bluuebunny/crossref_metadata_2025 huggingface dataset:
Revision of the International Neuroblastoma Pathology Classification
Authors: Peuchmaur Michel , d'Amore Emanuele S. G. , Joshi Vijay V. , Hata Jun‐ichi , Roald Borghild , Dehner Louis P. , Gerbing Robert B. , Stram Daniel O. , Lukens John N. , Matthay Katherine K. , Shimada Hiroyuki
Published: October 2003

BACKGROUND

Ganglioneuroblastoma, nodular (GNBn) comprises one of the categories of peripheral neuroblastic tumors. All tumors in this category, according to the original International Neuroblastoma Pathology Classification, are classified into an unfavorable histology group. Subsequently, it has been reported that GNBn can be divided into two prognostic subsets, a favorable subset (FS) and an unfavorable subset (US).

METHODS

Histology slides from 70 patients who were enrolled in Children's Cancer Group studies 3881 and 3891 and who had a diagnosis of GNBn were reviewed jointly by the members of International Neuroblastoma Pathology Committee (INPC): 1) to confirm the diagnosis of GNBn, 2) to identify the FS and US by applying the same age‐linked criteria that were used to distinguish the favorable histology group and unfavorable histology group in conventional neuroblastoma tumors from the neuroblastomatous component of GNBn tumors, and 3) to verify the significant prognostic difference between these two subsets. The patients had been used in a previous study, and survival data for the patients were updated since the time of their last report.

RESULTS

The review clarified and illustrated morphologic characteristics of classical GNBn and it variants. The diagnosis of GNBn was confirmed in 67 of 70 patients. There were 22 patients with GNBn in the FS and 45 patients with GNBn in the US. The estimated survival differences between the FS and US patients with GNBn were statistically significant (8‐year event free survival rate: 86.1% vs. 32.2%; P = 0.0003; overall survival rate: 90.5% vs. 33.2%; P = 0.0003).

CONCLUSIONS

This study confirmed the recently defined prognostic subsets of GNBn. The INPC proposes to modify the International Neuroblastoma Pathology Classification by distinguishing the FS and the US among patients with GNBn tumors. Cancer 2003. © 2003 American Cancer Society.
Outcome in 34 patients with juvenile‐onset mycosis fungoides
Authors: Wain E. Mary , Orchard Guy E. , Whittaker Sean J. , Spittle M.Sc. Margaret F. , Russell‐Jones Robin
Published: October 2003

BACKGROUND

Mycosis fungoides (MF) is predominantly a disease of older patients, but occasionally occurs in children. The aims of the current study were to describe the clinical presentation, pathologic features, and disease progression (DP) in patients who developed MF before age 16 years.

METHODS

A retrospective study was performed. Patients with juvenile‐onset MF were identified from our databases. Clinical features were determined from the medical records and patient interviews. Histologic, immunohistochemical, and T‐cell receptor (TCR) gene analysis was performed.

RESULTS

Thirty‐four patients were identified: 50% had Stage IA disease, 47% had Stage IB disease, and 3% had Stage IIA disease. The male‐to‐female ratio was 2:1. Clinical features included hypopigmented lesions (24%), poikiloderma (26%), pilotropic disease (9%), and disease associated with lymphomatoid papulosis (18%). Twenty‐eight patients had diagnostic histology, and six patients were included on the basis of compatible histology and a TCR clone in lesional skin. A cytotoxic immunophenotype was observed in 38%, including 71% of patients with hypopigmented lesions. Overall disease‐specific survival (DSS) rates at 5 and 10 years were 95% and 93%, respectively. DP rates were 5% at 5 years and 29% at 10 years. Subgroup analysis demonstrated improved DSS and reduced DP in patients with Stage IA disease, those with hypopigmented or poikilodermatous lesions, and those with associated lymphomatoid papulosis.

CONCLUSIONS

The prognosis for juvenile‐onset MF is similar to that of adult‐onset disease. There was an overrepresentation of a cytotoxic phenotype, which was most marked in hypopigmented variants. Widespread cutaneous disease (Stage IB) indicated a less favorable outcome. Cancer 2003. © 2003 American Cancer Society.
Clear cell ependymoma: A clinicopathologic and radiographic analysis of 10 patients
Authors: Fouladi Maryam , Helton Kathleen , Dalton James , Gilger Elizabeth , Gajjar Amar , Merchant Thomas , Kun Larry , Newsham Irene , Burger Peter , Fuller Christine
Published: October 2003

BACKGROUND

Clear cell ependymoma (CCE) is an uncommon central nervous system tumor with a predilection for the supratentorial region in children. Histologically, it may mimic oligodendroglioma, central neurocytoma, hemangioblastoma, and renal cell carcinoma.

METHODS

The authors reviewed the clinical, radiographic, and pathologic features, therapy, and outcome in 10 children with CCE who were treated at St. Jude Children's Research Hospital (1984–2003). Fluorescence in situ hybridization (FISH) was performed using 1p/1q, 19p/19q, CEP18/DAL1, and bcr/NF2 probe pairs.

RESULTS

The median patient age at diagnosis was 7.5 years (range, 1–19 years). Tumors occurred supratentorially in 9 of 10 patients. All tumors had rounded nuclei with surrounding, clear halos and at least focal perivascular pseudorosettes. Seven tumors had anaplastic features. No deletions involving 1p, 19q, or NF2 were detected. The tumors from 5 of 7 patients, all with anaplasia, had losses of both CEP18 and DAL‐1. Radiographically, all tumors were enhanced, and 9 tumors had associated cysts with enhancing walls. Seven patients underwent gross total resection, which was near total in one patient and subtotal in two patients. Five patients received immediate postoperative local radiotherapy. Three patients were diagnosed initially with pilocytic astrocytoma (one patient) and oligodendroglioma (two patients) and were observed. The progression‐free survival and overall survival rates at 5 years were 34% ± 20% and 75% ± 19%, respectively. The median follow‐up was 37 months (range, 5–239 months). Five patients developed local recurrence within a median of 9 months after diagnosis. Two patients developed extracranial soft tissue and lymph node metastases.

CONCLUSIONS

CCEs were found to have a predilection for extraneural metastases and early recurrence and demonstrate characteristic radiographic features, anaplastic histologic features, and chromosome 18 losses. The authors recommend resection followed by local radiotherapy as the treatment of choice in children. Cancer 2003. © 2003 American Cancer Society.
The role of late/slow replication of the FRA16D in common fragile site induction
Authors: Palakodeti Aparna , Han Yu , Jiang Yanwen , Le Beau Michelle M.
Published: October 2003

The FRA16D, at 16q23, spans the WWOX gene and is one of the most highly expressed common fragile sites observed when DNA replication is perturbed by aphidicolin. Several lines of evidence suggest that fragile sites are regions of DNA that are unusually sensitive to interference during replication. We have determined that the FRA16D alleles replicate in a synchronous fashion and that replication of these sequences occurs primarily in late S phase extending into G2 phase. Exposure to aphidicolin, an inhibitor of DNA polymerase α, results in a modest increase in cells with replication of FRA16D sequences in early S phase. This may represent initiation of replication in early S phase coupled with slow replication progression, or, alternatively, these cells may have passed through mitosis, entered the G1‐S phase of the next cell cycle, and initiated replication/repair. Our results support a model in which common fragile sites are sequences that may initiate replication in early‐mid S phase but are slow to complete replication, and the chromosomal breaks and gaps observed in metaphase cells result from unreplicated DNA. © 2003 Wiley‐Liss, Inc.
Frequent amplification and rearrangement of chromosomal bands 6p12‐p21 and 17p11.2 in osteosarcoma
Authors: Lau Ching C. , Harris Charles P. , Lu Xin‐Yan , Perlaky Laszlo , Gogineni Sheila , Chintagumpala Murali , Hicks John , Johnson Mark E. , Davino Nelson A. , Huvos Andrew G. , Meyers Paul A. , Healy John H. , Gorlick Richard , Rao Pulivarthi H.
Published: October 2003

Osteosarcoma (OS) is a highly malignant bone neoplasm of children and young adults. It is characterized by chaotic karyotypes with complex marker chromosomes. We applied a combination of molecular cytogenetic techniques including comparative genomic hybridization (CGH), spectral karyotyping (SKY), and fluorescence in situ hybridization (FISH) to decipher the chromosomal complexity in a panel of 25 tumors. Combined SKY and G‐banding analysis identified several novel recurrent breakpoint clusters and 9 nonrecurrent reciprocal translocations. CGH identified several recurrent chromosomal losses including 2q, 3p, 9, 10p, 12q, 13q, 14q, 15q, 16, 17p, and 18q, gains including Xp, Xq, 5q, 6p, 8q, 17p, and 20q, and high‐level chromosomal amplifications at Xp11.2, 1q21‐q22, 4p11, 4q12, 5p15, 6p12.1, 8q13, 8q23, 10q11, 10q22, 11q13, 11q23, 12q13‐q14, 13q21‐q34, 16q22, 17p11.2, 17q21‐q22, 18q22, 20p11.2, and 20q12. Frequent amplification and rearrangement involving chromosomal bands at 6p12‐p21 and 17p11.2 were found in 28% and 32% of cases, respectively. In an attempt to identify the genes involved in these amplicons, we used three nonoverlapping BAC clones contained within each amplicon as probes for FISH analysis, leading to a more detailed characterization and quantification of the 6p and 17p amplicons. © 2004 Wiley‐Liss, Inc.
Novel regions of amplification on 8q distinct from the MYC locus and frequently altered in oral dysplasia and cancer
Authors: Garnis Cathie , Coe Bradley P. , Ishkanian Adrian , Zhang Lewei , Rosin Miriam P. , Lam Wan L.
Published: October 2003

Genetic studies aimed at identifying key alterations in oral cancers have focused on analysis of tumors, with few such studies using early oral premalignant lesions (OPLs) because of limitations in both sample availability and size. In this study, we used a randomly amplified polymorphic DNA (RAPD)‐PCR approach to fingerprint DNA from microdissected normal and dysplastic cells and identified two recurrent genetic alterations on the long arm of chromosome 8 in OPLs, one mapping to 8q22 and the other to 8q24 near the MYC locus. We constructed a high‐resolution bacterial artificial chromosome (BAC) comparative genomic hybridization array consisting of 166 overlapping BAC clones that spans about 52 Mbp, from 8q21 to 8q24. Hybridization of DNA from microdissected oral tumors to the array revealed alteration at 8q24, with amplification of the BAC containing MYC. Strikingly, at least two other novel regions of amplification at 8q22 were identified. Microsatellite analysis of 93 oral dysplasias and tumors confirmed the presence of one of the alterations at 8q22. Loss of heterozygosity (LOH) at D8S1830, mapping within one of the regions of amplification, was observed in high frequency in both OPLs and tumors. Of the 37 cases with LOH at D8S1830, 23 (62%) showed retention at D8S1793, which maps 1.6 Mbp centromeric to MYC. This is further support for the alteration at 8q22 being distinct from MYC. These data raise the possibility of additional oncogenes on 8q near the MYC locus that are potentially involved in OPL disease progression. © 2003 Wiley‐Liss, Inc.
Contrasting in vivo and in vitro fates of glioblastoma cell subpopulations with amplified EGFR
Authors: Pandita Ajay , Aldape Kenneth D. , Zadeh Gelareh , Guha Abhijit , James C. David
Published: October 2003

Despite the high incidence of EGFR amplification in patient glioblastoma multiforme (GBM) tissues, only a single GBM cell line, of the many described in the literature, is known to contain and maintain amplified EGFR. Because EGFR mutations in GBM manifest primarily, if not exclusively, in amplified form, it follows that the availability of cell lines with mutation of endogenous EGFR would also be in short supply. In fact, there are no GBM cell lines harboring the common EGFR mutants described in patient GBMs. These observations suggest that in vivo environments select for EGFR amplification, whereas in vitro environments, specifically cell cultures, select against this gene alteration. To contrast directly the fates of EGFR amplification in vivo and in vitro, as well as to examine potential relationships between EGFR amplification and mutation, we have established and maintained GBM explants as xenografts by serial passaging in nude mice. Analysis of EGFR copy number and EGFR mutation status in 11 patient tumors and their corresponding xenografts, as well as the monitoring of EGFR copy number during the establishment of a GBM cell line from a xenograft with amplified EGFR, indicated that selection for EGFR amplification is an in vivo phenomenon. Furthermore, our data indicated that EGFR mutation occurs only in tumors with EGFR amplification and showed that the selection of amplified mutant EGFR over amplified wild‐type EGFR as a xenograft occurred rapidly and completely during tumor propagation. © 2003 Wiley‐Liss, Inc.
A nonredundant multicolor bar code as a screening tool for rearrangements in neoplasia
Authors: Müller Stefan , Eder Verena , Wienberg Johannes
Published: October 2003

A chromosome bar code describes the colored pattern of chromosome segments and is derived by multicolor fluorescence in situ hybridization (FISH) of defined molecular probes. Published approaches to the simultaneous differentiation of whole karyotypes with bar codes have not allowed the unequivocal identification of all chromosome segments because of color redundancy of the patterns from a multitude of identically colored segments. Here, we present a chromosome bar code approach in which the problem of color redundancy has been overcome. It allows the detailed description of translocations, including breakpoints as well as intrachromosomal rearrangements in the karyotype of tumor cells. The resolution of discernable bars was increased to 100 bars per haploid chromosome set by including human chromosome–specific probes and more well‐defined subregional probes such as chromosome arm‐ and segment‐specific probes. Technically, no limitation to further increase in the resolution of the pattern became apparent. The approach was validated by the analysis of four established tumor cell lines widely used as models in cell biology, revealing numerous inter‐ and intrachromosomal rearrangements. Chromosome bar coding as presented here may provide further useful information for the subregional assignment of chromosomal breakpoints in complex chromosome aberrations, as found in various neoplasms that cannot be obtained by chromosome painting or classical banding techniques alone. © 2003 Wiley‐Liss, Inc.
Acoustic noise concerns in functional magnetic resonance imaging
Authors: Moelker Adriaan , Pattynama Peter M.T.
Published: October 2003

Magnetic resonance (MR) acoustic scanner noise may negatively affect the performance of functional magnetic resonance imaging (fMRI), a problem that worsens at the higher field strengths proposed to enhance fMRI. We present an overview of the current knowledge on the effects of confounding acoustic MR noise in fMRI experiments. The principles and effectiveness of various methods to reduce acoustic noise in fMRI are discussed, practical considerations are addressed and recommendations are made. Hum. Brain Mapping 20:123–141, 2003. © 2003 Wiley‐Liss, Inc.
Physiological evidence of interaction of first‐ and second‐order motion processes in the human visual system: A magnetoencephalographic study
Authors: Sofue Ayako , Kaneoke Yoshiki , Kakigi Ryusuke
Published: October 2003

Humans have several mechanisms for the visual perception of motion, including one that is luminance‐based (first‐order) and another that is luminance‐independent (second‐order). Recent psychophysical studies have suggested that significant interaction occurs between these two neural processes. We investigated whether such interactions are represented as neural activity measured by magnetoencephalography (MEG). The second‐order motion of a drifting sinusoidal grating, which is defined by the speed of the dot motion, did not generate a response. Apparent motion (AM) of the square area, defined by the speed of randomly moving dots, evoked a magnetic response whose latency and amplitude changed with the distance that the area moved (a second‐order characteristic), though the response properties were significantly different from those for the first‐order AM. AM, defined by both first‐ and second‐order attributes, evoked an MEG response and the latencies and the amplitudes were distributed between those for the first‐ and second‐order motions. The cortical source of the response was estimated to be around MT+. The results show a distinct difference in the neural processing of the second‐order motion that cannot be explained by the difference in visibility, and they indicate that the interaction of the neural processes underlying first‐ and second‐order motion detection occurs before the MEG response. Our study provides the first physiological evidence of a neural interaction between the two types of early motion detection. Hum. Brain Mapp. 20:158–167, 2003. © 2003 Wiley‐Liss, Inc.
If the price is right: vagueness and values clarification in contingent valuation
Authors: Shiell Alan , Gold Lisa
Published: October 2003

The use of willingness to pay to value the benefits of health care is increasing. Much of this work assumes that health preferences are well formed or ‘complete’ and readily revealed if the right question is asked in the right way. We examined this assumption, seeking evidence in a mixed‐methods study that explored the meaning and implications of vague responses to a payment‐scale based willingness to pay exercise.

One‐half of the sample said that their vagueness meant that their maximum willingness to pay was actually greater than the amount that they had previously said it was. Thirty percent agreed that they would probably pay £10 more than a sum that they had previously said they would most definitely not pay, if they found this to be the cost of the vaccine. Interview data supported the view that the payment scale had failed to elicit the maximum willingness to pay and that some participants used the information on cost to help clarify their values, in contrast to the theory underpinning willingness to pay. The results suggest a need to consider values‐clarification in health economic evaluations. Copyright © 2002 John Wiley & Sons, Ltd.
A view from the bridge: agreement between the SF‐6D utility algorithm and the Health Utilities Index
Authors: O'Brien Bernie J. , Spath Marian , Blackhouse Gordon , Severens J.L. , Dorian Paul , Brazier John
Published: October 2003

Background: The SF‐6D is a new health state classification and utility scoring system based on 6 dimensions (‘6D’) of the Short Form 36, and permits a “bridging” transformation between SF‐36 responses and utilities. The Health Utilities Index, mark 3 (HUI3) is a valid and reliable multi‐attribute health utility scale that is widely used. We assessed within‐subject agreement between SF‐6D utilities and those from HUI3.

Methods: Patients at increased risk of sudden cardiac death and participating in a randomized trial of implantable defibrillator therapy completed both instruments at baseline. Score distributions were inspected by scatterplot and histogram and mean score differences compared by paired t‐test. Pearson correlation was computed between instrument scores and also between dimension scores within instruments. Between‐instrument agreement was by intra‐class correlation coefficient (ICC).

Results: SF‐6D and HUI3 forms were available from 246 patients. Mean scores for HUI3 and SF‐6D were 0.61 (95% CI 0.60–0.63) and 0.58 (95% CI 0.54–0.62) respectively; a difference of 0.03 (p<0.03). Score intervals for HUI3 and SF‐6D were (‐0.21 to 1.0) and (0.30–0.95). Correlation between the instrument scores was 0.58 (95% CI 0.48–0.68) and agreement by ICC was 0.42 (95% CI 0.31–0.52). Correlations between dimensions of SF‐6D were higher than for HUI3.

Conclusions: Our study casts doubt on the whether utilities and QALYs estimated via SF‐6D are comparable with those from HUI3. Utility differences may be due to differences in underlying concepts of health being measured, or different measurement approaches, or both. No gold standard exists for utility measurement and the SF‐6D is a valuable addition that permits SF‐36 data to be transformed into utilities to estimate QALYs. The challenge is developing a better understanding as to why these classification‐based utility instruments differ so markedly in their distributions and point estimates of derived utilities. Copyright © 2003 John Wiley & Sons, Ltd.
Illustrating the impact of including future costs in economic evaluations: an application to end‐stage renal disease care
Authors: Manns Braden , Meltzer David , Taub Ken , Donaldson Cam
Published: October 2003

There are strong theoretical arguments for including future costs for related and unrelated medical care and non‐medical expenditures within economic evaluations. Nevertheless, there is limited data on how inclusion of such costs affects the cost effectiveness of medical interventions in practice. For a low‐cost intervention that improves survival in end‐stage renal disease (ESRD) patients, we sought to determine how the inclusion of future costs for related medical care (i.e. dialysis and transplantation) and for unrelated medical care and non‐medical expenditure would affect the magnitude of the cost per QALY ratio. We performed a cost‐utility analysis comparing hemodialysis using a synthetic dialyser (the current treatment of choice in Canada) with the historical gold‐standard treatment (use of a cellulose dialyser). We contrasted the results of the analysis including and excluding various measures of future costs. While the inclusion of future costs for unrelated medical care and non‐medical expenditures had a significant impact on the cost per QALY ratio, the size of the cost per QALY ratio was most sensitive to inclusion of future costs for related medical care. Our analysis shows that even relatively inexpensive interventions that extend survival of dialysis patients may not be cost‐effective since, by extending survival, the extra outpatient dialysis costs that are incurred are large. Inclusion of such costs (which, in and of itself, is methodologically correct) in economic evaluations in this area may mitigate against the acceptance of interventions that are relatively inexpensive themselves but which improve patient survival. Copyright © 2003 John Wiley & Sons, Ltd.
Maxillary obturators: The relationship between patient satisfaction and speech outcome
Authors: Rieger Jana M. , Wolfaardt John F. , Jha Naresh , Seikaly Hadi
Published: October 2003

Background.

Patient satisfaction with a maxillary obturator has been studied in relation to extent of surgical defect, sociodemographic characteristics, scores on mental health inventories, and psychosocial adjustment to illness scales. However, review of the literature reveals limited study of the relationship between patient satisfaction with an obturator and clinical speech outcome measures. The purpose of this study is to relate patient satisfaction scores obtained by questionnaire with those obtained by means of clinical speech measurements.

Methods.

Acoustical, aeromechanical, and perceptual measurements of speech were collected for 20 patients after receiving a definitive obturator. Patient satisfaction with their obturator was later measured with the Obturator Functioning Scale (OFS).

Results.

Results reveal that poorer aeromechanical speech results were associated with patient‐reported avoidance of social events, whereas lower speech intelligibility outcomes were related to overall poorer perception of speech function on the OFS. Several background patient characteristics were significantly related to several responses on the OFS and to the aeromechanical assessment outcomes.

Conclusions.

Results from instrumental assessments of speech seem to be informative regarding not only speech outcome but also a patient's satisfaction with the obturator. Consideration of background patient characteristics is important when interpreting both clinically obtained and patient‐perceived outcomes. © 2003 Wiley Periodicals, Inc. Head and Neck 25: 000–000, 2003
Validation of tissue array technology in head and neck squamous cell carcinoma
Authors: Chen Bei , van den Brekel Michiel W. M. , Buschers Wim , Balm Alfons J. M. , van Velthuysen Marie‐Louise F.
Published: October 2003

Background.

The recent development of tissue array technology has potentiated large‐scale retrospective cohort studies using archival formalin‐fixed, paraffin‐embedded tissues. This study evaluates the potential for using archival head and neck cancer tissue in such arrays.

Method.

Tissue array blocks were made from 184 head and neck cancer specimens. Three core tissue biopsies (0.6 mm × 3–4 mm) were taken from individual “donor” paraffin‐embedded tumor blocks and arrayed into a new “recipient” paraffin block. Immunohistochemical (IHC) analyses were performed using antibodies recognizing cyclin‐D1, Rb, and EGFR. IHC was scored on a 6‐point scale for extent and a 3‐point scale for intensity. We compared the staining of tissue array disks with staining of full tissue sections.

Results.

Seventy‐four percent (475 of 640) of samples placed into tissue arrays were confirmed to represent tumor tissue. The remaining samples were lost during processing or contained too few tumor cells. Only 6% of cases were completely lost, whereas 55%, 28%, and 11% of cases were judged on 3, 2, or 1 disk, respectively. Cohen's κ coefficient was 0.66 for cyclin‐D1, 0.40 for EGFR, and 0.41 for Rb.

Conclusions.

Tissue array technology is a rapid and efficient method for retrospective analysis of protein expression and is a promising tool for validation of prognostic markers in large series of head and neck squamous cell carcinomas. The agreement in scoring of the full section and the tissue arrays is reasonable. Discordance is probably due to intraobserver variation and lack of robustness of the scoring inherent of the proteins studied. © 2003 Wiley Periodicals, Inc. Head and Neck 25: 000–000, 2003
The utility of tolonium chloride rinse in the diagnosis of recurrent or second primary cancers in patients with prior upper aerodigestive tract cancer
Authors: Epstein Joel B. , Feldman Roy , Dolor Rowena J. , Porter Stephen R.
Published: October 2003

Background.

The clinical utility of tolonium chloride rinse was compared with unaided visual examination alone in the diagnosis of oral carcinoma in patients previously treated for carcinoma of the upper aerodigestive tract.

Methods.

A total of 668 patients were enrolled in this multicenter study. At each site, an oral clinical visual examination was completed by one investigator followed by tolonium chloride rinse and examination by a second investigator blinded to the examination findings of the first investigator. If a lesion was considered suspicious (urgent biopsy required at the first visit), the lesion was biopsied after tolonium chloride rinsing. Patients with lesions characterized at the first visit as not suspicious (biopsy not urgent) or that stained with tolonium chloride were asked to return for a second visit. At the second visit, any residual lesion or lesions that retained tolonium chloride were biopsied.

Results.

A total of 96 biopsies was performed in 81 of the 668 patients (12.1%), of which 30 (31.3%) were diagnosed histologically as carcinoma/carcinoma in situ (CIS) and the remainder as inflammation (31.3%), keratosis (26.6%), dysplasia (21.9%), ulcer (2.1%), other (3.1%), or no abnormality (1.0%). Of the 30 lesions with the diagnosis of carcinoma/CIS, 12 (sensitivity 40.0%) were considered to be clinically suspicious (CS+), whereas 29 (sensitivity 96.7%) retained tolonium chloride (p = .0002). The predictive values of a positive test for clinical examination and tolonium chloride staining were similar (36.4% vs 32.6%; p = .5871), indicating that the greater sensitivity of tolonium chloride was not associated with an excessive number of unnecessary biopsies (false positives).

Conclusions.

Tolonium chloride rinse is more sensitive than clinical examination alone in detecting lesions that might be found on biopsy to be carcinoma or CIS. The increased sensitivity is largely attributed to lesions that stain but were not detected clinically on visual examination. © 2003 Wiley Periodicals, Inc. Head Neck 25: 000–000, 2003
A new technique for multiparameter imaging microscopy: Use of long decay time photoluminescent labels enables multiple color immunocytochemistry with low channel‐to‐channel crosstalk
Authors: Soini Aleksi E. , Kuusisto Ari , Meltola Niko J. , Soini Erkki , Seveus Lahja
Published: October 2003

In this report, we describe luminescence imaging microscopy using five different photoluminescent dyes in a single specimen. We combined the long decay time luminophores, europium(III) chelate, terbium(III) chelate, palladium(II) coproporphyrin, and platinum(II) coproporphyrin, with a green nuclear stain, Syto 25™, that emits conventional fast decaying fluorescence. The luminescence emissions from the five different luminophores were separated from each other by the differences in spectra and decay times using time‐resolved detection. Applicability of this dye‐combination for multiparameter analysis of a biological object was verified in a mixed population of peripheral blood leukocytes. Leukocyte cytocentrifugates were incubated in one step with a cocktail of luminophore‐conjugated antibodies recognizing neutrophil‐ and lymphocyte‐specific markers, followed by rapid staining with a mixture of nuclear stain and Pt‐porphyrin as an eosinophil stain. The results show that multiple luminescent dyes with long decay time can be used together, and in combination with a conventional fluorophore. The separation of the signals of the long decay time labels was distinctive and enabled reliable identification of different leukocyte types, as well as an automated cell count. The long decay time luminophores together with time‐resolved luminescence imaging microscopy (TR‐LIM) provide a unique tool for studies of simultaneous expression of multiple antigens at the level of a single cell. In comparison with other multiparameter imaging techniques, the described technique offers increased accuracy of results, simplification of preparation procedure, and dramatic shortening of the total processing time. To our knowledge, this is the first time that simultaneous fivefold labeling/staining and analysis in a single specimen has been performed in the field of immunocytochemistry. Microsc. Res. Tech. 62:396–407, 2003. © 2003 Wiley‐Liss, Inc.
Induced maturation of frog mast cells by nerve growth factor during ontogenesis
Authors: Baccari Gabriella Chieffi , Raucci Franca , Di Fiore Maria M. , Monteforte Rossella
Published: October 2003

The effect of nerve growth factor (NGF) on ontogenesis of frog mast cells was investigated in vivo by histochemical, morphometric, and ultrastructural analysis. Three groups of tadpoles at various stages of development were used. In the first group, the larvae received i.p. injections of 1 ng NGF/g; the second group received 10 ng NGF/g, while the control group received only the vehicle. The first recognizable mast cells arose symmetrically in the tongue at stage 26 of Witschi's standard table. At stages 26 and 29, the mast cell number in the NGF‐injected tadpoles was significantly higher than the control group. From stage 29 onward, the mast cell number rapidly increased in all groups. No significant differences in mast cell number were observed between the control group and the NGF‐injected groups at stages 31 and 33. Electron microscopy revealed that at metamorphic climax (stage 33), the mast cells in the NGF‐treated groups were more mature than those in the control group. Therefore, nerve growth factor at early stages of tadpole development is likely to induce differentiation of mast cell precursors, while at later stages it is likely to induce maturation of immature mast cells. The close anatomical association between mast cells and perineurium, observed during nerve development, is intriguing. Already in the early stages of nerve development, the mast cells form a network around Schwann cell–axon complexes, together with the perineurial cells. At climax, the mast cells are located between the perineurial layers, suggesting that they may play a role in the tissue‐nerve barrier of the perineurium. Nerve growth factor also seems to induce perineurial cell maturation. Microsc. Res. Tech. 62:439–450, 2003. © 2003 Wiley‐Liss, Inc.
Immunoelectron microscopic studies on protein gene product 9.5 and calcitonin gene‐related peptide in vallate taste cells and related nerves in the guinea pig
Authors: Huang Yi‐Jen , Wu Yu‐Hung , Lu Kuo‐Shyan
Published: October 2003

On the basis of our previous report that protein gene product 9.5 (PGP 9.5)‐immunoreactive nerve fibers and taste cells and calcitonin gene‐related peptide (CGRP)‐immunoreactive nerve fibers are found in guinea pig vallate papillae [Huang and Lu (1996b) Arch. Histol. Cytol. 59:433–441]. We speculated that PGP 9.5 might be a marker for taste receptor cells and that CGRP might play an important role in taste transmission. We, therefore, performed an immunohistochemical and ultrastructural analysis of taste cells and related nerves in guinea pig vallate papillae. In the connective tissue of the vallate papilla, the ultrastructural data revealed that the PGP 9.5‐immunoreactive nerve fibers were both myelinated and unmyelinated. The CGRP‐immunoreactive nerve fibers were unmyelinated and surrounded by the cytoplasm of Schwann cells as were the non‐immunoreactive fibers. In the vallate taste buds, only type III cells, which make synaptic contacts with intragemmal nerves, were PGP 9.5‐immunoreactive, while the nerve terminals making synaptic contact with the underlying type III cells were CGRP‐immunoreactive. From these observations, we conclude that: (1) PGP 9.5 might be a useful specific marker for type III cells in guinea pig vallate taste buds; and (2) CGRP‐containing nerve fibers might be primarily involved in the neural transmission of taste stimuli. Microsc. Res. Tech. 62:383–395, 2003. © 2003 Wiley‐Liss, Inc.
Effects of aging on neurons and glial cells from the superficial layers of the superior colliculus in rats
Authors: Díaz Florentina , Moreno Paloma , Villena Alicia , Vidal Lourdes , Pérez De Vargas Ignacio
Published: October 2003

The aim of this study was to investigate the effect of aging on glial cells and neurons from the superficial layers of the superior colliculus in rats. We used stereological methods to estimate the volume of the superficial layers, neuron size, and the number of neurons and glial cells in Wistar male rats aged 3, 24, 26, and 28 months. A 32.6% volume increase was found in the stratum griseum superficiale between the ages of 3 and 26 months, while in the 28‐month‐old animals a 19% decrease was observed. The stratum opticum did not show any changes in volume with age. Also, our analysis revealed a process of somatic and nuclear atrophy in the neurons of the superficial layers in animals aged 26 and 28 months. On the other hand, no statistically significant differences were found in the numbers of neurons. The number of glial cells in the stratum griseum superficiale showed an increase between the 3rd and 26th month, while the stratum opticum suffered no change. Microsc. Res. Tech. 62:431–438, 2003. © 2003 Wiley‐Liss, Inc.
Three‐dimensional computer reconstruction of large tissue volumes based on composing series of high‐resolution confocal images by GlueMRC and LinkMRC software
Authors: Karen Petr , Jirkovská Marie , Tomori Zoltán , Demjénová Erna , Janáček Jiří , Kubínová Lucie
Published: October 2003

Computer‐based visualization of large tissue volumes with high resolution based on composing series of high‐resolution confocal images is presented. GlueMRC and LinkMRC programs are introduced, implementing composition of overlapping series of optical sections captured by a confocal microscope, registration and subsequent composition of successive confocal stacks. Both programs are using an interactive approach in combination with automatic algorithms for image registration. Further, the method for obtaining surface renderings of microscopical structure under study is described. For this purpose, structure contours visible in the sections are interactively digitized using a Colon plug‐in module running in Ellipse environment. Then the coordinates of the contours are processed by special modules in the graphic programming environment IRIS Explorer and the structure surface is rendered. The method is shown on the 3‐D reconstruction of the capillary bed of human placental villi and chick embryonic gut and its vascular bed. Microsc. Res. Tech. 62:415–422, 2003. © 2003 Wiley‐Liss, Inc.
Transmission electron microscopic observations of membrane effects of antibiotic cecropin B on Escherichia coli
Authors: Chen Hueih Min , Chan Shiu‐Chiu , Lee Jao‐Chang , Chang Chia‐Ching , Murugan Marudhamuthu , Jack Ralph W.
Published: October 2003

The pathway of cell membrane lysis by the peptide antibiotic cecropin B (CB), which contains both a hydrophobic and an amphipathic α‐helix, was analysed by assessing the morphological changes of Escherichia coli following treatment with the peptide. Exposure of green fluorescent protein (GFP)‐expressing E. coli to CB does not lead to an efflux of GFP. Moreover, transmission electron microscopic (TEM) examination of cecropin B‐treated cells showed that severe swelling precedes cell death and that the outer membrane becomes distended away from the plasma membrane. Using immuno‐gold staining and TEM of E. coli expressing the maltose‐binding protein in the cytoplasm, it was apparent that the protein remains restricted to the cytoplasmic compartment. These observations suggest that CB causes gross disruption of the outer membrane of Gram‐negative bacteria. Circular dichroism measurements of CB in the presence of cell membrane‐mimicking liposomes showed that CB forms secondary structure dependent on the ratio of [lipid]/[peptide]. These observations from this study are important for the future design of custom antimicrobial peptides. Microsc. Res. Tech. 62:423–430, 2003. © 2003 Wiley‐Liss, Inc.
A soluble CAR‐SCF fusion protein improves adenoviral vector‐mediated gene transferto c‐Kit‐positive hematopoietic cells
Authors: Itoh Akira , Okada Takashi , Mizuguchi Hiroyuki , Hayakawa Takao , Mizukami Hiroaki , Kume Akihiro , Takatoku Masaaki , Komatsu Norio , Hanazono Yutaka , Ozawa Keiya
Published: October 2003

Background

Although adenoviral vectors primarily derived from the adenovirus serotype 5 (Ad5) are widely used for many gene transfer applications, they cannot efficiently infect hematopoietic cells, since these cells do not express the coxsackie‐adenoviral receptor (CAR).

Methods

We have developed a soluble fusion protein that bridges adenoviral fibers and the c‐Kit receptor to alter Ad5 tropism to immature hematopoietic cells. The CAR‐SCF fusion protein consists of the extracellular domains of CAR and stem cell factor (SCF). The human megakaryoblastic leukemia cell lines UT‐7 and M07e, human chronic myelogenous leukemia cell line K‐562, and erythroleukemia cell line TF‐1 were used to assess CAR‐SCF‐assisted Ad5‐mediated gene transfer. Hematopoietic cell lines were infected with an Ad5 vector (Ad5‐eGFP) or a fiber‐mutant Ad5/F35 (Ad5/F35‐eGFP) expressing the enhanced green fluorescent protein gene in the presence or absence of CAR‐SCF.

Results

Twenty‐four hours after infection, more than 80% of M07e cells infected in the presence of CAR‐SCF were eGFP‐positive, compared with very few eGFP‐positive cells following Ad5‐eGFP infection in the absence of CAR‐SCF. The enhancement of Ad5‐eGFP infection by CAR‐SCF was greater than that caused by Ad5/F35‐eGFP (50%). The ability of CAR‐SCF to enhance Ad5‐eGFP infectivity was highly dependent on cellular c‐Kit expression levels. Furthermore, CAR‐SCF also enhanced Ad5‐mediated gene transfer into human primary CD34+ cells.

Conclusions

The CAR‐SCF fusion protein assists Ad5‐mediated transduction to c‐Kit+ CAR− hematopoietic cells. The use of this fusion protein would enhance a utility of Ad5‐mediated hematopoietic cell transduction strategies. Copyright © 2003 John Wiley & Sons, Ltd.
Efficient gene transfer of a simian immuno‐deficiency viral vector into cardiomyocytes derived from primate embryonic stem cells
Authors: Nagata Mihoko , Takahashi Masafumi , Muramatsu Shin‐ichi , Ueda Yasuji , Hanazono Yutaka , Takeuchi Koichi , Okada Koji , Suzuki Yutaka , Kondo Yasushi , Suemori Masafumi , Ikeda Uichi , Nakano Imaharu , Kobayashi Eiji , Hasegawa Mamoru , Ozawa Keiya , Nakatsuji Norio , Shimada Kazuyuki
Published: October 2003

Background

Embryonic stem (ES) cells continually proliferate and can generate large numbers of differentiated cells. Genetic manipulation of transplantable cells derived from primate ES cells offers considerable potential for development research and regenerative cell therapy. However, protocols for efficient gene transfer into primate ES‐cell‐derived cells have not yet been established.

Methods

Spontaneously contracting areas were derived from cynomolgus monkey ES cells. Features of cardiomyocytes in the area were analyzed according to gene expression (RT‐PCR), morphology (immunostaining and electron microscopy), and function (intracellular calcium transience). Beating cells were transduced using a simian immunodeficiency virus (SIV) vector expressing enhanced green fluorescence protein (EGFP), then transplanted into ischemic rat myocardium.

Results

Beating cells derived from monkey ES cells displayed gene expression, ultrastructural and functional properties of early‐stage cardiomyocytes. Highly efficient (97% cardiac phenotype) and stable transduction of these ES‐cell‐derived cardiomyocytes was achieved using SIV vector without altering contractile function. In addition, transduced cardiomyocytes survived in the myocardium of a rat myocardial infarction model.

Conclusions

A lentiviral vector system based on SIV represents a useful vehicle for genetic modification of cardiomyocytes derived from primate ES cells, and can extend the application of primate ES cells to gene therapy. Copyright © 2003 John Wiley & Sons, Ltd.
Mice body weight gain is prevented after naked human leptin cDNA transfer into skeletal muscle by electroporation
Authors: Wang Xue‐Dong , Liu Jian , Yang Ji‐Chun , Chen Wei‐Qiang , Tang Jian‐Guo
Published: October 2003

Background

In this investigation, the feasibility of gene therapy for obesity by electroporational transfer of naked plasmid with leptin cDNA into skeletal muscle was tested. Both young and adult mice were studied.

Methods

Human leptin cDNA was attached to the human insulin precursor secretion signal peptide gene. The fused gene was then inserted into the mammalian expression vector pcDNA3.1(−) and transferred into skeletal muscle of normal female mice using electroporation.

Results

During the time of exogenic gene expression, daily food intake of leptin cDNA‐treated mice was observed to be lower than the control. The body weight gain was prevented efficaciously regardless of if they were young or adult. At the 7th week after gene transfer, the body weight of both young and adult leptin cDNA‐treated mice was about 20% lighter than the control. Although the body weight of pair fed controlled adult mice was close to the leptin cDNA‐treated mice at the 8th week, they were always heavier than the leptin cDNA‐treated mice before this time. The levels of retroperitoneal fats and serum TG of leptin cDNA‐treated mice were markedly lower than that of the control. The relative serum hyperleptinemic level could last for about 2 months. The expression of leptin cDNA in muscle cells was also detected by RT‐PCR. The levels of serum insulin and glucose of leptin cDNA‐treated mice decreased slightly. Our data also showed that the hyperleptinmia resulted in uterus expansion in young mice, but not in the adults.

Conclusion

The present study provides evidence of successful electroporation of naked plasmid DNA transfer for prevention or treatment of mice obesity. Copyright © 2003 John Wiley & Sons, Ltd.
Study of the metal dusting behaviour of high‐temperature alloys
Authors: Di Gabriele F. , Bernstein J. R. , Al‐Qhatani M. M. , Liu Z. , Jordan M. P. , Richardson J. A. , Stott F. H.
Published: October 2003

The corrosion phenomenon named metal dusting has been observed in many high‐temperature industrial plants. An experimental research programme is being carried out into the degradation resistance of wrought and cast commercial and development high‐temperature alloys in H2/CO gas mixtures at temperatures of 550°C to 750°C. Emphasis is placed on very high carbon activities, consistent with the next generation of steam‐reforming and similar plants that are susceptible to metal dusting. The overall programme is concerned with the mechanisms of initiation and propagation of dusting and the sensitivity to damage of the more resistant alloys, as a function of environmental parameters.

Initial tests have been carried out on a number of commercial alloys: Alloy 600, 693, 602CA, 601, 603 XL, 671, 617, 690 (wrought), and H46M (cast). The specimens were exposed to a gas mixture of high carbon activity at 650°C for a total of 1000 hours. Many of the alloys showed at least the initial stages of metal dusting. Preliminary analysis using electron microscopy revealed that initiation of metal dusting is influenced by microstructure, stress state and composition. In some cases, attack was enhanced at stress points, such as corners and edges. Sample holders were found to influence strongly the length of the initiation period for the onset of the corrosion phenomenon. The reaction layers in the alloy beneath areas of damage were analysed by EDX and EPMA. Mechanical characterisation of such areas has been carried out using nanoindentation methods. These early results are discussed in terms of the effectiveness of oxide scales in inhibiting the onset of damage and presence of impurities in the ceramic holder in initiating the onset of damage.
The influence of carbonyl sulphide on the inhibition of filamentary carbon deposition on stainless steel
Authors: Millward G. R. , Evans H. E. , Jones I. P. , Eley C. D. , Mowforth C. W.
Published: October 2003

This paper examines the role that COS plays in the inhibition of carbon deposition onto a Si‐free 20Cr‐25Ni‐Nb‐stabilized austenitic stainless steel. We have shown that the addition of just 240 vppb of COS to a standard gas deposit mixture of high carbon activity (1000 vppm C2H4, 1% CO, balance CO2) yields a dramatic fall in the amount of filamentous carbon deposited. The inhibitive effect persists when treatment is continued after COS has been subsequently removed from the treatment gas. Clearly, in these experiments the phenomenon is a result of substrate surface modification by the COS rather than changes in gas phase kinetics. Deposition onto surfaces where filamentous carbon has already been laid down is similarly inhibited.