bluuebunny/crossref_metadata_2025 huggingface dataset:
Anticonvulsant teratogenesis 5: Observer bias in a cohort study
Authors: Harvey Elizabeth A. , Coull Brent A. , Holmes Lewis B.
Published: June 2003

BACKGROUND

The most common physical effects of prenatal exposure to the anticonvulsant drugs phenytoin and phenobarbital are midface and digit hypoplasia. These features are subjective in nature, which makes their detection vulnerable to any bias the examiner has, such as an increased expectation that the infant being examined has been exposed prenatally to an anticonvulsant drug. The purpose of this analysis was to determine whether or not there was evidence of observer bias in the detection of presumed anticonvulsant drug‐related physical features.

METHODS

The presence or absence of three groups of features were compared: 1) midface hypoplasia, such as anteverted nostrils; 2) digit hypoplasia, e.g., tapered fingers, and 3) other features, e.g., prominent occiput, not known to be affected by exposure to anticonvulsants. These infants were examined by one masked physician examiner, using a study protocol, in three time periods: in two similar time periods, the masked examiner knew that one in four (25%) of the infants to be examined had been exposed to an anticonvulsant drugs; in the third time period, the chance was 50%.

RESULTS

The frequencies of the subjective features of midface (odds ratio [OR] = 2.6) and digit hypoplasia (OR = 3.9), but not the other features, were significantly more common in the time periods when the masked examiner knew that there was a 50% chance of exposure to anticonvulsant drugs, compared to the time periods when there was a 25% chance.

CONCLUSIONS

The findings suggest the effect of observer bias. In addition, there was evidence of context bias, meaning that a study that focuses on determining the presence of specific features will identify them more often. Birth Defects Research (Part A) 67:452–456, 2003. © 2003 Wiley‐Liss, Inc.
Omphalocele‐exstrophy‐imperforate anus‐spinal defects (OEIS) in triplet pregnancy after IVF and CVS
Authors: Shanske Alan L. , Pande Sumati , Aref Karim , Vega‐Rich Carlos , Brion Luc , Reznik Sandra , Timor‐Tritsch Ilan E.
Published: June 2003

BACKGROUND

Omphalocele‐exstrophy‐imperforate anus‐spinal defects (OEIS) complex is a rare sporadic condition.

CASE

We identified an infant with major malformations resembling OEIS. He was the product of a 30‐week triplet pregnancy conceived by in vitro fertilization (IVF) and evaluated by chorionic villi sampling (CVS). In this article, we review the possible pathogenetic mechanisms in this case, including IVF, multiple gestation, trauma to the uterus or uterine vessels following CVS, and placenta accreta.

CONCLUSIONS

We conclude that the cumulative effects of all or some of these factors may have resulted in uteroplacental insufficiency adequate to produce this phenotype. This case provides additional evidence for the uterine vascular pathogenesis of OEIS complex in humans. Birth Defects Research (Part A) 67467–471, 2003. © 2003 Wiley‐Liss, Inc.
Cyclophosphamide, methotrexate, and cytarabine embropathy: Is apoptosis the common pathway?
Authors: Vaux Keith K. , Kahole Nathanial C.O. , Jones Kenneth Lyons
Published: June 2003

BACKGROUND

Cyclophosphamide (CP) is an alkylating agent primarily used for the treatment of autoimmune disease and cancer. The purpose of this article is two‐fold: first, to indicate that CP is a recognized human teratogen based on the features seen in a child prenatally exposed to this agent, as well as features seen in the previously reported cases; second, to suggest a common pathway to explain the similarity in the pattern of malformation seen in infants prenatally exposed to CP, in infants prenatally exposed to methotrexate (MTX), and in infants prenatally exposed to cytosine arabinoside (CA).

METHODS

Case report and review of the literature of an infant prenatally exposed to CP during the first trimester with a specific pattern of malformation. Features are compared to seven previous reports.

RESULTS

A common pattern of malformation is delineated including growth deficiency, hypoplasia of the calvarial and facial bones, and oligodactyly.

CONCLUSIONS

The finding of a similar pattern of malformation among eight infants prenatally exposed to CP suggests that CP is a human teratogen. MTX and CA produce similar patterns of malformation in prenatally exposed infants despite very different pharmocologic profiles and metabolism. We speculate that the phenotype is a consequence of apoptosis in certain cells which are susceptible to the effects of the teratogen at specific stages of development. Birth Defects Research (Part A) 67403–408, 2003. Published 2003 Wiley‐Liss, Inc.
Substrate aggregation due to aerial hyphae during discontinuously mixed solid‐state fermentation with Aspergillus oryzae: Experiments and modeling
Authors: Schutyser M. A. I. , de Pagter P. , Weber F. J. , Briels W. J. , Boom R. M. , Rinzema A.
Published: June 2003

Solid‐state fermentation (SSF) is prone to process failure due to channeling caused by evaporative cooling and the formation of an interparticle mycelium network. Mixing is needed to break the mycelium network and to avoid such failure. This study presents the first attempt to quantify and predict the effect of mycelium bonds on particle mixing and vice versa. We developed a novel experimental set‐up to measure the tensile strength of hyphal bonds in SSF: Aspergillus oryzae was cultivated between two wheat‐dough disks and the tensile strength of the aerial mycelium was measured with a texture analyzer. Tensile strength at different incubation times was related to oxygen consumption, to allow a translation to a rotating drum with A. oryzae cultivated on wheat grain. We performed several discontinuously mixed solid‐state fermentations in the drum fermentor and measured the number and size of grain‐aggregates remaining after the first mixing action. We integrated data on mycelium tensile strength into a previously developed two‐dimensional discrete‐particle model that calculates forces acting on individual substrate particles and the resulting radial‐particle movements. The discrete‐particle model predicted the quantity and size of the aggregates remaining after mixing successfully. The results show that the first mixing event in SSF with A. oryzae is needed to break mycelium to avoid aggregate formation in the grain bed, and not to distribute water added to compensate for evaporation losses, or smooth out temperature gradients. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 503–513, 2003.
CFD‐aided design of a dynamic filter for mammalian cell separation
Authors: Castilho Leda R. , Anspach F. Birger
Published: June 2003

In the present work, a rotating disk filter was designed for mammalian cell separation with the aim of avoiding both cell damage and membrane fouling. Different geometric and operational variables of the rotating disk filter were studied using computational fluid dynamics (CFD) by varying rotor radius, rotor angle, membrane–rotor distance, and angular velocity. The combinations of these variables followed a statistical design, so that an analysis of the CFD results provided correlations describing the average shear stress on the membrane surface and the maximum shear stress in the whole module as a function of the variables studied. Based on these correlations, and on the shear resistance levels of Chinese hamster ovary (CHO) and baby hamster kidney (BHK) cell lines, which were investigated using a cone‐and‐plate viscosimeter, it was possible to determine the geometry and angular velocity that would minimize both cell damage and membrane fouling. After construction, the filter was tested in filtration experiments at increasing permeate fluxes. Cell viability remained >90% for the duration of the experiments (2.5 h), and no indication of fouling was observed. It was shown that the designed dynamic filter is able to effectively avoid both cell damage and membrane fouling, and thus can be used for mammalian cell harvesting and perfusion. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 514–524, 2003.
Effect of membrane morphology on system capacity during normal flow microfiltration
Authors: Zydney Andrew L. , Ho Chia‐Chi
Published: June 2003

Understanding the effects of membrane fouling on system capacity is critical for the successful design and scale‐up of microfiltration systems. The underlying morphology and structure of the microfiltration membrane can have a significant effect on system capacity by altering the rate and extent of fouling. Experimental data were obtained for system capacity during protein microfiltration using several model membranes with both homogeneous and composite structures. Data were compared with predictions of a new model that can account for both pore blockage and cake formation, and also includes the effects of membrane morphology on internal flow profiles within the membrane. Membranes with highly interconnected pores have a significantly higher capacity due to the reduction in flux decline arising from the fluid flow under and around any surface blockage. The model calculations are in good agreement with the flux decline data, allowing far more accurate predictions of system capacity than for the commonly used Vmax analysis. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 537–543, 2003.
Combined use of proteomic analysis and enzyme activity assays for metabolic pathway analysis of glycerol fermentation by Klebsiella pneumoniae
Authors: Wang Wei , Sun Jibin , Hartlep Michael , Deckwer Wolf‐Dieter , Zeng An‐Ping
Published: June 2003

The fed‐batch fermentation of glycerol to 1,3‐propanediol by Klebsiella pneumoniae displayed an unusual dynamic behavior that can be clearly divided into four distinct phases according to cell growth and CO2 evolution rate. Metabolism changed significantly during the different phases as reflected by the varied specific rates of substrate consumption and product formation. An assay of activities of the three initial enzymes of glycerol metabolism, namely glycerol dehydratase (GDHt), glycerol dehydrogenase (GDH), and 1,3‐propanediol‐oxidoreductase (PDOR), showed apparently different patterns of expression. To understand the culture dynamics and patterns of enzyme formation at a more systemic level we analyzed the expression patterns of intracellular proteins of K. pneumoniae from different phases of the fed‐batch fermentation using two‐dimensional gel electrophoresis (2DE). Two new enzymes, namely a phosphoenolpyruvate‐dependent dihydroxyacetone kinase (DHAK II) and a hypothetical oxidoreductase (HOR), which are directly related to glycerol metabolism and 1,3‐propanediol formation, were identified among the highly expressed proteins. The changes in expression of these new enzymes and several other proteins identified from the 2DE analysis helped to understand not only the dynamic behavior of the fed‐batch fermentation reported in this work but also some previously insufficiently understood phenomena related to this fermentation process. In particular, we demonstrated the combined use of proteomic analysis and enzyme activity assay data for metabolic pathway analysis and for a better identification of targets for bioprocess improvement. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 525–536, 2003.
Isolation of monocytes from human peripheral blood using immuno‐affinity expanded‐bed adsorption
Authors: Ujam L. B. , Clemmitt R. H. , Clarke S. A. , Brooks R. A. , Rushton N. , Chase H. A.
Published: June 2003

A novel technique for the separation of monocytes from human peripheral blood preparations has been developed. The technique is based on the use of expanded‐bed adsorption and a solid perfluorocarbon derivatized with avidin or streptavidin for the indirect positive or negative capture of cells labeled with biotinylated monoclonal antibodies. The perfluorocarbon support was prepared and characterized and the contactor design and operating conditions, that enable cells to be selectively isolated, were investigated. Experiments consisted of applying an immunolabeled pulse of 1 × 108 peripheral blood mononuclear cells (PBMCs), isolated by density gradient centrifugation, directly onto a refrigerated expanded bed. The major cell types remaining were T‐lymphocytes, B‐lymphocytes, and monocytes. Monocytes could be positively adsorbed, following labeling with anti‐CD14 mAb, with a clearance of up to 89% and a depletion factor of 7.6. They could also be “eluted” using mechanical shear, with a 77% yield of the applied cells at a purity of 90% and ≥ 65% viability. Negative isolation of monocytes, following labeling of the other cells present with anti‐CD2, CD7, CD16, CD19, and CD56 mAbs, resulted in lymphocyte depletions of up to 81% with a monocyte enrichment factor of 3.8 and purity of 71%. The monocyte viability in the flowthrough was assessed to be > 95%. This combination of expanded‐bed adsorption and fluidizable affinity supports shows significant potential for the intensification of cell separations. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 554–566, 2003.
Precipitation of RNA impurities with high salt in a plasmid DNA purification process: Use of experimental design to determine reaction conditions
Authors: Eon‐Duval Alex , Gumbs Karlene , Ellett Christopher
Published: June 2003

The use of high salt solution to precipitate RNA in a pharmaceutical‐grade plasmid DNA purification process was investigated. Five antichaotropic salts were tested for their potential to precipitate RNA. Calcium chloride was by far the best precipitant with high RNA removal in a very short incubation time. Calcium chloride precipitation conditions were investigated at two stages of a plasmid purification process using experimental design techniques. The effect of up to five factors on RNA precipitation and plasmid recovery was assessed by statistical modeling. Optimized conditions for calcium chloride precipitation were then introduced to the plasmid purification process resulting in the efficient removal of most impurities (RNA, chromosomal DNA, proteins, and endotoxins). © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 544–553, 2003.
A new kinetic scheme for lysozyme refolding and aggregation
Authors: Buswell A. Mark , Middelberg Anton P. J.
Published: June 2003

The competing first‐ and third‐order reaction scheme for lysozyme is shown to not predict fed‐batch lysozyme refolding when the model is parameterized using independent batch experiments, even when variations in chemical composition during the fed‐batch experiment are accounted for. A new kinetic scheme is proposed that involves rapid partitioning between the alternative fates of refolding and aggregation, and which allows for aggregation via a sequential mechanism. The model assumes that monomeric lysozyme in different states, including native, is able to aggregate with intermediates, accounting for recent experimental evidence that native protein can be incorporated into aggregates and explaining why native protein in the refolding buffer reduces yield. Stopped‐flow light‐scattering measurements were used to measure the association rate for the sequential aggregation mechanism, and refolding rate constants were determined in a series of batch experiments designed to be “snapshots” of the composition during a fed‐batch experiment. The new kinetic scheme gave a good a priori prediction of fed‐batch refolding performance. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 567–577, 2003.
Magnitude and kinetics of rehydration influence the viability of dehydrated E. coli K‐12
Authors: Mille Yannick , Beney Laurent , Gervais Patrick
Published: June 2003

The influence of rehydration conditions on the recovery of Escherichia coli K‐12 was studied. The results showed that the osmotic pressure gradient of rehydration shock realized before plating greatly affected cell viability. When rehydration occurred quickly from an hyperosmotic level of 133 MPa in glycerol solution before slow rehydration by plating on an agar surface to reach initial osmotic pressure (1.4 MPa), bacterial viability was strongly related to the intensity of the hypo‐osmotic gradient used. Rehydration to 107 MPa resulted in a survival ratio of 41%, whereas strong rehydration to 1.4 MPa resulted in only 0.7% survival. These studies also demonstrated the influence of the rehydration kinetic on cell recovery. An optimal rehydration rate of 0.136 MPa · s−1 increased cell recovery by a factor of 40 when compared with the faster and slower rates of 131.6 MPa · s−1 and 0.006 MPa · s−1, respectively. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 578–582, 2003.
Hydrodynamic characteristics of immobilized cell beads in a liquid–solid fluidized‐bed bioreactor
Authors: Wu Jane‐Yii , Chen Kuo‐Cheng , Chen Chun‐Ting , Hwang Sz‐Chwun John
Published: June 2003

This study examined the hydrodynamic characteristics of a liquid–solid fluidized‐bed bioreactor using elastic particles (PVA gel beads) of various diameters as carriers. The drag coefficient‐Reynolds number, velocity‐voidage, and expansion index‐Reynolds number relationships observed during fluidization of PVA gel beads in a fluidized bed in our experiments were compared with the published results. Predictions made from previous correlations were examined with our new experimental findings, revealing the inadequacy of most of these correlations. Thus, new correlations describing the above‐mentioned relationships are suggested. The drag coefficient of immobilized cell beads is larger than that of free cell ones at the same Reynolds number because the surface of the immobilized cell beads is rougher. For multiparticle systems, the correction factor, f(ϵ), is a function of the falling gel bead properties (Reynolds number) as well as the fluidized gel bead properties (Archimedes number), and depend strongly on the bed voidage (ϵ). A new simple relation was developed to predict easily the ϵ value from 0.5–0.9 at 4,986 < Ar < 40,745 or 34 < Ret < 186. For all the immobilized cell beads used in this study, the prediction error of the bed voidage was less than 5% at ϵ > 0.5. The prediction equations in this study can be further applied to analyzing the hydrodynamic characteristics of a fluidized‐bed reactor using similar entrapped elastic particles as carriers. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 583–594, 2003.
Bidirectional expression units enable streptogramin‐adjustable gene expression in mammalian cells
Authors: Fux Cornelia , Fussenegger Martin
Published: June 2003

Serious initiatives in gene therapy and tissue engineering require a sophisticated molecular toolbox combining DNA transfer technologies, human‐compatible transcription control systems, as well as compact and robust expression configurations. We have designed several versatile bidirectional expression cassettes that enable coadjustable expression of two desired transgenes in response to clinically licensed antibiotics of the streptogramin class (pristinamycin, Pyostacin®, Synercid®). The bidirectional expression modules consist of a central operator (PIR) that is specific for the pristinamycin‐dependent transactivator (PIT). Streptogramin‐adjustable binding of PIT to PIR transactivates two divergently oriented promoters and initiates transcription of the desired transgenes. The bidirectional expression module can be equipped with different minimal promoters and configured for expression of (1) two functional effector genes, (2) one effector gene and a reporter gene, (3) PIT and an effector gene to form a highly compact one‐vector expression arrangement. We have validated the streptogramin‐adjustable bidirectional expression technology in different basic and autoregulated expression configurations in a variety of mammalian and human cell lines. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 618–625, 2003.
Fed‐batch production of recombinant β‐galactosidase using the universal stress promoters uspA and uspB in high cell density cultivations
Authors: Prytz Ingela , Sandén Anna Maria , Nyström Thomas , Farewell Anne , Wahlström Åsa , Förberg Cecilia , Pragai Zoltan , Barer Mike , Harwood Colin , Larsson Gen
Published: June 2003

A high‐level production system using the universal stress promoters uspA and uspB in a fed‐batch cultivation based on minimal medium was designed. In development it was shown that a standard industrial fed‐batch protocol could not be used for this purpose since it failed to induce the levels of product as compared to the basal level. Instead, a batch protocol followed by a low constant feed of glucose was shown to give full induction. The levels of the product protein, β‐galactosidase, corresponded to approximately 25% of the total protein. Higher levels were found using the uspA than uspB vectors where uspA showed considerably higher basal level. The data indicate that the σ70 regulated promoter, uspA, although affected by the alarmone guanosine tetraphosphate, ppGpp, worked partly in a similar manner to constitutive promoters. An industrial high cell density fed‐batch cultivation on the basis of the suggested fed‐batch protocol and the uspA promoter gave a final β‐galatosidase concentration of 7 g/L and a final cell concentration of 65 g/L. The heterogeneity in production of the individual cell was measured by fluorescence microscopy. The data show that there is a process time independent heterogeneity in production, which is suggested to be caused by heterogeneity in the substrate uptake rate of the individual cell. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 595–603, 2003.
Effect of oxygen concentration on nitrification and denitrification in single activated sludge flocs
Authors: Satoh Hisashi , Nakamura Yoshiyuki , Ono Hideki , Okabe Satoshi
Published: June 2003

Simultaneous nitrification and denitrification (SND) was investigated in the single aeration tank of a municipal wastewater treatment plant. Microelectrode measurements and batch experiments were performed to test for the presence of SND. Microelectrodes recorded the presence of O2 concentration gradients in individual activated sludge flocs. When the O2 concentration in the bulk liquid was <45 μM, anoxic zones were detected within flocs with a larger diameter (approximately 3000 μm). The O2 penetration depth in the floc was found to be dependent on the O2 concentration in the bulk liquid. Nitrification was restricted to the oxic zones, whereas denitrification occurred mainly in the anoxic zones. The nitrification rate of the activated sludge increased with increasing O2 concentration in the bulk liquid, up to 40 μM, and remained constant thereafter. SND was observed in the aerated activated sludge when O2 concentration was in the range of 10 to 35 μM. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 604–607, 2003.
Lipase‐mediated acidolysis of butteroil with free conjugated linoleic acid in a packed bed reactor
Authors: Sehanputri Prima S. , Hill Charles G.
Published: June 2003

Acidolysis of butteroil with free conjugated linoleic acid (CLA) was studied in a packed bed reactor containing an immobilized Candida antarctica (fraction B) lipase. Kinetic data were used to develop quantitative reversible rate expressions of the general Michaelis–Menten form that also incorporate a term for first‐order deactivation of the enzyme. The extent of incorporation of CLA in the triacylglycerols of butteroil was characterized for reactions carried out at several temperatures (namely 45°, 50°, and 55°C) with different weight ratios of butteroil to CLA (namely 10:1 and 2:1). At the optimum operating temperature of 50°C, similar levels of incorporation of CLA (60% to 85%) were achieved at low space times (<3 h) for both 10:1 and 2:1 (w/w) ratios of butteroil to CLA. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 83: 608–617, 2003.
Telomere length variation and maintenance in hepatocarcinogenesis
Authors: Yokota Takashi , Suda Takeshi , Igarashi Masato , Kuroiwa Takashi , Waguri Nobuo , Kawai Hirokazu , Mita Yusaku , Aoyagi Yutaka
Published: June 2003

BACKGROUND

Despite the recent discovery of interchromosomal telomere length variation, a role for heterogeneity in telomere maintenance has yet to be established. This study investigated the significance of telomere length variation between chromosomes with respect to the association of cancer progression and telomere length regulation.

METHODS

Terminal restriction fragment (TRF) was evaluated in 20 surgically resected hepatocellular carcinoma specimens (HCC), corresponding noncancerous liver tissue specimens (NCL), and in 10 liver tissue specimens with chronic liver diseases devoid of cancer (DOC). Average TRF length (TRF‐A) was defined as the point of maximum intensity. Shorter and longer TRF lengths (TRF‐S and TRF‐L) were defined as the length above which 90% of TRF distribution was involved. A ratio, (TRF‐L−TRF‐S)/TRF‐A, was defined as telomere length dispersion.

RESULTS

The dispersion was significantly larger in HCC than in NCL specimens (P = 0.012) and in NCL than in DOC (P = 0.048). TRF‐A and TRF‐S were significantly shorter in HCC than in NCL (P = 0.0026, P = 0.0010). In seven patients in whom HCC recurred within 1 year, TRF‐A and TRF‐S were significantly shorter than in 10 patients in whom recurrence occurred after 1 year (P = 0.018, P = 0.0097). Telomeric repeat binding factor 1 was up‐regulated in HCC with elongated TRF‐L, whereas expression of human telomerase reverse transcriptase was greater in HCC with a shorter TRF‐S.

CONCLUSIONS

These results suggest that telomere length varied through chronic liver diseases by preferentially increasing shorter telomeres, whose length is a good indicator for malignant potential of HCC. Telomere length variation may be a crucial code in telomere maintenance through hepatocarcinogenesis. Cancer 2003;98:110–8. © 2003 American Cancer Society.

DOI 10.1002/cncr.11428
Endoscopic management of upper urinary tract transitional cell carcinoma
Authors: Daneshmand Siamak , Quek Marcus L. , Huffman Jeffry L.
Published: June 2003

BACKGROUND

The efficacy and long‐term results of endoscopic management of upper tract transitional cell carcinoma (TCC) were examined. The authors evaluated the accuracy of endoscopic biopsy in determining tumor grade in the subset of patients who underwent open surgical excision.

METHODS

Between 1987 and 2001, 50 patients (17 with a solitary kidney) underwent ureteroscopy and biopsy of upper tract TCC. Eleven patients underwent ureterectomy or nephroureterectomy shortly after endoscopic biopsy. There was no follow‐up for nine patients. Thirty patients underwent endoscopic ablation of their primary tumor with laser or electrofulguration at the time of the initial biopsy and were followed with close endoscopic surveillance at 3–4‐month intervals.

RESULTS

For the 30 patients who underwent endoscopic ablation, mean follow‐up was 38 months (range, 4–106 months). There was an average of 3.4 recurrences, with an average time to first recurrence of 7 months. Ten of the 30 patients underwent open resection during follow‐up. Six patients exhibited tumor progression at follow‐up. During the follow‐up period, one patient died of recurrent disease, and six died of other causes. Endoscopic biopsy accurately predicted the tumor grade for 8 of the 9 patients who had open tumor resection within 2 months of their last biopsy and for 10 of the 11 patients who had open resection shortly after their initial endoscopic biopsy (overall accuracy, 18 of 20 [90%]).

CONCLUSIONS

Endoscopic treatment of focal low‐grade TCC of the upper urinary tract is feasible and safe, provided that vigilant follow‐up and endoscopic surveillance are performed. Endoscopic biopsy provides accurate information regarding tumor grade. Cancer 2003;98:55–60. © 2003 American Cancer Society.

DOI 10.1002/cncr.11446
Central nervous system metastases in neuroblastoma
Authors: Matthay Katherine K. , Brisse Hervé , Couanet Dominique , Couturier Jerome , Bénard Jean , Mosseri Veronique , Edeline Véronique , Lumbroso Jean , Valteau‐Couanet Dominique , Michon Jean
Published: June 2003

BACKGROUND

Central nervous system (CNS) metastases rarely occur in patients with neuroblastoma, although recent reports suggest an increase in the rate. CNS recurrence may represent a different mechanism of spread than bone and bone marrow metastases and may be associated with unique genetic determinants. Further definition of the radiologic, clinical, and biologic features may provide clues to the predisposing factors and mechanisms of CNS dissemination.

METHODS

A retrospective analysis of all children ages 0–21 years with Stage IV neuroblastoma who were diagnosed at the Institut Curie and the Institut Gustave‐Roussy between 1985 and 2000 was performed with direct review of medical records and magnetic resonance images, computed tomography scans, and iodine‐123 or iodine‐131 metaiodobenzylguanidine scintiscans (MIBG scans). When tumor tissue was available, genetic analysis was performed using comparative genomic hybridization (CGH).

RESULTS

Of 434 patients with Stage 4 disease, 23 children had the CNS as their site of first recurrence. The estimated risk of CNS recurrence was 8.0% at 3 years, with no significant change in risk over the 15‐year period. Eleven patients had isolated CNS recurrences, and the remaining patients developed recurrences concomitantly in other sites. The sites of recurrences were parenchymal (n = 8 patients), parenchymal with meningeal (n = 7 patients), and meningeal alone (n = 8 patients). MIBG scans detected CNS lesions in only 43% of patients. Significant risk factors for CNS recurrence included lumbar puncture at diagnosis, ages 2–3 years, and MYCN gene amplification. Abnormalities that were identified using CGH, in addition to 2p24 amplification in 5/7, included gains of 17q and 18q and losses of 1p, 3p, 10q25‐26, and 11q.

CONCLUSIONS

The risk of CNS recurrence in patients with neuroblastoma is 8% at 3 years after diagnosis and has not increased in the last 15 years. Because the CNS often is the sole site of recurrence, either it may be a sanctuary site, or the biologic determinants of CNS metastasis may be unique. Elucidation of risk factors and pathogenesis may allow prevention of this fatal event. Cancer 2003;98:155–65. © 2003 American Cancer Society.

DOI 10.1002/cncr.11448
Detecting the expression of human endogenous retrovirus E envelope transcripts in human prostate adenocarcinoma
Authors: Wang‐Johanning Feng , Frost Andra R. , Jian Bixi , Azerou Ricardo , Lu Danielle W. , Chen Dung‐Tsa , Johanning Gary L.
Published: June 2003

BACKGROUND

The expression of human endogenous retrovirus (HERV) mRNA and proteins was associated recently with diseases that include human malignancies. The authors report that, in the current study, transcripts encoding the envelope region of an HERV family, HERV‐E, were expressed in human prostate carcinoma.

METHODS

RNA was isolated from various prostate tissues and was tested for the expression of various HERV envelope (env) genes by reverse transcriptase–polymerase chain reaction (RT‐PCR) analysis, RNA in situ hybridization (ISH), and Northern blot analysis. Variants of HERV that appeared in prostate carcinoma tissues were sequenced, and HERV‐E was expressed in prokaryotic and eukaryotic systems.

RESULTS

In the current study, the authors found that the mRNA of the env gene of one particular family of HERVs, HERV‐E, was expressed in some prostate carcinoma tissues (38.8% positive; n = 49 specimens) but not in normal prostate tissues using RT‐PCR, RNA ISH, and Northern blot assays. The expression of HERV‐E transcripts in prostate tumor epithelial cells was confirmed further by ISH using an HERV‐E specific antisense probe. Approximately 50% of the cDNA of HERV‐E obtained from prostate carcinoma specimens contained no stop codon and expressed proteins in prokaryotic or eukaryotic expression systems. Furthermore, the expression of both HERV‐E and ERV3 (another class of HERV) was detected in the same prostate carcinoma tissues.

CONCLUSIONS

The expression and distribution of multiple HERV‐E endogenous retroviral elements in prostate carcinoma, but not in normal control specimens, suggests that they may serve as novel tumor markers for the early diagnosis and immunotherapy of patients with prostate carcinoma. Cancer 2003;98:187–97. © 2003 American Cancer Society.

DOI 10.1002/cncr.11451
Gemcitabine and cisplatin as induction chemotherapy for patients with unresectable Stage IIIA‐bulky N2 and Stage IIIB nonsmall cell lung carcinoma
Authors: Cappuzzo Federico , Selvaggi Giovanni , Gregorc Vanesa , Mazzoni Francesca , Betti Maura , Rita Migliorino Maria , Novello Silvia , Maestri Antonio , De Marinis Filippo , Darwish Samir , De Angelis Verena , Nelli Fabrizio , Bartolini Stefania , Scagliotti Giorgio V. , Tonato Maurizio , Crinò Lucio
Published: June 2003

BACKGROUND

The objective of this trial was to evaluate the activity and safety of one of the newer platinum‐based doublets as a neoadjuvant regimen in patients with unresectable Stage IIIA‐bulky N2 and Stage IIIB nonsmall cell lung carcinoma (NSCLC).

METHODS

From June 1996 to April 2000, 129 consecutive patients with locally advanced NSCLC were treated with gemcitabine, 1000 mg/m2 on Days 1 and 8 and cisplatin, 70 mg/m2 on Day 2 (GC) of a 21‐day treatment cycle, for 4 cycles, as part of a combined‐modality approach.

RESULTS

After induction chemotherapy, 80 patients (62%; 95% confidence interval, 53.6–70.4%) achieved a partial response, 43 patients (33%) had stable disease, and 6 patients (5%) had disease progression during chemotherapy. Forty patients (31%), were considered resectable and underwent thoracotomy. Complete resectability was obtained in 38 patients (29%), with 2% of patients achieving a pathologic complete response. After surgery, 9 patients with Mountain Classification Stage IIIA NSCLC and 9 patients with Stage IIIB NSCLC received definitive adjuvant radiotherapy. Forty‐six of 52 patients with Stage IIIB disease and 24 of 37 patients with Stage IIIA disease who were not considered suitable for surgery received definitive radiotherapy. The median time to disease progression was 11.4 months, the median survival was 19.4 months (range, 1.2–55.2 + months), and the 1‐year survival rate was 74%. The lungs (33%) and the brain (21%) were the main sites of recurrence. Major toxicity was comprised of Grade 3–4 thrombocytopenia, which occurred in 34 patients (27%).

CONCLUSIONS

GC administered according to a 3‐week schedule was a highly active and safe regimen in patients with primary, unresectable, locally advanced NSCLC. Cancer 2003;98:128–34. © 2003 American Cancer Society.

DOI 10.1002/cncr.11460
Expression ofbcl‐2 protein is associated with shorter survival in nonsmall cell lung carcinoma
Authors: Huang Chris I. , Neuberg Donna , Johnson Bruce E. , Wei Jeanne Y. , Christiani David C.
Published: June 2003

BACKGROUND

The aim of this study was to assess the relationship between the expression of the 27‐kilodalton (kD)bcl‐2 protein and survival among nonsmall cell lung carcinoma (NSCLC) patients.

METHODS

Paired tissue samples of histologically confirmed tumor and uninvolved lung of 91 randomly selected NSCLC patients were used in this study. The expression of 27‐kDbcl‐2 proteins in uninvolved lung and tumor tissue specimens was analyzed using Western blot analysis. Overall survival was estimated using the Kaplan–Meier method. The relation between patient survival and expression of the 27‐kDbcl‐2 protein in uninvolved lung, tumor, and in uninvolved lung and/or tumor tissue specimens was assessed using log‐rank tests and Cox proportional hazards multiple regression.

RESULTS

The 27‐kDbcl‐2 protein was expressed in 54% of the uninvolved lung tissue specimens, in 53% of the tumor tissue specimens, and in 70% of the uninvolved lung and/or tumor tissue specimens. When NSCLC patient survival was considered with respect to the expression of the 27‐kDbcl‐2 protein, a statistically significant shorter survival was observed for patients whose tissue samples expressed the 27‐kDbcl‐2 protein in the uninvolved lung and in the uninvolved lung and/or tumor (log‐rank test,P= 0.008 andP= 0.001, respectively). Cox proportional hazards multiple regression models also showed statistically significant associations between shorter survival and positive 27‐kD protein expression in the uninvolved lung (hazard ratio of 2.27;P= 0.05) and in the uninvolved lung and/or tumor (hazard ratio of 5.58,P= 0.008) after controlling for tumor size, stage, the type of surgery received, smoking status, age, and cell type.

CONCLUSIONS

In the current study, patients with nonsmall cell lung carcinoma with positive expression of the 27‐kD proteinbcl‐2, either in the uninvolved lung or in uninvolved lung and/or tumor, were found to have significantly poorer postresection survival. Cancer 2003;98:135–43. © 2003 American Cancer Society.

DOI 10.1002/cncr.11461
Phase I study of intranodal delivery of a plasmid DNA vaccine for patients with Stage IV melanoma
Authors: Tagawa Scott T. , Lee Peter , Snively Jolie , Boswell William , Ounpraseuth Song , Lee Sandra , Hickingbottom Barbara , Smith John , Johnson Denise , Weber Jeffrey S.
Published: June 2003

BACKGROUND

Based on the likelihood of transfecting large numbers of local antigen‐presenting cells, a Phase I study in patients with Stage IV melanoma was conducted to determine the practicality, toxicity of, and immune responses to repeated infusions into a groin lymph node of escalating doses of a DNA plasmid encoding tyrosinase epitopes.

METHODS

Cohorts of 8 patients each received 200 μg, 400 μg, or 800 μg of DNA intranodally by pump over 96 hours every 14 days for 4 cycles. Blood was collected for immunologic assays and to measure plasmid in serum prior to treatment, 4 weeks later, and 8 weeks later. Scans and X‐rays were performed at baseline and after 8 weeks.

RESULTS

Treatment was tolerated well, with only five patients demonstrating Grade 1–2 toxicity. Vaccine delivery by 96‐hour infusions of plasmid into a groin lymph node resulted in only 1 episode of catheter leakage in 107 cannulations. Detection of plasmid in serum was rare and transient in two patients. Immune responses by peptide‐tetramer assay to tyrosinase 207–216 were detected in 11 of 26 patients. No clinical responses were seen. Survival of the heavily pretreated patients on this trial was unexpectedly long, with 16 of 26 patients alive at a median follow‐up of 12 months.

CONCLUSIONS

Infusion of a DNA plasmid vaccine into a groin lymph node was practical and well tolerated. Immune responses to a novel tyrosinase epitope were noted. Overall survival in this trial of heavily pretreated patients was unexpectedly long, with 16 of 26 patients alive after a follow‐up of 12 months, favoring immune responders. Cancer 2003;98:144–54. © 2003 American Cancer Society.
The relationship between psychologic distress and cancer‐related fatigue
Authors: Tchekmedyian N. Simon , Kallich Joel , McDermott Anne , Fayers Peter , Erder M. Haim
Published: June 2003

BACKGROUND

The authors examined the relationship between changes in depression and anxiety levels with changes in fatigue levels among anemic patients with lung cancer who participated in a randomized, double‐blind, placebo‐controlled clinical trial of darbepoetin alfa for the treatment of anemia.

METHODS

Patients completed the Brief Symptom Inventory (BSI) Depression and Anxiety subscales and the Functional Assessment of Cancer Therapy (FACT) Fatigue subscale during the trial. Pearson correlation coefficients were used to compare changes in the BSI scores with changes in the FACT Fatigue scores. Multiple regression models were used to explore sociodemographic and clinical explanatory variables.

RESULTS

At baseline, 25% and 35% of 250 patients reported high levels (normed BSI scores ≥ 65) of anxiety and depression, respectively. Correlations of changes in normed BSI Anxiety and Depression subscale scores with changes in FACT Fatigue scores had coefficients of −0.45 (P< 0.001) and −0.44 (P< 0.001), respectively. In the multiple regression models, change in the FACT Fatigue score was the only significant explanatory variable (P< 0.001). For every unit improvement in FACT Fatigue score, there was a corresponding improvement of 0.7 points and 0.8 points in anxiety and depression levels, respectively.

CONCLUSIONS

Improvements in fatigue were associated significantly with reductions in anxiety and depression. For patients with anemia, fatigue can be improved or reversed with darbepoetin alfa therapy. Thus, less fatigued patients also may benefit from reduced levels of anxiety and depression. Cancer 2003;98:198–203. © 2003 American Cancer Society.

DOI 10.1002/cncr.11463
Favorable cardiac risk among elderly breast carcinoma survivors
Authors: Lamont Elizabeth B. , Christakis Nicholas A. , Lauderdale Diane S.
Published: June 2003

BACKGROUND

There are two reasons why women who survive breast carcinoma may be at a lower risk of developing coronary heart disease (CHD) compared with women without a history of breast carcinoma. First, estrogens may be etiologic in the development of breast carcinoma and protective of CHD. Second, a common therapy for breast carcinoma (tamoxifen) may be associated with cardiac protection.

METHODS

In this population‐level cohort study, the authors analyzed data from the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER)‐Medicare program to study the cardiac risk of elderly female Medicare beneficiaries with and without a history of breast carcinoma. Using the SEER file, the authors identified elderly women survivors of Stage 0, I, or II breast carcinoma (n = 5980) diagnosed between the ages of 55 and 64. Using the Medicare 5% noncancer file, the authors also identified elderly women without a history of cancer (n = 23,165). They followed women from age 67 for up to 5 years for hospitalization for acute myocardial infarction (AMI) through a review of Medicare claims. The authors controlled the analyses for race, socioeconomic status, geographic location, cohort entry year, and medical comorbidity.

RESULTS

The hazard of hospitalization for AMI for breast carcinoma survivors relative to comparison patients was 0.66 (95% confidence interval, 0.49–0.88). This apparent cardioprotective effect of breast carcinoma survivorship was stronger in breast carcinoma survivors with documented cardiac risk factors.

CONCLUSIONS

Survivors of early‐stage postmenopausal breast carcinoma are at a significantly lower risk of hospitalization for AMI than women who do not have a history of breast carcinoma. That survivors' risk varies with previous cardiac risk factors may be consistent with effects of selective estrogen receptor modulators. This phenomenon should be evaluated further with individual‐level data containing information on patient cardiac risk factors and tamoxifen use to help clarify the mechanism behind the risk reduction. Cancer 2003;98:2–10. © 2003 American Cancer Society.

DOI 10.1002/cncr.11467
Detection, treatment, and outcome of isolated supraclavicular recurrence in 42 patients with invasive breast carcinoma
Authors: van der Sangen Maurice J.C. , Coebergh Jan‐Willem W. , Roumen Rudi M.H. , Rutten Harm J.T. , Vreugdenhil Gerard , Voogd Adri C.
Published: June 2003

BACKGROUND

There still is debate as to whether breast carcinoma patients with isolated supraclavicular recurrence should be considered to be patients with disseminated disease or patients for whom aggressive treatment with curative intent is justified.

METHODS

In the period 1984–1994, 4669 patients with invasive breast carcinoma underwent axillary dissection in 1 of 8 community hospitals in the southeastern part of the Netherlands. During follow‐up, 42 patients with isolated supraclavicular recurrence, without other sites of distant disease, were identified.

RESULTS

The median interval between treatment of the primary tumor and diagnosis of the supraclavicular recurrence was 2.5 years (range, 0.2–11.5 years). Radiotherapy was administered to 25 patients (60%), 5 of whom also underwent surgery and 16 of whom also received chemotherapy or hormonal therapy. Eleven patients received hormonal therapy only, and four received chemotherapy only. One patient received surgical treatment only, and one patient remained untreated. Complete remission was achieved in 35 patients (83%), but a second supraclavicular recurrence occurred in 12 (34% of patients who achieved complete remission). Overall, 6 patients (14%) were alive without evidence of disease after a follow‐up period of 4.4–8.3 years. The 5‐year actuarial overall survival and distant disease–free survival rates, based on the date of diagnosis of supraclavicular recurrence, were 38% (95% confidence interval [CI], 23–53%) and 22% (95% CI, 8–35%), respectively. The distant disease–free survival rate was somewhat better for the 25 patients who underwent radiotherapy as part of the treatment for supraclavicular recurrence than it was for the 17 patients who did not receive radiotherapy (P = 0.06); the difference became more pronounced after the exclusion of 8 patients who had received axillary and supraclavicular radiotherapy as part of treatment for the primary tumor (P = 0.002).

CONCLUSIONS

Although complete remission can be obtained in most patients with isolated supraclavicular recurrence, the prognosis for these patients is poor. Involved field radiotherapy appears to play an important role in the treatment of supraclavicular recurrence and may improve the distant recurrence–free survival rate. Cancer 2003;98:11–7. © 2003 American Cancer Society.

DOI 10.1002/cncr.11469
Candidemia in women with breast carcinoma treated with high‐dose chemotherapy and autologous bone marrow transplantation
Authors: Gottfredsson Magnús , Vredenburgh James J. , Xu Jianping , Schell Wiley A. , Perfect John R.
Published: June 2003

BACKGROUND

Invasive fungal infections, including candidemia, pose a major threat to patients with impaired immune defenses, including bone marrow transplantation (BMT) recipients. During 1992–1997, 845 women with multiple lymph node positive or metastatic breast carcinoma underwent high‐dose chemotherapy (HDC) and subsequent autologous BMT at Duke University Medical Center. No systemic antifungal prophylaxis was administered. The purpose of the current study was to evaluate the risk and long‐term outcome of candidemia in this patient population.

METHODS

Clinical data were collected on patients with candidemia, and a group of age‐matched control patients were identified who underwent HDC and BMT for breast carcinoma in the same time period. The difference in crude mortality between these two groups was used to calculate the attributable mortality of candidemia. The genetic relatedness of the fungal blood stream isolates was investigated by DNA fingerprinting. Antifungal susceptibility testing was performed using serial microdilution.

RESULTS

Twenty‐nine of 845 women developed candidemia (3.4%). The crude mortality of women with candidemia was 35% at 90 days after transplantation but 11% among women in the matched control group who were without infection (P = 0.01), for an attributable mortality rate of 24%. The most common pathogen was Candida tropicalis (50%), followed by Candida albicans (23%). The mortality was highest for women who were infected with C. albicans, followed by C. tropicalis, and other Candida species (P = 0.037). DNA fingerprinting of the yeasts revealed genetic heterogeneity in all species. However, 9 of 15 C. tropicalis isolates had identical DNA fingerprint profiles, suggesting spread of this genotype from a common source. All yeast isolates were susceptible to amphotericin B, and 20 of 30 isolates were susceptible to ≤ 8 μg/mL of fluconazole.

CONCLUSIONS

Candidemia was relatively infrequent after HDC and autologous BMT in women with for multiple lymph node positive or metastatic breast carcinoma. This was true even in the absence of systemic antifungal prophylaxis. The mortality attributable to candidemia in this patient population was 24% and was higher among patients who were infected by C. albicans compared with patients who were infected by other Candida, non‐albicans species. Cancer 2003;98:24–30. © 2003 American Cancer Society.

DOI 10.1002/cncr.11470