bluuebunny/crossref_metadata_2025 huggingface dataset:
Review of cytologic criteria of bacterial vaginosis: Examination of 2,841 Papanicolaou‐stained vaginal smears
Authors: Demirezen Şayeste
Published: September 2003

To review the cytologic criteria used in the diagnosis of bacterial vaginosis cases [Bv (+)], 2,841 Papanicolaou‐stained vaginal smears were examined cytologically. Clue cells and other cytologic findings were observed in 94 of 2,841 (3.30%) vaginal smears. To detect Bv(+) cases, 94 vaginal smears were screened again. Clue cells, profuse free cocci among cornified type epithelial cells, absence of PMNLs, and vaginal Lactobacilli were detected in 43 of 94 (45.74%). The observation of free cocci which resembled a nebulous appearance was detected first in this study. These four cytologic criteria strongly suggested Bv(+) cases (n = 43). Although clue cells were seen in 51 of 94 (54.25%) vaginal smears, due to the presence of abundant PMNLs, metaplastic cells, vaginal Lactobacilli, and some infectious agents, mainly Trichomonas vaginalis and yeasts, they were considered bacterial vaginosis negative [Bv(–)]. It was postulated that 51 vaginal smears might be aerobic vaginitis. In conclusion, if vaginal smears contain profuse PMNLs, some infectious agents, and metaplastic cells, and even clue cells have been observed they are not accepted as Bv(+) cases. Diagn. Cytopathol. 2003;29:156–159. © 2003 Wiley‐Liss, Inc.
Extraction of Juniperus oxycedrus ssp. oxycedrus essential oil by supercritical carbon dioxide: inﬂuence of some process parameters and biological activity
Authors: Marongiu Bruno , Porcedda Silvia , Caredda Alessandra , De Gioannis Barbara , Vargiu Laura , Paolo
Published: September 2003

Supercritical carbon dioxide extraction of the leaves and berries of Juniperus oxycedrus L. ssp. oxycedrus, coupled with a two‐stage separation, allowed the entrapment of cuticular waxes in the ﬁrst separator (90 bar, −10 °C), thereby allowing pure essential oils to be collected in the second separator (15 bar, 10 °C). An extraction carried out on a charge of leaves, at 90 bar and 50 °C, gave a yield of 0.8% (w/w) with respect to the charged material, of an oil whose major constituents were germacrene D (15.9%), manoyl oxide (10.2%) and 1‐epi‐cubenol (5.4%). The berry oil obtained, at the same pressure and temperature as those for the leaves, gave a yield of 0.45% (w/w) and was composed chieﬂy of germacrene D (13.8%), α‐pinene (10.5%) and β‐myrcene (8.1%). At a constant temperature of 50 °C, different extraction pressures (80, 90 and 100 bar) were tested in order to obtain the juniper wood essential oil. The extraction yield depended strongly on the extraction pressure, reaching a maximum of 14.7% (w/w) at 100 bar. The main constituents in the extract were δ‐cadinene, calamenene, cubenol and 1‐epi‐cubenol. Hydrodistillation of the wood of J. oxycedrus gave a yield of 11.0% (w/w) of essential oil, with a reduced level of sesquiterpene hydrocarbons and an enhanced amount of oxygenated sesquiterpenes, with respect to the oil obtained by supercritical carbon dioxide extraction. The extracts obtained at different pressures were tested for cytotoxicity, antiviral and antimicrobial activities. The results showed that the extracts of leaves and berries obtained at 200 bar were cytotoxic against different cell lines used to support virus growth. As far as antiviral activiy is concerned, some of the extracts were active against a single‐stranded RNA+ virus (Poliovirus‐1). When tested for antimicrobial activity, none of the samples were shown to be active. Copyright © 2003 John Wiley & Sons, Ltd.
Supercritical carbon dioxide separation of bergamot essential oil by a countercurrent process
Authors: Poiana Marco , Mincione Antonio , Gionfriddo Francesco , Castaldo Domenico
Published: September 2003

The efﬁciency of separation of bergamot essential oil, performed by a countercurrent column ﬁlled with Raschig rings and using supercritical carbon dioxide as partition solvent, is affected by various parameters. In the experiments explained in this work, the direct effect of CO2 density was shown and the ratio between the amount of oil loaded to on the column and the amount of CO2 used were discussed. The conditions that produced extracts with a similar volatile fraction composition of starting material and with a high yield (more than 80% of recovery) were those with a low feed:solvent ratio; the lowest bergaptene content was obtained at low CO2 density or at high feed:solvent ratio. A good result was observed at a CO2 density of 206 g/dm3 (8 MPa of pressure and a temperature gradient of 46–50–54 °C) and a feed:solvent ratio of 9.4–9.6; in this separation, a yield of 74–77% and a bergaptene content lower than 0.01% was measured. Copyright © 2003 John Wiley & Sons, Ltd.
Chemical composition of the leaf oil of Altingia excelsa Nornha
Authors: Kanjilal P. B. , Kotoky Rumi , Singh R. S.
Published: September 2003

The volatile components obtained by hydrodistillation from the leaves of Altingia excelsa Nornha, collected from the foothills of Arunachal Pradesh, were analysed by GC and GC–MS. A total of 52 compounds were identiﬁed, representing 99% of the total oil. The major components identiﬁed were α‐pinene (19.8%); β‐pinene (16.0%); α‐phellandrene (15.9%); limonene (10.9%) and β‐phellandrene (8.1%). Other components present in small or trace amounts were also identiﬁed. Copyright © 2003 John Wiley & Sons, Ltd.
Volatile constituents of the rhizome oil of Curcuma caesia Roxb. from central India
Authors: Pandey Ashok K. , Chowdhury Ashim R.
Published: September 2003

The volatile rhizomes oil of Curcuma caesia was analysed by GC–MS, which resulted in the identification of 30 components, representing 97.48% of the oil, with camphor (28.3%), ar‐turmerone (12.3%), (Z)‐β‐ocimene (8.2%), ar‐curcumene (6.8%), 1,8‐cineole (5.3%), β‐elemene (4.8%), borneol (4.4%), bornyl acetate (3.3%) and γ‐curcumene (2.82%) as the major constituents. Copyright © 2003 John Wiley & Sons, Ltd.
Generation of a floxed allele of Smad5 for cre‐mediated conditional knockout in the mouse
Authors: Umans Lieve , Vermeire Liesbeth , Francis Annick , Chang Hua , Huylebroeck Danny , Zwijsen An
Published: September 2003

Summary: Smad5 is a member of the Smad family of intracellular mediators of BMP signals and in endothelial cells of TGF‐β signals. We and others previously showed that loss of Smad5 in the mouse results in embryonic lethality (between E9.5–E11.5) due to multiple embryonic and extraembryonic defects. To circumvent the early embryonic lethality and to allow tissue‐ and time‐specific Smad5 inactivation, we created a conditional Smad5 allele in the mouse. Floxed Smad5 (Smad5flE2,Neo/flE2,Neo) mice were generated in which both exon2 and the Neo‐cassette were flanked by loxP sites. Here we demonstrate that embryos with ubiquitous Cre‐mediated deletion of Smad5 (Smad5flΔE2/flΔE2) phenocopy the conventional Smad5 knockout mice. Smad5flE2/flE2 mice are now available and will be a valuable tool to analyze the role of Smad5 beyond its crucial early embryonic function throughout development and postnatal life. genesis 37:5–11, 2003. © 2003 Wiley‐Liss, Inc.
Atrial cardiomyocyte‐specific expression of Cre recombinase driven by an Nppa gene fragment
Authors: de Lange Frederik J. , Moorman Antoon F.M. , Christoffels Vincent M.
Published: September 2003

Summary: To study the development of the atria, we produced a transgenic mouse line that expresses Cre under the regulatory control of a 7 kbp fragment of the Natriuretic peptide precursor type A gene (Nppa), from −3 kbp to +4 kbp relative to the transcription start site. Crossing this line with the R26R and Z/EG reporter lines revealed recombinase activity specifically in the cardiomyocytes of the atria and to a lesser extent the inflow tract from E10.5 onwards. At E14.5 recombination in the atria is almost complete. No recombination was observed outside the heart. These mice provide a tool to study gene function in the atria. genesis 37:1–4, 2003. © 2003 Wiley‐Liss, Inc.
VAChT‐Cre.Fast and VAChT‐Cre.Slow: Postnatal expression of Cre recombinase in somatomotor neurons with different onset
Authors: Misawa Hidemi , Nakata Kazuko , Toda Kyoko , Matsuura Junko , Oda Yoshio , Inoue Haruhisa , Tateno Minako , Takahashi Ryosuke
Published: September 2003

Summary: The cholinergic gene locus (CGL) consists of the genes encoding the choline acetyltransferase (ChAT) and the vesicular acetylcholine transporter (VAChT). To establish a cholinergic‐specific Cre‐expressing mouse, we constructed a transgene expression vector (VAChT‐Cre) with 11.3 kb human CGL in which a Cre‐IRES‐EGFP unit was inserted in the VAChT open reading frame. The activity of Cre, whose expression was driven by the VAChT promoter, was examined by crossing a reporter mouse (CAG‐CAT‐Z) in which expression of LacZ is activated upon Cre‐mediated recombination. Transgenic lines with the VAChT‐Cre construct displayed the restricted Cre expression in a subset of cholinergic neurons in the somatomotor nuclei and medial habenular nucleus, but absent in visceromotor and other central and peripheral cholinergic neurons. Cre expression was first observed at postnatal day 7 and later detected in ∼40–60% of somatomotor neurons. Based on the onset of Cre expression, we generated two mouse lines (two alleles; VAChT‐Cre.Fast and VAChT‐Cre.Slow) in which Cre expression reaches maximal levels fast and slow, respectively. The use of VAChT‐Cre mice should allow us to deliver Cre to a subset of postnatal motor neurons, thereby bypassing lethality and facilitating analysis of gene function in adult motor neurons. genesis 37:44–50, 2003. © 2003 Wiley‐Liss, Inc.
Reduced cell adhesion during mitosis by threonine phosphorylation of β1 integrin
Authors: Suzuki Katsuo , Takahashi Kazuhide
Published: September 2003

Cell shape and adhesion of cultured mammalian cells change dramatically during mitosis, however, how cell cycle‐dependent alterations in cell adhesion are regulated remain to be elucidated. We show here that normal human mammary epithelial (HME) cells which became less adhesive and adopted the rounded morphology during the G2/M phase of the cell cycle significantly reduced their dependence on β1 integrin‐mediated adhesion to laminin, by using function blocking antibody to β1 integrin. In G2/M cells, both total and cell surface expressions of β1 integrin were comparable with those in G1 cells but it was phosphorylated at threonines 788–789 within its cytoplasmic domain and coimmunoprecipitated Ca2+/calmodulin‐dependent protein kinase (CaMK) II. The threonine phosphorylated β1 integrin significantly reduced its intracellular linkage with actin, with no significant reduction in the actin expression. In contrast, β1 integrin in G1 cells was not threonine phosphorylated but formed a link with actin and coimmunoprecipitated the core enzyme of the serine/threonine protein phosphatase (PP) 2A. The results suggest that reduced β1 integrin‐mediated cell adhesion of HME cells to the substratum during mitosis may be induced by β1 integrin phosphorylation at threonines 788–789 and its reduced ability to link with the actin cytoskeleton. J. Cell. Physiol. 197: 297–305, 2003. © 2003 Wiley‐Liss, Inc.
How the other half lives, the amino‐terminal domain of the retinoblastoma tumor suppressor protein
Authors: Goodrich David W.
Published: September 2003

The retinoblastoma tumor suppressor gene (RB1) is currently the only known gene whose mutation is necessary and sufficient for the development of a human cancer. Mutation or deregulation of RB1 is observed so frequently in other tumor types that compromising RB1 function may be a prerequisite for malignant transformation. Identifying the molecular mechanisms that provide the basis for RB1‐mediated tumor suppression has become an important goal in the quest to understand and treat cancer. The lion's share of research on these mechanisms has focused on the carboxy‐terminal half of the RB1 encoded protein (pRB). This focus is with good reason since this part of the protein, now called the “large pocket,” is required for most of its known activities identified in vitro and in vivo. Large pocket mediated mechanisms alone, however, cannot account for all observed properties of pRB. The thesis presented here is that the relatively uncharacterized amino‐terminal half of the protein makes important contributions to pRB‐mediated tumor suppression. The goals of this review are to summarize evidence indicating that an amino‐terminal structural domain is important for pRB function and to suggest a general hypothesis as to how this domain can be integrated with current models of pRB function. J. Cell. Physiol. 197: 169–180, 2003© 2003 Wiley‐Liss, Inc.
STAT proteins: From normal control of cellular events to tumorigenesis
Authors: Calò Valentina , Migliavacca Manuela , Bazan Viviana , Macaluso Marcella , Buscemi Maria , Gebbia Nicola , Russo Antonio
Published: September 2003

Signal transducers and activators of transcription (STAT) proteins comprise a family of transcription factors latent in the cytoplasm that participate in normal cellular events, such as differentiation, proliferation, cell survival, apoptosis, and angiogenesis following cytokine, growth factor, and hormone signaling. STATs are activated by tyrosine phosphorylation, which is normally a transient and tightly regulates process. Nevertheless, several constitutively activated STATs have been observed in a wide number of human cancer cell lines and primary tumors, including blood malignancies and solid neoplasias. STATs can be divided into two groups according to their specific functions. One is made up of STAT2, STAT4, and STAT6, which are activated by a small number of cytokines and play a distinct role in the development of T‐cells and in IFNγ signaling. The other group includes STAT1, STAT3, and STAT5, activated in different tissues by means of a series of ligands and involved in IFN signaling, development of the mammary gland, response to GH, and embriogenesis. This latter group of STATS plays an important role in controlling cell‐cycle progression and apoptosis and thus contributes to oncogenesis. Although an increased expression of STAT1 has been observed in many human neoplasias, this molecule can be considered a potential tumor suppressor, since it plays an important role in growth arrest and in promoting apoptosis. On the other hand, STAT3 and 5 are considered as oncogenes, since they bring about the activation of cyclin D1, c‐Myc, and bcl‐xl expression, and are involved in promoting cell‐cycle progression, cellular transformation, and in preventing apoptosis. J. Cell. Physiol. 197: 157–168, 2003© 2003 Wiley‐Liss, Inc.
Hypothyroidism and Parkinson's disease and the issue of diagnostic confusion
Authors: García‐Moreno José M. , Chacón‐Peña José
Published: September 2003

Development of hypothyroidism may easily be overlooked when occurring together with Parkinson's disease (PD), because many of the symptoms of the two disorders are similar. We report on a case of a woman suffering from both PD and hypothyroidism and review the literature on the subject. © 2003 Movement Disorder Society
Efficacy of propranolol in hyperthyroid‐induced chorea: A case report
Authors: Hayashi Ryoichi , Hashimoto Takao , Tako Kenji
Published: September 2003

We describe a thyrotoxic patient with choreic movements. The latter gradually receded and disappeared as the patient was treated with methimazole and β‐adrenoceptor blocker, and the improvement deteriorated during a β‐adrenoceptor challenge test. Our findings suggest that the pathogenesis of thyrotoxic chorea seems more likely to be of biochemical origin rather than due to structural changes in the basal ganglia. © 2003 Movement Disorder Society
Dopa‐responsive dystonia presenting with prominent isolated bilateral resting leg tremor: Evidence for a role of parkin?
Authors: Postuma Ronald B. , Furukawa Yoshiaki , Rogaeva Ekaterina , St. George‐Hyslop Peter H. , Farrer Matt J. , Lang Anthony E.
Published: September 2003

We report on a GTP cyclohydrolase 1 mutation‐confirmed case of dopa‐responsive dystonia who presented with an isolated parkinsonian rest tremor starting at the age of 15 years. There was no dystonia or other features of parkinsonism. Sequencing of the parkin gene demonstrated polymorphisms in a compound heterozygous state. The spectrum of unusual clinical presentations of dopa‐responsive dystonia is discussed. © 2003 Movement Disorder Society
Frequencies of single nucleotide polymorphism in alcohol dehydrogenase7 gene in patients with multiple system atrophy and controls
Authors: Kim Hyun Sook , Lee Myung Sik
Published: September 2003

A polymerase chain reaction and direct sequencing of the ADH7 gene were carried out in 50 controls and 50 patients with probable multiple system atrophy (MSA). Seven SNPs, one insertion, and one mismatch were found in patients with MSA and controls. There was no significant difference in the frequencies of each SNP between the patients and the controls (P > 0.05). Interpretation of this negative finding should be cautious in view of the relatively small number of cases. © 2003 Movement Disorder Society
Vernier operation of series‐coupled optical microring resonator filters
Authors: Schwelb Otto , Frigyes István
Published: September 2003

Nonsymmetric Vernier type series‐coupled optical microring filters have been found to be free of the spurious responses that commonly occur in those of symmetric design. Large free spectral range (FSR) is obtained without significant decrease of the waveguide‐bending radius. The tradeoff between coupling coefficients, filter bandwidth, and the suppression of interstitial resonances has been investigated. Results on filter sensitivity to waveguide and coupler loss are also reported. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 257–261, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11185
Analysis of electromagnetic scattering of three‐dimensional dielectric bodies using Krylov subspace FFT iterative methods
Authors: Chen R. S. , Fan Z. H. , Yung Edward K. N.
Published: September 2003

In this paper, electromagnetic wave scattering is formulated in terms of the electric‐field integral equation (EFIE) for a dielectric body of general shape, inhomogeneity, and anisotropy. Applying the pulse‐function expansion and the point‐matching technique, the integral equation can be solved by the efficient Krylov subspace iterative fast Fourier transform (FFT) technique. Several typical Krylov subspace iterative FFT methods are used to solve the problems and their convergence behaviors are compared. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 261–267, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11186
Three‐dimensional reconstruction of a shallowly buried mine using time‐domain data
Authors: Zhou Hui , Takenaka Takashi , Tanaka Toshiyuki
Published: September 2003

A two‐dimensional (2D) time‐domain inverse scattering technique for reconstructing lossy media is extended to three‐dimensional (3D) cases. Forward and inversion algorithms of this approach are carried out by the finite‐difference time‐domain (FDTD) method. The reconstruction of a mine buried in soil illustrates that this method is one of useful techniques to detect buried mines. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 276–280, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11189
Near‐field imaging by negative permittivity media
Authors: Lu W. T. , Sridhar S.
Published: September 2003

The imaging of the near field by a slab of negative permittivity medium is discussed. The performance of this flat lens is limited by the working wavenumber k0 and the slab thickness d. For k0d ≪ 0.937, the resolution of the flat lens can be smaller than the wavelength and depends logarithmically on 1/k0d. The electric field is finite everywhere, but will diverge algebraically as one reduces k0d → 0. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 282–286, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11191
Finite‐element analysis of the dominant‐mode cutoff wavelength in dielectric‐loaded single‐ridge waveguides
Authors: Lu Mai , Leonard Paul J.
Published: September 2003

As a new kind of broadband microwave component, the cutoff characteristic of a dielectric‐loaded single‐ridge waveguide is analysed by the finite‐element method. Design data for cutoff wavelength of dominant mode for different values of relative permittivity εr equal to 1.0, 5.0, 10.0, and 15.0 with varied ridge dimensions have been presented in tabulated forms. Numerical results in this paper provide an extension to the existing design data of empty single‐ridge waveguides and are considered useful in practical applications. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 298–300, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11196
Relationship between root‐mean‐square and peak‐to‐peak jitter measurements
Authors: Babić Dubravko , Plombon John
Published: September 2003

The statistical relationship between peak‐to‐peak and root‐mean‐square (RMS) measurements of random and deterministic jitter is discussed. An analytic expression relating the two quantities is derived for normally distributed random jitter. This work is applicable to a production test of electronic or optical components that uses sampling oscilloscopes for jitter characterization. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 323–326, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11204
Studying 120° PBG waveguide bend using FDTD
Authors: Uusitupa Tero , Kärkkäinen Kimmo , Nikoskinen Keijo
Published: September 2003

In this paper, photonic‐bandgap‐waveguide (PBG‐WG) structures are considered. The WGs are based entirely on linear dielectric media. Analysis is based on numerical field computation by FDTD and post‐processing of the field data. A 120° PBG‐WG bend is investigated as an example. Power transmission results are given. Various issues related to simulation arrangements and PBG‐WG component design are discussed. © 2003 Wiley Periodicals, Inc. Microwave Opt Technol Lett 39: 326–333, 2003; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/mop.11205
Developmental potential of bovine nuclear transfer embryos and postnatal survival rate of cloned calves produced by two different timings of fusion and activation
Authors: Akagi Satoshi , Adachi Noritaka , Matsukawa Kazutsugu , Kubo Masanori , Takahashi Seiya
Published: September 2003

We compared developmental potential of somatic cell nuclear transfer (NT) embryos and postnatal survivability of cloned calves produced by two different fusion and activation protocols. As donor cells for NT, bovine cumulus cell‐derived cultured cells of passage 5 were used following culture in serum‐starved medium for 5–7 days. Enucleated oocytes were fused with donor cells at 21 or 24 hr post maturation. NT embryos fused at 21 hr were activated chemically 3 hr after fusion (DA group) and embryos fused at 24 hr were activated chemically immediately after fusion (FA group). Chemical activation was accomplished by calcium ionophore for 5 min and cytochalasin D + cycloheximide for 1 hr then cycloheximide alone for 4 hr. After in vitro culture in IVD101 medium for 7 days, embryo transfer was performed. Fusion rates were 86 and 84% in the DA and FA groups, respectively. Developmental rate to the blastocyst stage of NT embryos in the DA group was higher than in the FA group (42% vs. 28%). Pregnancy rate did not differ significantly between the DA and FA groups (11/13 and 5/7 at day 35), and 13 cloned calves (including 1 set of twins from a single embryo transfer) were born. High rates of postnatal mortality were observed in both groups. These results suggest that the DA method improves in vitro developmental potential of NT embryos, but the timing of fusion and chemical activation does not affect the pregnancy rate and the survivability of cloned calves. Mol. Reprod. Dev. 66: 264–272, 2003. © 2003 Wiley‐Liss, Inc.
Nuclear localization of NORs and centromeres in mouse oocytes during folliculogenesis
Authors: Longo Frank , Garagna Silvia , Merico Valeria , Orlandini Guido , Gatti Rita , Scandroglio Renato , Redi Carlo Alberto , Zuccotti Maurizio
Published: September 2003

Mouse oocytes at the germinal vesicle stage are characterized by one of two nuclear morphologies: surrounded nucleolus (SN), in which the nucleolus is surrounded by a rim of Hoechst positive chromatin and not surrounded nucleolus (NSN), in which this rim is essentially absent. This morphological difference has a biological relevance as NSN oocytes are transcriptionally active, yet incapable of development beyond the two‐cell stage. Whereas SN oocytes, which are transcriptionally inactive, are capable of development to the blastocyst stage. To further our understanding of the nuclear organization of the mouse oocyte during folliculogenesis, we have conducted a series of investigations employing silver methods that stain nucleolus organizer region (NOR), centromeres, and heterochromatin, as well as, specific antibodies for centromeres. Results obtained by a variety of microscopic methods (light, electron, immunochemical, and confocal) demonstrate: (1) a changing pattern of NOR staining during folliculogenesis that is specific to follicular type, and (2) significant differences in the organization of NORs and centromeres of isolated, antral NSN, and SN oocytes. These observations suggest possible means by which, chromosomes of mature, germinal vesicle oocytes are organized with respect to the nucleolus. Mol. Reprod. Dev. 66: 279–290, 2003. © 2003 Wiley‐Liss, Inc.
Apoptosis in sea bream Sparus aurata eggs
Authors: Carnevali O. , Polzonetti V. , Cardinali M. , Pugnaloni A. , Natalini P. , Zmora N. , Mosconi G. , Polzonetti‐Magni A.M.
Published: September 2003

In the present study, the sea‐bream Sparus aurata, a pelagic egg spawner, was used as experimental model, in order to establish the occurrence of apoptosis in vertebrates with external reproduction. The same female ovulates floating and nonfloating eggs, but only the former, after fertilization, proceed to embryo development. The eggs were divided into floating and nonfloating and both were analyzed for the presence of several apoptosis markers. The results here reported provide evidence that the nonfloating cells present severe shrinkage and highly express both FAS receptor and FAS ligand on their surface. Furthermore, DNA fragmentation and mitochondria swelling were found, suggesting that the nonfloating eggs were cells programmed to die. Mol. Reprod. Dev. 66: 291–296, 2003. © 2003 Wiley‐Liss, Inc.
Near‐infrared analysis of fennel (Foeniculum vulgare Miller) on different spectrometers—basic considerations for a reliable network
Authors: Steuer Boris , Schulz Hartwig
Published: September 2003

The aim of this study was to investigate the accuracy and transferability of near‐infrared (NIR) calibrations for estimating the content and composition of the volatile fraction in fennel fruits (Foeniculum vulgare Miller) as an example of medicinal and spice plants. A master calibration with spectra obtained on a scanning monochromator was generated using 345 samples from three different harvests (1997–1999). A subset of 70 samples from 1999 was also measured on a dispersive grating and a scanning diode array system to gain an insight into the inﬂuence of sample presentation and scanning techniques. For all instruments, calibrations with standard errors in the range of the reference method were achieved. Furthermore the inﬂuence of storage on NIR spectra and, additionally, the potential of transferring spectra between both scanning monochromators was studied. Copyright © 2003 John Wiley & Sons, Ltd.
Analysis of anthraquinones in cell cultures of Cinchona ‘Robusta’ by HPLC with photodiode array and mass spectrometry detection
Authors: Han Ying‐Shan , Hofte Bertil , der Heijden Robert van , Verpoorte Robert
Published: September 2003

An HPLC‐PAD‐ESI/MS method has been developed for the analysis of anthraquinones in cell cultures of Cinchona ‘Robusta’. Using a C18 column and gradient elution with a mobile phase system containing acetonitrile, water and triﬂuoroacetic acid, a satisfactory separation of both anthraquinone glycosides and aglycones in a crude dichloromethane extract could be obtained. Robustaquinone B was identiﬁed as a major anthraquinone in the extracts, and another ﬁve anthraquinones were tentatively identiﬁed from spectroscopic data. A number of minor unknown compounds were detected and were distinguished from the known anthraquinones. An isocratic system for the quantitative determination of robustaquinone B has also been developed. Copyright © 2003 John Wiley & Sons, Ltd.
Fast high‐performance liquid chromatographic analysis of naphthodianthrones and phloroglucinols from Hypericum perforatum extracts
Authors: Tolonen Ari , Hohtola Anja , Jalonen Jorma
Published: September 2003

Hypericum perforatum L. (St. John's Wort) has been used in modern medicine for treatments of depression and neuralgic disorders. An HPLC method with photodiode array detection for the rapid determination of the major active compounds, naphthodianthrones and phloroglucinols, has been developed. The method permits the determination of hypericin, protohypericin, pseudohypericin, protopseudohypericin, hyperforin and adhyperforin in an extract in less than 5 min. Good linearity over the range 0.5–200 µg/mL for hyperforin and 0.02–100 µg/mL for hypericin was observed. Intra‐assay accuracy and precision varied from 0.1 to 17% within these ranges. Lower levels of quantitative determination were 2 µg/mL for hyperforin and 0.5 µg/mL for hypericin, while detection limits were 0.1 and 0.02 µg/mL, respectively. Copyright © 2003 John Wiley & Sons, Ltd.
Determination of the triacylglycerol composition of coffee beans by reverse‐phase high‐performance liquid chromatography
Authors: Jham Gulab N. , Nikolova‐Damyavova Boryana , Viera Mirtes , Natalino Ricardo , Rodrigues Augusto Cezar
Published: September 2003

Reverse‐phase HPLC with refractive index and light scattering detectors in isocratic and gradient elution modes, respectively, was applied for the separation of the major triacylglycerols (TAG) in coffee lipids. Twelve TAG species could be identiﬁed and determined using a linear gradient of acetonitrile in dichloromethane: dichloroethane. The quantitative evaluation was based on the relative area percentages derived directly from a data‐station. The procedure was applied to determine the TAG composition of three types of coffee beans harvested in two coffee producing areas in Brazil and dried by two commonly used procedures. No signiﬁcant differences in the TAG compositions due to the type, origin and drying procedure were found. Copyright © 2003 John Wiley & Sons, Ltd.
Characterization of an unsaturated low‐density polyethylene
Authors: Smedberg A. , Hjertberg T. , Gustafsson B.
Published: September 2003

This study concerns a new group of low‐density polyethylenes (LDPEs)—unsaturated LDPE. The new LDPE is a copolymer between ethylene and 1,9‐decadiene and was polymerized in a commerical high‐pressure tubular reactor. The diene copolymerized with one double bond, leaving the other unreacted as a pendant side group. This yielded a copolymer containing a higher number of vinyl groups than ordinary LDPE. Fractionation of the copolymer and determination of the number of unsaturated structures in the different fractions by Fourier transform infrared spectroscopy revealed that the diene is homogeneously incorporated along the molar‐mass distribution curve. It is also possible to obtain copolymers with a varying vinyl content, without drastic changes in molar mass or molar‐mass distribution, by a controlled addition of 1,9‐decadiene to the reactor. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 2974–2984, 2003
Synthesis and properties of biodegradable copolymers based on 4,4′‐(adipoyldioxy)dicinnamic acid, 1,6‐hexanediol, and poly(ethylene glycol)s
Authors: Nagata Minoru , Hizakae Sigeki
Published: September 2003

4,4′‐(Adipoyldioxy)dicinnamic acid (CAC) was synthesized by a condensation of adipoyl chloride and 4‐hydroxycinnamic acid. The CAC6 copolymers were prepared by a high‐temperature solution polycondensation of a diacyl chloride of CAC, 1,6‐hexanediol (6), and poly(ethylene glycol) (PEG) in which the molecular weights of PEG are 1000, 2000, and 8300. Differential scanning calorimetric curves of the copolymers exhibited a glass‐transition temperature because of PEG moiety and two melting endotherms (Tm's); the one at the higher Tm was due to CAC6 moiety, and the other at the lower Tm was due to PEG moiety, suggesting that these copolymers are the block type. The incorporation of the PEG component decreased the tensile strength and initial modulus, but increased the elongation extremely. The enzymatic degradation was performed in phosphate buffer solution (pH 7.2) with Ps. cepacia lipase at 37 °C. The degradation rate of the copolymers increased significantly with an increasing content of PEG, which was correlated to the water absorption of the copolymers. All copolymers could undergo photocuring by ultraviolet (UV) light irradiation (λ > 280 nm) at ambient temperature, as examined by UV spectroscopy and solubility. The CAC6/E2000(50/50) film photocured for 3 min exhibited a good elastic property with a maximum tensile strength of 3.7 MPa and maximum elongation of 640%. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 2930–2938, 2003
Synthesis and cleavage of core‐labile poly(alkyl methacrylate) star polymers
Authors: Kilian Lars , Wang Zhen‐He , Long Timothy E.
Published: September 2003

Core‐cleavable star polymers were synthesized by the coupling of living anionic poly(alkyl methacrylate) arms with either dicumyl alcohol dimethacrylate (DCDMA) or 2,5‐dimethyl‐2,5‐hexanediol dimethacrylate (DHDMA). This synthetic methodology led to the formation of star polymers that exhibited high molecular weights and relatively narrow molecular weight distributions. The labile tertiary alkyl esters in the DCDMA and DHDMA star polymer cores were readily hydrolyzed under acidic conditions. High‐molecular‐weight star polymer cleavage led to well‐defined arm polymers with lower molecular weights. Hydrolysis was confirmed via 1H NMR spectroscopy and gel permeation chromatography. Thermogravimetric analysis (TGA) of the star polymers demonstrated that the DCDMA and DHDMA star polymer cores also thermally degraded in the absence of acid catalysts at 185 and 220 °C, respectively, and the core‐cleavage temperatures were independent of the arm polymer composition. The difference in the core‐degradation temperatures was attributed to the increased reactivity of the DCDMA‐derived cores. TGA/mass spectrometry detected the evolution of the diene byproduct of the core degradation and confirmed the proposed degradation mechanism. The DCDMA monomer exhibited a higher degradation rate than DHDMA under identical reaction conditions because of the additional resonance stabilization of the liberated byproduct, which made it a more responsive cleavable coupling monomer than DHDMA. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 3083–3093, 2003
Topological polymer chemistry by electrostatic self‐assembly
Authors: Tezuka Yasuyuki
Published: September 2003

Recent developments in topological polymer chemistry are outlined. First, nonlinear polymer topologies are systematically classified on the basis of topological considerations of constitutional isomerism in a series of alkanes (CnH2n+2), monocycloalkanes (CnH2n), and polycycloalkanes (CnH2n−2, CnH2n−4, etc.). Various pairs of topological isomers are identified in randomly coiled, flexible polymer molecules with cyclic and branched structures. An electro‐ static self‐assembly and covalent fixation strategy has subsequently been developed for the efficient synthesis of a variety of topologically unique polymers, including monocyclic and polycyclic polymers, topological isomers, and topological block copolymers. In this process, new telechelics with moderately strained cyclic onium salt groups carrying multifunctional carboxylate counteranions have been designed as key polymeric precursors. Further extensions of topological polymer chem‐ istry have been achieved by the use of cyclic telechelics (kyklo‐telechelics) and cyclic macromonomers, obtainable also by means of the electrostatic self‐assembly and covalent fixation process. © 2003 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 41: 2905–2917, 2003
The Jackknife applied to incomplete blood sampling models
Authors: Singer Júlia , Berger Marion
Published: September 2003

A nonparametric alternative to the extended parametric Satterthwaite method taking into account the correlation between samples obtained from the same animal is presented. Confidence intervals are computed using a modified jackknife standard error. The performance of the method is studied by means of simulation. This nonparametric method has a smaller coverage rate than the parametric one, but still close to the nominal confidence level. In addition, its confidence interval is shorter than that of the parametric method using the generalized Satterthwaite approximation. Copyright © 2003 John Wiley & Sons, Ltd.
A fixed sequence Bonferroni procedure for testing multiple endpoints
Authors: Wiens Brian L.
Published: September 2003

A method for controlling the familywise error rate combining the Bonferroni adjustment and fixed testing sequence procedures is proposed. This procedure allots Type I error like the Bonferroni adjustment, but allows the Type I error to accumulate whenever a null hypothesis is rejected. In this manner, power for hypotheses tested later in a prespecified order will be increased. The order of the hypothesis tests needs to be prespecified as in a fixed sequence testing procedure, but unlike the fixed sequence testing procedure all hypotheses can always be tested, allowing for an a priori method of concluding a difference in the various endpoints. An application will be in clinical trials in which mortality is a concern, but it is expected that power to distinguish a difference in mortality will be low. If the effect on mortality is larger than anticipated, this method allows a test with a prespecified method of controlling the Type I error rate. Copyright © 2003 John Wiley & Sons, Ltd.
Multimarker approach in transplanted mussels for evaluating water quality in Charentes, France, coast areas exposed to different anthropogenic conditions
Authors: Roméo M. , Mourgaud Y. , Geffard A. , Gnassia‐Barelli M. , Amiard J. C. , Budzinski H.
Published: September 2003

An active biomonitoring experiment was performed using mussels collected at a clean site, Fier d'Ars, and transplanted to two locations, outside the harbor of La Rochelle and in the Baie de L'Aiguillon along the coast of Charentes (French Atlantic coast) beginning in April for several months. Mussels were collected in June and October. The cadmium, copper, and zinc concentrations of all resident and transplanted mussel samples and the polycyclic aromatic hydrocarbon and polychlorinated biphenyl concentrations in some mussel samples and in the sediment samples were determined. Mussel response was evaluated for several biochemical biomarkers: concentrations of metallothionein, activities of glutathione S‐transferase and acetylcholinesterase (AChE) and levels of thiobarbituric reactive substance (TBARS). The physiological status of the animals was assessed using the condition index. A principal component analysis performed with the chemical and biochemical results of the evaluations of the resident and transplanted mussels collected in June allowed them to be separated into three groups: resident mussels from la Rochelle with high metal and TBARS levels, resident mussels from Baie de L'Aiguillon with a very high condition index, and resident mussels from Fier d'Ars and transplanted mussels at La Rochelle and Baie de L'Aiguillon with low TBARS and AChE activities. Strong seasonal variation from June to October of all parameters was noted. Mussels transplanted to La Rochelle appeared to be the most “polluted” in their pollutant concentrations and biochemical responses; moreover, the La Rochelle site had the highest concentration of organics in sediments of all the sites. The choice of Fier d'Ars as a reference site may be questionable because some of the biomarker responses of the mussels were higher than expected there, although these pollutants in mussels and sediment were present at the lowest concentrations measured. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 295–305, 2003.
PbCl2‐induced hyperpolarization of rat thymocytes: Involvement of charybdotoxin‐sensitive K+ channels
Authors: Nishizaki Yasutaka , Oyama Yasuo , Sakai Yoshiro , Hirama Seigo , Tomita Kazuyoshi , Nakao Hiromi , Umebayashi Chisato , Ishida Shiro , Okano Yoshiro , Carpenter David O.
Published: September 2003

The effect of PbCl2 on membrane potential and intracellular divalent metal cation concentrations of rat thymocytes was examined by flow cytometry. PbCl2 at concentrations of 0.3 μM or higher (up to 10 μM) produced persistent, dose‐dependent hyperpolarization (decrease in the intensity of di‐BA‐C4 fluorescence). Removal of external Ca2+ did not significantly affect the PbCl2‐induced hyperpolarization. Charybdotoxin, a specific antagonist of Ca2+‐dependent K+ conductance, greatly attenuated the PbCl2‐induced hyperpolarization. PbCl2 increased the intensity of fluo‐3 fluorescence under both normal Ca2+ and nominally Ca2+‐free conditions. These results suggest that Pb2+ enters thymocytes, causing an increase in fluo‐3 fluorescence, and activates Ca2+‐dependent K+ channels, resulting in hyperpolarization. The persistent activation of K+ channels by Pb2+, leading to persistent hyperpolarization, may be one mechanism whereby Pb2+ alters immune function, as membrane potential changes influence physiological functions of lymphocytes. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 321–326, 2003.
Comparative toxicity and biochemical responses of certain pesticides to the mature earthworm Aporrectodea caliginosa under laboratory conditions
Authors: Mosleh Yahia Y. , Ismail Saad M. M. , Ahmed Mohamed T. , Ahmed Yousery M.
Published: September 2003

This study was conducted to investigate the toxicity of aldicarb, cypermethrin, profenofos, chlorfluazuron, atrazine, and metalaxyl toward mature Aporrectodea caliginosa earthworms. The effects of the LC25 values of these pesticides on the growth rate in relation to glucose, soluble protein, and activities of glutamic‐oxaloacetic transaminase (GOT), glutamic‐pyruvic transaminase (GPT), acid phosphatase (AcP), and alkaline phosphatase (AIP) were also studied. The results showed that aldicarb was the most toxic of the tested pesticides, followed in order by cypermethrin, profenofos, chlorfluazuron, atrazine, and metalaxyl. A reduction in growth rate was observed in all pesticide‐treated worms, which was accompanied by a decrease in soluble protein and an increase in transaminases and phosphatases. Relationships between growth rate, protein content, transaminases, and phosphatases provided strong evidence for the involvement of pesticidal contamination in the biochemical changes in earthworms, which can be used as a bioindicator of soil contamination by pesticides. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 338–346, 2003.
Cyst‐based toxicity tests XV—Application of ostracod solid‐phase microbiotest for toxicity monitoring of contaminated soils
Authors: Chial Belgis , Persoone Guido
Published: September 2003

A new “culture/maintenance‐free” microbiotest has recently been developed for “direct‐contact” toxicity determination of contaminated sediments. The 6‐day Ostracodtoxkit F™ makes use of the neonates of the ostracod crustacean Heterocypris incongruens hatched from dormant eggs (cysts). The new low‐cost assay has already been applied in three studies on river sediment from Flanders (Belgium) and from Canada and was found to perform comparably to the 10‐day Hyalella azteca and Chironomus riparius tests in detecting and quantifying sediment toxicity. Taking into account that sediments are in fact underwater soils, the ostracod microbiotest was tentatively applied on 15 contaminated soils from Flanders (Belgium), and its sensitivity (based on mortality as the test criterion) was evaluated in comparison to the 28‐day reproduction inhibition assay with the springtail Folsomia candida. The results revealed that the ostracod test species was as sensitive as or, in several samples, even more sensitive than the springtails. Leachate experiments on the same soil samples also showed that in most cases ostracod mortality was a result of the (nonsoluble) toxicants bound to the solid‐phase particles, rather than of those that had dissolved in the water phase. Providing confirmation of these first findings through additional studies, the new culture/maintenance‐free ostracod microbiotest seems to have good potential as a low‐cost and user‐friendly tool for routine toxicity monitoring of contaminated soils. © 2003 Wiley Periodicals, Inc. Environ Toxicol 18: 347–352, 2003.
2‐Cyclopenten‐1‐one and prostaglandin J2 reduce restenosis after balloon angioplasty in rats: role of NF‐κB
Authors: Ianaro Angela , Maffia Pasquale , Cuzzocrea Salvatore , Mazzon Elena , Santoro Maria Gabriella , Di Rosa Massimo , Ialenti Armando
Published: September 2003

The aim of this study was to evaluate, using a rat model of balloon angioplasty, whether prostaglandin (PG) J2 and 2‐cyclopenten‐1‐one are able to reduce restenosis. We found that both PGJ2 and 2‐cyclopenten‐1‐one, administered by local application on carotid arteries, caused a dose‐dependent inhibition of neointimal formation. Furthermore, both agents prevented vascular negative remodeling. The effect of these compounds on restenosis was correlated with an inhibition of nuclear factor‐κB (NF‐κB) activation as well as of intercellular adhesion molecule‐1 (ICAM‐1) protein expression in injured carotid arteries of control animals. Our results show that cyclopentenone PGs and their derivatives reduce restenosis and may have therapeutic relevance for the prevention of human restenosis.
Conserved signature proposed for folding in the lipocalin superfamily
Authors: Greene Lesley H , Hamada Daizo , Eyles Stephen J , Brew Keith
Published: September 2003

We systematically identify a group of evolutionarily conserved residues proposed for folding in a model β‐barrel superfamily, the lipocalins. The nature of conservation at the structural level is defined and we show that the conserved residues are involved in a network of interactions that form the core of the fold. Exploratory kinetic studies are conducted with a model superfamily member, human serum retinol‐binding protein, to examine their role. The present results, coupled with key experimental studies conducted with another lipocalin β‐lactoglobulin, suggest that the evolutionarily conserved regions fold on a faster folding time‐scale than the non‐conserved regions.