bluuebunny/crossref_metadata_2025 huggingface dataset:
Halogenated monoterpene production by microplantlets of the marine red alga Ochtodes secundiramea within an airlift photobioreactor under nutrient medium perfusion
Authors: Polzin Jason P. , Rorrer Gregory L.
Published: March 2003

Macrophytic marine red algae are a unique source of novel and bioactive terpenoids, including halogenated monoterpenes. Biomass and halogenated monoterpene production by regenerated microplantlet suspension cultures derived from the red alga Ochtodes secundiramea were studied within a perfusion airlift photobioreactor. Photobioreactor cultivations were carried out at 26°C, 140 μE m−2s−1 light intensity, 0.3 air L−1 culture min−1 aeration (3500 ppm CO2), and ESS/seawater medium perfusion rate of 0.2 L medium L−1 culture d−1. Macronutrient concentrations in the perfusion medium were adjusted to provide nitrate delivery rates of 0.0063, 0.077, and 0.74 mmol L−1 d−1 at a fixed N:P ratio of 19:1. Growth was maximized at the highest nutrient delivery rate, where 10 g dry biomass L−1 culture was achieved after 30 days of cultivation. GC‐MS analysis of dichloromethane extracts from cell biomass revealed that O. secundiramea microplantlets produced myrcene, three acyclic halogenated monoterpenes (10‐bromomyrcene, 10‐bromo‐7‐chloromyrcene, 3,10‐dibromomyrcene), and one cyclic halogenated monoterpene (6‐bromo‐1,2,8‐trichloro‐3,4‐ochtodene). 10E‐bromomyrcene levels were much higher than those of its isomer 10Z‐bromomyrcene, demonstrating stereoselective halogenation. Maximum yields of 10E‐bromomyrcene and 6‐bromo‐1,2,8‐trichloro‐3,4‐ochtodene were 15 and 13 μmol/g dry cell mass, respectively. Increasing the rate of nutrient delivery increased the accumulation of myrcene and 10‐bromomyrcene during the first 14 days in culture. Furthermore, the yield selectivity toward higher halogenated monoterpenes increased as the rate of nutrient delivery decreased. From this data, a biogenic scheme was proposed where cyclic and acyclic halogenated monoterpenes are derived from sequential halogenation of myrcene, their common precursor. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 82: 415–428, 2003.
Business conceptions of sustainable development: a perspective from critical theory
Authors: Springett Delyse
Published: March 2003

The discourse of sustainable development reflects the particular interests and ‘stakes in the ground’ of groups, including corporates, who contest the definition of the concept. Perspectives from critical theory, supplemented by Foucauldian concepts, expose the role of power and hegemony in that contestation. Applied to understandings of sustainable development held by corporate managers, they reveal conceptions largely based in eco‐modernism and ‘political sustainability’. The preliminary outcomes of research in progress that is taking a dialectical approach to the examination of the discourse of sustainable development in New Zealand companies are discussed. A case is made for maintaining greater discursivity in corporations and the broader setting, since, in a Foucauldian sense, it is the discourse that is producing sustainable development. It is argued that the predominating ‘green business’ discourse engages only superficially with sustainable development, and that what is now needed is a discourse of business and sustainable development framed in critical theory. Copyright © 2003 John Wiley & Sons, Ltd. and ERP Environment
Corporate environmental reporting: what's in a metric?
Authors: Scott Marshall R. , Brown Darrell
Published: March 2003

Although there has been increased attention to corporate environmental reports (CERs), there has yet to be a close examination of the metrics used in these reports. Metrics do not address the content of CERs, but, perhaps more importantly, metrics provide the means for conveying the content. In this paper, we analyze metrics used in 79 corporations' recent CER reports. We define and use an 'environmental sustainability' lens, and apply two environmental metrics taxonomies to CER metrics. We also consider the implications of key internal and external firm factors on CER metrics. Our findings suggest that (i) firms' compliance with ISO 14001 increases the presence of future oriented metrics, (ii) a majority of CER content uses lagging metrics with descriptive and operational performance information, (iii) larger firms are more likely than smaller firms to use future oriented metrics and (iv) there are noticeable differences across countries/regions in terms of CER metrics. Several important issues seem evident from the study. First, the metrics most commonly used in CERs provide little information about future performance. Second, the majority of metrics describe operations performance rather than environmental impact. Third, even though the sample was chosen based on a priori indicators of corporate environmental awareness, only about half of the companies sampled had a CER available. Copyright © 2003 John Wiley & Sons, Ltd. and ERP Environment
D‐Tyrosine as a Chiral Precusor to Potent Inhibitors of Human Nonpancreatic Secretory Phospholipase A2 (IIa) with Antiinflammatory Activity
Authors: Hansford Karl A. , Reid Robert C. , Clark Chris I. , Tyndall Joel D. A. , Whitehouse Michael W. , Guthrie Tom , McGeary Ross P. , Schafer Karl , Martin Jennifer L. , Fairlie David P.
Published: March 2003

Few reported inhibitors of secretory phospholipase A2 enzymes truly inhibit the IIa human isoform (hnpsPLA2‐IIa) noncovalently at submicromolar concentrations. Herein, the simple chiral precursor D‐tyrosine was derivatised to give a series of potent new inhibitors of hnpsPLA2‐IIa. A 2.2‐Å crystal structure shows an inhibitor bound in the active site of the enzyme, chelated to a Ca2+ ion through carboxylate and amide oxygen atoms, H‐bonded through an amide NH group to His48, with multiple hydrophobic contacts and a T‐shaped aromatic‐group–His6 interaction. Antiinflammatory activity is also demonstrated for two compounds administered orally to rats.
Effect of Organic Additives on Electrochemical Surface Precipitation and Polymorphism of CaCO3
Authors: Granit N.F. , Korin E. , Bettelheim A.
Published: March 2003

The inhibition effect of three organic additives on the precipitation and polymorphism of CaCO3 deposited on gold surfaces was investigated using electrochemical and microscopic techniques. Additives, two polyacrylic acid (PAA) polymers with different molecular weights (Mw 2100, Mw 30,000), and 1,2,4,5‐benzenetetracarboxylic acid (BTCA), were either added to the solution before or during deposition. In the presence of 100 ppm of one of the three additives in solution, almost no scale was observed on the surface for at least 24 hours. In the presence of lower concentrations of PAA Mw 2100, only distorted calcite crystals were obtained while with PAA Mw 30,000 the polymorph was spherical vaterite. A mixture of calcite and vaterite was observed with the BTCA additive. Addition of the polymers inhibits further nucleation and growth even if added after partial deposition of CaCO3 while BTCA has no effect once nucleation has started. The results indicate that the inhibit
ion effect of the PAA polymers is due to adsorption on the electrode surface while the effect of BTCA is related to chelation of calcium ions in solution.
COVPROC method: strategy in modeling dynamic systems
Authors: Reinikainen Satu‐Pia , Höskuldsson Agnar
Published: March 2003

A new procedure to select subsets in sequential dynamic systems is presented. Subsets of variables and samples to be included in a PLS model are considered. The approach is a combination of PLS analysis and standard regression methods. It is based on the H‐principle of mathematical modeling, and therefore the aim is to balance improvement in fit and precision. One of the main aspects in the subset selection procedure is to keep the score space as large and as sensible as possible to gain a stable model. The procedure is described mathematically and demonstrated for a dynamic industrial case. The method is simple to apply and the motivation of the procedure is obvious for industrial applications. It can be used e.g. when modeling on‐line systems. Copyright © 2003 John Wiley & Sons, Ltd.
Aerosolabscheidung aus Rauchgasen zukünftiger Kraftwerke – Grundlagenuntersuchungen mittels Magnetic Resonance Imaging
Authors: van Buren V. , Reimert R.
Published: March 2003

No abstracts.
Selbstzündverhalten von Heizöl EL/Luft‐Gemischen bei atmosphärischem Druck
Authors: Steinbach N. , Lucka K. , Köhne H.
Published: March 2003

No abstracts.
Low molecular weight inhibitors of matrix metalloproteinases can enhance the expression of matrix metalloproteinase‐2 (gelatinase A) without inhibiting its activation
Authors: Kerkvliet Erika H. M. , Jansen Ineke D. C. , Schoenmaker Ton A. M. , Docherty Andy J. P. , Beertsen Wollter , Everts Vincent
Published: March 2003

BACKGROUND

In the current study, the authors investigated the effects of synthetic low molecular weight inhibitors of matrix metalloproteinases (MMPs) on the expression and activation of MMP‐2 in a three‐dimensional tissue system.

METHODS

Rabbit periosteal explants were cultured with or without various concentrations of the MMP inhibitors CT1166, CT1399, or CT1746, and conditioned media and tissue extracts were analyzed for the expression and activity of MMP‐2.

RESULTS

The data showed that blocking the activity of all MMPs with relatively high inhibitor concentrations completely prevented the conversion of pro‐MMP‐2 into its active form and that the level of protein was decreased. Selective inhibition of the activity of gelatinases (MMP‐2 and MMP‐9) by using low inhibitor concentrations, however, induced a higher level of active MMP‐2 and increased its expression significantly.

CONCLUSIONS

The current observations indicate that selective inhibitors of MMPs affect the expression and activity of MMP‐2, thus providing clues regarding the differing effects such inhibitors appear to have when applied in vivo. Cancer 2003;97:1582–88. © 2003 American Cancer Society.

DOI 10.1002/cncr.11193
Tumor mitotic rate is a more powerful prognostic indicator than ulceration in patients with primary cutaneous melanoma
Authors: Azzola Manuela F. , Shaw Helen M. , Thompson John F. , Soong Seng‐jaw , Scolyer Richard A. , Watson Geoffrey F. , Colman Marjorie H. , Zhang Yuting
Published: March 2003

BACKGROUND

The current study was performed to determine whether tumor mitotic rate (TMR) is a useful, independent prognostic factor in patients with localized cutaneous melanoma.

METHODS

From the Sydney Melanoma Unit database, 3661 patients with complete clinical information and details of primary tumor thickness, ulcerative state, and TMR were studied. TMR was expressed as mitoses per mm2in the dermal part of the tumor in which most mitoses were seen, as recommended in the 1982 revision of the 1972 Sydney classification of malignant melanoma. To determine which was the more prognostically useful method of grouping TMR, two separate methods (A and B) were used. Factors predicting melanoma‐specific survival were analyzed using the Cox proportional hazards regression model.

RESULTS

Patients with a TMR of 0 mitoses/mm2had a significantly better survival than those with 1 mitosis/mm2(P< 0.0001) but no significant survival differences were recorded for the stepwise increases from 1–2, 2–3, 3–4, and 4–5/mm2. Tumor thickness, ulceration, and TMR were closely correlated, whether TMR was grouped using Method A (0, 1–4, 5–10, and ≥ 11 mitoses/mm2) or Method B (0–1, 2–4, and ≥ 5 mitoses/mm2). However, Cox regression analysis indicated that the TMR was a highly significant independent prognostic factor, particularly when grouped according to Method A, in which it was second only to tumor thickness as the most powerful predictor of survival (P< 0.0001).

CONCLUSIONS

TMR is an important independent predictor of survival for melanoma patients. If confirmed by studies from other centers, it has the potential to further improve the accuracy of melanoma staging, as well as to define more rigidly the risk categories for patients entering clinical trials. Cancer 2003;97:1488–98. © 2003 American Cancer Society.

DOI 10.1002/cncr.11196
Down‐regulation of fragile histidine triad expression in prostate carcinoma
Authors: Fouts Rebecca L. , Sandusky George E. , Zhang Shaobo , Eckert George J. , Koch Michael O. , Ulbright Thomas M. , Eble John N. , Cheng Liang
Published: March 2003

BACKGROUND

The fragile histidine triad (FHIT) gene is a tumor suppressor gene that belongs to the histidine triad family of nucleoside binding proteins. The gene encompasses the common human chromosomal fragile site, the FRA3B locus at chromosome 3p14.2, and is expressed in most normal adult tissues and tumor cell lines. Numerous studies have indicated that the FHIT gene on chromosome 3p may play an important role in human neoplasia, although very few studies have investigated the FHIT gene in prostate carcinoma.

METHODS

Using immunohistochemical analyses, the authors studied the expression of FHIT in prostate tumors from 84 radical prostatectomy specimens to determine whether there were any correlations between FHIT expression and various clinicopathologic characteristics.

RESULTS

The percentages of cells stained with antibody to FHIT were significantly lower overall for tumor cells compared with normal cells (P = 0.0001). FHIT immunostaining intensity also was significantly lower for tumor cells compared with normal cells (P = 0.0001). A weak but statistically significant correlation (P = 0.045) was demonstrated with the presence of extraprostatic extension in the patient samples. No other significant correlation was seen between the percentage of cells stained for FHIT or FHIT immunostaining intensity and Gleason grade, tumor stage, tumor size, lymph node metastasis, surgical margins, vascular invasion, perineural invasion, or the presence of high‐grade prostatic intraepithelial neoplasia.

CONCLUSIONS

The data presented indicate a down‐regulation of the FHIT tumor suppressor gene in prostate carcinoma and, thus, propose a potential target for therapeutic intervention. Cancer 2003;97:1447–52. © 2003 American Cancer Society.

DOI 10.1002/cncr.11201
Heterogeneous expression of SNAP‐25 and synaptic vesicle proteins by central and peripheral inputs to sympathetic neurons
Authors: Gibbins Ian L. , Jobling Phillip , Teo Ee Hiok , Matthew Sue E. , Morris Judy L.
Published: March 2003

Neurons in prevertebral sympathetic ganglia receive convergent synaptic inputs from peripheral enteric neurons in addition to inputs from spinal preganglionic neurons. Although all inputs are functionally cholinergic, inputs from these two sources have distinctive neurochemical and functional profiles. We used multiple‐labeling immunofluorescence, quantitative confocal microscopy, ultrastructural immunocytochemistry, and intracellular electrophysiologic recordings to examine whether populations of inputs to the guinea pig coeliac ganglion express different levels of synaptic proteins that could influence synaptic strength. Boutons of enteric intestinofugal inputs, identified by immunoreactivity to vasoactive intestinal peptide, showed considerable heterogeneity in their immunoreactivity to synaptosome‐associated protein of 25 kDa (SNAP‐25), synapsin, synaptophysin, choline acetyltransferase, and vesicular acetylcholine transporter. Mean levels of immunoreactivity to these proteins were significantly lower in terminals of intestinofugal inputs compared with terminals of spinal preganglionic inputs. Nevertheless, many boutons with undetectable levels of SNAP‐25 immunoreactivity formed morphologically normal synapses with target neurons. Treatment with botulinum neurotoxin type A (20–50 nM for 2 hours in vitro) generated significant cleavage of SNAP‐25 and produced similar dose‐ and time‐dependent inhibitions of synaptic transmission from all classes of inputs, regardless of their mean level of SNAP‐25 expression. The simplest interpretation of these results is that only synaptic boutons with detectable levels of SNAP‐25 immunoreactivity contribute significantly to fast cholinergic transmission. Consequently, the low synaptic strength of intestinofugal inputs to final motor neurons in sympathetic pathways may be due in part to the low proportion of their boutons that express SNAP‐25 and other synaptic proteins. J. Comp. Neurol. 459:25–43, 2003. © 2003 Wiley‐Liss, Inc.
Neurochemically distinct classes of myenteric neurons express the μ‐opioid receptor in the guinea pig ileum
Authors: Ho Anthony , Lievore Annarita , Patierno Simona , Kohlmeier Sean E. , Tonini Marcello , Sternini Catia
Published: March 2003

The μ‐opioid receptor (μOR), which mediates many of the opioid effects in the nervous system, is expressed by enteric neurons. The aims of this study were to determine whether 1) different classes of myenteric neurons in the guinea pig ileum contain μOR immunoreactivity by using double‐ and triple‐labeling immunofluorescence and confocal microscopy, 2) μOR immunoreactivity is localized to enteric neurons immunoreactive for the endogenous opioid enkephalin, and 3) μOR immunoreactivity is localized to interstitial cells of Cajal visualized by c‐kit. In the myenteric plexus, 50% of μOR‐immunoreactive neurons contained choline acetyltransferase (ChAT) immunoreactivity, whereas about 43% of ChAT‐immunoreactive neurons were μOR immunoreactive. Approximately 46% of μOR myenteric neurons were immunoreactive for vasoactive intestinal polypeptide (VIP), and about 31% were immunoreactive for nitric oxide synthase (NOS). μOR immunoreactivity was found in about 68% of VIP‐containing neurons and 60% of NOS‐immunoreactive neurons. Triple labeling showed that about 32% of μOR neurons contained VIP and ChAT immunoreactivities. The endogenous opioid enkephalin (ENK) was observed in about 30% of μOR neurons; conversely, 48% of ENK neurons contained μOR immunoreactivity. μOR was not detected in neurons containing calbindin, nor in interstitial cells of Cajal. μOR‐immunoreactive fibers formed a dense network around interstitial cells of Cajal in the deep muscular plexus. This study demonstrates that μOR is expressed by neurochemically distinct classes of myenteric neurons that are likely to differ functionally, is colocalized with the endogenous opioid ENK, and is not expressed by interstitial cells of Cajal. J. Comp. Neurol. 458:404–411, 2003. © 2003 Wiley‐Liss, Inc.
A current‐mode 1kHz/10kHz low‐voltage integrated analogue PLL for lock‐in portable applications
Authors: Ferri Giuseppe , Massa Ernesto
Published: March 2003

We present a low‐supply voltage (2V) low‐power consumption (500W) analogue phase‐locked loop (PLL), working at two low frequencies (1 and 10kHz), to be used in an integrated lock‐in amplifier. An externally settable control bit allows the switching operation between the two different frequencies. The circuit has been designed in a standard 0.6–m CMOS technology and differs from the standard analogue PLL architectures for the current mode implementation of both the loop filter and of the oscillator. Three different locked waveforms (sinusoidal, triangular, squared) can be obtained at the PLL output. Simulation results, obtained through the use of PSPICE and using accurate transistor models, will be proposed. The pull‐in ranges are about ±250Hz around 1 and ±1.3kHz around 10kHz, with pull‐in times of about 10 and 4ms, respectively. Copyright © 2003 John Wiley & Sons, Ltd.
Detection of human papillomavirus in sanitary napkins: A new paradigm in cervical cancer screening
Authors: Tong Tommy R. , Chan Olivia W.H. , Chow Tat‐Chong , Yu Vivian , Leung Kai‐Man , To Shui‐Him
Published: March 2003

Human papillomavirus was successfully detected by polymerase chain reaction (PCR) in menstrual blood or vaginal discharge collected in sanitary napkins in 100% of 17 women having koilocytosis, cervical intraepithelial neoplasia, or squamous carcinoma. We advocate this form of cervical cancer screening because of its high sensitivity and acceptance by patients. Diagn. Cytopathol. 2003;28:140–141. © 2003 Wiley‐Liss, Inc.
Development of transgenic chickens expressing bacterial β‐galactosidase
Authors: Mozdziak P.E. , Borwornpinyo S. , McCoy D.W. , Petitte J.N.
Published: March 2003

Replication‐defective retroviral vectors are efficient vehicles for the delivery of exogenous genes, and they may be used in the generation of transgenic animals. The replication‐defective retroviral SNTZ vector carrying the lacZ gene with a nuclear localized signal was injected into the subgerminal cavity of freshly laid eggs. Subsequently, the eggs were allowed to hatch, and the chickens were screened for the lacZ gene by using the polymerase chain reaction. Eight of 15 male chickens that survived to sexual maturity contained the lacZ gene in their semen. Subsequently, these males were mated with wild‐type female chickens. From one of the eight lacZ‐positive G0 males, two lacZ‐positive male chickens were produced from a total of 224 G1 progeny for a germline transmission rate of 0.89%. Both G1 male chickens carrying the lacZ gene were mated with wild‐type female chickens and 46.5% of the G2 progeny contained the lacZ gene, which is consistent with the expected Mendelian 50% ratio for a heterozygous dominant allele. The product of the lacZ gene, nuclear localized β‐galactosidase, was expressed in primary myoblast cultures derived from G2 chickens, and it was also expressed in whole G2 chicken embryos. Developmental Dynamics, 2003. © 2003 Wiley‐Liss, Inc.
Determination of retinal cell fates is affected in the absence of extraocular striated muscles
Authors: Kablar Boris
Published: March 2003

Neural retinas of genetically modified mouse embryos and fetuses entirely lacking extraocular striated muscles (designated as Myf5‐/‐:MyoD‐/‐ or amyogenic) are used to study in vivo the role of extraocular muscle (i.e., fetal ocular movements) in the genesis of retinal cell diversity. Although retinal lamination and the total number of cells per retinal layer appeared unaffected in amyogenic fetuses, electron microscopy and histochemistry revealed the absence of cholinergic amacrine cell type. By contrast, the amounts of other amacrine cell subpopulations (calretinin‐, tyrosine hydroxylase‐, and parvalbumin‐expressing) were increased, whereas the amounts of Islet1/2‐expressing retinal ganglion cells were decreased. Surprisingly, it was not possible to detect any change in proliferation or cell death. Consistently, the number of progenitors for retinal ganglion cells (nestin‐expressing precursors) were increased, whereas the amounts of precursors for amacrine cells (syntaxin‐ and VC1.1‐expressing precursors) were decreased in the mutant retinas. The difference in requirements for extraocular muscle support in regulation of precise ratios of retinal neuronal cell types suggests an essential role of extrinsic cues in the determination of retinal cell fates. Taken together, it appears that patterning mechanisms intrinsic to the neural retina specify the basic organization of retinal spatial organization (e.g., retinal layers and total number of cells). However, extrinsic cues seem to change intrinsic properties (e.g., competence) of retinal progenitor cells and influence the ratios of the differentiated cell types (i.e., cell fate choice) they produce. Developmental Dynamics 226:000–000, 2003. © 2003 Wiley‐Liss, Inc.
Preoccupation, food, and failure: An investigation of cognitive performance deficits in dieters
Authors: Jones Nicola , Rogers Peter J.
Published: March 2003

Objective

This study tested two explanations of the cognitive performance deficits of dieters. First, these deficits are due to the metabolic consequences of food deprivation. Second, they are psychological in origin (i.e., preoccupation with dieting‐related thoughts during dieting depletes the cognitive resources available for nondieting tasks).

Method

Dieters and nondieters performed a battery of cognitive tasks before and after consumption of a high‐energy chocolate bar. Eating the chocolate bar was expected to reverse the effects of food deprivation but, as a “diet‐threatening” food, it could increase dieting‐related preoccupation.

Results and Conclusions

The results supported the latter prediction. Performance on a memory task was further impaired, rather than improved, after food consumption in dieters. In addition, dieters experienced a significant increase in the number of food and dieting‐related thoughts. Some dieters attributed their poorer performance to distraction by these thoughts. Marked individual differences in thought content accounted for some of the variability in the effects on performance. © 2003 by Wiley Periodicals, Inc. Int J Eat Disord 33: 185–192, 2003.
Eating behaviors and attitudes in women with infertility: A controlled study
Authors: Fassino Secondo , Garzaro Lorenzo , Pierò Andrea , Daga Giovanni Abbate , Bulik Cynthia
Published: March 2003

Objective

Infertility arises from a complex pathogenic process in which it is often difficult to identify etiology. Psychological and behavioral factors may play a role in some cases of infertility. The extent to which eating behaviors and attitudes contribute to infertility is unknown.

Method

In this study, 120 subjects with infertility, 80 fertile women, and 90 patients suffering from anorexia nervosa, restricting subtype (AN‐R), were assessed with the Eating Disorder Inventory (EDI‐2).

Results

The EDI‐2 highlights differences among patients with AN‐R and fertile and infertile subjects. Infertile patients had higher scores on the Interpersonal Distrust, Interoceptive Awareness, and Maturity Fears EDI‐2 subscales than fertile subjects. Logistic regression identifies the independent variables of interpersonal distrust, interoceptive awareness, maturity fears, and asceticism as predictors of infertility.

Discussion

Infertile patients without eating disorders share some psychological features of women with AN. These features do not include disturbed eating attitudes and behaviors, but rather feelings of inadequacy, insecurity, and maturity fears. © 2003 by Wiley Periodicals, Inc. Int J Eat Disord 33: 178–184, 2003.
Perfect linear phase IIR filter with flat passband amplitude and equiripple stopband attenuation characteristics
Authors: Tamura Keimei , Matsumura Shuitsu , Kato Kyoko , Takebe Tsuyoshi
Published: March 2003

It is known that several techniques for real‐time implementation of perfect linear phase IIR filters have been developed, achieving higher computational efficiency than conventional FIR filters with similar performance. This paper presents new methods for designing transfer function of perfect linear phase IIR filters with flat passband and equiripple stopband amplitude characteristics. The proposed methods are based on the formulation of an eigenvalue problem for low‐pass filters and a set of linear equations for bandpass filters. In wide‐band, narrow transition cases, a bump in amplitude may appear near the band edge. However, it can be removed by specifying amplitude to unity at several frequencies. The methods are computationally efficient. Design examples of low‐pass and bandpass filters are shown. © 2003 Wiley Periodicals, Inc. Electron Comm Jpn Pt 3, 86(7): 15–29, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/ecjc.10071
Weak agonist self‐peptides promote selection and tuning of virus‐specific T cells
Authors: Saibil Samuel D. , Ohteki Toshiaki , White Forest M. , Luscher Mark , Zakarian Arsen , Elford Alisha , Shabanowitz Jeffery , Nishina Hiroshi , Hugo Patrice , Penninger Josef , Barber Brian , Hunt Donald F. , Ohashi Pamela S.
Published: March 2003

Recent progress has begun to define the interactions and signaling pathways that are triggered during positive selection. To identify and further examine self‐peptides that can mediate positive selection, we searched a protein‐database to find peptides that have minimal homology with the viral peptide (p33) that activates a defined P14 transgenic TCR. We identified four peptides that could bind the restriction element H‐2Db and induce proliferation of P14 transgenic splenocytes at high concentration. Two of the four peptides (DBM and RPP) were able to positively select thevirus‐specific TCR in fetal thymic organ culture but were unable to induce clonal deletion. Reverse‐phase HPLC and mass spectrometry demonstrated that these peptides were presented by H‐2Db molecules on thymic epithelial cell lines. We also examined whether the selecting ligands altered T cell responsiveness in vitro. DBM‐selected T cells lost their ability to respond to the positively selecting ligand DBM, whereas RPP‐selected T cells only retainrd their ability to respond to high concentrations of RPP. These results demonstrate that self‐peptides that mediate positiveselection can differentially "tune" the activation threshold of T cells and alter the functional repertoire of T cells.
Toll‐like receptor‐mediated tyrosine phosphorylation of paxillin via MyD88‐dependent and ‐independent pathways
Authors: Hazeki Kaoru , Masuda Naoyo , Funami Kenji , Sukenobu Naoe , Matsumoto Misako , Akira Shizuo , Takeda Kiyoshi , Seya Tsukasa , Hazeki Osamu
Published: March 2003

Toll‐like receptor (TLR)‐mediated recognition of pathogens represents one of the most important mechanisms of innate immunity. A proximal signaling event of TLR is the direct binding of an adaptor protein MyD88 to TLR and recruitment of the IL‐1R‐associated kinase (IRAK). In the present study, we examined the effect of several TLR ligands on protein tyrosine phosphorylation in rat macrophages. Macrophage‐activating lipopeptide‐2 kDa (MALP2) and lipoarabinomannan were used as activators of TLR2, while lipopolysaccharides (LPS) and lipoteichoic acid were used as TLR4 ligands. All these ligands induced tyrosine phosphorylation of proline‐rich tyrosine kinase 2 (Pyk2) and its substrate paxillin, an integrin‐associated focal adhesion adaptor protein, in the macrophages. PP2, an inhibitor of Src family tyrosine kinases, prevented the TLR‐induced phosphorylation of paxillin and Pyk2 without affecting TLR‐induced IRAK activation. MALP2 failed to induce paxillin phosphorylation in the macrophages from MyD88‐knockout mice. In contrast, the effect of LPS weakened, but was still observed even in the MyD88‐deficient cells. Thus, TLR regulate the function of paxillin in an Src family‐dependent mechanism through both MyD88‐dependent and MyD88‐independent pathways.
Superoxide activates very late antigen‐4 on an eosinophil cell line and increases cellular binding to vascular cell adhesion molecule‐1
Authors: Chuang Kuo‐Pin , Tsai Wei‐Shi , Wang Ying‐Jan , Shieh Chi‐Chang
Published: March 2003

The recruitment of eosinophils to the airway is a key event in the pathogenesis of allergy. Very late antigen‐4 (VLA‐4), an integrin ligand for vascular cell adhesion molecule‐1 (VCAM‐1), is expressed on eosinophils. VLA‐4‐mediated adhesion of eosinophils to VCAM‐1 may contribute to their selective recruitment to tissues in allergy. Reactive oxygen species (ROS), including nitric oxide (NO), are abundant in the airway of allergic patients, but their role in pathogenesis of allergy is unclear. In this investigation, we studied the effects of ROS on integrin‐mediated eosinophil adhesion. Recombinant soluble VCAM‐1 and ICAM‐1 were used to test the effects of ROS on the integrin‐mediated adhesion of an eosinophil cell line. We used phorbol 12‐myristate 13‐acetate‐stimulated neutrophils and hypoxanthine to generate superoxide, NO donors as sources of NO, and a static cell‐to‐protein adhesion assay to analyze cellular adhesion. Stimulated neutrophils significantly increased eosinophil binding to VCAM‐1, which was reversed in the presence of superoxide dismutase. Neutrophils from a chronic granulomatous disease patient lacked this activity in enhancing eosinophil adhesion. Our results suggest that the balance between ROS molecules in different tissue microenvironments may change the integrin‐mediated leukocyte adhesion and is likely to be a key factor in leukocyte recruitment in allergic inflammation.
Characterization of antigen‐specific repertoire diversity following in vitro restimulation by a recombinant adenovirus expressing human cytomegalovirus pp65
Authors: Hamel Yamina , Rohrlich Pierre , Baron Véronique , Bonhomme Delphine , Rieux‐Laucat Frederic , Necker Antje , Lemonnier François , Ferradini Laurent , Fischer Alain , Cavazzana‐Calvo Marina
Published: March 2003

Human cytomegalovirus (HCMV) and adenovirus cause significant morbidity and mortality in immunocompromised hosts undergoing allogeneic stem cell transplantation. We have previously establisheda procedure for the generation of polyclonal CTL with specificity against adenovirus and HCMV using a recombinant adenovirus encoding the HCMV pp65 protein (RAdpp65). However, specific CTL expandedafter in vitro culture steps were subjected to several in vitro restimulations and, depending on the protocol adopted, this could lead to a selection bias, compromising the clinical benefit. To determine which part of the memory repertoire is selected after in vitro restimulation, we have followed the specificity and clonal composition of pp65‐peptide‐specific CD8+ T cells in HLA‐A*201 individuals before and after repeated in vitro restimulation of cells with RAdpp65, combining HLA tetrameric complexes and immunoscope analysis. Tetramer staining showed that, after in vitro restimulation, up to 60% of CD8+ T cells were virus‐specific. Immunoscope analysis showed that the predominant TCRBV diversity of pp65‐specific clones was conserved, demonstrating that the memory repertoire was preserved all along the procedure. Altogether, these results suggest that the use of RAdpp65 to induce CMV‐ and adenovirus‐specific CTL maybe appropriate for immunotherapy.
Design and Structural Analysis of Metallamacrocycles Based on Zinc Halides and a V‐Shaped Bismonodentate Ligand of the Cyclophane Type
Authors: Klein Cédric , Graf Ernest , Hosseini Mir Wais , De Cian André , Kyritsakas Nathalie
Published: March 2003

The ligand 1, based on the [1,1,1,1]cyclophane skeleton and bearing two pyridine units as monodentate coordination sites on the same face of the backbone, leads, in the presence of ZnX2 (X = Cl, Br or I) complexes, to three isostructural neutral metallamacrocycles with an M/L ratio of 2:2. All three binuclear complexes have been characterised in the solid state by single‐crystal X‐ray diffraction. All three metallamacrocycles possess a roof‐shaped geometry affording a pocket with a single opening. The latter is occupied by solvent molecules in the solid state. A correlation between the size of the halide and dimensions of the pocket is described. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Bis‐Aminals of Linear Tetraamines: Kinetic and Thermodynamic Aspects of the Condensation Reaction
Authors: Chuburu Françoise , Tripier Raphaël , Baccon Michel Le , Handel Henri
Published: March 2003

The kinetic and thermodynamic aspects of the condensation reaction of dicarbonyl compounds with linear tetraamines were examined in the light of identification of intermediates and DFT calculations. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Xe@cryptophane Complexes with C2 Symmetry: Synthesis and Investigations by 129Xe NMR of the Consequences of the Size of the Host Cavity for Xenon Encapsulation
Authors: Brotin Thierry , Dutasta Jean‐Pierre
Published: March 2003

New cryptophanes‐223 (1), ‐233 (2), and ‐224 (3) with C2 symmetry, bearing different linkers connecting the two cyclotriveratrylene units were synthesized following a multi‐step procedure. The formation of the xenon@cryptophane complexes was investigated by 129Xe NMR spectroscopy. Cryptophanes‐223 (1) and ‐233 (2) complex xenon efficiently in 1,1,2,2‐[D2]tetrachloroethane solution with somewhat lower binding constants (K = 2810 M−1 and 810 M−1 respectively at 278 K) than that observed with cryptophane‐A (K = 3900 M−1). The free and bound guests were observed at room temperature under slow‐exchange conditions on the 129Xe NMR timescale. A linear relationship was observed between the binding constants and the internal volume of the hosts. Under the same conditions, the Xe@cryptophane‐224 [Xe3] complex underwent a fast‐exchange process on the NMR timescale. The decomplexation activation energies Ea, and the associated parameters ΔH‡ and ΔS‡, were determined for the Xe1 and Xe2 complexes from variable temperature 1D‐EXSY experiments. These investigations emphasized the importance of the size of the internal cavity of the cryptophanes for guest encapsulation, and the extreme sensitivity of the 129Xe chemical shift toward slight structural modifications of the atom’s environment. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Synthesis of Polysubstituted Alkenes by Heck Vinylation or Suzuki Cross‐Coupling Reactions in the Presence of a Tetraphosphane−Palladium Catalyst
Authors: Berthiol Florian , Doucet Henri , Santelli Maurice
Published: March 2003

Through the use of [PdCl(C3H5)]2/cis,cis,cis‐1,2,3,4‐tetrakis[(diphenylphosphanyl)methyl]cyclopentane as a catalyst, a range of vinyl bromides undergo Suzuki cross‐couplings with arylboronic acids in good yields. Furthermore, the catalyst can be used at low loadings, even for reactions of sterically hindered substrates. Mediated by this catalyst, stilbene and 1,1‐diphenylethylene undergo Heck reactions with aryl halides to give triarylethylene derivatives. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003)
Ingroup identification as a moderator of positive–negative asymmetry in social discrimination
Authors: Hodson Gordon , Dovidio John F. , Esses Victoria M.
Published: March 2003

Group members typically favour ingroups over outgroups, particularly when distributing positive rather than negative resources. The present investigation examined whether the positive–negative discrimination asymmetry in the minimal group paradigm varies as a function of ingroup identification. After being categorized into arbitrary groups, participants expressing low to high ingroup identification allocated positive, neutral, or negative outcomes on the basis of group membership (i.e. ingroup versus outgroup recipients). The interaction between ingroup identification and outcome valence revealed that identification influenced the magnitude of discrimination asymmetry. Specifically, increases in identification led to discrimination in favour of the ingroup for positive but not negative outcomes. The implications for intergroup behaviour more generally are considered. Copyright © 2002 John Wiley & Sons, Ltd.
Characterization of pharmaceutical drugs by a modified nonaqueous capillary electrophoresis – mass spectrometry method
Authors: Peri‐Okonny Unita L. , Kenndler Ernst , Stubbs R. John , Guzman Norberto A.
Published: March 2003

A simple method for the separation and characterization of a group of nine basic compounds, comprising seven tricyclic antidepressant and two bronchodilator drugs, by nonaqueous capillary electrophoresis (NACE) employing ultraviolet and mass spectrometry detection is described. After optimization of the electrophoresis separation conditions, including the compositions of the electrolyte and the organic solvent, a reliable separation of all nine basic analytes was achieved in 80 mM ammonium formate dissolved in a methanol‐acetonitrite (80:20 v/v) mixture, having an apparent pH of 8.7. The volatile nonaqueous electrolyte system used with a normal electroosmotic flow polarity also provided an optimal separation condition for the characterization of the analytes by mass spectrometry. When results were compared with reversed‐phase gradient and isocratic high‐performance liquid chromatography (HPLC) methods, the NACE method provided greater efficiency, achieving baseline resolution for all nine basic compounds in less than 30 min. The NACE method is suitable for use as a routine procedure for the rapid separation and characterization of basic compounds and is a viable alternative to HPLC for the separation of a wide range of pharmaceutical drugs.
Single‐nucleotide polymorphism analysis by allele‐specific extension of fluorescently labeled nucleotides in a microfluidic flow‐through device
Authors: Russom Aman , Ahmadian Afshin , Andersson Helene , Nilsson Peter , Stemme Göran
Published: March 2003

We describe a microfluidic approach for allele‐specific extension of fluorescently labeled nucleotides for scoring of single‐nucleotide polymorphism (SNP). The method takes advantage of the fact that the reaction kinetics differs between matched and mismatched configurations of allele‐specific primers hybridized to DNA template. A microfluidic flow‐through device for biochemical reactions on beads was used to take advantage of the reaction kinetics to increase the sequence specificity of the DNA polymerase, discriminating mismatched configurations from matched. The volume of the reaction chamber was 12.5 nL. All three possible variants of an SNP site at codon 72 of the p53 gene were scored using our approach. This work demonstrates the possibility of scoring SNP by allele‐specific extension of fluorescently labeled nucleotides in a microfluidic flow‐through device. The sensitive detection system and easy microfabrication of the microfluidic device enable further miniaturization and production of an array format of microfluidic devices for high‐throughput SNP analysis.
An interface chip connection between capillary electrophoresis and thermal lens microscope
Authors: Uchiyama Kenji , Hibara Akihide , Sato Kiichi , Hisamoto Hideaki , Tokeshi Manabu , Kitamori Takehiko
Published: March 2003

A thermal lens microscope (TLM) detection of capillary electrophoresis (CE) utilizing microchip technology was developed. Fused‐silica capillaries with an inner diameter of 50 μm were directly connected to a microchannel in a microchip. The detection limit by TLM was estimated as 2.8×10‐7 absorbance by measuring pure water. The detection limit of derivatized amino acids determined by CE‐TLM was estimated as 2.4×10‐8 M, which was 100 times lower than that of conventional absorbance detection.
The mitochondrial proteins of the neuroblastoma cell line IMR‐32
Authors: Fountoulakis Michael , Schlaeger Ernst‐Jürgen
Published: March 2003

Mitochondrial proteins exert important functions in biological pathways, particularly they are involved in apoptotic processes. We applied proteomics technologies to analyze the mitochondrial proteins of the neuroblastoma cell line IMR‐32, which is often used in apoptosis studies. The proteins were analyzed by two‐dimensional (2‐D) electrophoresis followed by matrix‐assisted laser desorption/ionization‐mass spectrometry (MALDI‐MS). 185 different gene products were identified, of which approximately 55% were enzymes with a broad spectrum of catalytic activities. Sixteen proteins were detected only in this preparation, the others have been detected in two or more protein samples analyzed by MS in our laboratory. The 16 unique gene products were represented by one spot each, whereas most of the frequently detected proteins were represented by multiple spots. In average, approximately 5–10 spots corresponded to one gene product. For two thirds of the proteins identified, an annotation exists in the SWISS‐PROT database about their subcellular location. They are mainly described as mitochondrial, 8 as endoplasmic reticulum, 3 as peroxisomal and only 12 low‐abundance proteins are described as cytosolic proteins. The list includes about 30 unknown, hypothetical or poorly described gene products. Some of them are represented by strong spots and the present study shows that they are indeed expressed and are localized in the mitochondria.
Capillary electrophoresis and fluorescence studies on molecular beacon‐based variable length oligonucleotide target discrimination
Authors: Ramachandran Akhilesh , Zhang Minquan , Goad David , Olah Glenn , Malayer Jerry R. , El Rassi Ziad
Published: March 2003

Molecular beacons (MBs) are oligonucleotide probes having a compact hairpin structure, with a fluorophore attached to one end and a quencher molecule attached to the other end. In its native state, the fluorophore is quenched by virtue of its proximity to the quencher molecule. Upon hybridization with its complementary oligonucleotide target, fluorescence is elicited due to a conformational change that results in separation of the fluorophore and quencher molecule. The present study describes the hybridization interaction of an MB to various complementary target sequences. The effects of temperature and length of complementary target sequences on hybridization were investigated using capillary electrophoresis and solution‐based fluorescence techniques. Hybridization efficiency was dependent on the ability of the target sequences to destabilize the stem region by binding directly to the stem region. Optimal hybridization occurred between 40 and 50°C for all targets tested, with the true target forming a more stable hybrid complex.
Insoluble eggshell matrix proteins – their peptide mapping and partial characterization by capillary electrophoresis and high‐performance liquid chromatography
Authors: Mikšík Ivan , Charvátová Jana , Eckhardt Adam , Deyl Zdeněk
Published: March 2003

Avian eggshell matrix proteins were studied by two analytical approaches. Peptide mapping was done by trypsin and pepsin followed by collagenase cleavage; analyses were carried out by capillary electrophoresis and reversed‐phase high‐performance liquid chromatography (HPLC). Comparison of peptide maps obtained by both methods revealed a complex mixture of peptides in the insoluble layers of the eggshell; it was concluded that there are at least three different insoluble protein/peptide layers in the avian eggshell (cuticle, palisade, and mammillary layer). Partial characterization of peptides in each layer was made by HPLC‐mass spectrometry analysis. There is an evidence that the eggshell insoluble proteins contain species susceptible to collagenase cleavage, however, the sequences split by this enzyme probably are not those typical for the main triple‐helical core of collagenous proteins. It is proposed that the action of collagenase upon eggshell proteins is caused by the side effect of collagenase described previously with synthetic peptides. Some of the proteins present are probably glycosylated. Fatty acid content in the insoluble eggshell layers (after decalcification) was in the range of 2–4% (which reflected both lipid and lipoproteins bound fatty acids). Porphyrin pigments are dominant in the cuticle layer.