the in situ system, preservation of the intracellular ion composition was better. Acceptable results were obtained with perfusion with Krebs‐Ringer's buffer without glucose for 30 minutes. In this case, the elemental content of the cell did not change appreciably during incubation. If glucose was added, the Na concentration increased in comparison to the control, both in crypt and villus cells. It is concluded that the intestinal epithelium is a sensitive system, very prone to disturbance of its homeostasis. However, the in situ system can be used in studies of agonist‐induced ion transport. Microsc. Res. Tech. 62:211–217, 2003. © 2003 Wiley‐Liss, Inc.
Junctional complex revisited by high‐resolution scanning electron microscopy
Authors: Testa Riva Francesca , Serreli Silvia , Loy Francesco , Riva Alessandro 
Published: September 2003

AbstractThe present study correlates the ultrastructural morphology of junctional complexes as revealed by transmission electron microscopy (TEM) with that observed by high‐resolution scanning electron microscopy (HRSEM), thanks to a new modification of the osmium tetroxide maceration technique. The removal of all cytoplasmic organelles by this technique allows the inspection of the inner side of the plasmalemma. With this treatment, a continuous band of tightly packed particles is observed at the most apical portion of lateral membranes. Just below this band, irregular clusters of apparently identical particles are placed all around the cellular contour. The topographical correspondence among these clusters and spot desmosomes seen by TEM identifies them as desmosomes. The continuous band seems to represent the combination of both zonulae, occludens and adherens. Regarding the nature of the particles, we suppose that they probably consist of peripheral membrane proteins clustered at the cytoplasmic surface of intercellular junctions and involved in the linkage between cytoskeleton and plasmalemma. Microsc. Res. Tech. 62:225–231, 2003. © 2003 Wiley‐Liss, Inc.
Dynamics of subcellular organelles, growth hormone, Rab3B, SNAP‐25, and syntaxin in rat pituitary cells caused by growth hormone releasing hormone and somatostatin
Authors: Matsuno Akira , Itoh Johbu , Takekoshi Susumu , Itoh Yoshiko , Ohsugi Yoshitaka , Katayama Haruko , Nagashima Tadashi , Osamura R. Yoshiyuki 
Published: September 2003

AbstractRab3B is involved in the exocytosis of synaptic vesicles and secretory granules in the central nervous system and the anterior pituitary cells. The aim of this study was to elucidate both the role of rab3B in GH secretion and the mutual relationship of rab3B and SNARE proteins. Adult male rats were injected intravenously with 10 μg of growth hormone releasing hormone (GHRH) or 10 μg of somatostatin (SRIF). Untreated rats were used as controls, and their pituitary glands were sectioned for histochemical examination. Rab3B is localized on the limiting membrane of the secretory granules and the cytosol. Confocal laser scanning microscopic observation of immunohistochemical double staining of rab3B and GH revealed that immunoreactivity of rab3B increased in GHRH‐treated rats and decreased in SRIF‐treated rats. Confocal laser scanning microscopic observation of immunohistochemical double staining of SNAP‐25, syntaxin, and rab3B revealed the co‐localization of rab3B and these SNARE proteins in GHRH‐treated rats, and their dissociation in SRIF‐treated rats. These results suggest that rab3B plays a principal role in GH secretion in the anterior pituitary cells and that SNAP‐25 and syntaxin act as co‐workers with rab3B in the functional regulation of GH secretion. Microsc. Res. Tech. 62:232–239, 2003. © 2003 Wiley‐Liss, Inc.