bluuebunny/crossref_metadata_2025 huggingface dataset:
Visual scoring versus histogram subtraction of in vivo binding of immunoglobulins against platelets after transfusion
Authors: Levin M.‐D. , van der Holt B. , de Veld J. C. , Gratama J. W. , de Vries W. , van't Veer M. B.
Published: December 2003

BACKGROUND

We developed a technique, in vivo binding of immunoglobulins in the platelet immunofluorescence test (IVBI‐PIFT), that detects immunoglobulins bound in vivo to transfused platelets. The visually scored results of this technique, however, are susceptible to interobserver variation. We describe a more objective method to generate results in IVBI‐PIFT.

METHODS

We studied 201 samples in 120 patients with hematologic malignancies in the IVBI‐PIFT. Histogram subtraction, i.e., fluorescence (anti–immunoglobulin G and fluorescein isothiocyanate) histogram before platelet transfusion subtracted from the histogram after platelet transfusion, was compared with visual scoring (pattern 1: no enhanced fluorescence before and after transfusion; pattern 2: enhanced fluorescence before and after platelet transfusion; pattern 3: enhanced fluorescence before transfusion; pattern 4: enhanced fluorescence after transfusion, interpreted as alloimmunization). After histogram subtraction, the number of remaining events (events post substraction, EPS) and the mean amount of fluorescence of these remaining events (mean channel post substraction, MCPS) were used and compared with the visual scoring and with platelet survival after transfusion.

RESULTS

In 26 (13%) of the 201 samples studied in the IVBI‐PIFT, fewer than three of five observers agreed on the visually scored pattern. In the 175 (87%) remaining samples, histogram subtraction showed a significant differentiation between pattern 4 and patterns 1 and 2 by using EPS, whereas patterns 4 and 3 were distinguished by using MCPS. The combination of EPS and MCPS differentiated best between pattern 4 and patterns 1, 2, and 3 (73% sensitivity, 96% specificity, 79% positive predictive value, and 95% negative predictive value). In contrast, the predictive value for platelet recovery after 1 and 16 h of pattern 4 from the visual scoring method and the results of histogram subtraction were poor.

CONCLUSION

This objective method of histogram subtraction correlated well with the visual scoring method of IVBI‐PIFT. © 2003 Wiley‐Liss, Inc.
Monitoring hyperproliferative disorders in human skin: Flow cytometry of changing cytokeratin expression
Authors: Franssen M. E. J. , Boezeman J. B. M. , van de Kerkhof P. C. M. , van Erp P. E. J.
Published: December 2003

BACKGROUND

Monitoring dynamics of different cell populations in solid tissues using flow cytometry has several limitations. The interaction and changes in epidermal subpopulations in hyperproliferative skin disorders such as psoriasis, a very common chronic inflammatory skin disease, may, however, elucidate the role of different cell populations in the pathogenesis and the effect of therapy in these disorders. We describe a new, simple method for identifying and quantifying different epidermal subpopulations in hyperproliferative skin disorders and an in vivo model for epidermal hyperproliferation by using six‐parameter assessment and three‐color flow cytometry.

METHODS

Epidermal single‐cell suspensions were derived from normal human skin before and after standardized injury and from untreated and treated psoriatic lesions. Samples were stained with anti‐cytokeratins 6 (K6) and 10 (K10), anti‐vimentin, and 4′,6‐diamidino‐2‐phenylindole. With three‐color flow cytometry, different epidermal subpopulations were further defined by six different parameters.

RESULTS

Correlations were found for mild psoriasis and K10+K6− cells and for moderate psoriasis and K10−K6+ cells. Treatment of mild psoriasis resulted in a 70% increase of the K10+K6− cells and an 18% decrease of the K10−K6− cells, whereas treatment of more severe psoriasis resulted in a 77% increase of the K10+K6− cells and a 33% decrease of the K10−K6+ cells. The tape‐stripping model showed changes in suprabasal keratin expression before proliferative activity.

CONCLUSIONS

The present flow cytometric assay permits simultaneous quantification of epidermal differentiation, inflammation, proliferation, and hyperproliferation‐associated keratinization and provides information on pathogenesis and therapy of hyperproliferative skin disorders. © 2003 Wiley‐Liss, Inc.
Carcinoid tumors of the lung: A diagnostic challenge in bronchial washings
Authors: Aron Manju , Kapila Kusum , Verma Kusum
Published: December 2003

Bronchial washings are used routinely in the diagnosis of lung tumors. However, unlike other tumors, the diagnosis of bronchial carcinoids on bronchial washings is difficult. We reviewed 17 cases of histologically proven bronchial carcinoids from the files of the cytology laboratory over a period of 15 yr (1986–2001). The bronchial washings and histology sections of all the cases were reviewed separately by two independent observers and the results tabulated. Two cases had inadequate bronchial washings for evaluation and were excluded from the study. A growth was identified on bronchoscopy in 13 of 15 cases. Initial cytologic diagnoses were ?adenocarcinoma/?carcinoid and suspicious of carcinoid in one case each. However, on review, tumor was identified in 10 of 13 cases initially considered to be negative. The possible reasons for a false‐negative report on initial cytology include the paucity of tumor cell fragments in the bronchial washings (5 of 12 cases showing only one to two tumor fragments) and their bland appearance, often being mistaken for benign columnar cells. This study highlights the potential pitfalls in the diagnosis of bronchial carcinoids on bronchial washings and underlines the importance of a diligent search in cases with high clinical suspicion and positive bronchoscopic findings. Diagn. Cytopathol. 2004;30:62–66. © 2004 Wiley‐Liss, Inc.
Microglandular hyperplasia has a cytomorphological spectrum overlapping with atypical squamous cells—cannot exclude high‐grade squamous intraepithelial lesion (ASC‐H)
Authors: Shidham Vinod B. , Rao R. Nagarjun , Machhi Jinobya , Chavan Ashwini
Published: December 2003

We retrospectively evaluated cytological interpretations of conventional cervical smears in 48 cases of biopsy‐proven microglandular hyperplasia and compared them with 15 negative controls. A glandular pattern was noted in all 48 cases but was predominant in 73% (35/48) of the cases. Immature metaplastic pattern was present in 71% (34/48) and was predominant in 27% (13/48). These features were not observed in negative control smears. 85% of cases (41/48) were interpreted as negative for epithelial cell abnormality. Two cases with predominantly glandular pattern (6%, 2/35) were interpreted as atypical glandular cells. Five cases with predominantly immature metaplastic pattern (38%, 5/13) showed checkerboard arrangement or rows of single cells with slightly larger atypical nuclei leading to interpretation as “atypical squamous cells—cannot exclude high‐grade squamous intraepithelial lesion.” The metaplastic cells with normoblast‐like karyorrhectic apoptotic debris confined to the nuclear area were scattered among these cells in microglandular hyperplasia with metaplastic pattern. Diagn. Cytopathol. 2004;30:57–61. © 2004 Wiley‐Liss, Inc.
Diagnostic value of HepPar1, pCEA, CD10, and CD34 expression in separating hepatocellular carcinoma from metastatic carcinoma in fine‐needle aspiration cytology
Authors: Saad Reda S. , Luckasevic Todd M. , Noga Christine M. , Johnson Douglas R. , Silverman Jan F. , Liu Yulin L.
Published: December 2003

Differentiating primary and metastatic hepatic malignancies can be diagnostically challenging in fine‐needle aspiration cytology (FNAC). We compared four immunohistochemical (IHC) markers, pCEA, CD10, HepPar1, and CD34, in differentiating hepatocellular carcinoma (HCC) from metastatic carcinoma (MC) in FNAC specimens. Sixty cases of liver FNAC with their corresponding cell blocks were retrieved from the hospital computer system, including 30 HCC and 30 MC (15 colon, 10 breast, and 5 pancreas). The diagnoses were confirmed by clinical follow‐up and surgical resection or core needle biopsy. The direct cytologic smears were air‐dried and Diff‐Quik‐stained, and alcohol‐fixed and Papanicolaou‐stained. Cell block sections from the aspirates were immunostained for pCEA, CD10, HepPar1, and CD34. IHC on cytologic smears for HCC was performed on 10 cases and compared with the cell block results. In HCC, CD10, and pCEA demonstrated the characteristic canalicular staining in 23/30 (77%) and 24/30 (80%) of the cases, respectively; however, none of the MC showed a canalicular staining pattern. HepPar1 was positive in 26/30 (87%) of the HCC cases and one MC. CD34 stained sinusoidal endothelial cells in 27/30 (90%) cases of HCC and six MC. Our results demonstrate that the canalicular staining pattern for CD10 and sinusoidal staining pattern of CD34 are very specific, in addition to the high specificity and sensitivity of HepPar1 for HCC. Cell blocks were more informative in demonstrating the characteristic architecture and immunostaining pattern of the malignancy than the cytologic smears. An IHC panel consisting of pCEA, CD10, HepPar1, and CD34 is useful for confirming HCC in FNAC of the liver. Diagn. Cytopathol. 2004;30:1–6. © 2004 Wiley‐Liss, Inc.
ThinPrep® for cytologic evaluation of follicular thyroid lesions: Correlation with histologic findings
Authors: Tulecke Mark A. , Wang Helen H.
Published: December 2003

Fine needle aspiration (FNA) is often the first step in management of a thyroid nodule. Although papillary carcinomas have distinctive features on conventional smears as well as ThinPrep® preparations, cytopathologists rely on the architectural arrangement of cells to classify follicular lesions. The accuracy of ThinPrep® in this regard has not been determined. We reviewed all thyroidectomy specimens from a 22‐mo period that showed a follicular lesion and had one or more preoperative FNA. The architecture of the lesions on histology was classified as predominantly (>70%) macrofollicular (MA), predominantly microfollicular (MI), mixed (MX), or cystic (C). The presence of colloid, Hürthle cell features, cystic change, Hashimoto's thyroiditis, and nonspecific lymphocytic thyroiditis were also recorded. All FNA specimens were processed with the ThinPrep® method and were categorized as nondiagnostic, benign (MA or MX), indeterminate (due to suboptimal cellularity), or suspicious (consistent with a microfollicular or Hürthle cell neoplasm) based on cellularity, architectural arrangement of the follicular cells, and the presence or absence of colloid on FNA. Those cytologic specimens in the indeterminate category were subcategorized as suggestive of an MA, MX, or MI lesion if possible. Histocytologic findings were correlated using Fisher's exact test. A total of 95 patients with 115 FNAs were included in the study (mean age: 48 yr; 75 females and 20 males). Seven of the FNAs were nondiagnostic, 23 benign, 69 indeterminate (5, 8, and 30 favor MA, MX, and MI, respectively, 6 cyst contents and 20 not subclassifiable) and 16 suspicious. The cytohistologic correlation for architecture was significant (P = 0.003). The cytohistologic correlation of cystic change, Hürthle cell change, and the presence of colloid (large fragments of colloid and “tissue‐paper‐like material” on cytology vs. abundant colloid on histology) was highly significant (P < 0.001, < 0.001, and < 0.03, respectively). In conclusion, thyroid FNA with ThinPrep® is useful in predicting the architectural pattern of follicular lesions. It is also reliable in predicting cystic change, Hürthle cell change, and the presence of colloid. Watery colloid is probably present as “tissue‐paper‐like material” in thyroid FNA ThinPrep® preparations. Diagn. Cytopathol. 2004;30:7–13. © 2004 Wiley‐Liss, Inc.
Comparison of cytologic features of giant‐cell tumor and giant‐cell tumor of tendon sheath
Authors: Gupta Kirti , Dey P. , Goldsmith Ritalin , Vasishta R.K.
Published: December 2003

The cytologic features in twelve cases of giant‐cell tumor (GCT) and five cases of giant‐cell tumor of tendon sheath (GCTTS) diagnosed by fine‐needle aspiration cytology (FNAC) are described. All of these cases were histopathologically confirmed. The aspirates of GCT are composed of a dual population of mononucleated spindle cell and multinucleated giant cells. The peripheral adherence of giant cells to the spindle cell is the feature of diagnostic significance in GCT. In GCTTS, the aspirate consists of a polymorphic population composed of mononuclear histiocyte‐like cells, hemosiderin laden macrophages, foamy macrophages, and a few multinucleated giant cells. FNAC can be used as a diagnostic tool for an early and accurate detection of these two giant cell–rich lesions, since the cytologic features when evaluated in conjunction with the clinical and radiologic features are sufficiently diagnostic. Diagn. Cytopathol. 2004;30:14–18. © 2004 Wiley‐Liss, Inc.
Cytology of low‐grade endocrine neoplasms involving liver: A series of 24 specimens, including 4 with hepatoid or glandular features
Authors: Crapanzano John P.
Published: December 2003

Low‐grade endocrine neoplasms (LGENs) involving the liver usually show typical endocrine features. Occasionally these tumors display cytologic characteristics that mimic hepatocellular carcinoma (HCC) or adenocarcinoma (ACA). Twenty‐four liver FNABs from 22 patients were reviewed. Twenty specimens showed cytologic characteristics typical of LGENs, including small to medium‐sized cells with salt‐and‐pepper or granular chromatin, moderate to high N/C ratios, and inconspicuous or absent nucleoli. Plasmacytoid cells were observed in 19 cases. Immunocytochemical stains (ICCs) confirmed endocrine differentiation in 9 cases. 4 FNABs were comprised of larger tumor cells that displayed cytologic features that overlapped with HCC or ACA, including transgressing or wrapping endothelium, moderate to abundant cytoplasm, and conspicuous nucleoli. LGENs involving the liver typically demonstrate characteristic cytologic features. Cases comprised of larger cells with abundant cytoplasm, conspicuous nucleoli, and transgressing or wrapping endothelium may pose diagnostic difficulties. ICCs and plasmacytoid morphology are beneficial in classifying the endocrine origin of these tumors. Diagn. Cytopathol. 2004;30:31–38. © 2004 Wiley‐Liss, Inc.
Steady‐state ranked set sampling for replicated environmental sampling designs
Authors: Barabesi Lucio , Pisani Caterina
Published: December 2003

In replicated sampling protocols, each design replicate is independently and randomly placed onto the study region. Subsequently, in order to estimate the objective parameter, the Horvitz–Thompson (HT) estimator is usually considered for each design replicate and an overall estimator results from the average of the single estimators. Obviously, the procedure gives rise to a sample of HT estimators under simple random sampling (SRS) in such a way that the objective parameter is estimated by the corresponding sample mean. However, this procedure is likely to produce uneven coverage of the region and hence a large variability of the overall estimator. Therefore, to avoid such drawbacks, a quasi‐systematic protocol for the design replicates is proposed. In this case, the suggested procedure gives rise to a sample of HT estimators under steady‐state ranked set sampling (SRSS)–a generalization of well‐known ranked set sampling (RSS)–so as to estimate the objective parameter by the corresponding sample mean. The proposed method produces large efficiency gains and does not involve supplementary sampling costs or extra field work. Copyright © 2003 John Wiley & Sons, Ltd.
Trivial reductions of dimensionality in the propagation of uncertainties: a physical example
Authors: Bolado Ricardo , Mira José
Published: December 2003

When performing uncertainty analysis on a mathematical model of a physical process, some coefficients of the differential equations appear as a result of elementary operations of other coefficients. It is shown in this article that variance reduction techniques should be applied on the ‘final’ or ‘reduced’ coefficients and not on the original ones, thus reducing the variance of the estimators of the parameters of the output variable distribution. We illustrate the methodology with an application to a physical problem, a radioactive contaminant transport code. A substantial variance reduction is achieved for the estimators of the distribution function, the mean and the variance of the output. Copyright © 2003 John Wiley & Sons, Ltd.
Site scores and conditional biplots in canonical correspondence analysis
Authors: Graffelman Jan , Tuft Robyn
Published: December 2003

Canonical correspondence analysis is an important multivariate technique in community ecology. It produces an interesting biplot that summarizes the data matrices involved in the analysis. The method produces two sets of site scores that can be used in a biplot. One set concerns site scores that are weighted averages of the species scores (WA scores), and the other set represents site scores that are linear combinations of the environmental variables (LC scores). We show that the use of both sets of scores in a CCA biplot can be justified. The use of the WA scores leads to the best possible representation of the species data conditional on the representation of the weighted averages. Likewise, the LC scores lead to the best possible representation of the environmental variables, also conditional on the representation of the weighted averages and on the use of a Mahalanobis metric. The eigenvalues obtained in CCA indicate how well the species data are represented when LC scores are used. The quality of representation of the species data when WA scores are used can be computed from the CCA eigenvalues and the variances of the WA scores. Scalar products between WA scores and environmental variable vectors do not form a biplot of the environmental data. Theoretical results are illustrated with Australian data from freshwater ecology. Copyright © 2003 John Wiley & Sons, Ltd.
Translocation t(11;14) in multiple myeloma: Analysis of translocation breakpoints on der(11) and der(14) chromosomes suggests complex molecular mechanisms of recombination
Authors: Fenton James A. L. , Pratt Guy , Rothwell Dominic G. , Rawstron Andy C. , Morgan Gareth J.
Published: December 2003

We describe the characterization of the genomic DNA breakpoints of two multiple myeloma (MM) patients with t(11;14). IGH translocation events are present in many MM tumors, and it is proposed that they occur early in the pathogenesis, based on the assumption that the translocations are simple reciprocal events mediated by errors in class‐switch recombination (CSR). We provide evidence from two patients that the translocation event can be more complex, with DNA from chromosome band 11q13 joined to apparently already recombined hybrid (Sμ/Sγ) switch region sequences. In one case, there was also evidence that a further rearrangement had occurred at the t(11;14) recombination site, resulting in an inversion of 40 bp of the 5′Sμ flanking sequence. This suggests that primary IGH arrangements in MM may be more complex than previous myeloma models have suggested, but that they essentially occur through illegitimate CSR events. © 2003 Wiley‐Liss, Inc.
Clonal variation of the immunogenotype in relapsed ETV6/RUNX1‐positive acute lymphoblastic leukemia indicates subclone formation during early stages of leukemia development
Authors: Peham Martina , Konrad Marianne , Harbott Jochen , König Margit , Haas Oskar A , Panzer‐Grümayer E. Renate
Published: December 2003

Recent data suggest that late relapses evolve from an ancestral ETV6/RUNX1‐positive (also designated TEL/AML1‐positive) clone resulting from secondary changes (ETV6 deletion) that differ from those of the initial leukemia and, as a consequence, may also deviate in their clonotypic immunoglobulin/T‐cell receptor (IG/TCR) gene rearrangements. The aim of our study was to compare the immunogenotype and fluorescence in situ hybridization (FISH) patterns of the unrearranged ETV6 allele of matched diagnosis/relapse samples from 12 children with an early or late relapse. We identified varying degrees of differences in the IG/TCR in six of them. A clonal change or evolution of the unrearranged ETV6 allele was also observed in six children but remained unchanged in three. However, these two parameters were not in concordance, nor did the immunogenotype pattern correlate with the duration of the first remission. We therefore propose that the potential of the immunogenotype to diversify depends primarily on the stage of IG/TCR gene configuration of the cell in which the ETV6/RUNX1 gene fusion takes place. © 2003 Wiley‐Liss, Inc.
Mutational analysis of susceptibility genes RNASEL/HPC1, ELAC2/HPC2, and MSR1 in sporadic prostate cancer
Authors: Nupponen Nina N. , Wallén Mika J. , Ponciano Damaris , Robbins Christiane M. , Tammela Teuvo L.J. , Vessella Robert L. , Carpten John D. , Visakorpi Tapio
Published: December 2003

Three putative prostate cancer–susceptibility genes, RNASEL/HPC1 at 1q24, MSR1 at 8p22, and ELAC2/HPC2 at 17p11, have recently been identified. Our objective was to investigate somatic mutations in these genes in sporadic prostate cancer. We analyzed 39 clinical prostate cancer specimens, 10 prostate cancer xenografts (LuCaP series), and 4 prostate cancer cell lines (LNCaP, DU145, PC‐3, and MPC‐3) for genetic changes using denaturing high‐performance liquid chromatography and direct sequencing in order to screen the whole coding regions of RNASEL and MSR1, as well as exons 7 and 17 of ELAC2. The known 471delAAAG truncating mutation was found in the RNASEL gene in cell line LNCaP. The only new missense variation in RNASEL, Gly296Val, was found in cell line DU145, but not in any other samples. RNASEL and ELAC2 also showed the common missense polymorphic changes. A previously reported truncating mutation (Arg293X) was found in MSR1 in the germ line of one individual. Our results indicate that inactivation of the RNASEL, ELAC2, or MSR1 genes by somatic mutation is a rare phenomenon in sporadic prostate cancer. © 2003 Wiley‐Liss, Inc.
Activated BRAF targets proximal colon tumors with mismatch repair deficiency and MLH1 inactivation
Authors: Domingo Enric , Espín Eloi , Armengol Manel , Oliveira Carla , Pinto Mafalda , Duval Alex , Brennetot Caroline , Seruca Raquel , Hamelin Richard , Yamamoto Hiroyuki , Schwartz Simó
Published: December 2003

BRAF, a serine/threonine kinase of the RAF family, is a downstream transducer of the RAS‐regulated MAPK pathway and signals upstream of MEK1/2 kinases. Recently, activating mutations within BRAF have been reported in a high percentage of melanomas and colorectal carcinomas and shown to have oncogenic capabilities. Further, their association to mismatch‐repair‐deficient tumors has suggested the involvement of the RAS/RAF pathway in the tumorigenesis of microsatellite‐unstable colon cancers, and that RAS and RAF mutations are alternative genetic events. We determined whether colorectal mismatch‐repair‐deficient tumors with BRAF mutations show a specific genotype when compared with tumors with wild‐type BRAF, and whether they can be associated with a particular clinicopathological feature. Here, we report a striking association of BRAF, but not of APC, KRAS2, AXIN2, and TP53 mutations, with proximal mismatch‐repair‐deficient colon tumors and MLH1 hypermethylation. Our results support the hypothesis that proximal and distal colorectal tumors with mismatch repair deficiency harbor different genetic alterations, and we suggest that the involvement of the RAS/RAF pathway in colorectal tumorigenesis is differentially modulated according to tumor location and MLH1 inactivation. © 2003 Wiley‐Liss, Inc.
Expression of a naturally occurring constitutively active variant of the epidermal growth factor receptor in mouse fibroblasts increases motility
Authors: Pedersen Mikkel W. , Tkach Vadim , Pedersen Nina , Berezin Vladimir , Poulsen Hans S.
Published: December 2003

Tumor cell motility is one of the rate‐limiting steps of invasion, which defines progression toward a more malignant phenotype. Elevated expression of epidermal growth factor receptor (EGFR) in many cancers is associated with progression of superficial to invasive forms of the disease. The naturally occuring type III mutant epidermal growth factor receptor (EGFRvIII) is a tumor‐specific, ligand‐independent, constitutively active variant of the epidermal growth factor receptor. EGFRvIII is expressed frequently by a number of human solid tumours including those of the lung, breast, prostate, brain and ovary. Our study was designed to investigate the effect of EGFRvIII expression on cell motility and compare it to that of ligand‐activated EGFR using transfected fibroblasts. We show here using time‐lapse video recording that expression of EGFRvIII greatly enhances the motility of fibroblasts independently of ligand stimulation. In addition, expression of EGFRvIII caused a marked increase in the number of cellular protrusions (lamellipodia) and a reduction in the number of stress fibers and focal adhesions. The EGFR tyrosine kinase inhibitor, AG1478, and the MEK inhibitor, U0126, blocked these cellular effects of EGFRvIII. Two cell lines expressing different levels of EGFR were used for comparison. The low‐expressing cell line responded to EGF treatment by increasing motility in a manner very similar to the motility induced by EGFRvIII. In contrast, the high‐expressing cell line responded to EGF by detachment from the extracellular matrix and decreased motility. Cellular detachment was correlated to a high phosphorylation of PLC‐γ, whereas increased motility was correlated to a high level of ERK phosphorylation. Overall these results indicate that tumor‐associated EGFR mutations might be critical for tumor cell motility, invasion and thus progression of disease. © 2003 Wiley‐Liss, Inc.
Elevated levels and distinct patterns of expression of A‐type cyclins and their associated cyclin‐dependent kinases in male germ cell tumors
Authors: Liao Ching , Li Sherry Q. , Wang XiangYuan , Muhlrad Samantha , Bjartell Anders , Wolgemuth Debra J.
Published: December 2003

Aberrant expression of several key regulators controlling the G1/S phase of the cell cycle has been implicated in human male germ cell tumorigenesis. Given the critical role of cyclin A2 at both the G1/S and G2/M transitions and the essential role for cyclin A1 in male germ cell development, our present study focused on the involvement of the A‐type cyclins in the transformation and progression of male germ cell tumors (GCTs). The expression of the A‐type cyclins and their catalytic partners Cdk1 and Cdk2 was examined in all types and stages of human male GCTs, including carcinoma in situ(CIS), seminoma and non‐seminoma GCTs, along with normal testis samples. Elevated levels of cyclin A2, Cdk1 and Cdk2 were detected in the majority of GCTs and were correlated with the invasiveness of the tumors (p < 0.05). Cyclin A1 expression was virtually undetectable in CIS and seminoma, but was aberrantly expressed in all non‐seminomatous GCTs. Cyclin A2 expression was strongly correlated with that of its catalytic partners Cdk1 and Cdk2 in all types of testicular tumors examined (p < 0.05), whereas a strong correlation between cyclin A1 and Cdk1 or Cdk2 was only seen in non‐seminomatous GCTs (p < 0.05). Histone kinase activities of cyclin A1/Cdks and cyclin A2/Cdks were found to be elevated in tumors. Our data suggest that aberrant expression of A‐type cyclins and their Cdks is a significant factor in male germ cell tumorigenesis. The abundant ectopic expression of cyclin A1 in non‐seminomatous GCTs and its absence in CIS and seminomas is likely linked to the tumor transformation and progression and may be relevant to clinical prognosis. Supplementary material for this article can be found on the International Journal of Cancer website at http://www.interscience.wiley.com/jpages/0020‐7136/suppmat/index.html.© 2003 Wiley‐Liss, Inc.
hnRNP L enhances sensitivity of the cells to KW‐2189
Authors: Taguchi Fumiko , Kusaba Hitoshi , Asai Akira , Iwamoto Yasuo , Yano Keiichi , Nakano Hirofumi , Mizukami Tamio , Saijo Nagahiro , Kato Harubumi , Nishio Kazuto
Published: December 2003

Heterogeneous nuclear ribonucleoproteins (hnRNPs) are involved in several RNA‐related biological processes. We demonstrated hnRNP L as a candidate protein of DARP (duocarmycin‐DNA adduct recognizing protein) by gel shift assay and amino acid sequencing. Stable transfectants of hnRNP L showed high sensitivity of the cells to the growth inhibitory effect of KW‐2189, a duocarmycin derivative in vitro. Immunostaining of hnRNP L demonstrated differential intracellular localization of hnRNP L among human lung cancer cell lines. A transfection study using a series of deletion mutants of hnRNP L fused to indicated that the N‐terminal portions of RRM(RNA recognition motif)1, RRM3 and RRM2 are involved in localization of hnRNP L. We identified sequences in these portions that have high homology with the sequences of known NLS (nuclear localization signal) and NES (nuclear export signal). hnRNP L is a factor that determines the sensitivities of cancer cells to the minor groove binder, and overexpression and differential intracellular localization of hnRNP L are involved in its function in lung cancer. © 2003 Wiley‐Liss, Inc.
Stat5a is tyrosine phosphorylated and nuclear localized in a high proportion of human breast cancers
Authors: Cotarla Ion , Ren Shuxun , Zhang Ying , Gehan Edmund , Singh Baljit , Furth Priscilla A.
Published: December 2003

Signal transducers and activators of transcription (STATs) are latent cytoplasmic transcription factors that are activated and translocated into the nucleus after phosphorylation at a conserved tyrosine residue. Mouse model studies have demonstrated that activated Stat5a acts as a critical survival factor for normal, preneoplastic and malignant mammary epithelial cells. Very limited information is available, however, on the expression, tyrosine phosphorylation status and nuclear localization of Stat5a in human breast cancers. In our study, the pattern of Stat5a cellular localization was analyzed by immunohistochemistry in a series of 83 randomly selected primary human breast adenocarcinomas. Immunoprecipitation/Western blotting and immunohistochemistry assays employing different phospho‐specific antibodies verified Stat5a tyrosine phosphorylation status. Stat5a was nuclear localized and tyrosine phosphorylated in 59 of 78 (76%) breast cancers examined; 38 of 78 (49%) demonstrated Stat5a nuclear localization in more than 25% of the breast cancer cells within the adenocarcinomas. Nuclear localized Stat5a was associated positively with increased levels of histologic differentiation (p = 0.03). A statistically significant positive association with p27 nuclear localization also was identified (p = 0.05). No relationship was found between nuclear localized Stat5a and menopausal status, tumor size, ploidy, percentage of cells in S‐phase, lymph node metastases, ER, ErbB2, nuclear localized p21 or nuclear localized Stat5b/Stat3. As its role in human breast cancer progression and response to therapy is defined, Stat5a could become a new molecular target for breast cancer therapy. © 2003 Wiley‐Liss, Inc.
p66Shc protein is upregulated by steroid hormones in hormone‐sensitive cancer cells and in primary prostate carcinomas
Authors: Lee Ming‐Shyue , Igawa Tsukasa , Chen Siu‐Ju , Van Bemmel Dana , Lin Jamie S. , Lin Fen‐Fen , Johansson Sonny L. , Christman Judith K. , Lin Ming‐Fong
Published: December 2003

Members of Shc family conventionally serve as critical adaptors in tyrosine phosphorylation signal transduction pathways. p66Shc protein, a member of Shc family, is predominantly expressed in epithelial cells, whereas the regulation of its expression remains an enigma. We describe the effect of steroid hormones on the protein level of p66Shc and growth stimulation in hormone‐sensitive human prostate, testicular and breast cancer cells. In DHT‐treated androgen‐sensitive prostate cancer LNCaP C‐33 cells, the protein level of p66Shc was elevated by approximately 3‐fold, correlating with increased cell growth. This DHT effect on p66Shc protein level and growth regulation was also observed in another androgen‐sensitive prostate cancer cell line MDA PCa2b as well as 2 testicular cancer cell lines, Tera‐1 and Tera‐2 cells. Similarly, the female sex hormone estrogen had a stimulating effect on p66Shc protein level and proliferation in estrogen‐sensitive MCF‐7 breast cancer cells. The upregulation of p66Shc protein level by DHT was competitively abolished by Casodex, an androgen antagonist used to treat prostate cancer. Moreover, immunohistochemical analyses showed that the p66Shc protein level was significantly higher in primary prostate tumors than in adjacent non‐cancerous cells (p < 0.05). The data collectively indicate that p66Shc protein levels correlate with steroid hormone‐stimulated cell growth and prostate carcinogenesis. © 2003 Wiley‐Liss, Inc.
mRNA expression of leukemia‐associated antigens in patients with acute myeloid leukemia for the development of specific immunotherapies
Authors: Greiner Jochen , Ringhoffer Mark , Taniguchi Masanori , Li Li , Schmitt Anita , Shiku Hiroshi , Döhner Hartmut , Schmitt Michael
Published: December 2003

Specific immunotherapies for patients with acute myeloid leukemia (AML) using leukemia‐associated antigens (LAA) as target structures might be a therapeutic option to enhance the graft‐vs.‐leukemia effect observed after allogeneic stem cell transplantation or to prolong a complete remission (CR) achieved by chemotherapy. Significant mRNA expression of LAA is a prerequisite for such immunotherapies. Here, previously characterized antigens associated with solid tumors (TAA) and newly characterized LAA were investigated for their expression in up to 60 AML patients and in leukemia cell lines. To investigate their specificity for leukemic blasts, the mRNA expression was also characterized in PBMN and CD34 positive cells of healthy volunteers and in a panel of normal tissues. The following antigens showed high mRNA expression in AML patients: MPP11 was detected in 43/50 (86%), RHAMM in 35/50 (70%), WT1 in 40/60 (67%), PRAME in 32/50 (64%), G250 in 18/35 (51%), hTERT in 7/25 (28%) and BAGE in 8/30 (27%) of AML patients. Real‐time RT‐PCR showed a tumor‐specific expression of the antigens BAGE, G250 and hTERT, as well as highly tumor‐restricted expression for RHAMM, PRAME and WT1. The antigen MPP11 was overexpressed. These antigens might be candidates for immunotherapies of leukemia patients and, because of their simultaneous expression, also for polyvalent vaccines. © 2003 Wiley‐Liss, Inc.
Serum concentrations of IGF‐I, IGFBP‐3 and c‐peptide and risk of hyperplasia and cancer of the breast in postmenopausal women
Authors: Schairer Catherine , Hill Deirdre , Sturgeon Susan R. , Fears Thomas , Pollak Michael , Mies Carolyn , Ziegler Regina G. , Hoover Robert N. , Sherman Mark E.
Published: December 2003

Experimental evidence suggests that insulin and insulin‐related growth factors may play a role in breast pathology through their mitogenic and anti‐apoptotic effects on breast cells. Our objective was to assess the relationship between serum concentrations of insulin‐like growth factor‐I (IGF‐I), its major binding protein (IGFBP‐3), the ratio IGF‐I:IGFBP‐3, c‐peptide (a marker of insulin secretion) and the ratio c‐peptide:fructosamine (a marker of insulin resistance) and the risk of epithelial hyperplasia (an established breast cancer risk factor) and localized breast cancer among postmenopausal women. Study subjects were patients who provided serum before breast biopsy or mastectomy in 3 hospitals in Grand Rapids, MI between 1977 and 1987. Two case groups, 186 subjects with epithelial hyperplasia of the breast and 185 subjects with localized breast cancer, were compared to 159 subjects with nonproliferative breast changes that have not been associated with increased breast cancer risk. Serum concentrations of IGF‐I, IGFBP‐3 and the ratio IGF‐I:IGFBP‐3 were not related to risk of either hyperplasia or breast cancer. For women in the highest quartile of c‐peptide or of c‐peptide:fructosamine compared to those in the lowest quartile, the odds ratios (ORs) for hyperplasia were 3.0 (95% confidence interval [CI] 1.4–6.5) and 3.3 (95% CI 1.5–7.3), respectively (p trend = 0.02 and 0.02, respectively). The corresponding ORs for breast cancer were 1.5 (95% CI 0.7–3.0) and 1.6 (95% CI 0.8–3.2), respectively (p trend = 0.35 and 0.25, respectively). Our results suggest that insulin and insulin resistance may play a role in breast pathology in postmenopausal women. © 2003 Wiley‐Liss, Inc.
Identification of bladder cancer antigens recognized by IgG antibodies of a patient with metastatic bladder cancer
Authors: Ito Keiichi , Fujita Tomonobu , Akada Masanori , Kiniwa Yukiko , Tsukamoto Makoto , Yamamoto Aiko , Matsuzaki Yuriko , Matsushita Maiko , Asano Takako , Nakashima Jun , Tachibana Masaaki , Hayakawa Masamichi , Ikeda Hideyuki , Murai Masaru , Kawakami Yutaka
Published: December 2003

To identify tumor antigens useful for the diagnosis and treatment of patients with bladder cancer, a lambda phage cDNA library constructed from a high‐grade bladder cancer cell line was screened with autologous serum from a patient with metastatic bladder cancer. Forty‐eight distinct antigens were isolated. By evaluating the immunogenicity and the tissue‐specific expression, KU‐BL‐1 and KU‐BL‐2 were identified as immunogenic antigens with restricted tissue expression. KU‐BL‐1 was found to be a putative human lipoic acid synthetase with a metal‐binding site, CXXXCXXC, that was expressed in bladder cancer cell lines and most bladder cancer tissues, as well as normal bladder mucosa and testis tissues. Immunoglobulin (Ig)G antibody to KU‐BL‐1 was detected in 2 of 28 patients with bladder cancer, but not in 30 healthy individuals. KU‐BL‐2 was found to be a putative human kelch‐like protein that was homologous to Drosophila
kelch, with a BTB/POZ domain and kelch repeats. KU‐BL‐2 was expressed in bladder cancer cell lines, most bladder cancer tissues, testis and heart, but not in normal bladder mucosa. IgG antibody to KU‐BL‐2 was detected in 8 of 28 patients with bladder cancer, but not in 16 healthy individuals. Tumor reactive T cells were induced from peripheral blood mononuclear cells (PBMC) by stimulation with one of the HLA–A24 binding KU‐BL‐2 peptides. Therefore, KU‐BL‐1 and KU‐BL‐2, which showed preferential expression in bladder cancer with restricted expression in normal tissues, as well as immunogenicity in multiple patients with bladder cancer, may be useful for the development of diagnostic and therapeutic methods for patients with bladder cancer. © 2003 Wiley‐Liss, Inc.
The role of type of tobacco and type of alcoholic beverage in oral carcinogenesis
Authors: Castellsagué Xavier , Quintana Maria Jesús , Martínez Maria Carmen , Nieto Adoración , Sánchez Maria José , Juan Amparo , Monner Antoni , Carrera Marta , Agudo Antoni , Quer Miquel , Muñoz Nubia , Herrero Rolando , Franceschi Silvia , Bosch F. Xavier
Published: December 2003

Incidence rates of oral and oropharyngeal cancers (oral cancer) in Spain are among the highest in Europe. Spain has a population heavily exposed to various types of tobacco and alcoholic beverages but the role and impact of tobacco type and beverage type in oral carcinogenesis remain controversial. To estimate the independent and joint effects of tobacco smoking and alcohol drinking habits on the risk of developing oral cancer, we carried out a multicenter, hospital‐based, case‐control study in Spain. Data from 375 patients newly diagnosed with cancer of the oral cavity or oropharynx and 375 matched control subjects were analyzed using multivariate logistic regression procedures. All exposure characteristics of amount, duration and cessation of both tobacco smoking and alcohol drinking were strongly associated with cancer risk following a dose‐dependent relationship. At equal intake or duration levels, black‐tobacco smoking and drinking of spirits were both associated with a 2‐ to 4‐fold increase in cancer risk compared to blond tobacco smoking or drinking of wine or beer, respectively. While ever exposure to smoking only or drinking only was associated with a moderate and nonsignificant increase in cancer risk, a history of simultaneous exposure to both habits was associated with a 13‐fold increase that was compatible with a synergistic effect model (p‐value for interaction: 0.008). Exposure to black tobacco smoking and/or drinking of spirits may account for up to 77% of oral cancer occurrence in Spain. Both black tobacco smoking and drinking of spirits place individuals at a very high risk of developing oral cancer. Simultaneous exposure to tobacco and alcohol consumption increases oral cancer risk in a synergistic fashion, even when consumption levels are moderate. These results underline the importance of type of tobacco and alcohol concentration in oral carcinogenesis. © 2003 Wiley‐Liss, Inc.
Two new trifunctional antibodies for the therapy of human malignant melanoma
Authors: Ruf Peter , Jäger Michael , Ellwart Joachim , Wosch Susanne , Kusterer Elisabeth , Lindhofer Horst
Published: December 2003

Trifunctional antibodies are able to redirect T cells and Fcγ receptor+ accessory immune cells to tumor targets. The simultaneous activation of these different classes of effector cells results in efficient killing of the tumor cells by different mechanisms such as phagocytosis and perforin‐mediated cytotoxicity. Here, we introduce 2 new trifunctional antibodies specific for human melanoma. These trifunctional antibodies recognize with one binding arm CD3 on human T cells. The other binding arm is directed against melanoma‐associated proteoglycans or melanoma‐associated gangliosides (GD2 as well as GD3). They mediate specific lysis of various melanoma cell lines in correlation with the level of antigen expression in short‐term cytotoxicity experiments. A combination of the 2 trifunctional antibodies was equally or even more efficient. Moreover, they induced a strong Th1 cytokine pattern with high amounts of IFN‐γ and low or no IL‐4. Accordingly, CD4+ and especially CD8+ T cells expanded, whereas B cells, NK cells and monocytes decreased. The cytokine response was up to 16‐fold higher when tumor cells were present. IFN‐γ reached cytotoxic concentrations for SK‐MEL‐23 melanoma cells. The induction of a T‐cell‐activatory and melanoma cell‐inhibitory cytokine milieu together with the redirection of T‐cell‐ and accessory cell‐mediated cytotoxicity are interesting features of these trifunctional antibodies. They may be a new option for the therapy of human malignant melanoma. © 2003 Wiley‐Liss, Inc.
Do younger female breast cancer patients have a poorer prognosis? Results from a population‐based survival analysis
Authors: Chia Kee Seng , Du Wen Bo , Sankaranarayanan R. , Sankila Risto , Wang Hao , Lee Jeannette , Seow Adeline , Lee Hin Peng
Published: December 2003

Younger women who develop breast cancer are hypothesized to have poorer survival rates than women who develop it at a later stage in life. Several studies have suggested that differences in biologic characteristics of breast cancer in younger (premenopausal) and older (postmenopausal) women may account for the prognostic variation. This population‐based cohort study reports on survival rates of breast cancer in Singapore and examines the hypothesis that younger breast cancer patients have a poorer prognosis. A total of 6,397 breast cancer patients diagnosed from 1968 to 1992 were identified from the population‐based cancer registry and followed up through 1997. Outcome measures were relative survival rates (RSRs) calculated using Hakulinen's method and excess hazards ratios (HRs) derived from a regression model based on relative survival. The 2‐, 5‐ and 10‐year RSRs were worse among those aged > 75 (65%, 48% and 39%, respectively). The best survival rates were seen among those aged 40–44 (84%, 67% and 56%). Patients younger than 35 years faired reasonably well (79%, 60% and 50%). When the data were stratified according to clinical stage and calendar year, the highest risk of excess deaths was found in women ≥ 75 years old. In patients with localized cancer and/or regional metastases, those in the 35–39 age group had the lowest excess risk. In patients with distant metastases, those younger than 35 years of age had the lowest excess risk of death. At the population level, younger women (< 45 years) with breast cancer in Singapore have higher relative survival rates. © 2003 Wiley‐Liss, Inc.
Structural elucidation of natural 2‐hydroxy di‐ and tricarboxylic acids and esters, phenylpropanoid esters and a flavonoid from Autonoë madeirensis using gas chromatographic/electron ionization, electrospray ionization and tandem mass spectrometric techniques
Authors: Dias C. , Dias M. , Borges C. , Almoster Ferreira M. A. , Paulo A. , Nascimento J.
Published: December 2003

The chemical composition of Autonoë madeirensis bulbs was characterized as part of a systematic phytochemical study of this species. The compounds reported were mainly identified on the basis of gas chromatography/electron ionization, electrospray ionization and tandem mass spectrometry. The structures of the pure compounds were also characterized by means of other physical and spectroscopic data (m.p., IR, UV, NMR). The compounds identified were 2‐hydroxy di‐ and tricarboxylic acids and esters (malic acid, citric acid and their methyl and ethyl esters), cis‐ and trans‐hydroxycinnamic esters (methyl and ethyl p‐coumarate and methyl ferulate) and a new flavone diglucoside, 7‐O‐[β‐glucosyl‐(1→2)‐O‐β‐glucosyl]apigenin, the interglucosidic linkage (1→2) of which is, to the best of our knowledge, reported for the first time in a diglucoside of apigenin. The results may contribute to the chemotaxonomy of the Autonoë genus and lead to a rapid tool for the systematic characterization of these compounds in plant extracts. Copyright © 2003 John Wiley & Sons, Ltd.
Formation of solvated ions in the atmospheric interface of an electrospray ionization triple‐quadrupole mass spectrometer
Authors: Schlosser Gitta , Takáts Zoltán , Vékey Károly
Published: December 2003

A simple method capable of generating and investigating various solvent clusters and solvated ions was developed. The technique opens a door to studying these complexes on commercially available instruments. Formation of the desired solvated ion in the gas phase was achieved by introducing the appropriate volatile solvent vapour into the curtain gas stream. Capabilities of the technique are illustrated by generating alkali, alkaline earth and transition metal cations solvated by various volatile compounds such as water, methanol and acetonitrile. Depending on the ligands and on the experimental conditions, clusters of 2–100 molecules may be observed. Isotope labelling suggests that these are formed by a re‐solvation process in the curtain gas region. Copyright © 2003 John Wiley & Sons, Ltd.
Chemical cross‐linking and mass spectrometry for mapping three‐dimensional structures of proteins and protein complexes
Authors: Sinz Andrea
Published: December 2003

Chemical cross‐linking of proteins, an established method in protein chemistry, has gained renewed interest in combination with mass spectrometric analysis of the reaction products for elucidating low‐resolution three‐dimensional protein structures and interacting sequences in protein complexes. The identification of the large number of cross‐linking sites from the complex mixtures generated by chemical cross‐linking, however, remains a challenging task. This review describes the most popular cross‐linking reagents for protein structure analysis and gives an overview of the strategies employing intra‐ or intermolecular chemical cross‐linking and mass spectrometry. The various approaches described in the literature to facilitate detection of cross‐linking products and also computer software for data analysis are reviewed. Cross‐linking techniques combined with mass spectrometry and bioinformatic methods have the potential to provide the basis for an efficient structural characterization of proteins and protein complexes. Copyright © 2003 John Wiley & Sons, Ltd.