bluuebunny/crossref_metadata_2025 huggingface dataset:
Functionalization of poly(4‐vinyl pyridine) grafted cellulose by quaternization reactions and a study on the properties of postquaternized copolymers
Authors: Chauhan Ghanshyam S. , Singh Baljit , Dhiman Surya K.
Published: December 2003

Graft copolymers of 4‐vinyl pyridine (4‐VP) synthesized by using simultaneous gamma irradiation method were further functionalized by post polymer quaternization reactions at N: of the pyridine ring of the graft copolymers. Using the optimum grafting conditions reported earlier for the grafting of 4‐VP onto extracted cellulose, graft copolymer was synthesized in bulk and was further functionalized by quaternization with hexyl bromide (C6H13Br), benzyl chloride (C6H5CH2Cl), n‐butyl bromide (C4H9Br), and maleic anhydride (MAnh). The quaternized polymers were studied for treatment of hardness of water, antibacterial action, emulsification properties, metal ion uptake and stability toward thermal degradation, and enzymatic and alkaline hydrolysis to evaluate the suitability of these polymers in harsh chemical, microbial, and thermal environments. The functionalized polymers were also characterized for surface morphology (SEM), elemental analysis and FTIR for investigations of structural aspects, and to obtain evidence for quaternization. The functionalized copolymers exhibit a range of properties that can be used in many fields of water purification technologies including antibacterial agents and ion exchangers. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2454–2464, 2004
Structural material from waste plastic
Authors: Abdel‐Goad Mahmoud A.‐Halim
Published: December 2003

In efforts to contribute to community development, particularly in the context of Egyptian communities, waste plastics materials were successfully recycled without the difficult task of separation and reused to economically produce new structural material. Recycling was performed by mixing molten waste plastics with sand to produce these new materials. Samples with different percentages of plastics and different particle sizes of sand were used in the process. Materials showed acceptable density and high compressive strength, which was shown to be at a maximum with contents of about 30–40% waste plastic. Furthermore, certain types of sand having different colors were used to produce attractive materials, suitable for decorative uses. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2543–2547, 2004
Kinetics of heterogeneous polymerization of vinylidene chloride. I. Mechanism and modeling
Authors: Lu Qun , Weng Zhi‐Xue , Huang Zhi‐Ming , Tao Chun‐Yuan , Pan Zu‐Ren
Published: December 2003

The similarities and differences between the heterogeneous polymerization of vinylidene chloride (VDC) and vinyl chloride (VC) were compared and analyzed. The effect of the poly(vinylidene chloride)(PVDC) crystal morphology on the kinetics of VDC polymerization was discussed. Based on the assumption that polymerization occurs in the monomer phase and on the edges of the PVDC lamellar crystals, a mechanism of VDC polymerization is proposed and then a kinetic model for VDC bulk/suspension polymerization is developed according to the elementary reactions. The model can be used to predict polymerization rate and conversion. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2576‐2581, 2004
Adsorption and complexation of chitosan wet‐end additives in papermaking systems
Authors: Li Houbin , Du Yumin , Xu Yongmei
Published: December 2003

The adsorption of several chitosan samples with different degrees of deacetylation (65, 73, 85, and 92%) and their interactions with the main components of reed pulp suspensions were investigated with gel permeation chromatography, phenol/sulfuric acid spectrophotometry, microelctrophoresis, and retention/drainage methods. The results showed that the chitosan additive was almost completely adsorbed onto the surfaces of the cellulosic fibers, especially onto the surfaces of fines in a variety of cellulosic systems at low dosages corresponding to those used in industrial operations. This adsorption increased as the degree of deacetylation of chitosan increased. Moreover, the aggregation of the fine cellulosic particles was maximum at a dosage of about 10 mg/kg of chitosan. The uptake of dissolved and colloidal carbohydrates always occurred in the cellulose suspensions upon the addition of chitosan. These phenomena were related to the maximum retention of fines and drainage performance of the pulp suspensions; the optimum drainage dosage was found to be about 5–8 mg of chitosan per gram of oven‐dried pulp. The interactions between chitosan and the cellulosic substrates were dominated by a bridging mechanism at pH ≈ 7 for these experiments. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2642–2648, 2004
Preparation and characterization of modified‐clay‐reinforced and toughened epoxy‐resin nanocomposites
Authors: Zhang Kailiang , Wang Lixin , Wang Fang , Wang Guangjian , Li Zuobang
Published: December 2003

Epoxy–clay nanocomposites were prepared by the dispersion of an organically modified layered clay in an epoxy resin (diglycidyl ether of bisphenol A) and curing in the presence of methyl tetrahydro acid anhydride at 80–160°C. The nanometer‐scale dispersion of layered clay within the crosslinked epoxy‐resin matrix was confirmed by X‐ray diffraction and transmission electron microscopy, and the basal spacing of the silicate layer was greater than 100–150 Å. Experiments indicated that the hydroxyethyl groups of the alkyl ammonium ions, which were located in the galleries of organically modified clay, participated in the curing reaction and were directly linked to the epoxy‐resin matrix network. Experimental results showed that the properties of epoxy were improved, evidently because of the loading of organically modified clay. The impact strength and tensile strength of the nanocomposites increased by 87.8 and 20.9%, respectively, when 3 wt % organic clay was loaded, and this demonstrated that the composites were toughened and strengthened. The thermal‐decomposition and heat‐distortion temperatures were heightened in comparison with those of pure epoxy resin, and so were the dynamic mechanical properties, including the storage modulus and glass‐transition temperature. Moreover, experiments showed that most properties of the composites were ameliorated with low clay contents. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2649–2652, 2004
Preparation and performance testing of sulfonated poly(phenylene oxide) based composite membranes for nanofiltration
Authors: Nogami Nobuhiro , Chowdhury Geeta , Matsuura T.
Published: December 2003

Sulfonated brominated poly(2,6‐dimethyl‐1,4‐phenylene oxide) (SPPOBr) was synthesized by a sequence of bromination and sulfonation. A thin film of SPPOBr was coated on top of a commercial poly(ether sulfone) membrane. Pure butoxyethanol (BE) solvent or a BE/isopropyl alcohol (IPA) solvent mixture was used to dissolve SPPOBr in the coating process. The thin film composite membranes so prepared were then tested for the separation of carbohydrate and electrolyte solutes. We found that the flux and the carbohydrate separation both increased significantly with increasing IPA content in the solvent mixture. However, the separation of electrolyte solutes did not change significantly. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 91: 2624–2628, 2004
Automatic quantification of gene amplification in clinical samples by IQ‐FISH
Authors: Narath R. , Lörch T. , Rudas M. , Ambros P. F.
Published: December 2003

BACKGROUND

The reliable detection and quantification of gene amplifications is crucial to clinical practice. Although there are different detection techniques, the fluorescence in situ hybridization (FISH) method has become highly accepted over past years because it is a reliable, robust, and quick method. Unfortunately, automatic quantification of gene amplification based on fluorescence intensities has not been possible thus far. Because current spot counting methods are reliable only when analyzing low amplification rates, we attempted to establish another method, i.e., to quantify the intensity of different FISH signals using an automatic fluorescence microscopical device on interphase nuclei: interphase quantitative FISH (IQ‐FISH).

METHODS

We quantified the fluorescence intensities of the differently labeled FISH probes (MYCN and D2Z) hybridized to three different neuroblastoma cell lines, six peripheral blood (PB) samples, 10 spiked PB samples, and nine neuroblastoma samples using the Metafer4 system (MetaSystems, Altlussheim, Germany). To obtain the MYCN copy number per cell, the ratio between the fluorescence intensities of the MYCN gene and reference sequence (D2Z) was calculated. For automatic analysis of the HER‐2/neu status in tumor cells, labeled FISH probes specific for HER‐2/neu and a chromosome 17–specific probe were hybridized to peripheral blood and tumor specimens and analyzed using the automatic device.

RESULTS

When measuring the fluorescence intensity per cell for both probe pairs (MYCN/D2Z and HER‐2/17p), amplified and non‐amplified cells, showed distinct peaks with only little overlap. Whereas normal cells showed a fluorescence ratio peak for MYCN/D2Z between 200 and 800, cells with MYCN amplification clearly exceeded this ratio value (1000 to 25,000). When mixing a varying number of MYCN amplified cells (range 9–91%) to normal PB, the spiked tumor cells could be identified. Even one neuroblastoma tumor cell in 1000 mononucleated cells could reliably be detected using our device. In neuroblastoma patient samples, non‐amplified cells were distinguished from amplified cells. Automatically and manually counted signals gave matching results in amplified and non‐amplified samples. HER‐2/neu–amplified cells were automatically detected in the breast cancer samples analyzed.

CONCLUSION

The automatic measurement of fluorescence signal intensities not only allows a reliable discrimination between non‐amplified and amplified cells but also exact quantification of amplified sequences. This is the prerequisite for the following applications: detection of amplified cells in the bone marrow and second‐look specimens; comparison between primary and relapse or pre‐ and post‐chemotherapeutic specimens; detection of tumors with focal gene amplification; and quantification of elimination of amplified gene sequences. © 2003 Wiley‐Liss, Inc.
Flow cytometric quantitation of red blood cell vesicles in thalassemia
Authors: Pattanapanyasat Kovit , Noulsri Egarit , Fucharoen Suthat , Lerdwana Surada , Lamchiagdhase Pornvaree , Siritanaratkul Napadol , Webster H. Kyle
Published: December 2003

BACKGROUND

Thalassemia is a hereditary hemolytic anemia caused by mutations in the globin gene complex. Circulatory disturbances including arterial and venous thrombosis have also been noted in these patients. Aggregability of abnormal RBC and the high level of membrane‐derived microparticles stemming from activated platelets and other blood cells are thought to be responsible for the associated thrombotic risk. Destruction of RBC is also thought to be an important pathophysiological consequence, particularly through the formation of circulating vesicles. To our knowledge, there has been no attempt to quantitatively evaluate the number of RBC vesicles in thalassemia. This prompted us to study the level of RBC vesicles in the peripheral blood of thalassemia patients using quantitative flow cytometry.

METHODS

Whole blood from each subject was doubly stained for RBC and platelet or annexin V markers, together with the known density TruCount™ beads. RBC vesicles were gated according to their forward/side scatter and RBC marker. Percentage of RBC vesicles and their absolute number were analyzed by flow cytometry.

RESULTS

Our data indicated that RBC vesicles were annexin V–positive. The number of annexin V–positive events was higher than their intact RBCs. RBC vesicles were present in both normal and thalassemic blood samples, but the numbers of RBC vesicles were significantly higher in thalassemia. Both the percentage and the absolute number of RBC vesicles were especially marked in splenectomized subjects with β‐thalassemia/Hemoglobin E. When clinical and hematological indices were compared with RBC vesicles, there was an inverse relationship between the degree of severity in thalassemia patients and the number of RBC vesicles.

CONCLUSION

Flow cytometric quantitation of RBC vesicles is simple, reliable and may offer new insights in to study of the relationship between defective hemoglobin synthesis, RBC perturbation and pathophysiological complications in thalassemia. © 2003 Wiley‐Liss, Inc.
Cytologic diagnosis of ossifying fibromyxoid tumor of soft tissue: A case report
Authors: Mohanty Sambit K. , Srinivasan Radhika , Rajwanshi Arvind , Vasishta Rakesh Kumar , Vignesh P.S.
Published: December 2003

Ossifying fibromyxoid tumor (OFMT) of soft tissue is an uncommon, recently described neoplasm. It is usually, but not universally, a benign neoplasm and cases of metastatic so‐called “malignant OFMT” have been reported. Although the histopathologic features of OFMT are well characterized, the same is not entirely true so far as the cytology literature is concerned. There are only three cytologic descriptions of OFMT to date to the best of our knowledge. A 62‐year‐old woman presented with a subcutaneous well‐circumscribed nodular soft tissue mass at the upper end of the shoulder. X‐ray and computed tomography revealed an incomplete shell of calcification surrounding the nodular mass along with foci of ossification within it. Fine‐needle aspiration (FNA) cytology showed round to polygonal to spindle shaped neoplastic cells arranged in clusters, cords, and small aggregates and lying discretely in a mucoid background. Some osteoid‐like material was also seen. The tumor was completely excised and histopathologic examination confirmed the diagnosis of OFMT. OFMT should be considered as a possibility in FNA cytology of myxoid soft tissue tumor especially with radiologic evidence of ossification. Diagn. Cytopathol. 2004;30:41–45. © 2004 Wiley‐Liss, Inc.
Paratesticular congenital malignant rhabdoid tumor diagnosed by fine‐needle aspiration cytology. A case report
Authors: Salamanca Javier , Rodríguez‐Peralto José Luis , Azorín Daniel , Ballestín Claudio , De Agustín Pedro
Published: December 2003

We report the FNA features of a congenital malignant extrarenal rhabdoid tumor (MERT) located in the right paratesticular area of a newborn full‐term boy (39 wk gestation), with disseminated metastases in the liver and right parietal region. The diagnosis was suggested two days after birth by fine‐needle aspiration biopsy (FNAB) of the parietal mass, which demonstrated an atypical large cell proliferation with vesicular nuclei, prominent nucleoli, and abundant cytoplasm exhibiting paranuclear dense inclusions. The diagnosis was confirmed by histopathologic and immunohistochemical examination of the primary paratesticular tumor. To the best of our knowledge, this is the third MERT reported in the paratesticular region, one of the few congenital extrarenal non‐central nervous system cases, and the third congenital case (renal or extrarenal) primarily diagnosed by FNAB. We emphasize the characteristic cytologic features of a congenital rhabdoid tumor, which must be known by pathologists because of the clinical and prognostic implications. Diagn. Cytopathol. 2004;30:46–50. © 2004 Wiley‐Liss, Inc.
Monophasic synovial sarcoma: A cytologic spectrum
Authors: Ewing C. Alexander , Zakowski Maureen F. , Lin Oscar
Published: December 2003

Synovial sarcoma (SS) is a high‐grade sarcoma that can be diagnosed in cytology with certainty only when it presents with a biphasic pattern. Monophasic SS (MSS), however, is a diagnostic consideration when a uniform spindle cell population is present. The purpose of this study was to evaluate a series of cytologic cases of MSS and its cytologic presentation. Twenty‐one FNAs of histologically confirmed MSS were reviewed. Specimens consisted of tissue fragments and single cells containing scant granular cytoplasm, medium‐sized nuclei, and coarse chromatin. A monotonous spindle pattern with comma‐shaped nuclei was present in 5 cases. Sixteen cases contained oval and spindled nuclei. Eight of these specimens contained round nuclei, and 5 of these cases showed prominent nucleoli and cohesive clusters, reminiscent of biphasic SS. We conclude that a spectrum of cytologic findings can be seen in MSS, including a secondary population of cells with morphology usually typical of biphasic SS. Diagn. Cytopathol. 2004;30:19–23. © 2004 Wiley‐Liss, Inc.
Primary pulmonary adenoid cystic carcinoma: Report of a case diagnosed by fine‐needle aspiration cytology
Authors: Qiu Suimin , Nampoothiri Madhavan M. , Zaharopoulos Paul , Logroño Roberto
Published: December 2003

Adenoid cystic carcinoma of the lower respiratory tract is an uncommon tumor that can arise in the mainstem bronchus and often presents as an endobronchial mass lesion causing bronchial obstruction with post obstructive atelectasis and pneumonia. Exfoliative cytology is seldom useful in the diagnosis of primary bronchial adenoid cystic carcinoma, because these neoplasms usually have a submucosal location with often intact mucosa. Since most endobronchial adenoid cystic carcinomas are endoscopically visible, bronchoscope‐guided fine‐needle aspiration constitutes an excellent approach to establish a pathologic diagnosis. The fine‐needle aspiration cytology of primary pulmonary adenoid cystic carcinoma has been rarely described. We report a case of primary adenoid cystic carcinoma of the lung having characteristic cytologic features and correlate with computed tomography, bronchoscopic, and histological findings. Bronchoscope‐guided aspiration cytology provided a conclusive diagnosis of adenoid cystic carcinoma, which was further corroborated by histology in the pneumonectomy specimen. Diagn. Cytopathol. 2004;30:51–56. © 2004 Wiley‐Liss, Inc.
Developmental changes in eyeblink conditioning and simple spike activity in the cerebellar cortex
Authors: Nicholson Daniel A. , Freeman John H.
Published: December 2003

The activity of neurons in the cerebellum exhibits learning‐related changes during eyeblink conditioning in adult mammals. The induction and preservation of learning‐related changes in cerebellar neuronal activity in developing rats may be affected by the level of maturity in cerebellar feedback to its brainstem afferents, including the inferior olive. Developmental changes in cerebellar plasticity were examined by recording the activity of Purkinje cells in eye regions of cerebellar cortical lobule HVI (lobulus simplex) in infant rats during eyeblink conditioning. The percentage and amplitude of eyeblink conditioned responses increased as a function of age. Analyses of Purkinje cell simple spike activity revealed developmental increases in the number of units that exhibited stimulus‐evoked and learning‐related changes in activity. Moreover, the magnitude of these changes exhibited a substantial age‐related increase. The results support the view that the emergence of learning‐specific cerebellar plasticity and the ontogeny of eyeblink conditioning are influenced by developmental changes in the synaptic interactions within brainstem–cerebellum circuits. © 2003 Wiley Periodicals, Inc. Dev Psychobiol 44: 45–57, 2004.
The aims and accomplishments of comparative psychology
Authors: Lickliter Robert
Published: December 2003

Greenberg, Partridge, Weiss, and Pisula propose a new perspective of comparative psychology, based in large part on an anagenetic and dynamic systems approach to the development and evolution of behavior. Their view appreciates the probabilistic nature of behavioral development and promotes the value of an integrative levels concept for generating testable hypothesis regarding the complex relationship between biology, context, and developmental history underlying behavioral and psychological functioning. However, the authors fail to represent the full scope of contemporary comparative psychology by overlooking several core aims of the field, including (a) the use of animal models to shed light on human behavior and development and (b) understanding the role of behavior as a leading edge in the evolutionary process. © 2003 Wiley Periodicals, Inc. Dev Psychobiol 44: 26–30, 2004.
Finding large‐scale spatial trends in massive, global, environmental datasets
Authors: Johannesson G. , Cressie N.
Published: December 2003

As technology progresses, the availability of massive environmental data with global spatial coverage has become quite common. An example of such data is total column ozone (TCO) remotely sensed from a satellite. In their raw form, these data are often spatially (and temporally) dense, but irregular. The problem considered here is one of detecting a large‐scale spatial trend at a given time point (actually, in a given time interval). We propose a sequential aggregation method, producing different levels of coarser (spatial) resolution data and, at the same time, preserving both the local information content and the locations of the raw data. In estimating the large‐scale trend, we consider different types of smooth spatial trend surfaces on the sphere, all linear combinations of smooth basis functions and satisfying the topological constraints imposed by the sphere. These include trend surfaces based on tensor products of splines, spherical harmonic basis functions, smoothing spherical splines and a new trend surface that we call fixed‐rank smoothing (FRS). The FRS trend surfaces can be based on any set of smooth basis functions on the sphere and are estimated via penalized weighted‐least‐squares regression (ridge regression) using a data‐adaptive penalty term. In comparing the various trend surfaces considered, we look at data fidelity, trend‐surface consistency when fitted to data at different resolutions, and small‐ and large‐scale spatial behavior of the resulting detrended fine‐resolution data. An application to the TCO data reveals that the large‐scale spatial trend can be detected and effectively estimated using coarse‐resolution data. The FRS trend surfaces are seen to achieve better data fidelity than other trend surfaces considered, and, in terms of trend‐surface consistency and small‐ and large‐scale residual behavior, FRS is seen to have as good, and sometimes better, performance. Copyright © 2003 John Wiley & Sons, Ltd.
A novel gene, FGA7, is fused to RUNX1/AML1 in a t(4;21)(q28;q22) in a patient with T‐cell acute lymphoblastic leukemia
Authors: Mikhail Fady M. , Coignet Lionel , Hatem Nadia , Mourad Zeinab I. , Farawela Hala M. , Kaffash Dalal M. El , Farahat Nahla , Nucifora Giuseppina
Published: December 2003

AML1 is among the most frequent targets of chromosomal rearrangements in human leukemias. We report here the molecular analysis of a t(4;21)(q28;q22) that has disrupted AML1 in a patient with de novo T‐cell acute lymphoblastic leukemia. By using 3′‐RACE analysis, we show that this rearrangement results in the fusion of a novel gene immediately downstream of exon 5 or exon 6 of AML1, indicating that the AML1 breakpoint lies in intron 6 and that alternative fusion splice variants are generated. The sequence of the novel gene, located at 4q28, does not have any significant homology with any of the known genes in the human GenBank DNA database. However, the first 118 bases are identical to a part of a human ovarian EST. Also, its high homology with mouse and rat sequences suggests that this sequence most probably represents a part of a novel gene, which we named FGA7 (Fused Gene 7 to AML1). Following the AML1 open reading frame, the FGA7 sequence encodes an unknown protein of 27 amino acids. We isolated three bacterial artificial chromosome (BAC) clones that contain the FGA7 sequence and confirmed the breakpoint of the gene on the patient's metaphase spreads by fluorescence in situ hybridization using these BACs as probes. RT‐PCR and Northern blot analyses revealed that FGA7 is expressed in ovarian and skeletal muscle tissues. The predicted AML1‐FGA7 chimeric proteins contained a limited number of residues fused to AML1 in a situation similar to that reported for the AML1‐EAP fusion that is a product of t(3;21). It is possible that the expression of a constitutively shortened AML1 could compete with full‐length AML1 and act as a dominant negative inhibitor of the promoters that the core binding factor activates. © 2004 Wiley‐Liss, Inc.
Distinct MDM2 and P14ARF expression and centrosome amplification in well‐differentiated liposarcomas
Authors: Perucca‐Lostanlen D. , Rostagno P. , Grosgeorge J. , Marcié S. , Gaudray P. , Turc‐Carel C.
Published: December 2003

Well‐differentiated liposarcomas (WDLs) are common soft‐tissue tumors in adults. They are characterized by large marker chromosomes and/or ring chromosomes containing 12q‐derived sequences in which MDM2 is consistently amplified. WDLs are subdivided into two subtypes according to their karyotype. Type D cells exhibit a near‐diploid karyotype, with very few or no chromosome changes. Type H cells exhibit a near‐tetraploid karyotype and many structural changes. Expression of P14ARF, MDM2, and TP53 proteins was assayed in the two WDL subtypes to establish whether distinct expression profiles correlated with cell ploidy. Although a transcriptionally functional TP53 was present in most tumors independent of their karyotype, type H cells were characterized by high levels of P14ARF and MDM2 proteins. Although amplified within similar chromosome markers in type D tumors, MDM2 did not appear to be overexpressed. In addition, it was present as a C‐terminal truncated protein, indicative of alternatively spliced variants of MDM2 mRNA. As the existence of karyotypically distinct tumors could result from alterations of the mitotic machinery, we investigated the centrosome behavior in the two WDL subtypes. Centrosome amplification occurred in WDL tumors types H and D independent of their ploidy status. Moreover, no functional centrosome difference was found between the two tumor subtypes. © 2003 Wiley‐Liss, Inc.
Germ‐line ATM gene alterations are associated with susceptibility to sporadic T‐cell acute lymphoblastic leukemia in children
Authors: Liberzon Ella , Avigad Smadar , Stark Batia , Zilberstein Julia , Freedman Laurence , Gorfine Malka , Gavriel Herzel , Cohen Ian J. , Goshen Yacov , Yaniv Isaac , Zaizov Rina
Published: December 2003

A major feature of ataxia‐telangiectasia (A‐T) is an increased risk of cancer, particularly of lymphoid malignancies. We studied ATM gene involvement in leukemic cells derived from 39 pediatric T‐cell acute lymphoblastic leukemias (ALLs). Two types of sequence changes—truncating and missense—were identified in 8 T‐cell ALL samples: 3 truncating changes, all previously identified in A‐T (R35X, −30del215, 2284delCT), and 3 missense variants (V410A, F582L, F1463C) were found, none associated with loss of heterozygosity (LOH). In all patients studied, the mutation was present in the germ‐line. A‐T carriers, defined by the finding of truncating mutations, were found to be 12.9 times more frequent than in the normal population (P = 0.004). A normally ethnically matched population was screened for the 3 missense variants, and their frequency was significantly more prevalent (4.9‐fold excess) than in the normal population (P = 0.03). Our data suggest there is some evidence of an association between missense alterations in the ATM gene and T‐cell ALL. A significant difference in the mean age at diagnosis of T‐cell ALL was noted between patients harboring an ATM sequence change and those with no change, 5.4 years and 9.7 years, respectively (P = 0.001). No ATM alterations were identified in relapse samples, indicating that ATM does not play a role in disease progression. The high prevalence of germ‐line truncating and missense ATM gene alterations among children with sporadic T‐cell ALL suggests an association with susceptibility to T‐cell acute leukemia and supports the model of predisposition to cancer in A‐T heterozygotes. © 2003 Wiley‐Liss, Inc.
Amplification and overexpression of the IGF2 regulator PLAG1 in hepatoblastoma
Authors: Zatkova Andrea , Rouillard Jean‐Marie , Hartmann Wolfgang , Lamb Barbara J. , Kuick Rork , Eckart Markus , von Schweinitz Dietrich , Koch Arend , Fonatsch Christa , Pietsch Torsten , Hanash Sam M. , Wimmer Katharina
Published: December 2003

There is evidence that 8q amplification is associated with poor prognosis in hepatoblastoma. A previous comparative genomic hybridization analysis identified a critical region in chromosomal bands 8q11.2–q13. Using restriction landmark genomic scanning in combination with a virtual genome scan, we showed that this region is delineated by sequences within contig NT_008183 of chromosomal subbands 8q11.22–q11.23. A real‐time PCR–based genomic copy number assay of 20 hepatoblastomas revealed gain or amplification in this critical chromosomal region in eight tumors. The expression of four genes and expressed sequence tags (ESTs) within this newly defined region was assayed by real‐time reverse transcriptase polymerase chain reaction (RT‐PCR) in four tumors with and six tumors without gain or amplification. The PLAG1 oncogene was found to be highly expressed in all but one tumor compared to normal liver tissue. Furthermore, quantitative RT‐PCR revealed that the expression level of the developmentally regulated transcription factor PLAG1 was 3–12 times greater in hepatoblastoma tumors and cell lines compared to age‐matched normal liver and comparable to the expression in fetal liver tissue. PLAG1 has been shown be a transcriptional activator of IGF2 in other tumor types. Using luciferase reporter assays, we demonstrated that PLAG1 transactivates transcription from the embryonic IGF2 promoter P3, also in hepatoblastoma cell lines. Thus, our results provide evidence that PLAG1 overexpression may be responsible for the frequently observed up‐regulation of IGF2 in hepatoblastoma and therefore may be implicated in the molecular pathogenesis of this childhood neoplasia. © 2003 Wiley‐Liss, Inc.
T‐cell receptor Vβ gene usage by T cells reactive with the tumor‐rejection antigen SART‐1 in oral squamous cell carcinoma
Authors: Kumamaru Wataru , Nakamura Seiji , Kadena Tsutomu , Yamada Akira , Kawamura Eiji , Sasaki Masanori , Ohyama Yukiko , Toyoshima Takeshi , Hayashida Jun‐nosuke , Itoh Kyogo , Shirasuna Kanemitsu
Published: December 2003

We recently described that the SART‐1690–698 peptide could induce HLA‐A24‐restricted cytotoxic T lymphocytes (CTLs), which recognize the SART‐1 tumor cells from peripheral blood mononuclear cells (PBMCs) of HLA‐A24+ cancer patients. In our study, in 5 of 14 HLA‐A24+ patients with oral squamous cell carcinomas (SCCs), CTLs could be induced with the SART‐1690–698 peptide from the PBMCs. In 2 of the patients from whom the highest CTL activities were induced, the T‐cell receptor (TCR) Vβ repertoire expressed by the SART‐1690–698‐specific CTLs was found to be restricted and multiple Vβ families were predominantly expressed in each patient. Although the predominant Vβ families were different between the 2 patients, Vβ7 was highly and commonly predominant. The same predominant Vβ families were also detected in the tumor‐infiltrating lymphocytes (TILs) from each patient, and each Vβ family contained one or more unique T‐cell clonotypes. The unique T‐cell clonotypes were found to be common between the TILs and SART‐1690–698‐specific CTLs from each patient, and especially 2 T‐cell clonotypes with Vβ7 were identical even in the 2 patients. One of the 2 T‐cell clonotypes with Vβ7 was detected in the TILs from 11 of 14 HLA‐A24+ patients and another was found in those from 8 of HLA‐A24+ patients, while none of 10 HLA‐A24− patients demonstrated both T‐cell clonotypes. These results strongly suggest that the T‐cell clonotypes with Vβ7 are major TCR Vβ genes expressed by SART‐1690–698‐specific CTLs. Furthermore, autologous tumor cells from one of the HLA‐A24+ patients stimulated the PBMCs and regional lymph node cells (LNCs) to expand the same T‐cell clonotypes as those in the SART‐1690–698‐specific CTLs. These results strongly suggest that the SART‐1690–698‐specific CTLs clearly accumulate in vivo, especially in the TILs, as a consequence of in situ antigenic stimulation by autologous tumor cells. The identification of the unique TCR Vβ genes used by SART‐1259‐specific CTLs should help to improve the diagnosis of the specific immune response in patients with SART‐1 cancers, especially during anticancer immunotherapy. © 2003 Wiley‐Liss, Inc.
Improved immunogenicity of a self tumor antigen by covalent linkage to CD40 ligand
Authors: Huang Hsing‐I , Wu Ping‐Yi , Teo Chin‐Yee , Chen Min‐Nan , Chen Yi‐Chau , Silin Dmytro , Tao Mi‐Hua
Published: December 2003

The interaction between the CD40 ligand (CD40L) and CD40 on antigen‐presenting cells (APCs) is critical in promoting humoral and cellular immune responses. Agonistic anti‐CD40 monoclonal antibody and soluble CD40L can act as powerful adjuvants to promote vaccination, but usually require repeated high‐dose treatment. In this study, we demonstrate that the adjuvant effect of CD40L can be greatly improved by directly linking the antigen to CD40L. We constructed a fusion protein (Id‐CD40L) consisting of the extracellular domain of CD40L and the idiotype (Id) protein, a weakly immunogenic tumor‐specific antigen derived from the murine 38C13 B‐cell lymphoma. The soluble Id‐CD40L fusion protein retained CD40 binding activity and stimulated CD80 and CD86 upregulation and interleukin (IL)‐12 production by macrophages. Immunization of mice with Id‐CD40L without adjuvants resulted in high titers of anti‐Id Abs dominated by the IgG1 isotype and protected the mice from subsequent lethal tumor challenge. In a dose‐response study, we demonstrated that Id‐CD40L elicited anti‐Id antibody (Ab) responses in all immunized animals, even at a dose as low as 0.5 μg. Immunization with free Id and an IgG‐CD40L fusion protein, which was identical in structure to Id‐CD40L but lost the Id determinant, resulted in significant lower anti‐Id responses, indicating that physical linkage between the tumor antigen and CD40L was required for the optimal immune response. These results demonstrate that fusing CD40L to a candidate antigen can greatly improve the adjuvant activity of CD40L. This approach may be useful in developing vaccines for a variety of malignant and infectious diseases. © 2003 Wiley‐Liss, Inc.
Loading mitomycin C inside long circulating hyaluronan targeted nano‐liposomes increases its antitumor activity in three mice tumor models
Authors: Peer Dan , Margalit Rimona
Published: December 2003

The frequent overexpression of the hyaluronan receptors CD44 and RHAMM in cancer cells opens the door for targeting by the naturally‐occurring high‐Mr hyaluronan. This is the first time effective in vivo tumor targeting is reported for mitomycin C (MMC) loaded inside nano‐sized hyaluronan‐liposomes (denoted tHA‐LIP). The severe adverse effects of free MMC made it a rational candidate for an effective targeted carrier. In vitro, loading MMC inside tHA‐LIP increased drug potency 100‐fold, in cells overexpressing, but not in cells underexpressing, hyaluronan receptors. Both types of liposomes were non‐toxic and reduced MMC‐related toxicity in healthy C57BL/6 mice. In 3 tumor models, BALB/c bearing C‐26 solid tumors; C57BL/6 bearing B16F10.9 or (separately) D122 lung metastasis, tHA‐LIP were long‐circulating, 7‐fold and 70‐fold longer than nt‐LIP and free MMC, respectively. tHA‐LIP‐mediated MMC accumulation in tumor‐bearing lungs was 20% of injected dose, compared to 0.6% and 4% with free drug and nt‐LIP, respectively. Tumor‐free lungs showed low accumulation, irrespective of drug formulation. Key indicators of therapeutic responses, tumor progression, metastatic burden and survival, were superior (p < 0.001) in animals receiving MMC‐loaded tHA‐LIP, no treatment, MMC‐loaded nt‐LIP and free drug. In conclusion, tHA‐LIP perform as tumor‐targeted carriers, with promising prospects for treatment of tumors overexpressing hyaluronan receptors. © 2003 Wiley‐Liss, Inc.
Evaluation of the ability of digital infrared imaging to detect vascular changes in experimental animal tumours
Authors: Xie Wei , McCahon Pip , Jakobsen Karen , Parish Christopher
Published: December 2003

Infrared imaging has frequently been used in the past to detect changes in skin surface temperature associated with breast cancer. Usually a 1–2°C elevation in skin surface temperature is observed at the tumour periphery, and it has been proposed that this change is due to hypervascularity resulting from tumour‐associated angiogenesis. In our study, we used the rat mammary adenocarcinoma 13762 MAT, a tumour that has been used to identify antiangiogenic drugs, to investigate whether infrared imaging can detect angiogenesis in malignant tumours. If successful, it was hoped that this technique would represent a simple, noninvasive, procedure for monitoring the activity of antiangiogenic drugs. It was found that, unlike breast cancer patients, no tumour‐associated increase in skin surface temperature was observed, but a constant and highly significant reduction in temperature was noted that was independent of tumour size and was produced by relatively small tumours (≥ 0.5 cm in diameter). The explanation for this effect is unclear but it may be due to the poorly vascularised nature of rapidly growing tumours. Nevertheless, our study indicates that the peripheral temperature elevation reported in breast cancer patients is unlikely to be due to hypervascularity resulting from tumour‐induced angiogenesis. An alternative explanation is that the temperature increase is due to a chronic inflammatory response around developing breast tumours. With increasing evidence that inflammation can enhance tumour growth and is associated with a poor prognosis, this suggestion implies that infrared imaging may have considerable prognostic value. © 2003 Wiley‐Liss, Inc.
Grape seed extract inhibits advanced human prostate tumor growth and angiogenesis and upregulates insulin‐like growth factor binding protein‐3
Authors: Singh Rana P. , Tyagi Anil K. , Dhanalakshmi Sivanandhan , Agarwal Rajesh , Agarwal Chapla
Published: December 2003

Dietary intake of many fruits and vegetables has been shown to be associated with reduced risk of cancer. We investigated the in vivo efficacy of grape seed extract (GSE, patented as Traconol) against prostate cancer (PCA) and associated molecular events. Athymic nude mice were implanted with hormone‐refractory human prostate carcinoma DU145 cells and fed with 100 and 200 mg/kg/day (5 days/week) doses of GSE for 7 weeks. At the end of experiment, tumors were immunohistochemically analyzed for cell proliferation, apoptosis and angiogenesis. Our data show that GSE feeding strongly inhibited tumor growth that accounted for 59–73% (p < 0.001) inhibition in tumor volume and 37–47% (p < 0.05) decrease in tumor weight at the end of the experiment. It did not show any significant change in body weight gain profile and diet consumption. Immunohistochemical analysis of tumors showed that GSE decreases proliferation index by 51–66% (p < 0.001) and increases apoptotic index by 3–4‐fold (p < 0.001). CD31 staining for endothelial cells, showed decrease in intratumoral microvasculature in GSE‐fed group of mice. Control tumors showed 64.0 ± 1.6 CD31 positive cells/400× field compared to 23.2 ± 0.9 and 15.7 ± 0.08 (p < 0.001) CD31 positive cells in 100 and 200 mg/kg doses of GSE‐treated tumors, respectively. GSE strongly inhibited (47–70%, p < 0.05) vascular endothelial growth factor (VEGF) secretion in conditioned medium by DU145 cells. Recently, the circulating level of insulin‐like growth factor binding protein (IGFBP)‐3 is shown to inversely related with PCA risk, growth and prognosis. Consistent with this, we observed 6–7‐fold (p < 0.001) increase in tumor‐secreted levels of IGFBP‐3 after GSE feeding. In other immunohistochemical studies, compared to controls, tumor xenografts from GSE‐fed groups of mice showed a moderate decrease in VEGF but an increase in IGFBP‐3 levels. These findings suggest that GSE possesses in vivo anticancer efficacy against hormone‐refractory human PCA, which is associated with its antiproliferative, proapoptotic and antiangiogenic activities together with upregulation of IGFBP‐3. © 2003 Wiley‐Liss, Inc.
Non‐attendance in breast cancer screening is associated with unfavourable socio‐economic circumstances and advanced carcinoma
Authors: Zackrisson Sophia , Andersson Ingvar , Manjer Jonas , Janzon Lars
Published: December 2003

Our first objective was to assess changes in non‐attendance, proportion of advanced breast cancer and survival in Malmö Mammographic Service Screening Program, MMSSP, compared to a former trial, Malmö Mammographic Screening Trial, MMST. Our second objective was to describe non‐attenders in MMSSP in socio‐economic terms and risk for advanced breast cancer compared to attenders. Information from hospital and national registers was used to identify 33,800 women invited to service screening in MMSSP 1990–93. Attendance rates at first screening, the proportion of advanced breast cancers (Stage II–IV) and survival among non‐attenders in MMSSP were compared to the non‐attenders and with the control group of the former trial, MMST. Various socio‐economic factors were assessed as potential predictors of non‐attendance in MMSSP. Odds ratios (OR) and 95% confidence interval (CI) were computed. Incidence of breast cancer during a 10‐year‐period, relative risks and 95% CI among non‐attenders compared to attenders in MMSSP were computed. Attendance rates were significantly lower in MMSSP but a lower proportion of advanced breast cancers and a somewhat better survival among breast cancer cases (not significant) was seen in non‐attenders in MMSSP compared to MMST. In MMSSP non‐attendance was associated with being unmarried, being born abroad, being not currently employed, crowded housing conditions and low income. Incidence of advanced breast cancer was significantly higher among non‐attenders than among attenders. Attendance has decreased over time and potential reasons are discussed. Stage distribution and survival among non‐attenders seem to have improved. Several socio‐economic factors predict non‐attendance and non‐attenders are at higher risk for advanced breast cancer. © 2003 Wiley‐Liss, Inc.
Protein identification by peptide mass fingerprinting and peptide sequence tagging with alternating scans of nano‐liquid chromatography/infrared multiphoton dissociation Fourier transform ion cyclotron resonance mass spectrometry
Authors: Kosaka Toshiyuki , Yoneyama‐Takazawa Tomoko , Kubota Kazuishi , Matsuoka Tatsuji , Sato Ikuya , Sasaki Takamitsu , Tanaka Yorihisa
Published: December 2003

We have developed a method for protein identification with peptide mass fingerprinting and sequence tagging using nano liquid chromatography (LC)/Fourier transform ion cyclotron resonance mass spectrometry (FTICR‐MS). To achieve greater sensitivity, a nanoelectrospray (nano‐ES) needle packed with reversed‐phase medium was used and connected to the nano‐ES ion source of the FTICR mass spectrometer. To obtain peptide sequence tag information, infrared multiphoton dissociation (IRMPD) was carried out in nano‐LC/FTICR‐MS analysis. The analysis involves alternating nano‐ES/FTICR‐MS and nano‐ES/IRMPD‐FTICR‐MS scans during a single LC run, which provides sets of parent and fragment ion masses of the proteolytic digest. The utility of this alternating‐scan nano‐LC/IRMPD‐FTICR‐MS approach was evaluated by using bovine serum albumin as a standard protein. We applied this approach to the protein identification of rat liver diacetyl‐reducing enzyme. It was demonstrated that this enzyme was correctly identified as 3‐α‐hydroxysteroid dehydrogenase by the alternating‐scan nano‐LC/IRMPD‐FTICR‐MS approach with accurate peptide mass fingerprinting and peptide sequence tagging. Copyright © 2003 John Wiley & Sons, Ltd.
Identification of anthraquinone coloring matters in natural red dyes by electrospray mass spectrometry coupled to capillary electrophoresis
Authors: Puchalska Maria , Orlińska Magdalena , Ackacha Mohamed A. , Połeć‐Pawlak Kasia , Jarosz Maciej
Published: December 2003

Capillary electrophoresis with UV/visible diode‐array detection (DAD) and electrospray mass spectrometric (ESI‐MS) detection were used for the identification of anthraquinone color components of cochineal, lac‐dye and madder, natural red dyestuffs often used by ancient painters. For the purpose of such analysis, ESI‐MS was found to be a much more appropriate detection technique than DAD one owing to its higher sensitivity (detection limits in the range 0.1–0.5 µg ml−1) and selectivity. The method developed made it possible to identify unequivocally carminic acid and laccaic acids A, B and E as coloring matters in the examined preparations of cochineal and lac‐dye, respectively. In madder, EuropeanRubia tinctorum, alizarin and purpurin were found. The method allows the rapid, direct and straightforward identification and quantification of components of natural products used in art and could be very helpful in restoration and conservation procedures. Copyright © 2003 John Wiley & Sons, Ltd.
Neuroprotective functions of prion protein
Authors: Roucou Xavier , Gains Malcolm , LeBlanc Andréa C.
Published: December 2003

The normal function of prion protein (PrP) is usually disregarded at the expense of the more fascinating role of PrP in transmissible prion diseases. However, the normal PrP may play an important role in cellular function in the central nervous system, since PrP is highly expressed in neurons and motifs in the sequence of PrP are conserved in evolution. The finding that prion null mice do not have a significant overt phenotype suggests that the normal function of PrP is of minor importance. However, the absence of PrP in cells or in vivo contributes to an increased susceptibility to oxidative stress or apoptosis‐inducing insults. An alternative explanation is that the PrP normal function is so important that it is redundant. Probing into the characteristics of PrP has revealed a number of features that could mediate important cellular functions. The neuroprotective actions so far identified with PrP are initiated through cell surface signaling, antioxidant activity, or anti‐Bax function. Here, we review the characteristics of the PrP and the evidence that PrP protects against neurodegeneration and neuronal cell death. © 2003 Wiley‐Liss, Inc.
Short‐range axonal/dendritic transport by myosin‐V: A model for vesicle delivery to the synapse
Authors: Brown Jeremiah R. , Stafford Phillip , Langford George M.
Published: December 2003

Myosin‐V is a versatile motor involved in short‐range axonal/dendritic transport of vesicles in the actin‐rich cortex and synaptic regions of nerve cells. It binds to several different kinds of neuronal vesicles by its globular tail domain but the mechanism by which it is recruited to these vesicles is not known. In this study, we used an in vitro motility assay derived from axoplasm of the squid giant axon to study the effects of the globular tail domain on the transport of neuronal vesicles. We found that the globular tail fragment of myosin‐V inhibited actin‐based vesicle transport by displacing native myosin‐V and binding to vesicles. The globular tail domain pulled down kinesin, a known binding partner of myosin‐V, in affinity isolation experiments. These data confirmed earlier evidence that kinesin and myosin‐V interact to form a hetero‐motor complex. The formation of a kinesin/myosin‐V hetero‐motor complex on vesicles is thought to facilitate the coordination of long‐range movement on microtubules and short‐range movement on actin filaments. The direct interaction of motors from both filament systems may represent the mechanism by which the transition of vesicles from microtubules to actin filaments is regulated. These results are the first demonstration that the recombinant tail of myosin‐V inhibits vesicle transport in an in vitro motility assay. Future experiments are designed to determine the functional significance of the interaction between myosin‐V and kinesin and to identify other proteins that bind to the globular tail domain of myosin‐V. © 2003 Wiley Periodicals, Inc. J Neurobiol 58: 175–188, 2004