bluuebunny/crossref_metadata_2025 huggingface dataset:
Models of Visceral Pain: Colorectal Distension (CRD)
Authors: Jones R. Carter W. , Gebhart G.F.
Published: August 2004

The visceromotor response to balloon distension of the colon is a robust behavioral model of visceral nociception in rodents and is ideally suited for studying the visceral antinociceptive activity of drugs. This unit describes, in detail, quantification of this response with the use of electromyography in both rats and mice.
Measurement of Cell Death in Mammalian Cells
Authors: Cummings Brian S. , Schnellmann Rick G.
Published: August 2004

This unit presents methods used to assess cell death in mammalian cells. The unit is divided into five sections: (1) a brief overview of cytotoxicity and pathways of cell death, (2) an improved method to measure cell death using lactate dehydrogenase (LDH) release as a marker of membrane integrity, (3) a flow cytometry method that simultaneously measures two types of cell death, oncosis and apoptosis, (4) use of nuclear morphology to assess apoptosis and oncosis, and (5) a brief discussion of the use of cytotoxicity assays to determine the mechanisms of cell death.
Multidisciplinary Approaches for Characterizing Synaptic Vesicle Proteins
Authors: Leenders Miriam , Gerwin Claudia , Sheng Zu‐Hang
Published: August 2004

Investigation of synaptic vesicle membrane proteins using multidisciplinary approaches, particularly to characterize synaptic vesicle proteins in synapses, can greatly advance our knowledge of the molecular mechanisms involved in synaptic vesicle exocytosis and neurotransmission. Three approaches are presented in this unit to identify and characterize synaptic vesicle proteins. The first is a subcellular fractionation method used to isolate synaptic vesicles from rat brain synaptosomal preparations, which can then be used in a variety of biochemical studies on synaptic vesicle proteins. The second is a detailed procedure for pre‐embedding immunogold staining and electron microscopic observation, techniques that permit the morphological identification of synaptic vesicle proteins in individual vesicles at the intact synapse. Finally, a protocol for immunocytochemical staining of cultured hippocampal neurons for light microscopic examination is provided, which allows one to stain multisynaptic vesicle proteins and determine their localization in relation to other proteins or subcellular structures in synapses.
Macronutrient Selection in Experimental Animals
Authors: Kanarek Robin B.
Published: August 2004

To address the question of how animals choose diets from an array of nutritionally different foods, researchers have designed experiments in which animals are provided with separate sources of the three macronutrients, protein, fat and carbohydrate. These experiments are useful for investigating the neuroanatomical and neurochemical mechanisms involved in food choice, as well as for assessing the effects of changes in physiological status (e.g., pregnancy; lactation and increased energy expenditure) or disease states (e.g., diabetes; and obesity) on nutrient intakes.
Synthesis of Oligoribonucleotides Containing N6‐Alkyladenosine and 2‐Methylthio‐N6‐Alkyladenosine
Authors: Kierzek Elzbieta , Kierzek Ryszard
Published: August 2004

The N6‐alkyladenosines and 2‐methylthio‐N6‐alkyladenosines are the most common modified adenosine nucleosides, and transfer ribonucleic acids (tRNA) are particularly rich in these modified nucleosides. They are present at position 37 of the anticodon arm, and the contributions of these hypermodified nucleosides to codon‐anticodon interactions as well as to translation are significant, although they are not fully understood. This unit describes a new chemical synthesis method for oligoribonucleotides containing N6‐alkyladenosines and 2‐methylthio‐N6‐alkyladenosines via postsynthetic modifications of precursor oligoribonucleotides. To obtain oligoribonucleotides containing N6‐alkyladenosines, a precursor oligoribonucleotide carrying 6‐methylthiopurine riboside residues was used, whereas for the synthesis of oligoribonucleotides containing 2‐methylthio‐N6‐alkyladenosines, a precursor oligoribonucleotide carrying the 2‐methylthio‐6‐chloropurine riboside was applied. This allowed synthesis of modified oligoribonucleotides containing naturally occurring modified nucleosides such as N6‐isopentenyladenosine (i6A), N6‐methyladenosine (m6A), 2‐methylthio‐N6‐isopentenyladenosine (ms2i6A), and 2‐methylthio‐N6‐methyladenosine (ms2m6A), as well as several unnaturally modified adenosine derivatives.
Analysis of Telomere Length and Telomerase Activity
Authors: Hathcock Karen S. , Hodes Richard J. , Weng Nan‐Ping
Published: August 2004

Telomeres are specialized DNA‐protein structures present at the ends of all linear chromosomes and are characterized by (TTAGGG)n hexanucleotide repeats and associated proteins. Telomere length has been implicated in cell survival and replicative capacity of dividing somatic cells. In the absence of an active compensatory mechanism, telomere lengths shorten as a consequence of proliferation, both in vitro and in vivo. However, this loss of telomeric repeats can be compensated and telomere length maintained by the enzyme telomerase, which is capable of adding (TTAGGG) repeat sequences to the ends of telomeres. This unit describes methods that are used for the measurement of telomere length and telomerase activity in human and murine cells.
Culture of HIV in Monocytes and Macrophages
Authors: Crowe Suzanne , Maslin Clare , Ellery Philip , Kedzierska Katherine
Published: August 2004

Monocytes and macrophages constitute important cellular targets for macrophage‐tropic (M‐tropic) strains of HIV‐1 and are believed to play a major role in the pathogenesis of AIDS. This updated unit provides procedures for culturing HIV in these target cells under suspension and adherent cell culture conditions. The unit also provides methods for expanding M‐tropic primary isolates of HIV and determining the 50% tissue culture infectious dose (TCID50) of HIV stocks.
Isothermal Titration Calorimetry
Authors: Velázquez‐Campoy Adrián , Ohtaka Hiroyasu , Nezami Azin , Muzammil Salman , Freire Ernesto
Published: August 2004

In the last two decades, isothermal titration calorimetry (ITC) has become the preferred technique to determine the binding energetics of biological processes, including protein‐ligand binding, protein‐protein binding, DNA‐protein binding, protein‐carbohydrate binding, protein‐lipid binding, and antigen‐antibody binding. In this unit several protocols are presented, ranging from the basic ones that are aimed at characterizing binding of moderate affinity to advanced protocols that are aimed at determining very high or very low affinity binding processes. Also, alternate protocols for special cases (homodimeric proteins and unstable proteins) and additional information accessible by ITC (heat capacity and protonation/deprotonation processes coupled to binding) are presented.
Searching WormBase for Information About Caenorhabditis elegans
Authors: Schwarz Erich M. , Sternberg Paul W.
Published: August 2004

WormBase is the major public biological database for the nematode Caenorhabditis elegans. It is meant to be useful to any biologist who wants to use C. elegans, whatever his or her specialty. WormBase contains information about the genomic sequence of C. elegans, its genes and their products, and its higher‐level traits such as gene expression patterns and neuronal connectivity. Also, WormBase contains sequence and gene structures of C. briggsae, a closely related worm. These data are interconnected, so that a search beginning with one object (such as a gene) can be directed to related objects of a different type (such as the DNA sequence of the gene, or the cells in which the gene is active). One can also do searches for complex data sets. The WormBase developers group actively invites suggestions for improvement from users. WormBase's source code and underlying database are freely available for local installation and improvement.
Identifying Functional Sites Based on Prediction of Charged Group Behavior
Authors: Ondrechen Mary Jo
Published: August 2004

This protocol describes the implementation and interpretation of THEMATICS, a simple computational predictor of functional information for proteins from the three‐dimensional structure. This method is based on the computation of the electrical potential function for the protein and the calculation of the predicted titration curves for each of the titratable groups in the protein. While most of the titratable residues in a protein have predicted titration behavior that fits the Henderson‐Hasselbalch equation, the ionizable residues in the active site generally deviate dramatically from the typical behavior. From the calculated titration curves, one identifies those residues that deviate significantly from Henderson‐Hasselbalch behavior. A cluster of two or more of such deviant titratable residues in physical proximity is a reliable predictor of active‐site location.
The rate acceleration in solid‐state polycondensation of PET by nanomaterials
Authors: Yu Huimin , Han Keqing , Yu Muhuo
Published: August 2004

The effect of nanomaterials on the solid‐state polycondensation (SSP) of PET was investigated using intrinsic viscosity measurement, wide‐angle X‐ray diffraction, differential scanning calorimetry, and polarizing microscope. The results showed that the montmorillonite nanomaterials could greatly increase the rate of solid‐state polycondensation of PET, probably due to the nucleation of montmorillonite nanomaterials for PET crystallization, which resulted in lower crystallinity, more small crystals, and more surfaces of the crystals. The surfaces of microcrystal and richer amorphous regions benefitted the polycondensation reaction of PET and diffusion of volatile by‐products, which led to the higher rate of SSP. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 971–976, 2004
Synthesis and characterization of poly(propylene)/montmorillonite nanocomposites by simultaneous grafting‐intercalation
Authors: Xie Shaobo , Zhang Shimin , Wang Fosong
Published: August 2004

A novel process for the preparation of poly(propylene)/montmorillonite (PP/MMT) nanocomposites was developed via simultaneous solution grafting‐intercalation in the presence of a reactive ammonium cation that can be grafted onto poly(propylene). Partially introducing this reactive cation into long alkyl ammonium modified MMT interlayers can transfer a conventional microcomposite into intercalated/exfoliated nanocomposites, which was evidenced by X‐ray diffraction (XRD) and transmission electron microscopy (TEM). The PP chains were tethered onto the clay surface through the bridge of the reactive ammonium cations, which can be characterized by FTIR. The bridged chemical bonding also results in a good interface adhesion between PP and MMT, as confirmed by SEM investigation. The enhanced thermal properties of PP/MMT nanocomposites were characterized by thermogravimetric analysis. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1018–1023, 2004
Properties of silica/clay filled heavy‐duty truck tire thread formulation
Authors: Ogbebor O. J , Farid A. S. , Okwu U. N.
Published: August 2004

A tire thread formulation for heavy‐duty trucks containing SBR/BR rubber blend and varying proportions of silica/clay fillers including a silane‐coupling agent have been investigated. The various mixes were compounded in a Banbury ‘O’ mixer and vulcanized using the EV‐system. Silica/clay (80/0) served as the control mix. The oscillating disc rheometer (ODR) was used in determination of cure characteristics. Substitution of silica (80 phr) with china clay up to 40 phr increased the cure rate of the rubber blend mixes as well as their maximum torque level (Tmax). Tmax was observed to be highest at a filler blend ratio of 40/40 phr. Synergism between silica and clay at this filler blend mixture is suggested to be responsible for the observation. The heat buildup was reduced from 43 to 20°C as the clay content increased. Results also showed that the rubber blend compound containing silica/clay (60/20) filler blend in the stated ratio exhibited the best balance of properties in the critical parameters such as the absolute torque level (69.5 dNm), heat buildup (39°C), and abrasion resistance (0.574 mg.loss/1,000 rev). The rate of depreciation of abrasion resistance of rubber blend compound as the clay content increased was found to be 0.035 mg loss/1,000 rev as silica is substituted with one part of china clay phr. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1024–1028, 2004
Effect of proton implantation on the tribological properties of phenolphthalein poly(ether sulfone)
Authors: Pei Xianqiang , Wang Qihua , Wang Haijun , Mu Bo
Published: August 2004

Phenolphthalein poly(ether sulfone) (PES‐C) was implanted with 110 keV protons in four doses of 1 × 1014, 5 × 1014, 2.5 × 1015, and 1.25 × 1016 ions cm−2. The friction and wear properties of both implanted and unimplanted PES‐C were investigated with a THT07–135 high‐temperature tribometer against a steel ball. Results revealed that the friction coefficient and wear rate both decreased with increasing doses: especially when the dose reached 1.25 × 1016 ions cm−2, its wear rate decreased nearly two orders of magnitude. Scanning electron microscopy results revealed that the size of wear debris of unimplanted PES‐C was greater than that of the implanted one. The worn surface of unimplanted PES‐C showed plastic deformation, plastic flow, and adhesiveness, whereas the worn surface of the implanted one showed only some mild nicks. The carbon element distribution map revealed that much more carbon element transferred to the counterpart steel ball friction against the implanted PES‐C. The IR spectrum showed that partial degradation might occur when PES‐C was implanted with protons. Measurement of hardness revealed an increase with increasing doses. The reason for the increase might be the formation of carbon‐rich structures on the surface of the sample. From the above, conclusions can be drawn that the wear mechanisms of unimplanted samples were mainly related to adhesion, plastic deformation, and plastic flow, whereas that of implanted samples was mainly attributed to mild “three‐body” wear. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1043–1048, 2004
Effects of compatibilizer on the layered silicate/ethylene vinyl acetate nanocomposite
Authors: Suh Il Soon , Ryu Sung Hun , Bae Jong Hyun , Chang Young Wook
Published: August 2004

Dispersion behavior of monmorillonite (MMT) is investigated in ethylene vinyl acetate (EVA)/MMT nanocomposite with various vinyl acetate content. Maleic anhydride (MAH) grafted polyethylenes with various MAH contents are used as a compatibilizer to enhance the dispersion of MMT. DMA and XRD studies indicate that an intercalated/exfoliated structure is obtained and vinyl acetate content and the concentration of PEMA play a critical role in EVA/MMT nanocomposite. Higher vinyl acetate content and concentration of grafted maleic anhydride result in better dispersion of MMT. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 94: 1057–1061, 2004
Comparison of extra-anatomic bypass grafting with angioplasty for atherosclerotic disease of the supra-aortic trunks
Authors: Modarai B , Ali T , Dourado R , Reidy J F , Taylor P R , Burnand K G
Published: August 2004

Background

Symptomatic stenosis of the supra-aortic trunks (subclavian, innominate and common carotid arteries) can be treated by angioplasty/stenting or surgical bypass. The aim of this study was to compare the initial success and outcome of these two types of treatment.

Methods

A prospective database was used to collect information on the presentation, initial success, complications and outcome in 76 patients treated in a single centre between 1983 and 2003.

Results

Thirty-five surgical extra-anatomic bypasses were performed, 13 carotid to carotid, 14 carotid to subclavian, two carotid to axillary, three axillary to axillary, one subclavian to axillary and two subclavian to subclavian. One graft occluded after 19 years. No limbs were amputated and no patient had a stroke. The secondary patency rate was 97 per cent at a mean follow-up of 5 years. Forty-one angioplasties were attempted, 34 of the left subclavian, six of the right subclavian and one of the innominate artery. Angioplasty for six subclavian occlusions was unsuccessful. Twenty-seven of 33 arteries remained patent at a mean follow-up of 4 years after a successful endovascular procedure.

Conclusion

Extra-anatomic bypass for supra-aortic trunk disease has a better patency than angioplasty, with a comparable complication rate.
Elective sirolimus‐eluting stent implantation for multivessel disease involving significant LAD stenosis: One‐year clinical outcomes of 99 consecutive patients—the Rotterdam experience
Authors: Arampatzis Chourmouzios A. , Hoye Angela , Lemos Pedro A. , Saia Francesco , Tanabe Kengo , Degertekin Muzaffer , Sianos Georgios , Smits Pieter C. , van der Giessen Willem J. , McFadden Eugene , van Domburg Ron , de Feyter Pim , Serruys Patrick W.
Published: August 2004

The aim of this study was to evaluate the effectiveness of sirolimus‐eluting stent (SES) implantation for patients with multivessel disease, which included left anterior descending artery (LAD) treatment. Since April 2002, SES has been utilized as the device of choice for all interventions in our institution as part of the Rapamycin‐Eluting Stent Evaluated at Rotterdam Hospital (RESEARCH) registry. In the first 6 months of enrolment, 99 consecutive patients (17.6% of the total population) were treated for multivessel disease involving the LAD. The impact of SES implantation on major adverse cardiac events (MACE) was evaluated. All the patients received SES in the LAD. Additional stent implantation in the right coronary artery, the left circumflex, or in all three major vessels was attempted successfully in 32 (32%), 51 (52%), and 16 (16%) of the treated patients respectively. During a mean follow‐up of 360 ± 59 days (range, 297–472 days), we had one death, one non‐Q‐wave myocardial infarction, and eight patients required subsequent intervention. The event‐free survival of MACE at 1 year was 85.6%. SES implantation for multivessel disease in a consecutive series of patients is associated with low incidence of adverse events. The reported results are related predominantly to the reduction in repeat revascularization. Catheter Cardiovasc Interv 2004;63:57–60. © 2004 Wiley‐Liss, Inc.
Synthesis and Magnetic Behavior of La2‐xRxBaCuO5 Oxides (R: Pr and Nd; x = 1 and 2).
Authors: Sanchez‐Capuchino R. , Climent E. , de Paz J. Romero , Saez‐Puche R.
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Chemo‐ and Stereoselective Synthesis of Benzocycloheptene and 1‐Benzoxepin Derivatives as α‐Sympathomimetic and Anorexigenic Agents.
Authors: Tandon Vishnu K. , Singh Kunwar A. , Awasthi Anoop K. , Khanna J. M. , Lal Bansi , Anand Nitya
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
A New and Facile Method for Stereoselective Synthesis of (E)‐Styryl Bromides by the Reduction of 1,1‐Dibromoalkenes Using LiAlH4—EtOAc (1:1).
Authors: Horibe Hideo , Kondo Kazuhiro , Okuno Hiroaki , Aoyama Toyohiko
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Oxidative C—C Bond‐Forming Reaction of Electron‐Rich Alkylbenzyl Ether with Trimethylvinyloxysilane.
Authors: Ying Bai‐Ping , Trogden Bridget G. , Kohlman Daniel T. , Liang Sidney X. , Xu Yao‐Chang
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Synthesis and Characterization of 1,2‐bis(2′‐Pyrazineethynyl) Benzene Palladium(II) Dichloride and Its Catalysis of the Suzuki Coupling Reaction.
Authors: Schultheiss Nate , Barnes Charles L. , Bosch Eric
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Synthesis of Hydroxy‐7H‐benzo[c]fluoren‐7‐ones.
Authors: Coelho Paulo J. , Salvador Maria A. , Oliveira M. Manuel , Carvalho Luis M.
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Synthesis of N‐Protected Cyano Aziridines.
Authors: Fioravanti Stefania , Morreale Alberto , Pellacani Lucio , Tardella Paolo A.
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Selective Functionalization in Positions 2 and 3 of Indole via an Iodine—Copper Exchange Reaction.
Authors: Yang Xiaoyin , Althammer Andreas , Knochel Paul
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Synthesis and Biological Activity of N‐Aryl‐2‐aminothiazoles: Potent Pan Inhibitors of Cyclin‐Dependent Kinases.
Authors: Misra Raj N. , et al. et al.
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Orally Active Factor Xa Inhibitors: 4,5,6,7‐Tetrahydrothiazolo[5,4‐c]pyridine Derivatives.
Authors: Haginoya Noriyasu , Kobayashi Syozo , Komoriya Satoshi , Hirokawa Yumiko , Furugori Taketoshi , Nagahara Takayasu
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Efficient One‐Pot Synthesis of Secondary Cyclic Phosphanes with Easy Regeneration of the Phosphorus‐Donor Reagent Used.
Authors: Baccolini Graziano , Boga Carla , Galeotti Matteo
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Synthesis of Novel Unsymmetrically Bridgehead‐Substituted Phenylselenobicyclo[1.1.1]pentanes.
Authors: Marinozzi Maura , Fulco M. Carmela , Rizzo Rosa , Pellicciari Roberto
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Crystallization‐Induced Asymmetric Transformation. Application to Conjugate Addition of Benzylamine to Amides of Benzoylacrylic Acid.
Authors: Jakubec Pavol , Berkes Dusan , Povazanec Frantisek
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Enantiomeric Synthesis of the SPIKET‐P Enantiomers.
Authors: Sharma Anubha , Iyer Prema , Gamre Sunita , Chattopadhyay Subrata
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Progress in the Chemistry of Metallabenzynes
Authors: Jia Guochen
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Synthesis of Polyheterocyclic Nitrogen‐Containing Marine Natural Products.
Authors: Fernandez David , Ahaidar Abderaouf , Danelon Gerardo , Cironi Pablo , Marfil Marta , Perez Olga , Cuevas Carmen , Albericio Fernando , Joule John A. , Alvarez Mercedes
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Sulfoximines: Synthesis and Catalytic Applications
Authors: Okamura Hiroaki , Bolm Carsten
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Phosphinidenes
Authors: Lammertsma Koop
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Ozone in Arteriosclerotic Plaques: Searching for the “Smoking Gun”
Authors: Sies Helmut
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Gene Delivery Using Glycoclusters — Designing Viral (50 nm) Sizes
Authors: Aoyama Yasuhiro
Published: August 2004

For Abstract see ChemInform Abstract in Full Text.
Influence of rheumatoid arthritis in the enantioselective disposition of fenoprofen
Authors: Barissa Giuliano Rodrigo , Poggi Josiane Cristófani , Donadi Eduardo Antônio , dos Reis Marina Lemos , Lanchote Vera Lucia
Published: August 2004

To investigate the influence of rheumatoid arthritis on the stereoselective disposition of fenoprofen administered as a racemic mixture, eight patients with rheumatoid arthritis receiving calcium rac‐fenoprofen (200 mg/8 h) and 7 healthy volunteers given single oral dose (600 mg) were investigated. Serial blood samples and urine were collected from zero to 24 h after fenoprofen (FEN) administration. The following differences were observed between the (+)‐(S) and (−)‐(R)‐FEN in the patients with rheumatoid arthritis (means 95% CI, Wilcoxon test, P < 0.05): Cmax 14.1 (12.5–15.8) versus 3.6 (2.5–4.7) μg/ml; AUC 80.5 (67.3–93.7) versus 12.1 (8.8–15.4) μg·h/ml; ClT/f 1.3 (1.0–1.5) versus 9.1 (6.5–11.8) l/h; and t1/2 3.1 (2.3–3.9) versus 1.2 (0.8–1.6) h. The ClT/f of (−)‐(R)‐FEN was reduced in patients with rheumatoid arthritis when compared to healthy volunteers: 9.1 (6.5–11.8) versus 17.4 (13.9–20.9) l/h; P < 0.05 Mann–Whitney test. The administration of rac‐FEN as a single dose to healthy volunteers or multiple doses to patients with rheumatoid arthritis resulted in lower ClT/f for the (+)‐(S)‐FEN. The lower ClT/f of (−)‐(R)‐FEN observed for patients with rheumatoid arthritis is consistent with lower clearance by inversion, although other metabolic pathways, drug interactions, and bioavailability of the individual enantiomers may also contribute to the difference. Chirality 16:602–608, 2004. © 2004 Wiley‐Liss, Inc.
The neutrophil, not the tumor
Authors: Pentheroudakis George , Malamou‐Mitsi Vassiliki , Briasoulis Evangelos , Damala Konstantina , Vassou Amalia , Vartholomatos George , Kolaitis Nikos , Pavlidis Nikolas
Published: August 2004

BACKGROUND

Patients with resected breast carcinoma who received granulocyte—colony‐stimulating factor (G‐CSF)–supported adjuvant chemotherapy exhibited an increase in their serum CA 15‐3 levels. The authors investigated the role of G‐CSF–induced neutrophil MUC1 expression in this setting.

METHODS

Twenty‐two patients with resected early breast carcinoma and 6 patients with high‐grade lymphoma received chemotherapy cycles with or without G‐CSF support. When given, G‐CSF was administered for either 5 or 10 days per cycle. Immunocytochemical staining and flow cytometric analysis of peripheral blood neutrophils and bone marrow myeloid cells for MUC1 epitopes were performed during treatment.

RESULTS

At baseline, the median serum CA 15‐3 was 16 U/mL, with weak MUC1 expression in peripheral neutrophils (median immunocytochemical score [ICCS] = 40, flow cytometric score [FCS] = 211 antibody molecules per neutrophil). For patients receiving chemotherapy cycles with 5‐day G‐CSF support, median CA 15‐3 levels increased moderately (median = 28 U/mL; P = 0.016) and absolute neutrophil counts (ANC) did not increase, whereas ICC staining showed a moderate increase (median ICCS = 105; P = 0.015). For patients receiving chemotherapy cycles with 10‐day G‐CSF, serum CA 15‐3 levels increased 2–4‐fold from baseline levels and reached abnormal levels (median = 47; P < 0.0005) and the ANC increased (median = 21,400/mm3; P = 0.007), whereas significant induction of MUC1 expression occurred in the cell membrane and mostly in the cytoplasm of neutrophils (median ICCS = 162; P = 0.001). Flow cytometry detected increased cytoplasmic, but not surface, neutrophil MUC1 expression in the 10‐day G‐CSF group only (baseline median FCS = 3975, 4th cycle median = 6327 molecules per cell; P = 0.028). In the bone marrow, induction of MUC1 expression was observed in the 10‐day G‐CSF group only in band forms and neutrophils, but not in more immature myeloid cells. Serum CA 15‐3 levels and ICC score were found to demonstrate a linear relation. When ICCS and ANC were incorporated in a combined score, its relation with serum CA 15‐3 levels demonstrated a parabolic (cubic) pattern. Serum CA 15‐3 levels, ANC, and neutrophil MUC1 staining returned to baseline after the completion of therapy. No excess of malignant recurrences were observed.

CONCLUSIONS

Women with resected breast carcinoma who received G‐CSF–primed chemotherapy showed serum CA 15‐3 elevation due to an increase in peripheral blood neutrophil number and induced neutrophil cytoplasmic MUC1 expression, which was caused by G‐CSF. Physicians should be aware of this interaction. Cancer 2004. © 2004 American Cancer Society.
2‐Fluoro‐2‐deoxy‐D‐glucose positron emission tomography imaging is predictive of pathologic response and survival after preoperative chemoradiation in patients with esophageal carcinoma
Authors: Swisher Stephen G. , Erasmus Jeremy , Maish Mary , Correa Arlene M. , Macapinlac Homer , Ajani Jaffer A. , Cox James D. , Komaki Ritsuko R. , Hong David , Lee Hoon K. , Putnam Joe B. , Rice David C. , Smythe W. Roy , Thai Linh , Vaporciyan Ara A. , Walsh Garrett L. , Wu Tsung‐Teh , Roth Jack A.
Published: August 2004

BACKGROUND

The current study was performed to assess the value of 2‐fluoro‐2‐deoxy‐d‐glucose positron emission tomography (FDG‐PET) in predicting the pathologic response and survival of patients with esophageal carcinoma treated with preoperative chemoradiation (CRT) and tumor resection. Preliminary reports suggest that FDG‐PET may be predictive of the response of esophageal carcinoma patients to preoperative CRT.

METHODS

Eighty‐three patients with resectable esophageal carcinoma who underwent preoperative CRT and FDG‐PET and tumor resection were evaluated for pathologic response to CRT, percent residual tumor, and survival.

RESULTS

The majority of patients in the current study were men (74 of 83 patients; 89%). Most tumors were adenocarcinomas (73 of 83 tumors; 88%) and clinical EUST3/4 (69 tumors; 83%) or N1 (46 tumors; 55%). FDG‐PET after preoperative CRT identified pathologic responders but failed to rule out microscopic residual tumor in 13 of 73 cases (18%). Pathologic response was found to correlate with the post‐CRT FDG‐PET standardized uptake value (SUV) (P = 0.03) and a post‐CRT FDG‐PET SUV of ≥ 4 was found to be the only preoperative factor to correlate with decreased survival (2‐year survival rate of 33% vs. 60%; P = 0.01). On univariate Cox regression analysis, only post‐CRT FDG‐PET was found to be correlated with post‐CRT survival (P = 0.04).

CONCLUSIONS

Post‐CRT FDG‐PET was found to be predictive of pathologic response and survival in patients with esophageal carcinoma who undergo preoperative CRT. Esophagectomy should still be considered even if the post‐CRT FDG‐PET scan is normal because microscopic residual disease cannot be ruled out. Cancer 2004. © 2004 American Cancer Society.
Fluorescence‐intensity multiplexing: Simultaneous seven‐marker, two‐color immunophenotyping using flow cytometry
Authors: Bradford Jolene A. , Buller Gayle , Suter Michael , Ignatius Michael , Beechem Joseph M.
Published: August 2004

Background

Conventional immuno‐based multiparameter flow cytometric analysis has been limited by the requirement of a dedicated detection channel for each antibody‐fluorophore set. To address the need to resolve multiple biological targets simultaneously, flow cytometers with as many as 10–15 detection channels have been developed. In this study, a new Zenon immunolabeling technology is developed that allows for multiple antigen detection per detection channel using a single fluorophore, through a unique method of fluorescence‐intensity multiplexing. By varying the Zenon labeling reagent‐to‐antibody molar ratio, the fluorescence intensity of the antibody‐labeled cellular targets can be used as a unique identifier. Although demonstrated in the present study with lymphocyte immunophenotyping, this approach is broadly applicable for any immuno‐based multiplexed flow cytomety assay.

Methods

Lymphocyte immunophenotyping of 38 clinical blood specimens using CD3, CD4, CD8, CD16, CD56, CD19, and CD20 antibodies was performed using conventional flow cytometric analysis and fluorescence‐intensity multiplexing analysis. Conventional analysis measures a single antibody‐fluorophore per photomultiplier tube (PMT). Fluorescence‐intensity multiplex analysis simultaneously measures seven markers with two PMTs, using Zenon labeling reagent‐antibody complexes in a single tube: CD19, CD4, CD8, and CD16 antibodies labeled with Zenon Alexa Fluor®488 Mouse IgG1 labeling reagent and CD56, CD3, and CD20 antibodies labeled with Zenon R‐Phycoerythrin (R‐PE) Mouse IgG1 or IgG2b labeling reagents.

Results

The lymphocyte immunophenotyping results from fluorescence‐intensity multiplexing using Zenon labeling reagents in a single tube were comparable to results from conventional flow cytometric analysis.

Conclusions

Simultaneous evaluation of multiple antigens using a single fluorophore can be performed using antibodies labeled with varying ratios of a Zenon labeling reagent. Labeling two sets of antibodies with different Zenon labeling reagents can generate characteristic and distinguishable multivariate patterns. Combining multiple antibodies and fluorescent labels with fluorescence intensity multiplexing enables the resolution of more cellular targets than detection‐channels, allowing sophisticated multiparameter flow cytometric studies to be performed on less complex 2‐ or 3‐detection‐channel flow cytometers. For typical biological samples, approximately 2–4 cellular targets per detection channel can be resolved using this technique. © 2004 Wiley‐Liss, Inc.
Single‐cell gel electrophoresis assay monitors precise kinetics of DNA fragmentation induced during programmed cell death
Authors: Chandna Sudhir
Published: August 2004

Background

Single‐cell gel electrophoresis, or the comet assay, a technique widely used for DNA damage analysis, has been used recently for detecting DNA fragmentation in cells undergoing apoptosis. However, the number of variants of this assay used thus far primarily detected the late stages of DNA fragmentation. Therefore, monitoring the progression of DNA fragmentation, which could greatly improve the analysis of cell death induction and progression at the single‐cell level, has not been possible with this assay.

Methods

In the present study, a modification of the original neutral comet assay developed by Ostling and Johanson (Biochem Biophys Res Commun 123:291–298, 1984) was used to detect various stages of DNA fragmentation. This assay involves cell lysis with anionic detergents at nearly neutral pH (9.5) and does not include high salt concentration, unlike most other published methods. BMG‐1 human glioma cells were induced to undergo programmed cell death by treating with a large dose (100 μM) of etoposide, and comets were prepared after different durations (1–24 h) of treatment.

Results

In contrast to results of previously published studies, comets with different shapes reflecting progressive stages of DNA fragmentation were observed. Of these, six distinct shapes were identified and divided into three different categories based on the extent of fragmentation. Type A comets had a large head separated by a narrow “neck” region from an oval bulging tail that indicated initiation of fragmentation. Type B and C comets had a constantly diminishing head associated with a corresponding expansion of the tail and reflected intermediate and late stages of fragmentation, respectively. Type A and B comets appeared at a high frequency during early time points (1–6 h), whereas type C comets that indicated late stages of fragmentation were observed only after extended treatment (24 h). As a result, an elaborate kinetics of the progression of DNA fragmentation could be obtained.

Conclusion

The present single‐cell gel electrophoresis assay offers a significant improvement in monitoring the kinetics of DNA fragmentation induced during programmed cell death. Coupled with its simplicity and the ability to detect responses of small cell subpopulations, this method can be used for a reliable and sensitive analysis of the progression of cell death in different cell types and treatment conditions. © 2004 Wiley‐Liss, Inc.
The necessity of observing real life situations: Palestinian‐Israeli violence as a laboratory for learning about social behaviour
Authors: Bar‐Tal Daniel
Published: August 2004

Social psychology emerged in the early part of the last century as a distinct discipline that focused on the study of social behaviour of individuals and collectives. Over time, however, social psychology has relatively ignored the ‘social’ part of the equation and has become mainly concerned with individual behaviour. The major part of social psychological research was carried out in the artificial context of the experimental laboratory. Studying social behaviour in real life contexts is essential, not only to return social psychology to its roots, but also to ensure that our contributions are both theoretically rich and socially valuable. Observation of real life situations is essential if we want to advance our understanding of how individuals and collectives behave. To illustrate the importance of a contextually rich social psychology and the usefulness of natural observations, the recent violent confrontation between the Israelis and the Palestinians is described and analysed, focusing on social behaviours of Israeli Jews. In conclusion, it is argued that social psychology should strive towards equilibrium between natural and experimental approaches, between personal and contextual emphases and between micro and macro perspectives. Copyright © 2004 John Wiley & Sons, Ltd.
Implications for the future of service delivery system reform
Authors: Morrill William A.
Published: August 2004

This chapter discusses the interaction between policy, program development, and evaluation in the reform of the human services delivery system with specific focus on the evaluation challenges and approaches to them.
An Unusual Domino Retro‐Ene–Conia Reaction: Regio‐ and Stereoselective One‐Carbon Ring Expansion of Fenchol Derivatives
Authors: Rüedi Georg , Laikov Dimitri N. , Hansen Hans‐Jürgen
Published: August 2004

The 2‐exo‐substituted fenchol derivatives 1–7, easily prepared from (−)‐fenchone in good‐to‐excellent yields, were pyrolyzed by dynamic gas‐phase thermo‐isomerization (DGPTI). At temperatures of ca. 620°, the substrates with a hydroxyallyl (1–4) or a hydroxypropargyl moiety (6) underwent an initial retro‐ene reaction under cleavage of the C(2)C(3) bond to form enol‐ene intermediates with no loss of optical activity. These intermediates then experience either tautomerization to the corresponding α,β‐unsaturated ketones or subsequent Conia rearrangement under one‐carbon ring expansion of the fenchone system to a bicyclo[3.2.1]octane framework. In the case of the isopropenyl substrate 3, the sterically crowded Conia product underwent a new type of ‘deethanation’ reaction by stepwise loss of two Me radicals, giving rise to the thermodynamically favored enone 21. A similar relaxation behavior was observed in the case of the ethynyl substrate 6, which showed a remarkable 1,3‐Me shift after the Conia reaction, leading to the α,β‐unsaturated cyclic ketone 25. The homolytic cleavage of the weakest single bond in 1–3 turned out to be a competing reaction pathway. Intramolecular H‐abstraction within the generated diradical intermediates produced the monocyclic ketones 8, 16, and 19, besides the products obtained by tautomerization and Conia reaction. In contrast, a Ph substituent at C(2) in 7 allowed only the passage through a diradical species to provide phenone 26, which was converted by regioselective Baeyer–Villiger oxidation to the optically active cyclopentanol 29. Both reaction channels, the domino retro‐ene–Conia rearrangement and the diradical‐promoted H‐transfer, have been shown to proceed highly stereoselectively. The absolute configuration of the newly formed stereogenic centers in all compounds was assigned by 1H‐NOE experiments. The reaction mechanism of the novel domino retro‐ene–Conia reaction was established by both a series of 2H‐ and 13C‐labeling experiments, as well as by a detailed computational analysis.
Berchemins A and B: Novel Enzyme‐Inhibiting Dimeric Lignan Glycosides from Berchemia pakistanica
Authors: Mukhtar Naveen , Malik Abdul , Iqbal Kiran , Tareen Rasool Bakhsh , Khan Shamsun Nahar , Nawaz Sarfraz Ahmad , Choudhary M. Iqbal
Published: August 2004

Phytochemical investigation of Berchemia pakistanica by extensive bioassay‐guided fractionation resulted in the isolation of two novel dimeric lignan glycosides, berchemin A (1) and berchemin B (2), and of the four known compounds β‐sitosterol, β‐sitosterol 3‐O‐β‐D‐glucoside, vavain, and daphnetin 8‐β‐D‐glucoside. The structures of 1 and 2 were elucidated with the help of modern spectroscopic techniques, and the known compounds were identified by comparing their spectral data with those reported in the literature. Berchemins A and B were found to inhibit lipoxygenase and α‐glucosidase enzymes in a concentration‐dependent fashion.
Natural p‐Menthene Monoterpenes: Synthesis of the Enantiomeric Forms of Wine Lactone, Epi‐wine Lactone, Dill Ether, and Epi‐dill Ether Starting from a Common Intermediate
Authors: Serra Stefano , Fuganti Claudio
Published: August 2004

A concise synthesis of the enantiomeric forms of wine lactone (5), epi‐wine lactone (14), dill ether (6), and epi‐dill ether (15) was accomplished starting from the enantiomeric forms of p‐mentha‐1,8(10)‐diene‐3,9‐diol (8) (Scheme 3). The latter compounds were previously prepared in high optical purity by means of lipase‐mediated kinetic resolution, and they were the common building blocks for this divergent synthesis. The key steps were a number of chemo‐ and diastereoselective reactions of which the oxidation of 8 to the lactone 7 (see Table 1), the diastereoselective reduction of 7 to the lactones 14 and 5 (see Table 2), and the reduction of enol‐ether 16 to ether 15 were studied comprehensively. The effectiveness of this new synthetic approach allows the preparation of the title p‐menthane monoterpenes, which are of considerable interest to the perfume industry, in high yield and in a few simple experimental steps.
Expression of HYAL2 mRNA, hyaluronan and hyaluronidase in B‐cell non‐Hodgkin lymphoma: Relationship with tumor aggressiveness
Authors: Bertrand Philippe , Courel Marie‐Noëlle , Maingonnat Catherine , Jardin Fabrice , Tilly Hervé , Bastard Christian
Published: August 2004

Hyaluronidases and their substrate, hyaluronan (HA), were mainly explored in solid tumors but rarely in hematologic malignancies. While HA involvement was demonstrated in invasion and metastasis in most cases of solid tumors, the role of hyaluronidases in cancer progression remains controversial. One of the hyaluronidases, HYAL2, is suspected to be involved in the first step of HA degradation. In this work, HYAL2 mRNA, HA and total hyaluronidases expression were examined in lymphoma tissue extracts and correlated to the lymphoma subtype. Real‐time RT‐PCR was performed to evaluate HYAL2 mRNA. HA and hyaluronidase were assayed by enzyme‐linked sorbent assay. Our results showed that HYAL2 mRNA expression was correlated to lymphoma diagnosis (p = 6 × 10−3) and was significantly lower in high‐grade lymphoma, i.e., diffuse large B‐cell diffuse lymphomas (DLBCLs). Several forms of hyaluronidase were detected by zymography and total hyaluronidase activity detected in tissue extracts was not significantly different according to tumor grade. HA levels also correlated to lymphoma subtype (p = 1 × 10−5) and were higher in DLBCLs. Moreover, HYAL2 mRNA and HA expressions were inversely correlated (p = 0.035). HYAL2 gene is localized on chromosome 3p21, which contains candidates tumor suppressor genes. Our results suggest that HYAL2 may have a prognostic significance in lymphomas and an antioncogenic activity. Conversely, HA overexpression in high‐grade lymphomas is in favor of its involvement in tumor development and could provide a useful target for lymphoma therapy using HA‐binding peptides.
Seasonal Variation of the Macrozoobenthic Community Structure at Low Salinities in a Mediterranean Lagoon (Monolimni Lagoon, Northern Aegean)
Authors: Kevrekidis Theodoros
Published: August 2004

The macrozoobenthic community structure and dynamics at low salinities (0.3–6 psu) in a Mediterranean lagoon (Monolimni lagoon) were investigated. Samples were collected monthly from February 1998 to February 1999 at two sampling stations. Community structure was analyzed by means of uni‐ and multivariate methods. 21 taxa were collected; the amphipod Corophium orientale and the gastropod Ventrosia maritima dominated the assemblages. Total abundance peaked (50,000–60,000 individuals m–2) in mid or late autumn. Community structure showed an almost even seasonal periodicity; seasonal changes were mainly derived from the intense variation in abundance of most species and the non–occurrence of a few ones (e.g. Corophium insidiosum, Polydora ciliata) in spring and summer. Non– occurrence, which led to a depression of the most diversity indices, was possibly the only direct impact of the extremely low salinities (~0.3 psu) on community structure. The main structuring factors of the community in the deeper outer part of the lagoon were water temperature and depth, and in the innermost part, where a Ruppia maritima meadow occurred, were water temperature and predation pressure by crabs (Carcinus aestuarii) and gobies (Knipowitchia caucasica). A temporary decline in total abundance in summer followed an increase in abundance of these predators. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Adaptations of an Emergence Trap for Use in Tropical Streams
Authors: Freitag Hendrik
Published: August 2004

An emergence trap based on the MPI Schlitz model was designed for use in the tropics and it was tested over one year in Palawan, the Philippines. Instructions for construction and use are given here. Only commonly available materials were used, except for the collection assembly made of UV‐light permeable acrylic glass. Heavy and bulky assemblies were avoided to enable easy transportation in the field. A special modification allows a fast and easy replacement of the screen when damaged by flooding, as is often required when traps are used downstream of headwaters. This type of trap also allows sampling of a wide littoral strip. Problems concerning the use of emergence traps in the humid tropics are discussed based on experience at different longitudinal stream sections in Palawan. The results presented here suggest that this trap should be used especially for qualitative or semi‐quantitative approaches. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Composition and Longitudinal Patterns of Aquatic Insect Emergence in Small Rivers of Palawan Island, the Philippines
Authors: Freitag Hendrik
Published: August 2004

This study presents the first emergence trap samples from streams in the Philippines and Greater Sunda. Aquatic insect emergence from two small rivers and longitudinal patterns including estuaries are compared. A decline of total emergence towards estuaries was observed, affecting all major orders. Diptera, namely Chironomidae, dominated all sites. High abundances in Ceratopogonidae, Odonata, and Coleoptera were found, compared to other emergence studies from tropical and temperate latitudes. Ephemeroptera displayed a highly variable contribution to the emergence from Palawan as well as in other comparative studies either supported by the appropriate conditions for certain functional groups or limited by environmental variables such as pH. Trichoptera are likely to tolerate a wider range of environmental conditions and they are consequently able to fill further niches where Ephemeroptera are under‐represented. Except for scarce abundances of Plecoptera observed in this and other studies from the tropics, no substantial differences in emergence composition at order level existed between temperate and tropical rivers, however, with a remarkable local variation. Components of riparian and non‐aquatic insects and non‐emergent fauna contributing to the collections are discussed based on trap features. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)