bluuebunny/crossref_metadata_2025 huggingface dataset:
Modifications to Therapy for Multiple Myeloma: Pegylated Liposomal Doxorubicin in Combination With Vincristine, Reduced-Dose Dexamethasone, and Thalidomide
Authors: Hussein Mohamad A.
Published: August 2004

Learning Objectives

After completing this course, the reader will be able to:

Discuss the current issues surrounding the treatment of multiple myeloma. Describe how treatment modifications have improved the tolerability and efficacy of this multiple myeloma therapy. Explain recent treatment advances in this disease.

Access and take the CME test online and receive one hour of AMA PRA category 1 credit at CME.TheOncologist.com

The combination of vincristine, doxorubicin, and dexamethasone is an effective treatment for multiple myeloma that produces a more rapid response than other regimens, probably a function of the high-dose, intense steroid schedule. However, vincristine/doxorubicin/dexamethasone administration requires a 96-hour continuous infusion delivered via a central venous catheter, which necessitates hospitalization in a large number of patients and may increase the risk for infection. Moreover, the high dosages of corticosteroids required with this regimen can cause substantial toxicity. Therefore, a number of modifications to the regimen have been evaluated in an effort to improve its tolerability and efficacy. These include replacing doxorubicin with pegylated liposomal doxorubicin and using a reduced frequency of dexamethasone, and, later, the addition of thalidomide. The results of an ongoing study demonstrated that this latest regimen (including thalidomide) is associated with an improved response rate and a higher quality of response compared with previous regimens in patients with relapsed/refractory multiple myeloma. This modified regimen is well tolerated when prophylactic and supportive measures are incorporated. Although additional follow-up is required to determine the effect on survival, this modified regimen has significant potential in the management of advanced myeloma.
Signal Events: Cell Signal Transduction and Its Inhibition in Cancer
Authors: Rowinsky Eric K.
Published: August 2004

Learning Objectives

After completing this course, the reader will be able to:

Recognize the importance of identifying new molecular targets for cancer therapy and how they relate to the development of novel therapeutic agents with high therapeutic indices and low toxicity. Explain the process of signal transduction (specifically the ErbB family of receptors and the mTOR pathways) and how it relates to cell growth, differentiation, and survival, and describe the effects of aberrations in signal transduction on the development of neoplasms. Describe some of the novel targeted therapies that have been developed and explain the mechanisms by which signal transduction inhibitors inhibit tumor growth and induce tumor regression in patients with cancer.

Access and take the CME test online and receive one hour of AMA PRA category 1 credit at CME.TheOncologist.com

Signal transduction refers to communication processes used by regulatory molecules to mediate the essential cell processes of growth, differentiation, and survival. Signal transduction elements interact through complex biochemically related networks. Aberrations in signal transduction elements can lead to increased proliferative potential, sustained angiogenesis, tissue invasion and metastasis, and apoptosis inhibition. Most human neoplasms have aberrant signal transduction elements. Several compounds that target aberrant signal transduction elements, such as those in the ErbB family of tyrosine kinase receptors and mammalian target of rapamycin, are in development. To date, commercially available signal-transduction-targeting compounds include trastuzumab, a monoclonal antibody against the ErbB-2 receptor for the treatment of metastatic breast cancer overexpressing the ErbB-2 (HER-2) receptor, and gefitinib, an inhibitor of the ErbB-1 receptor tyrosine kinase that recently received regulatory approval for the treatment of patients with non-small cell lung cancer. In contrast to traditional cytotoxic treatments, although signal transduction inhibitors are capable of inducing tumor regression, particularly in malignancies that are principally driven by specific target aberrations, preclinical and early clinical investigations suggest that their predominant beneficial effects are growth inhibitory in nature; therefore, new clinical trial designs and evaluation end points may be required to ultimately assess their value. Prospective profiling of patients and tumors to determine treatment response is also essential to the success of these clinical trials. However, responsiveness to these novel therapies is dependent on a multitude of factors that ultimately determine the robustness and quality of the downstream response.
Use of Bronchoalveolar Lavage to Detect Lung Injury
Authors: Henderson Rogene F. , Muggenburg Bruce A.
Published: August 2004

This unit describes how bronchoalveolar lavage can be used in laboratory animals to sample the epithelial lining fluid of the lung for information on the degree of pulmonary inflammation induced by exposure to an airborne toxicant. The technique allows quantitative assessment of inflammatory responses and is valuable for providing dose‐response information in exposed animals. Lavage fluid samples may also be used for proteomic analyses, and the protein expression profiles may be used to address specific mechanistic questions.
Spectrophotometric Determination of Protein Concentration
Authors: Simonian Michael H.
Published: August 2004

This unit describes spectrophotometric and colorimetric methods for measuring the concentration of a sample protein in solution. Absorbance measured at 280 nm (A280) is used to calculate protein concentration by comparison with a standard curve or published absorptivity values for that protein (a280). Alternatively, absorbance measured at 205 nm (A205) is used to calculate the protein concentration. The A280 and A205 methods can be used to quantify total protein in crude lysates and purified or partially purified protein. A spectrofluorometer or a filter fluorometer can be used to measure the intrinsic fluorescence emission of a sample solution; this value is compared with the emissions from standard solutions to determine the sample concentration. The fluorescence emission method is used to quantify purified protein. This simple method is useful for dilute protein samples and can be completed in a short amount of time. There are two colorimetric methods: the Bradford colorimetric method, based upon binding of the dye Coomassie brilliant blue to the protein of interest, and the Lowry method, which measures colorimetric reaction of tyrosyl residues in the protein sample.
Social Transmission of Food Preference in Mice
Authors: Wrenn Craige C.
Published: August 2004

The social transmission of food preference (STFP) is an ethologically relevant test of olfactory memory that can be used in mice. In this test, “observer” mice interact with a “demonstrator” mouse that has recently eaten a novel food. When observer mice are presented with a choice between the food eaten by the demonstrator and some other novel food, observer mice prefer the food eaten by the demonstrator. This phenomenon depends on the observer mice detecting olfactory cues on the breath of the demonstrator mouse during their interaction. The subsequent food preference serves as a measure of memory for those olfactory cues. This unit describes a method for performing STFP in mutant mice. The method explains how to address potential confounding factors and avoid possible pitfalls.
Synthesis of 5′‐O‐Phosphoramidites with a Photolabile 3′‐O‐Protecting Group
Authors: Beier Markus , Hoheisel Jörg D.
Published: August 2004

This unit describes the chemical synthesis of phosphoramidite building blocks that carry a protecting group at the 3′ position. These inversely oriented synthons expose a 2‐(2‐nitrophenyl)propoxycarbonyl (NPPOC) group as the photolabile protecting group of choice. Among other applications, the building blocks can be employed for light‐controlled in situ synthesis of DNA microarrays, producing arrayed oligonucleotides that are attached to the support via their 5′ ends, leaving their 3′ termini available to act as substrates for polymerases.
Overview of mRNA Expression Profiling Using Microarrays
Authors: Katagiri Fumiaki , Glazebrook Jane
Published: August 2004

Microarray technology allows simultaneous measurement of the mRNA levels of thousands of genes. This powerful technology has applications in addressing many biological questions that were not approachable previously; however, the enormous size of microarray data sets leads to issues of experimental design and statistical analysis that are unfamiliar to many molecular biologists. The type of array used, design of the biological experiment, number of experimental replicates, and statistical method for data analysis should all be chosen based on the scientific goals of the investigator. This overview presents a discussion of the relative merits and limitations of various methods with respect to some common applications of microarray experiments.
Cytokine, Growth Factor, and Chemokine Ligand Database
Authors: Taub Dennis D.
Published: August 2004

Cytokines, chemokines, and growth factors are soluble proteins produced by leukocytes and other cell types, which act as chemical communicators between cells and tissues within the body. These proteins mediate a number of functions, ranging from effects on cell growth, differentiation, survival, and a number of effector activities. The main purpose of this unit is to provide an easy‐to‐use reference summarizing some of the current information on well‐established cytokine, growth factore, and chemokine ligands. The information provided is confined largely to alternative names, protein properties, genetic chracteristics, chromosomal localization, cellular expression patters, cellular‐tissue targets, stimuli regulating expression, functional properties, expression analysis, and clinical utility.
Clinical Validity and Clinical Utility of Genetic Tests
Authors: Burke Wylie
Published: August 2004

In considering the appropriate use of new genetic tests, clinicians and health care policymakers must consider the accuracy with which a test identifies a patient's clinical status (clinical validity) and the risks and benefits resulting from test use (clinical utility). Genetic tests in current use vary in accuracy and potential to improve health outcomes, and these test properties may be influenced by the clinical setting in which the test is used. In addition, the relative importance of clinical validity and utility varies according to the proposed use of the test. This unit defines clinical validity and clinical utility, provides examples, and considers the implications of these test properties for clinical practice.
Short Tandem Repeat Analysis for Human Identity Testing
Authors: Butler John M.
Published: August 2004

Short tandem repeat (STR) typing has become the workhorse of modern forensic DNA analysis. The most common form of STR typing uses laser‐induced fluorescence detection of dye‐labeled polymerase chain reaction (PCR) products following capillary electrophoresis (CE) size‐based separation. This unit describes the techniques and marker systems most widely used around the world in constructing criminal DNA databases and conducting forensic casework. In addition, these same STR loci and multiplex PCR assays are commonly used for paternity testing, missing persons, and mass disaster investigations. Both autosomal and Y‐chromosome STRs are discussed in the context of their various applications. Interpretation issues surrounding degraded or mixed DNA specimens as well as tri‐allelic patterns and variant alleles are also illustrated. Finally, future technologies and DNA marker systems are briefly explored relative to the state‐of‐the‐science today.
Labeling of Bacterial Pathogens for Flow Cytometric Detection and Enumeration
Authors: Harkins Kristi R. , Harrigan Kelley
Published: August 2004

Traditional uses of flow cytometry have been for research and diagnostic purposes, on mammalian cells. This unit focuses on using a flow cytometer for enumerating specific bacteria and parasites. Several different labeling methods are discussed, as well as how to set up a cytometer for detecting and enumerating bacteria. Labeling methods include direct detection with a primary fluorochrome‐conjugated antibody and indirect labeling with an unconjugated primary and a fluorochrome‐conjugated secondary antibody, as well as labeling with rRNA sequence‐specific peptide nucleic‐acid probes end‐labeled with a fluorochrome. Data and methods throughout this unit focus on detection of Salmonella in clean matrices; however, pertinent information is also provided on using these methods to label other pathogens. The Commentary covers critical aspects of the protocols, and also includes information and suggestions on the application of these methods for testing in the food and pharmaceutical industries, as well as in environmental water testing.
An Overview of Gene Identification: Approaches, Strategies, and Considerations
Authors: Baxevanis Andreas D.
Published: August 2004

Modern biology is on the verge of officially ushering in a new era in science with the completion of the sequencing of the human genome in April 2003. While often erroneously called the “post‐genome era”, this will actually truly mark the beginning of the “genome era,” a time in which the availability of sequence data for many genomes will have a significant effect on how science is performed in the 21st century. This unit offers an overview of many of the gene prediction methods that are currently available and offers a general assessment of how well the methods work for various problems.
An Overview of Spotfire for Gene‐Expression Studies
Authors: Kaushal Deepak , Naeve Clayton W.
Published: August 2004

Spotfire DecisionSite for Functional Genomics (referred to here as Spotfire) is a powerful data mining and visualization application with use in many disciplines. This unit provides an overview of Spotfire's utility in analyzing gene expression data obtained from DNA microarray experiments. Analysis of microarray data requires software‐based solutions able to handle and manipulate the enormous amount of data generated. Spotfire provides a solution for accessing, analyzing and visualizing data generated from microarray experiments. Spotfire is designed to allow biologists with little or no programming or statistical skills to transform, process, and analyze microarray data.
Loading and Preparing Data for Analysis in Spotfire
Authors: Kaushal Deepak , Naeve Clayton W.
Published: August 2004

This unit strictly focuses on data preparation within Spotfire. Microarray data exist in a variety of formats, which often depend on the particular array technology and detection instruments used. The first protocols in this unit describe loading Affymetrix and GenePix data into Spotfire. Once the data are loaded, it is necessary to filter and preprocess the data prior to analysis. Subsequently, the data transformation and normalization techniques presented here, are critical to correctly performing powerful microarray data mining expeditions. These steps extract or enhance meaningful data characteristics and prepare the data for the application of certain analysis methods such as statistical tests to compute significance and clustering methods—which mostly require data to be normally distributed. The unit outlines several methods for normalizing the data within an experiment and between multiple experiments.
Die Mechanismen der Stille‐Reaktion
Authors: Espinet Pablo , Echavarren Antonio M.
Published: August 2004

Vor achtzehn Jahren hat John K. Stille in der Angewandten Chemie über eine neuartige Kupplung von Organostannanen mit organischen Elektrophilen berichtet, die schließlich unter seinem Namen bekannt wurde. Seither hat sich diese Methode zu einem vielseitigen und facettenreichen Gebiet voller versteckter Möglichkeiten zum Forschen, Entdecken und Genießen entwickelt. Die neuesten Modifizierungen lassen Syntheseziele in greifbare Nähe rücken, von denen man vor einigen Jahren nur zu träumen wagte. Auch beim Verständnis der mechanistischen Details der Umsetzungen wurden große Fortschritte erzielt. Daher sollte einer gezielten Anwendung dieser wichtigen Reaktion und ihrer neuen Varianten, die sich auf mehr als nur empirische Befunde stützt, nichts mehr im Wege stehen. Dieser Aufsatz legt über diese Fortschritte kritisch Rechenschaft ab.
The Mechanisms of the Stille Reaction
Authors: Espinet Pablo , Echavarren Antonio M.
Published: August 2004

Eighteen years ago in Angewandte Chemie John K. Stille reviewed a novel methodology, which eventually became known by his name, for the coupling of organostannanes with organic electrophiles. Since then that seed has blossomed into a multifaceted methodology full of hidden possibilities to explore, discover, and enjoy. Very recent modifications are making synthetic wishes come true that were only dreamed of a few years ago. Moreover, as important advances are being made in the understanding of the mechanistic details of the process, it is becoming increasingly possible to apply this essential reaction and its new variants in a less empirical way. The purpose of this Review is to give a critical account of this progress.
Analysis of mechanical, thermal, and morphological behavior of polycaprolactone/wood flour blends
Authors: Wu Chin‐San
Published: August 2004

In the present study, the properties of polycaprolactone (PCL) and wood flour (WF) blends were examined by Fourier transform infrared (FTIR) spectroscopy, differential scanning calorimetry (DSC), Instron mechanical tester, and scanning electron microscopy (SEM). As for results, the mechanical properties of PCL were lowered obviously, due to the poor compatibility between the two phases, when it was blended with wood flours. A fine dispersion and homogeneity of wood flour in the polymer matrix could be obtained when the acrylic acid grafted PCL (PCL‐g‐AA) was used to replace PCL for manufacture of blends. This better dispersion is due to the formation of branched and crosslinked macromolecules since the PCL‐g‐AA copolymer had carboxyl groups to react with the hydroxyls. This is reflected in the mechanical and thermal properties of the blends. In comparison with pure PCL/WF blend, the increase in tensile strength at break was remarkable for PCL‐g‐AA/WF blend. The PCL‐g‐AA/WF blends are more easily processed than the PCL/WF ones since the former had lower melt viscosity than the latter. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1000–1006, 2004
Free‐radical bulk polymerization of styrene with a new trifunctional cyclic peroxide initiator
Authors: Sheng Wei‐Chen , Wu Jian‐Yi , Shan Guo‐Rong , Huang Zhi‐Ming , Weng Zhi‐Xue
Published: August 2004

The bulk free‐radical polymerization of styrene in the presence of a new cyclic trifunctional initiator, 3,6,9‐triethyl‐3,6,9‐trimethyl‐1,4,7‐triperoxonane, was studied. Full‐conversion‐range experiments were carried out to assess the effects of the temperature and initiator concentration on the polymerization kinetics, molecular weight, and polydispersity. Gel permeation chromatography was used to measure the molecular weight and the molecular weight distribution of polystyrene. When this multifunctional initiator was used for styrene polymerization at higher temperatures, it was possible to produce polymers with higher molecular weights and narrower molecular weight polydispersity at a higher rate. This showed that the molecular weight and polydispersity were influenced by the initiator concentration and the polymerization temperature in an unusual manner. Moreover, polystyrene, obtained with trifunctional peroxide, had OO bonds in the molecular chains and was investigated with differential scanning calorimetry and gel permeation chromatography. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1035–1042, 2004
Cure kinetics of epoxy resin and aromatic diamines
Authors: Ghaemy M. , Barghamadi M. , Behmadi H.
Published: August 2004

The kinetics of curing reaction of a diglycidyl ether of a bisphenol‐A based epoxy (DGEBA) with 4,4′‐diaminostillbene (DAS) and 4,4′‐diaminoazobenzene (DAAB) as curing agents are studied by differential scanning calorimetery (DSC) using the isothermal technique. The experimental data show that the cure reaction is autocatalytic in nature, and all kinetic parameters of the curing reaction are determined using a semiempirical equation. The reaction of DGEBA with DAS is faster than that with DAAB under the same conditions and the activation energies of both systems are higher than those reported for other aromatic diamines. With increasing isothermal temperature and concentration of curing agents the rate constants are increased by the increasing of probability collisions between epoxide and primary amine groups while the activation energies remain constant. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1049–1056, 2004
Angiographic adverse events during percutaneous coronary intervention fail to predict creatine kinase‐MB elevation
Authors: Blankenship James C. , Islam M. Ashequl , Wood G. Craig , Iliadis Elias A.
Published: August 2004

We attempted to determine if aggressive detection of angiographic adverse events during coronary intervention could predict subsequent creatine kinase (CK)‐MB elevations. During coronary intervention, both fluoroscopy and cine angiography were used to detect angiographic adverse events. At least one angiographic adverse event occurred in 133/251 (53%) of procedures. CK‐MB elevation occurred in 24% of procedures. Slow flow during the procedure (P = 0.002) and chest discomfort at the end of the procedure (P = 0.007) were the strongest predictors of CK‐MB elevation. Among procedures with no angiographic adverse events, CK‐MB elevation occurred in 15/121 (12%), accounting for 25% of CK‐MB elevations. We conclude that CK‐MB elevation occurs after angiographically uncomplicated coronary interventions even when angiographic adverse events are aggressively detected. Routine monitoring of cardiac enzymes is necessary to detect all patients who will experience myocardial injury after coronary intervention. Catheter Cardiovasc Interv 2004;63:31–41. © 2004 Wiley‐Liss, Inc.
Transthoracic access for cardiac catheterization
Authors: Maher Kevin O. , Murdison Kenneth A. , Norwood William I. , Murphy John D.
Published: August 2004

Patients who have undergone Fontan palliation for congenital heart disease have limited access to the pulmonary venous atrium. We describe a transthoracic approach to the pulmonary venous atrium for diagnostic and therapeutic cardiac catheterization. A total of 22 patients ranging in age from 14 months to 25 years and weighing 7–68 kg underwent transthoracic catheterization. With the aid of fluoroscopic and angiographic markers, a 20 gauge needle was inserted into the pulmonary venous atrium. A floppy‐tipped wire was inserted, and a 4 Fr sheath was placed, upsized as needed. Access to the pulmonary venous atrium was obtained for all patients. Therapeutic procedures were completed for all patients. Transthoracic access to the pulmonary venous atrium in patients who have undergone a Fontan operation is a reliable and rapid mode of access, allowing for complex therapeutic procedures to be performed in the catheterization laboratory, obviating the need for additional open heart surgery. Catheter Cardiovasc Interv 2004;63:72–77. © 2004 Wiley‐Liss, Inc.
Catheter‐based intracardiac echocardiography in the interventional cardiac laboratory
Authors: Liu Zheng , McCormick Daniel , Dairywala Ismail , Surabhi Satish , Goldberg Sheldon , Turi Zoltan , Vannan Mani A.
Published: August 2004

Recent advances in technology have engendered a renewed enthusiasm in the use of intracardiac echocardiography (ICE) to guide and assess cardiac interventions. AcuNav is a phased‐array sector imaging probe equipped with color and spectral Doppler capabilities. Previous‐generation imaging catheters yielded unfamiliar limited‐depth radial images with no flow information. Current imaging technology such as the AcuNav has not only consolidated the role of ICE but opened newer applications in the interventional laboratory. ICE has clear advantages over transesophageal echocardiography as the imaging modality of choice in the cardiac catheterization and electrophysiological laboratories. We review the technical evolution of ICE and describe the expanded utility of the AcuNav imaging catheter during cardiac interventions. Catheter Cardiovasc Interv 2004;63:63–71. © 2004 Wiley‐Liss, Inc.
Carotid stenting with a new system for distal embolic protection and stenting in high‐risk patients: The carotid revascularization with ev3 arterial technology evolution (CREATE) feasibility trial
Authors: Safian Robert D. , Bacharach J. Michael , Ansel Gary M. , Criado Frank J.
Published: August 2004

The purpose of this study was to evaluate the feasibility of carotid artery revascularization using a new system for carotid stenting and distal embolic protection in 30 patients with severe carotid stenosis and high risk for carotid endarterectomy (Carotid Revascularization With ev3 Arterial Technology Evolution, or CREATE). Previous studies suggest that patients with carotid stenosis and serious comorbid cardiopulmonary and anatomic conditions are at high risk for carotid endarterectomy. All patients underwent percutaneous revascularization using the Protégé GPS self‐expanding nitinol stent (ev3, Plymouth, MN) and the Spider distal embolic protection system (ev3). In‐hospital and 30‐day outcomes were analyzed. High‐risk features included age > 75 years (63%), left ventricular ejection fraction < 35% (20%), and restenosis after prior carotid endarterectomy (53%). Procedural success was 100%. In‐hospital complications included severe vasovagal reactions in six patients (20%) and a popliteal embolus in one patient (3.3%), treated by successful embolectomy. During 30 days of follow‐up, two patients (6.6%) experienced minor neurological deficits, including transient expressive aphasia that resolved without therapy in one patient and homonymous hemianopsia due to contralateral posterior circulation stroke in one patient. This study supports the feasibility of percutaneous carotid artery revascularization with the Protégé GPS self‐expanding stent and Spider distal embolic protection system, which will be evaluated in a large multicenter pivotal trial (CREATE Pivotal Trial). Catheter Cardiovasc Interv 2004;63:1–6. © 2004 Wiley‐Liss, Inc.
Vectorial Electron Relay at ITO Electrodes Modified with Self‐Assembled Monolayers of Ferrocene–Porphyrin–Fullerene Triads and Porphyrin–Fullerene Dyads for Molecular Photovoltaic Devices
Authors: Imahori Hiroshi , Kimura Makoto , Hosomizu Kohei , Sato Tomoo , Ahn Tae Kyu , Kim Seong Keun , Kim Dongho , Nishimura Yoshinobu , Yamazaki Iwao , Araki Yasuyuki , Ito Osamu , Fukuzumi Shunichi
Published: August 2004

Systematic series of indium tin oxide (ITO) electrodes modified covalently with self‐assembled monolayers (SAMs) of ferrocene–porphyrin–fullerene triads and porphyrin–fullerene dyads were designed to gain valuable insight into the development of molecular photovoltaic devices. The structures of SAMs on ITO have been investigated by UV/Vis absorption spectroscopy, atomic force microscopy, and cyclic voltammetry. The photoelectrochemical and photophysical (fluorescence lifetime and time‐resolved transient absorption) properties were also determined. The highest quantum yield of photocurrent generation (11 %) among donor–acceptor linked systems which are covalently attached to the surface of ITO electrodes was achieved with SAMs of ferrocene–zinc porphyrin–fullerene linked triad on ITO electrodes. The quantum yields of photocurrent generation correlate well with the charge‐separation efficiency and the lifetime of the charge‐separated state of the porphyrin–fullerene linked systems in solution. These results provide valuable information for the construction of photonic molecular devices and artificial photosynthetic systems on ITO electrodes.
Accurate and simple discrimination of mouse pulmonary dendritic cell and macrophage populations by flow cytometry: Methodology and new insights
Authors: Vermaelen Karim , Pauwels Romain
Published: August 2004

Background

The need to accurately discriminate dendritic cells (DCs) and macrophages (Mphs) in mouse lungs is critical given important biological differences. However, a validated flow cytometry–based method is still lacking, resulting in much confusion between both cell types.

Methods

Single‐cell suspensions freshly obtained from collagenase‐digested lung tissue were stained with a CD11c‐specific monoclonal antibody, detected using a PE‐Cy5 or APC‐conjugated secondary reagent. Cellular immunophenotype was simultaneously explored using a panel of PE‐conjugated markers. The FL1 or FITC‐detection channel was reserved for the assessment of autofluorescence.

Results

CD11c‐bright cells were heterogeneous and displayed a bimodal distribution with regard to autofluorescence (AF). CD11c+/low‐AF cells were lineage‐negative and showed features compatible with myeloid DCs. This was confirmed by morphology, potent T‐cell stimulatory function in a mixed‐leukocyte reaction, surface expression of MHCII and costimulatory molecules, and further immunophenotypical criteria, including the expression of Mac‐1 and absence of CD8α. In contrast, CD11c+/high‐AF cells displayed the features of pulmonary Mphs, including typical Mph morphology, very weak induction of T‐cell proliferation, low to absent expression of MHCII and costimulatory molecules, and very low levels of Mac‐1 as well as F4/80. We also show that only CD11c+/high‐AF cells strongly expressed the macrophage marker MOMA‐2, while interestingly Mac‐3 was expressed at high levels by CD11c+/high‐AF and low‐AF alike.

Conclusions

This study shows that the combination of CD11c‐expression and autofluorescence is necessary and sufficient to accurately separate DCs from macrophage subpopulations in mouse lungs. © 2004 Wiley‐Liss, Inc.
Flow cytometric kinetic assay of the activity of Na+/H+ antiporter in mammalian cells
Authors: Dolz María , O'Connor José‐Enrique , Lequerica Juan L.
Published: August 2004

Background

The Na+/H+ exchanger (NHE) of mammalian cells is an integral membrane protein that extrudes H+ ion in exchange for extracellular Na+ and plays a crucial role in the regulation of intracellular pH (pHi). Thus, when pHi is lowered, NHE extrudes protons at a rate depending of pHi that can be expressed as pH units/s.

Methods

To abolish the activity of other cellular pH‐restoring systems, cells were incubated in bicarbonate‐free Dulbecco's modified Eagle's medium buffered with HEPES. Flow cytometry was used to determine pHi with 2′,7′‐bis‐(2‐carboxyethyl)‐5‐(and‐6)‐carboxyfluorescein acetoxymethyl ester or 5‐(and‐6)‐carboxy SNARF‐1 acetoxymethyl ester acetate, and the appropriate fluorescence ratios were measured. The calibration of fluorescence ratios versus pHi was established by using ionophore nigericin. The activity of NHE was calculated by a kinetic flow cytometric assay as the slope at time 0 of the best‐fit curve of pHi recovery versus time after intracellular acidification with a pulse of exogenous sodium propionate.

Results

The kinetic method allowed determination of the pHi‐dependent activity of NHE in cell lines and primary cell cultures. NHE activity values were demonstrated to be up to 0.016 pH units/s within the pHi range of 7.3 to 6.3. The inhibition of NHE activity by the specific inhibitor ethyl isopropyl amiloride was easily detected by this method.

Conclusions

The assay conditions can be used to relate variations in pHi with the activity of NHE and provide a standardized method to compare between different cells, inhibitors, models of ischemia by acidification, and other relevant experimental or clinical situations. © 2004 Wiley‐Liss, Inc.
Circadian‐ and temperature‐specific effects of early deprivation on rat maternal care and pup development: Short‐term markers for long‐term effects?
Authors: Rüedi‐Bettschen Daniela , Feldon Joram , Pryce Christopher R.
Published: August 2004

We compare the effects on pup body weight and on maternal care of 4‐hr separation from dam and littermates on postnatal Days 1 to 14 (early deprivation, ED) under different thermal and circadian conditions. ED was performed at either 21°C (Cold), or 32°C (Warm), and either during the light or dark phase. The comparison group was nonhandling (NH), either under a nonreversed (Light) or reversed (Dark) cycle. At weaning, Cold ED pups were of lower body weight than Warm ED pups, and Warm ED pups were of lower body weight than NH pups. Light and Dark ED pups received high care at reunion relative to NH, and Cold ED pups received higher care at several hours postreunion relative to Warm ED and NH pups. We propose that reduced pup weight and increased maternal care are short‐term markers for the severity of Cold ED, and that this manipulation could therefore impact negatively on emotionality and cognition in adulthood. © 2004 Wiley Periodicals, Inc. Dev Psychobiol 45:59‐71, 2004.
Development of play fighting in kindling‐prone (FAST) and kindling‐resistant (SLOW) rats: How does the retention of phenotypic juvenility affect the complexity of play?
Authors: Reinhart Christine J. , Pellis Sergio M. , McIntyre Dan C.
Published: August 2004

Rats selectively bred for susceptibility to amygdala kindling (FAST) have been shown to retain neural and behavioral features of the juvenile phase into adulthood. In contrast, rats selectively bred for resistance to amygdala kindling (SLOW) are neurobehaviorally more typically adult. The development of play fighting in male and female rats of both selected lines was studied. Given the apparent association of juvenility and play often noted in the literature for mammals in general, it was predicted that the FAST rats should be more playful and be more likely to retain the juvenile tactics of play that lead to more prolonged and complex patterns of social contact. As expected, FAST rats initiated more playful attacks and were more likely to defend against attacks than SLOW rats as both juveniles and adults. Unexpectedly, however, both selected lines exhibited patterns of defense that reduced the likelihood of complex and prolonged social contact. Importantly, the two selected lines did so by very different means. The FAST rats did so by avoiding contact whereas the SLOW rats did so by responding in an adult‐typical manner that blocks contact. That is, the FAST rats exaggerated the changes typically occurring at puberty whereas the SLOW rats, at all ages, responded in a more adult manner. These data suggest that the different components of play fighting do not change uniformly with changes in the neurobehavioral underpinnings of juvenility. © 2004 Wiley Periodicals, Inc. Dev Psychobiol 45:83‐92, 2004.
Influence of object spatial location and task complexity on children's use of their preferred hand depending on their handedness consistency
Authors: Leconte Pascale , Fagard Jacqueline
Published: August 2004

The goal of the present study was to compare the use of the preferred versus the nonpreferred hand in children performing two tasks of different levels of complexity, in various positions in space. Right‐handed and left‐handed consistent children were compared to their inconsistent counterparts. The results showed that the general tendency to use one's preferred hand to grasp an object is more or less pronounced depending on the object's spatial location and the task's complexity. The children used their preferred hand more often even when the object was presented contralaterally, and did so to a greater extent during the more complex task. Inconsistent‐handers were more likely to shift to using their nonpreferred hand when the object was presented to this hand's side. Finally, we found that the influence of handedness consistency on hand use for the spatial positions varied with the complexity of the task. These results favor a view of handedness as a dynamic process in which motor preferences interact with task demand, probably through task‐related attention. © 2004 Wiley Periodicals, Inc. Dev Psychobiol 45:51‐58, 2004.
A New Diterpenoid Alkaloid from Aconitum episcopale
Authors: Li Zi Yan , Zhao Jingfeng , Yang Jing‐Hua , Zhang Hong Bin , Li Liang
Published: August 2004

A new diterpenoid alkaloid with an epoxy ring between C(3) and C(17) was isolated from Aconitum episcopale. The structure was elucidated on the basis of spectral and chemical evidence.
A Pair of Novel Heptentriol Stereoisomers from the Ascomycete Daldinia concentrica
Authors: Wang Fei , Liu Ji‐Kai
Published: August 2004

A pair of novel heptentriol stereoisomers, hept‐6‐ene‐2,4,5‐triols 2 and 3, were isolated from the culture broth of the ascomycete Daldinia concentrica (Bolton : Fries) Cesati & De Notaris, besides three known compounds, i.e., 2,3‐dihydro‐5‐hydroxy‐2‐methyl‐4H‐1‐benzopyran‐4‐one (1), 3,5‐dihydroxy‐2‐(1‐oxobutyl)‐cyclohex‐2‐en‐1‐one (4), and pyroglutamic acid (=5‐oxo‐L‐proline; 5). Their structures were determined by spectroscopic means, including 2D‐NMR (HMQC, HMBC, 1H,1H‐COSY).
Obesity and risk of cancer in Japan
Authors: Kuriyama Shinichi , Tsubono Yoshitaka , Hozawa Atsushi , Shimazu Taichi , Suzuki Yoshinori , Koizumi Yayoi , Suzuki Yoko , Ohmori Kaori , Nishino Yoshikazu , Tsuji Ichiro
Published: August 2004

We conducted a population‐based prospective cohort study in Japan to examine the relationship between body mass index (BMI) and the risk of incidence of any cancer and of cancer at individual sites. Body mass index was calculated from self‐administered body weight and height at baseline. Relative risks (RR) and 95% confidence intervals (CI) were calculated in multivariate proportional‐hazards models. Among 27,539 persons (15,054 women and 12,485 men) aged 40 years or older who were free of cancer at enrollment in 1984, 1,672 (668 women and 1,004 men) developed cancer during 9 years of follow‐up. In women, after adjustment for potential confounders, the RR of all cancers associated with different BMI, relative to a BMI of 18.5–24.9, were 1.04 (95% CI = 0.85–1.27) for BMI = 25.0–27.4, 1.29 (1.00–1.68) for BMI = 27.5–29.9 and 1.47 (1.06–2.05) for BMI ≥30.0 (p for trend = 0.007). Higher BMI was also significantly associated with higher risk of cancers of the colorectum, breast (postmenopausal), endometrium and gallbladder in women. In men, we observed significantly increased all‐cancer risk among only never‐smokers. Overweight and obesity could account for 4.5% (all subjects) or 6.2% (never‐smokers) of the risk of any cancer in women and −0.2% (all subjects) or 3.7% (never‐smokers) in men. The value for women was within the range among women reported from Western populations (3.2%–8.8%). Our data demonstrate that excess weight is a major cancer risk among Japanese women.
Dietary factors and truncating APC mutations in sporadic colorectal adenomas
Authors: Diergaarde Brenda , Tiemersma Edine W. , Braam Hanneke , van Muijen Goos N.P. , Nagengast Fokko M. , Kok Frans J. , Kampman Ellen
Published: August 2004

Inactivating mutations in APC are thought to be early, initiating events in colorectal carcinogenesis. To gain insight into the relationship between diet and inactivating APC mutations, we evaluated associations between dietary factors and the occurrence of these mutations in a Dutch case–control study of sporadic colorectal adenomas (278 cases; 414 polyp‐free controls). Direct‐sequencing was used to screen adenomas for mutations in the mutation cluster region of APC; truncating mutations were detected in 161 (58%) of the adenomas. Red meat consumption was significantly differently related to polyps with truncating APC mutation (APC+ polyps) compared to polyps without truncating APC mutation (APC− polyps) (highest vs. lowest tertile, odds ratio [OR] = 0.5, 95% confidence interval [CI] = 0.3–1.0). High intake of red meat and fat seemed to increase the risk of APC− polyps only (APC+ vs. controls: red meat, OR = 1.0, 95% CI = 0.6–1.6; fat, OR = 1.1, 95% CI = 0.6–1.9; APC− vs. controls: red meat, OR = 1.8, 95% CI = 1.0–3.1; fat, OR = 1.9, 95% CI = 1.0–3.7). Intake of carbohydrates was inversely associated with both polyp groups, most noticeably with APC− polyps. Most other evaluated dietary factors were not distinctively associated with a specific APC status. None of the dietary factors was specifically associated with a particular type of truncating APC mutation. Our data suggest that red meat and fat may increase the risk of APC− polyps in particular, whereas carbohydrates may especially decrease the risk of APC− polyps. However, most examined dietary factors do not appear to be specifically associated with the occurrence of truncating APC mutations in colorectal adenomas but seem to affect both pathways equally.
p21Waf1/Cip1 and p53 are downstream effectors of protein kinase C delta in tumor suppression and differentiation in human colon cancer cells
Authors: Perletti Gianpaolo , Marras Emanuela , Dondi Davide , Osti Daniela , Congiu Terenzio , Ferrarese Roberto , de Eguileor Magda , Tashjian Armen H.
Published: August 2004

We have previously demonstrated that the delta isoform of protein kinase C (PKCδ) is importantly involved in cell growth inhibition and tumor suppression in colon cancer cells. To investigate further the activity and mechanism of action of PKCδ, we have retrovirally transduced a PKCδ cDNA in HCT116 human colon cancer cells. PKCδ‐overexpressing cells (HCT116/PKCδ) were growth‐inhibited, showed marked morphologic changes and underwent multinucleation and phenotypic changes characteristic of mitotic catastrophe. Compared to controls, HCT116/PKCδ cells showed a highly attenuated tumorigenic profile and poor anchorage‐independent growth. In addition, transfected cells established junction‐coordinated intercellular communications, expressed cell surface microvilli and overexpressed the colon differentiation marker alkaline phosphatase. HCT116/PKCδ cells also produced the 89 kDa, carboxy‐terminal catalytic domain of PARP. In HCT116/PKCδ cells, p21Waf1/Cip1 and p53 were transiently upregulated for 48 hr after PKCδ transduction. In a p21 null subline of HCT116 cells (HCT116/p21null), overexpression of PKCδ did not affect tumorigenicity or differentiation, indicating that p21 is essential for the antitumorigenic activity of PKCδ. Similarly, overexpression of PKCδ caused no significant phenotypic changes in HCT116/E6 cells, an HCT116 subline in which the p53 protein is downregulated by the human papillomavirus E6 gene product. We conclude that overexpression of PKCδ in human colon cancer cells induces multiple antineoplastic effects that depend on the activities of p21Waf1/Cip1 and p53.
CYP1A1 variants and smoking‐related lung cancer in San Francisco bay area Latinos and African Americans
Authors: Wrensch Margaret R. , Miike Rei , Sison Jennette D. , Kelsey Karl T. , Liu Mei , McMillan Alex , Quesenberry Charles , Wiencke John K.
Published: August 2004

We examined CYP1A1 T6235C (M1) and A4889G (M2) polymorphisms in San Francisco Bay Area African Americans and Latinos who were newly diagnosed with primary lung cancer from September 1998 to November 2002 and in age‐gender‐ethnicity frequency‐matched controls. Owing mainly to rapid mortality of cases, overall percentages of cases genotyped were 26% and 32% for Latinos and African Americans, respectively. CYP1A1 variants were genotyped for Latinos (104 cases, 278 controls) and African Americans (226 cases, 551 controls). M1 and M2 frequencies in controls were 0.23 and 0.02 for African Americans and 0.38 and 0.29 for Latinos. In Latinos, the overall inverse odds ratio (OR) of 0.51 (95% CI = 0.32–0.81) for M1 variant genotype resulted from an inverse interaction with smoking. Nonsmokers with M1 genotype had a slight elevated OR (1.5; 0.59–3.7), but those with less than 30 or 30 or more pack‐year history had 0.20 (0.06–0.70) and 0.21 (0.06–0.81) times (about 1/5) the odds expected if smoking and genotype were independent lung cancer risk factors. African Americans had interactions of similar magnitude that were not statistically significant. Results for M2 were very similar. Inverse interactions of CYP1A1 variants and smoking‐associated lung cancer risk in Latinos might be causal, due to undetected bias or confounding, or represent a unique linkage disequilibrium between a new lung cancer locus and CYP1A1 in this highly admixed population.
Longitudinal and Vertical Spatial Gradients in the Distribution of Fish within a Canyon‐shaped Reservoir
Authors: Vašek Mojmír , Kubečka Jan , Peterka Jiří , Čech Martin , Draštík Vladislav , Hladík Milan , Prchalová Marie , Frouzová Jaroslava
Published: August 2004

The large‐scale spatial distribution of fish was investigated within a morphometrically simple canyon‐shaped reservoir with a single major tributary and a longitudinal trophic gradient (Římov Reservoir, Czech Republic). Samples of fish were taken by Nordic survey gill nets (several mesh sizes from 8 to 70 mm knot to knot) installed as surface nets at several offshore areas located along the longitudinal axis of the reservoir. Surveys were carried out in late summer during 1999–2003. An obvious distribution gradient of fish was revealed along the longitudinal axis of the Římov Reservoir. The total relative fish abundance and biomass (catch per unit effort) decreased considerably from the upstream end of the reservoir toward the dam. Roach (Rutilus rutilus), bleak (Alburnus alburnus) and bream (Abramis brama) comprised the bulk of catches at all areas. Enhanced dominance of bream was observed in the fish assemblage at the uppermost, more eutrophic area of the reservoir. The highest number of fish species and the highest abundance of young‐of‐the‐year fish were also observed in the tributary area. In the downstream part of the reservoir, gill net surveys along the vertical depth profiles indicated that offshore fish occupied mostly the epilimnion. Extreme flood events affected the Římov Reservoir, however, it seemed they had no significant impact on the gradients described. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Novel fabricated matrix via electrospinning for tissue engineering
Authors: Khil Myung‐Seob , Bhattarai Shanta Raj , Kim Hak‐Yong , Kim Sung‐Zoo , Lee Keun‐Hyung
Published: August 2004

Electric field‐driven fiber formation (electrospinning) is developing into a practical means for preparing novel porous filament with unusual structures and affordable mechanical properties. Polycaprolactone (PCL) was dissolved in solvent mixtures of methylene chloride/N,N‐dimethyl formamide with ratios of 100/0, 75/25, and 50/50 (v/v) for electrospinning. The filament was formed by coagulation of the spinning solution following the well‐known principle of phase separation in polymer solutions valid in other wet shaping processes. A strand of electrospun porous filament consisted of fibers ranging from 0.5 to 12 μm in diameter. To evaluate the feasibility of three‐dimensional fabric as scaffold matrices, the plain weave, which is the simplest of the weaves and the most common, was prepared with porous PCL filament. The growth characteristics of MCF‐7 mammary carcinoma cells in the woven fabrics showed the important role of matrix microstructure in proliferation. This study has shown that woven fabrics, consisting of porous filaments via electrospinning, may be suitable candidates as tissue engineering scaffolds. © 2004 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 72B: 117–124, 2005