bluuebunny/crossref_metadata_2025 huggingface dataset:
‘The doctor knows best’ revisited: physician perspectives
Authors: Freedman Tovia G.
Published: October 2002

An ethnographic study was undertaken to observe in vivo medical encounters between oncology physicians and breast cancer patients. The aim of the study was to gain an inside view of how physicians and patients conceptualize the decision making process during cancer diagnosis and treatment. The confluence of the changes in the role of women in society, the shifting requirements for the delivery of medical services and a complex array of medical options have all contributed to changes in the role of the physician from ‘the doctor knows best’ to a more reciprocal role. The study revealed physician–patients perspectives that may facilitate future medical encounters. In addition, some troubling practices were observed that raise dilemmas about the parameters of informed consent and ethical concerns regarding how medical options are presented. Copyright © 2002 John Wiley & Sons, Ltd.
Palaeoclimate reconstruction on Big Lyakhovsky Island, north Siberia—hydrogen and oxygen isotopes in ice wedges
Authors: Meyer Hanno , Dereviagin Alexander , Siegert Christine , Schirrmeister Lutz , Hubberten Hans‐W.
Published: October 2002

Late Quaternary permafrost deposits on Big Lyakhovsky Island (New Siberian Islands, Russian Arctic) were studied with the aim of reconstructing the palaeoclimatic and palaeoenvironmental conditions of northern Siberia. Hydrogen and oxygen stable isotope analyses are presented for six different generations of ice wedges as well as for recent ice wedges and precipitation. An age of about 200 ka BP was determined for an autochtonous peat layer in ice‐rich deposits by U/Th method, containing the oldest ice wedges ever analysed for hydrogen and oxygen isotopes. The palaeoclimatic reconstruction revealed a period of severe winter temperatures at that time. After a gap in the sedimentation history of several tens of thousands of years, ice‐wedge growth was re‐initiated around 50 ka BP by a short period of extremely cold winters and rapid sedimentation leading to ice‐wedge burial and characteristic ice‐soil wedges (‘polosatics’). This corresponds to the initial stage for the Late Weichselian Ice Complex, a peculiar cryolithogenic periglacial formation typical of the lowlands of northern Siberia. The Ice Complex ice wedges reflect cold winters and similar climatic conditions as around 200 ka BP. With a sharp rise in δ18O of 6‰ and δD of 40‰, the warming trend between Pleistocene and Holocene ice wedges is documented. Stable isotope data of recent ice wedges show that Big Lyakhovsky Island has never been as warm in winter as today. Copyright © 2002 John Wiley & Sons, Ltd.
Enhanced expression of vimentin in motile prostate cell lines and in poorly differentiated and metastatic prostate carcinoma
Authors: Lang Shona H. , Hyde Catherine , Reid Ian N. , Hitchcock Ian S. , Hart Claire A. , Gordon Bryden A. A. , Villette Jean‐Marie , Stower Michael J. , Maitland Norman J.
Published: October 2002

BACKGROUND

The metastatic potential of a series of prostate cell lines was analysed by measuring motility and invasiveness, and further correlated to the expression of epithelial differentiation markers.

METHODS

Invasion and motility were measured using in vitro assays. Immunohistochemistry of cell lines and tissues was used to identify expression of cytokeratins 8 and 1, 5, 10, 14, vimentin, prostate specific antigen, prostate specific membrane antigen, androgen receptor, desmoglein, E‐cadherin, β1 integrin, CD44, hmet, vinculin and actin.

RESULTS

Expression of vimentin was the only marker to correlate with motility, no markers correlated to invasion. Lower vimentin expression was observed in cells with low motility (PNT2‐C2) and high expression in cells with high motility (P4E6, PNT1a, PC‐3). Vimentin expression was not detected in well differentiated tumours, moderately differentiated tumours contained vimentin positive cells (1/9 bone scan negative, 2/5 bone scan positive), but the majority of poorly differentiated cancers (4/11 bone scan negative, 9/14 bone scan positive) and bone metastases (7/8) had high vimentin expression in tumour cells.

CONCLUSIONS

Motile prostate cancer cell lines express vimentin. In tissue sections, the presence of vimentin positive tumour cells correlated positively to poorly differentiated cancers and the presence of bone metastases. Prostate 52: 253–263, 2002. © 2002 Wiley‐Liss, Inc.
Polymorphisms in the androgen receptor and type II 5α‐reductase genes and prostate cancer prognosis
Authors: Shibata Atsuko , Garcia Maria Isabel , Cheng Iona , Stamey Thomas A. , McNeal John E. , Brooks James D. , Henderson Stavonnie , Yemoto Cheryl E. , Peehl Donna M.
Published: October 2002

BACKGROUND

Cytosine‐adenine‐guanine repeat length of the androgen receptor gene and the A49T and V89L polymorphisms of the 5α‐reductase (SRD5A2) gene have been associated with prostate cancer.

METHODS

We investigated the relationship of the three genetic polymorphisms to tumor grade among 211 men who had undergone radical prostatectomy. Subjects had prostate cancer <3 cm3 with a percentage of cancer represented by Gleason grade 4 or 5 (% Gleason grade 4/5) of either ≥20% or ≤5%. We also examined the association between those genetic markers and prostate specific antigen (PSA) failure among 112 subjects with ≥20% Gleason grade 4/5.

RESULTS

In cross‐sectional analysis, none of the polymorphisms was a significant predictor of % Gleason grade 4/5. In longitudinal analysis, the LL genotype at the V89L site was associated with statistically significant four‐ to sixfold increase in PSA failure risk after adjustment for clinicopathologic variables.

CONCLUSIONS

We observed poorer prognosis among men with the LL genotype at codon 89 of the SRD5A2 gene. Lack of consistency between studies must be resolved before clinical utility of this marker is established. Prostate 52: 269–278, 2002. © 2002 Wiley‐Liss, Inc.
Susceptibility to proteolysis of triosephosphate isomerase from two pathogenic parasites: Characterization of an enzyme with an intact and a nicked monomer
Authors: Reyes‐Vivas Horacio , Martínez‐Martínez Eduardo , Mendoza‐Hernández Guillermo , López‐Velázquez Gabriel , Pérez‐Montfort Ruy , Tuena de Gómez‐Puyou Marietta , Gómez‐Puyou Armando
Published: October 2002

The susceptibility to subtilisin of homodimeric triosephosphate isomerase from Trypanosoma brucei (TbTIM) and Trypanosoma cruzi (TcTIM) was studied. Their amino sequence and 3D structure are markedly similar. In 36 h of incubation at a molar ratio of 4 TIM per subtilisin, TcTIM underwent extensive hydrolysis, loss of activity, and large structural alterations. Under the same conditions, only about 50% of the monomers of TbTIM were cleaved in two sites. The higher sensitivity of TcTIM to subtilisin is probably due to a higher intrinsic flexibility. We isolated and characterized TbTIM that had been exposed to subtilisin. It exhibited the molecular mass of the dimer, albeit it was formed by one intact and one nicked monomer. Its kcat with glyceraldehyde 3‐phosphate was half that of native TbTIM, with no change in Km. The intrinsic fluorescence of nicked TbTIM was red‐shifted by 5 nm. The association between subunits was not affected. The TbTIM data suggest that there are structural differences in the two monomers or that alterations of one subunit change the characteristics of the other subunit. In comparison to the action of subtilisin on TIMs from other species, the trypanosomal enzymes appear to be unique. Proteins 2002;48:580–590. © 2002 Wiley‐Liss, Inc.
Automatic breakpoint detection for retrospective cumulative sum charts
Authors: Taylor A.L. , Tait S.P. , Porter M.A. , Perry M.J. , Nicolson R.W.
Published: October 2002

Cumulative sum (cusum) methods can be used for monitoring processes and for retrospective (historical) data analysis. Most software only provides the former. The comment by Williamson that retrospective cusum analysis is a neglected area is still true. Though not in vogue, retrospective cusum analysis is useful for investigations such as benchmarking of processes, identifying causes of process decay, selecting reference data sets for typicality studies, and reporting of historical data. Even those texts which cover retrospective analyses, usually ignore the question of identifying multiple points of change (breakpoints), and present essentially manual methods for assessing single breakpoints. Most users of statistical methods want software solutions that are easy to use and require little user intervention or interpretation. Direct implementation of manual method does not give a user robust solution. Problems are illustrated. Attempts to use monitoring CuSums in retrospective analysis can also lead to errors. A practical recursive method is presented for breakpoint identification and significance assessment, which can be automated. Copyright © 2002 John Wiley & Sons, Ltd.
An antiviral principle present in a purified fraction from Melia azedarach L. leaf aqueous extract restrains herpes simplex virus type 1 propagation
Authors: Alché Laura E. , Barquero Andrea A. , Sanjuan Norberto A. , Coto Celia E.
Published: October 2002

Meliacine (MA), an antiviral principle isolated from leaves of Melia azedarach L., exhibits potent antiviral activity against herpes simplex virus type 1 (HSV‐1) by inhibiting specific infected‐cell polypeptides (ICPs) produced late in infection. Some of these are involved in DNA synthesis and in the assembly of nucleocapsids. The present report provides additional evidence to elucidate the mode of action of MA against HSV‐1. Time‐of‐addition experiments confirmed that MA affects a late event in the multiplication cycle of HSV‐1. We showed that MA diminished the synthesis of viral DNA and inhibited the spread of infectious viral particles when HSV‐1 that expresses β‐galactosidase activity was used. In addition, the lack of a protein with an apparent MW of 55 KD was detected in MA‐treated cell extracts. Ultrastructural analysis of infected cells showed that, in the case of MA treatment, a large number of unenveloped nucleocapsids accumulated in the cytoplasm and a minor proportion of mature virus was found in cytoplasmic vesicles.

These findings suggest that MA exerts an antiviral action on both the synthesis of viral DNA and the maturation and egress of HSV‐1 during the infection of Vero cells. Copyright © 2002 John Wiley & Sons, Ltd.
Parametric optimization of magnetic‐field‐assisted abrasive flow machining by the Taguchi method
Authors: Singh Sehijpal , Shan H. S. , Kumar Pradeep
Published: October 2002

Some hybrid‐machining processes have been developed in the recent past with a view to devising composite machining processes, which are able to overcome the limitations of one process with the help of advantageous features of another similar process. The present paper identifies the parameters of abrasive flow machining (AFM) that significantly affect the material removal when a magnetic field is applied around the workpiece. The Taguchi method has been adopted for studying the effect of magnetic‐field‐assisted AFM parameters, individually, on the abrasion rate of work materials. Optimization of the process parameters has been carried out for the purpose of off‐line monitoring of the process. Copyright © 2002 John Wiley & Sons, Ltd.
Complex symmeterized analysis of benzene vibrations
Authors: Rashev Svetoslav
Published: October 2002

The aim of this work is to introduce specific complex coordinates and wave functions (Hamiltonian eigenfunctions) for the description of vibrational motion in benzene. When suitably chosen, these complex functions are shown to possess interesting transformation behavior under the symmetry operations of the molecular symmetrical top point group D6. This behavior is analyzed and classified as “complex symmetry types” (CSTs). CSTs can be defined for all symmetrical top point groups. The description in terms of complex symmeterized coordinates and wave functions allows for the construction of a separable symmeterized vibrational basis set for benzene. New results from calculations on vibrational energies and redeterminations of harmonic force constants in benzene, obtained using the CST description, are presented. © 2002 Wiley Periodicals, Inc. Int J Quantum Chem, 2002
Best Available Treatment Technologies Applied to Groundwater Circulation Wells
Authors: Elmore Andrew Curtis , Graff Thomas
Published: October 2002

Groundwater circulation wells (GCWs) are a quasi‐in‐situ method for remediating
groundwater in areas where remediation techniques that limit the water available for municipal, domestic,
industrial, or agricultural purposes are inappropriate. The inherently resource‐conservative nature of
groundwater circulation wells is also philosophically appealing in today's culture, which is supportive of
green technologies. Groundwater circulation wells involve the circulation of groundwater through a
dual‐screen well, with treatment occurring between the screens. The wells are specifically designed so that
one well screen draws in groundwater and the second returns the groundwater after it has been treated within the
well. Historically, the treatment has been performed with specialized equipment proprietary to GCW vendors. Two
full‐scale pilot systems at a formerly used Defense Superfund site in Nebraska used best available
technologies for treatment components. A multiple‐tray, low‐profile air stripper typically used for
pump‐and‐treat remediation systems was successfully adapted for the GCW pilot system located in a
trichloroethylene (TCE) hot spot. An ultraviolet water disinfection system was successfully adapted for
the GCW pilot system located in a hot spot contaminated with the explosive compound
hexhydro‐1,3,5‐trinitro‐1,3,5‐triazine (RDX). The pilot systems showed that
GCW technology is competitive with a previously considered pump‐and‐treat alternative for focused
extraction, and the regulatory community was supportive of additional GCW applications. A remedial design for the
site includes 12 more GCW systems to complete focused remediation requirements. © 2002 Wiley Periodicals,
Inc.
A Pearson‐type goodness‐of‐fit test for stationary and time‐continuous Markov regression models
Authors: Aguirre‐Hernández R. , Farewell V. T.
Published: October 2002

Markov regression models describe the way in which a categorical response variable changes over time for subjects with different explanatory variables. Frequently it is difficult to measure the response variable on equally spaced discrete time intervals. Here we propose a Pearson‐type goodness‐of‐fit test for stationary Markov regression models fitted to panel data. A parametric bootstrap algorithm is used to study the distribution of the test statistic. The proposed technique is applied to examine the fit of a Markov regression model used to identify markers for disease progression in psoriatic arthritis. Copyright © 2002 John Wiley & Sons, Ltd.
Gender and stress: is gender role stress? A re‐examination of the relationship between feminine gender role stress and eating disorders
Authors: Bekker Marrie H. J. , Boselie Kirsten A. H. M.
Published: October 2002

The present study was, first, aimed at examining the relationship between eating disorders, feminine gender role stress and other types of stress. In addition, we investigated whether eating disordered women compared to non‐clinical controls use depressogenic coping more often. We hypothesized that women with eating disorders would, compared to controls, suffer from more stress, irrespective of the type of stress, and that they would use depressogenic coping more frequently. Participants were 36 women suffering from eating disorders (mean age 25.8 years) and 53 controls (mean age 21.2 years). Questionnaires were administered reflecting the presence (or absence) of anorexia nervosa and/or bulimia nervosa, feminine gender role stress, perceived life stress, and coping styles. Compared with controls, eating disordered women reported higher levels of feminine gender role stress, but also higher levels of masculine gender role stress as well as recently experienced stress. Stress in general rather than femininity‐related stress exclusively was related to eating disorders. In addition, women suffering from eating disorders used emotional coping more often than the control group. We tend to conclude that not their relatively high stress levels are constitutive for eating disorders, but rather a specific way of coping with negative emotions, emotion‐focused coping by means of emotional eating. Copyright © 2002 John Wiley & Sons, Ltd.
Lessons learned becoming a framework developer
Authors: Carey James , Carlson Brent
Published: October 2002

When we began developing frameworks we were naive about what was involved in developing a framework versus other software. We had experience in object‐oriented development, but even that did not prepare us for some of what we learned as we developed frameworks. This paper captures some of the lessons that we learned as we became framework developers. If you have already developed frameworks these will validate what you have already learned, but for someone just starting out these are the areas where we believe you will be surprised. For example, are you ready to become comfortable exposing your work completely, since almost everything might be part of the framework product? Are you ready to become more disciplined so you can manage and deliver all these pieces? Other lessons will validate and enhance what you learned developing object‐oriented software; for example, the value of patterns, especially to define customization points. All are part of the biggest lesson of all, that as a framework developer you are no longer independently developing an application for an end‐user, instead you are on the many diverse customer teams using the framework—helping them deliver their applications. Copyright © 2002 John Wiley & Sons, Ltd.
Phenotypic, ethologically based resolution of spontaneous and D2‐like vs D1‐like agonist‐induced behavioural topography in mice with congenic D3 dopamine receptor “knockout”
Authors: McNamara Fergal N. , Clifford Jeremiah J. , Tighe Orna , Kinsella Anthony , Drago John , Fuchs Sara , Croke David T. , Waddington John L.
Published: October 2002

Uncertainty as to the functional role of the D3 dopamine receptor, due primarily to a paucity of selective agonists or antagonists, is being addressed in mice with targeted gene deletion (“knockout”) thereof. This study describes, for the first time, the phenotype of congenic D3‐null mice. Initially, 129/Sv × C57BL/6 D3‐null mice were backcrossed 14 times onto C57BL/6; they were then assessed using an ethologically based approach which resolves all topographies of behaviour within the mouse repertoire. The ethogram of D3‐null mice, on comparison with wildtypes, was characterised by no alteration in any topography of behaviour over an initial period of exploration; subsequent assessment over several hours revealed only increased rearing among females due to delayed habituation. Low doses of the selective D2‐like agonist RU 24213 (0.016–0.25 mg/kg) inhibited topographies of exploratory behaviour; this effect was diminished in D3‐null mice only when investigated following prolonged habituation, and then only for certain topographies of behaviour, primarily sniffing and rearing. High doses of RU 24213 (0.1–12.5 mg/kg) induced stereotyped sniffing and “ponderous” locomotion, while the selective D1‐like agonist SK&F 83959 (0.016–2.0 mg/kg) promoted characteristic grooming syntax; these effects did not differ materially between the genotypes. When examined topographically on an essentially congenic C57BL/6 background (<0.005% 129/Sv), the resultant phenotype indicated essential conservation of the mouse ethogram, high‐dose D2‐like stimulatory effects, and D1‐like stimulatory effects in the absence of D3 receptors. A role for D3 receptors in inhibitory processes appeared topographically circumscribed and only when baseline levels of behaviour were low. Synapse 46:19–31, 2002. © 2002 Wiley‐Liss, Inc.
Incision of AP sites in lung cancer patients: a pilot study
Authors: Rossi Ottavio , Filiberti Rosangela , Neri Monica , Biggi Stefano , Satragno Livia , Puntoni Riccardo , Frosina Guido
Published: October 2002

DNA base excision repair (BER) removes frequent DNA lesions of either endogenous or exogenous origin. Some indications point to BER defects in lung cancer patients. We have investigated the ability of ten lung cancer patients to repair natural AP sites, the most frequent genetic lesion, using an in vitro assay in which peripheral blood lymphocytes (PBL) extracts incise randomly depurinated plasmid DNA. The median value of repair activity in patients was lower than that of matched controls but the difference did not reach significance. Unlike other BER enzymatic steps, marked defects in incision of AP sites may not be associated with lung carcinogenesis. © 2002 Wiley‐Liss, Inc.
Study of chromosome damage in patients with breast cancer treated by two antineoplastic treatments
Authors: Silva L.M. , Takahashi C.S. , Carrara H.H.A.
Published: October 2002

The frequencies of chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) were determined in peripheral blood lymphocyte cultures from women with breast cancer treated by chemotherapy (CT) with FEC (5‐fluorouracil, epirubicin, and cyclophosphamide) or CMF (cyclophosphamide, methotrexate, and 5‐fluorouracil) cocktail in six CT cycles. The number of patients in each CT cycle were from 1 to 3 for SCE and 2 to 5 for CA. Samples were collected before and 48 h after CT. Although the size of the sample was limited and interindividual variability was wide, it appears that a 21‐day interval between CT sessions is sufficient for cell recovery. This fact was demonstrated by the reduction in CA and SCE frequencies between cycles in parallel with the unchanged mitotic index and proliferative index values. © 2002 Wiley‐Liss, Inc.
Effects of methionine on selenium embryotoxicity in cultured rat embryos
Authors: Usami Makoto , Tabata Hirobumi , Ohno Yasuo
Published: October 2002

Effects of methionine, an essential amino acid, on the embryotoxicity of selenium (Se) were examined using the rat embryo culture. Rat embryos at day 9.5 of gestation were cultured for 48 h in the presence of sodium selenite at 10 and 20 μM or sodium selenate at 30 and 100 μM with or without the addition of 1 mM DL‐methionine. Selenite at 20 μM or selenate at 100 μM alone increased the incidence of embryonic malformation and inhibited the embryonic growth. The addition of methionine increased the incidence of embryonic malformation at 10 μM of selenite but decreased the incidence of embryonic malformation at 100 μM of selenate. On the other hand, the addition of methionine partially restored the inhibited embryonic growth at 20 μM of selenite or at 100 μM of selenate. It was considered from these results that the methionine availability in the embryonic environment and the oxidation state of Se are critical in Se embryotoxicity. © 2002 Wiley‐Liss, Inc.
The Use of Hydrophilic Groups in Aqueous Organic Reactions
Authors: Itami Kenichiro , Yoshida Jun‐Ichi
Published: October 2002

The representative examples of beneficial effects of hydrophilic groups in aqueous organic reactions are described, including the Diels–Alder reactions, hetero Diels–Alder reactions, Claisen rearrangement, radical reactions, and transition metal–catalyzed reactions. Although the low solubility of organic molecules in water has been a bane in aqueous organic reactions, the incorporation of hydrophilic groups into the substrate structure can overcome the solubility problem and at the same time enhance the hydrophobic effect. In some cases, interesting micellar effects are observed because of the amphiphilic natures of such molecules. The emerging concept of “removable hydrophilic group,” in which the solubility problems have been alleviated, yet the initial product can still be transformed into a variety of products with the removal of the hydrophilic groups, is also described. © 2002 The Japan Chemical Journal Forum and Wiley Periodicals, Inc. Chem Rec 2: 213–224, 2002: Published online in Wiley InterScience (www.interscience.wiley.com) DOI 10.1002/tcr.10021
Acute toxicity and bioaccumulation of pesticide Diazinon in red tilapia (Oreochromis niloticus x Mossambicus albina)
Authors: Palacio Jaime A. , Henao Beatriz , Vélez Jorge H. , González Jairo , Parra Carlos M.
Published: October 2002

Young red tilapias were exposed for 96 h to each one of 6 concentrations of the pesticide Diazinon in order to determine the pesticide's acute toxicity level. After the ascertaining the lethal concentration (LC50) at 96 h, a level 10 times lower was selected for the bioaccumulation study of the pesticide in male and female specimens exposed for 9 days. The elimination process was carried out for 10 days beginning right after the conclusion of the accumulation process. Analytical procedures were developed and used for the studies of acute toxicity and bioaccumulation of Diazinon in red tilapia. A lethal concentration [LC50 (96 h)] of 3.85 mg/L was found, and steady‐state accumulation, at a concentration of 28.45 mg/kg, was reached at 7.72 days. In the elimination process a concentration of 0.29 mg/kg was found in tilapia tissue by the sixth day after the fish were moved to clean water, and it continued to decrease quickly toward nondetectable levels. © 2002 Wiley Periodicals, Inc. Environ Toxicol 17: 334–340, 2002; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/tox.10063
L1–L3 Coster–Kronig and Li (i = 1,2,3) sub‐shell fluorescence yields for Th and U
Authors: Kumar Ajay , Puri Sanjiv , Mehta D. , Garg M. L. , Singh Nirmal
Published: October 2002

The L1–L3 Coster–Kronig yield, f13, for Th and U was determined using measured intensities of the Lα x‐rays emitted following selective ionization of the L3 sub‐shell by Mo K x‐rays (B < Einc. < B) and of those emitted following ionization of the Li (i = 1,2,3) sub‐shells by the 59.54 keV γ‐rays (B < Einc. < BK), where B is the K shell/Li sub‐shell ionization threshold of the target element. The measurements were performed using an energy‐dispersive x‐ray fluorescence set up involving an Si(Li) detector and photon sources consisting of an 241Am radioactive annular source used in (i) the direct excitation mode and (ii) the secondary excitation mode together with the Mo secondary exciter. In addition, the Li (i = 1,2,3) sub‐shell fluorescence yields (ωi) for Th and U were deduced using our earlier measured x‐ray fluorescence cross‐sections at incident photon energies in the range 17–60 keV and the theoretical photoionization cross‐sections. The measured f13 and ωi yields were compared with the available semi‐empirical values and the theoretical values based on relativistic Hartree–Dirac–Slater (RDHS) calculations. Copyright © 2002 John Wiley & Sons, Ltd.
The SCR1 gene from Schwanniomyces occidentalis encodes a highly hydrophobic polypeptide, which confers ribosomal resistance to cycloheximide
Authors: Hoenicka Janet , Fernández Lobato María , Marín Dolores , Jiménez Antonio
Published: October 2002

In Saccharomyces cerevisiae, the SCR1 gene from Schwanniomyces occidentalis is known to induce ribosomal resistance to cycloheximide (cyh). A 2.8 kb DNA fragment encoding this gene was sequenced. Its EMBL Accession No. is AJ419770. It disclosed a putative tRNAAsn (GUU) sequence located downstream of an open reading frame (ORF) of 1641 nucleotides. This ORF was shown to correspond to SCR1. It would encode a highly hydrophobic polypeptide (SCR1) with 12 transmembrane domains. SCR1 is highly similar to a variety of yeast proteins of the multidrug‐resistance (MDR) family. However, SCR1 only conferred resistance to cyh but not to benomyl or methotrexate. The cyh‐resistance phenotype induced by SCR1 was confirmed in several S. cerevisiae strains that expressed this gene to reside at the ribosomal level. In contrast, a β‐galacosidase‐tagged SCR1 was found to be integrated in the endoplasmic reticulum (ER). It is proposed that the ribosomes of yeast cells expressing SCR1 undergo a conformational change during their interaction with the ER, which lowers their affinity for cyh‐binding. If so, these findings would disclose a novel ribosomal resistance mechanism. Copyright © 2002 John Wiley & Sons, Ltd.
Staurosporine‐induced morphological changes in the rat osteoblasts.
Authors: Yang Rong‐Sen , Lu Kuo‐Shyan , Fu Wen‐Mei , Liu Tang‐Kue , Lin‐Shiau Shoei‐Yn
Published: October 2002

Treatment of cultured rat osteoblasts with staurosporine caused a rapid outgrowth of long slender cellular processes and the formation of stellate cells. The number of stellate cells increased with higher concentrations of and longer incubation with staurosporine. Scanning electron microscopy (SEM) revealed that the smooth surface of control polygonal cells became ruffled with many long slender cellular processes, thus increasing the cell surface area. Transmission electron microscopy (TEM) of the stellate cells showed a rich accumulation of large lipid droplets in the cytoplasm. Some lipid droplets had coalesced under the cytoplasmic membrane. We suggest that staurosporine has an effect on the differentiation of cultured rat osteoblasts.
Subcellular localization and antiviral activity of carminic acid/poly r(A‐U) combinations
Authors: Krabill Keith , Jamison James M. , Gilloteaux Jacques , Summers Jack L.
Published: October 2002

Carminic acid (CAR) enhances the antiviral activity of poly r(A‐U) twelve‐fold without increasing interferon induction, inactivating the vesicular stomatitis virus or inducing host cell cytotoxicity. Phase contrast photomicrographs of human foreskin fibroblasts (HSF) incubated with CAR alone, poly r(A‐U) alone or with a CAR/poly r(A‐U) combination illustrate that the CAR/poly r(A‐U) combinations display altered subcellular distribution with the CAR being localized in the nucleoli and chromatin. Phase contrast and fluorescence photomicrographs of adriamycin (ADR)‐treated and ADR/poly r(A‐U)‐treated HSF cells corroborate these findings. These results suggest that modulation of one or more nucleolar processes may be responsible for the enhanced antiviral activity.
Comparative expression of endogenous retrotransposons in adult mouse mammary gland during normal development and differentiation
Authors: Asch Harold L. , Vinci Robert Z. , Chang Jae Y. , Natoli Farah , Stoler Daniel L. , Anderson Garth R. , Asch Bonnie B.
Published: October 2002

Amplified expression of the endogenous retrotransposons, intracisternal A particles (IAPs) and murine leukemia virus‐related elements (MLVEs), along with decreased expression of VL30 elements frequently occurs during mouse mammary tumorigenesis. We have now analyzed the expression of these retroelements during the normal developmental and differentiation cycle of the mammary gland as found in virgin, pregnant, lactating, and post‐lactation adult female BALB/c mice. Retrotransposon expression was either unchanged or decreased during the progressive stages of the cycle compared to virgin tissue. Likewise, growth of mammary epithelial cells in primary culture had little or no effect on expression of IAPs, MLVEs and VL30 sequences. Thus, the dramatic changes involving these retrotransposons in many mouse mammary tumors appear unrelated to any normal state.
The type III‐9 repeat of human fibronectin is encoded by a single exon which is not alternatively spliced.
Authors: Gardella Rita , Barlati Sergio , Ferraboli Sergio , Colombi Marina
Published: October 2002

Type III homologies of human fibronectin are generally encoded by two exons, with the exception of the ED‐A and ED‐B repeats which are encoded by a single exon undergoing alternative splicing. We report that also the type III‐9 homology is encoded by a single exon. Further more, RT‐PCR analysis, performed on mRNA purified from fetal and adult tissues and from normal and tumor‐derived cell types, showed that the III‐9 region is not undergoing alternative splicing in all samples tested.
Expression of the 30 kD cysteine endoprotease B in germinating barley seeds
Authors: Marttila Salla , Porali Ilkka , Ho T.‐H. David , Mikkonen Anita
Published: October 2002

The expression of a 30 kD cysteine endoprotease (EP‐B) was studied by in situ hybridization and immunomicroscopy to clarify its role in germinating barley grains. At the beginning of germination, EP‐B mRNA was expressed in the scutellar epithelium and aleurone cells next to the embryo. Later, mRNA levels were highest in the aleurone layer proceeding to the distal end of the grain. During the first day of germination, EP‐B protein was strongly localized to the germ aleurone and scutellar epithelium from where the secretion into the starchy endosperm began. Secretion was also observed to proceed along the aleurone layer to the distal end. These results show that EP‐B is differentially localized during germination, and both scutellum and aleurone layer are able to synthesize and secrete EP‐B protein.
Surface labelling of plant protoplasts with fluorochromes for discrimination of heterokaryons by microscopy and flow cytometry
Authors: van Kesteren W. J. P. , Tempelaar M. J.
Published: October 2002

Surface labelling of plant protoplasts was tested for use in mass fusion systems and heterokaryon detection. Parameters have been established for biotinylation and subsequent incubation with avidin‐coupled fluorochromes. The procedure is rapid (less than 3 hours) and does not affect viability. Fusion responses were the same as with unlabelled protoplasts. From a range of fluorochromes tested, fluorescein and phycoerythrin proved best suited for detection experiments with protoplasts of both suspension and leaf origin. With this standard combination of labels, as applied in experiments with animal cells, heterokaryons from fused plant protoplasts could clearly be discriminated from other protoplasts by means of fluorescence microscopy or flow cytometry with a single combination of filters and wavelengths.
The survival of cytochalasin‐induced polykaryons following exposure to cytotoxic agents.
Authors: Court J. B. , Burn C. , Louis D. S. , Moore J. L.
Published: October 2002

Using CHO‐K1, HeLa S3 and two Walker lines (WR and WS) differentially sensitive to cis‐diamminedichloroplatinum(II) (cisplatin), the survival after exposure to cisplatin, mitomycin C, vinblastine, vincristine or cytosine arabinoside has been determined either of clonogens or of cells rendered polyploid by post‐exposure incubation in the presence of cytochalasin B (CB). It is suggested that the inhibition of cytokinesis by CB permits an assessment to be made of the fraction of damage whose expression is cell division‐related, possibly including that resulting from a loss or malsegregation of genetic material. It was found that the response of polykaryons in comparison to clonogens was both agent‐ and cell line‐dependent. After cisplatin exposure, polykaryon survival (defined as the ability to accumulate at least 16C DNA) declined exponentially with dose and was qualitatively, and to some extent quantitatively, similar to that observed previously after irradiation. In HeLa S3, giant cells induced by 10‐20Gy irradiation in the absence of CB exhibited a radiation dose‐dependent reduction in the relative frequency of highly polyploid cells which was similar to that observed in CB‐induced polykaryons.
The increase in cytosolic levels of protein kinase C precedes the growth factor requirements during the transition stem cells to differentiated progeny in the MCF‐7 breast tumor cells.
Authors: Gomez Maria L. , Resnicoff Mariana , Medrano Estela E. , Tellez‐Inon Maria T.
Published: October 2002

The expression of regulatory genes within the context of a differentiation program can have profound long‐term consequences in tissues with permanent renewing populations. The breast tumor cell line MCF‐7 retains in culture some of the characteristics of a unidirectional differentiation pathway. We show that the cytosolic activity of the regulatory enzyme protein kinase C (PKC) precedes and continues the sequence of maturation in pre‐differentiated subpopulations derived from a stem cell fraction. However, the activity declines in the most differentiated, post‐mitotic fraction. These results indicate that PKC may be considered among the regulatory genes in MCF‐7 cells that specify maturation of the stem cell progeny.
The effect of spermine on the disaccharidase activities in suckling rats of different age.
Authors: Shimizu Kazuo , Mushiake Sotaro , Yoshimura Norikazu , Harada Tokuzo , Okada Shintaro
Published: October 2002

The influence of the age of the rat on the maturation of disaccharidase activities induced by spermine was studied. Three‐day‐ and 9‐day‐old rats were used in the experiment. Spermine was administered orally or directly into the stomach using thin tubing daily for 3 days, and disaccharidase activities in the jejunum were measured. While spermine caused maturation of not only lactase activity but also maltase and sucrase activities in 9‐day‐old rats, it only caused maturation of lactase activity in 3‐day‐old rats. Histological studies showed no significant changes in the jejunum of 3‐day‐old rats treated with spermine.
The influence of maternal nicotine exposure on neonatal lung metabolism. Protective effect of ascorbic acid.
Authors: Maritz G. S.
Published: October 2002

The aim of the present study was to establish whether ascorbic acid supplementation (1 mg/kg/body mass/day) during pregnancy and lactation will prevent the effect of maternal nicotine exposure (1 mg/kg body weight/day) on neonatal lung carbohydrate, DNA and protein metabolism. The data show that the adult lung ascorbic acid content was reduced by 76% after exposure to nicotine. In contrast, maternal nicotine exposure during pregnancy and lactation has no effect on neonatal lung ascorbic acid content. However, ascorbic acid supplementation during pregnancy and lactation prevented the adverse effects of maternal nicotine exposure on neonatal lung carbohydrate, DNA and protein metabolism.
Absence of high levels of DNA polymerase α activity in the nuclear matrix prepared from mouse erythroleukemia cells
Authors: Martelli Alberto M. , Neri Luca M. , Billi Anna Maria , Bareggi Renato , Cocco Lucio
Published: October 2002

We have examined the association of DNA polymerase α activity with the nuclear matrix prepared by different techniques from mouse erythroleukemia cells. At variance with the data obtained using other cell types we have found that only a small amount (less than 2%) of nuclear DNA polymerase α activity resisted extraction with high‐ionic strength buffers, even if nuclei were heat‐stabilized by incubation at 37°C for 45 min prior to subfractionation. The recovery of DNA polymerase α activity bound to the matrix was unaffected by the type of extracting agent used (NaCl or (NH4)2 SO4), by the extraction sequence or by the method employed for obtaining nuclei. These results could indicate that in some types of cells the nuclear matrix is not involved in DNA replication.
A comparison of bovine serum albumin and chicken ovalbumin as supplements for the serum‐free growth of chinook salmon embryo cells, CHSE‐214.
Authors: Barlian A. , Ganassin R. C. , Tom D. , Bols N. C.
Published: October 2002

Chicken ovalbumin and bovine serum albumin (BSA) were compared as supplements to the basal medium, L‐15, for the serum‐free cultivation of the Chinook Salmon Embryo cell line, CHSE‐214. Unlike L‐15 alone, ovalbumin and some commercial BSA preparations allowed cell proliferation and development of confluent monolayer cultures. However, only a fatty acid‐free BSA (2%) supported continuous proliferation for two years through approximately 15 subcultivations. For this, subcultivation was achieved with non enzymatic cell dissociation solutions. Also, new serum‐free subcultivation techniques were developed that utilized avian egg white trypsin inhibitors to terminate the action of either bovine or cod trypsin. Finally, CHSE‐214 were successfully cryopreserved in 2% BSA, allowing all cell cultivation steps to be performed in the absence of FBS.
Electron microprobe analysis in periodontal guided tissue regeneration.
Authors: Campos A. , González‐Jaranay M. , Moreu G. , Sánchez‐Quevedo M. C.
Published: October 2002

Electron microprobe analysis was used to determine the evolution of Ca, P and S in regenerated tissue surrounding incisors roots after periodontal treatment with guided tissue regeneration. Our results, which showed increased Ca and P, and decreased S are discussed in relation to the process of mineralization electron probe microanalysis with potentially provided an accurate means of assessing the degree of mineralization in extremely small tissue samples.
Estrogen‐regulated expression of a growth arrest specific gene (gas‐1) in rat uterus
Authors: Ferrero Marina , Cairo Gaetano
Published: October 2002

gas‐1 belongs to a family of growth arrest specific genes negatively regulated after growth induction of arrested cells. We report the expression of gas‐1 in an in vivo system of cell proliferation. gas‐1 mRNA accumulates progressively in the uterus of ovariectomized rats with a peak at three weeks after surgery. After estrogen treatment gas‐1 mRNA levels decrease within two hours, at a time when c‐myc expression is greatly increased, and return to pretreatment levels at 48 hours. Treatment with cycloheximide does not prevent estrogen‐induced down‐regulation of gas‐1 mRNA levels. The present results show that: i) estrogen affects the uterine growth state by regulating the expression of both positively and negatively acting genes, ii) gas‐1 expression is controlled by cellular growth state also in vivo.
Correction of the defective extracellular matrix of Ehlers‐Danlos syndrome skin fibroblasts by dexamethasone.
Authors: Moro Laura , Colombi Marina , Zoppi Nicoletta , Ghinelli Annalisa , Barlati Sergio
Published: October 2002

Skin fibroblasts derived from Ehlers‐Danlos syndrome (EDS) patients lack an organized extracellular matrix (ECM) of fibronectin (FN) and often show an accumulation of cytoplasmic FN. The treatment of EDS cells of different types (I to VIII) with 10‐7 M dexamethasone (dex), as well as cocultivation with control fibroblasts, induced in most cases the assembly of a FN‐like ECM. The study of FN mRNA expression by dot‐blot hybridization and of FN released into the culture media of EDS cells showed that the correction of the defective FN‐ECM of EDS cells by dex treatment is associated in most cases with an increase of FN mRNA synthesis and of secreted FN.
4‐Hydroxynonenal induces membrane perturbations and inhibition of basal prostacyclin production in endothelial cells, and migration of monocytes.
Authors: Moldovan Nicanor I. , Lupu Florea , Moldovan Leni , Simionescu Nicolae
Published: October 2002

Cultured bovine aortic endothelial cells (BAEC) were incubated for 5 days with 10−5 4‐hydroxynonenal (HN). HN treated BAEC and controls were either (i) further incubated with 125I‐polymyxin B (IPxB) or with radioiodinated, inactivated coagulation factor Xa (IFXai) as markers of membrane phospholipid perturbation, or (ii) assayed for the synthesis of prostacyclin (PGI2) and thromboxane A2 (TXA2). Rabbit blood mononuclear cells enriched in monocytes (MC) were isolated and assayed for chemotactic response to HN. The results showed six ‐ fold increases of IPxB and IFXai binding to BAEC treated with HN, as compared to untreated controls. We also found in HN treated cells a marked inhibition of PGI2 synthesis, but an unmodified TXA2 production. In addition, HN in the 10‐5‐10‐10 M range induced oriented migration of MC.
Studies on 17‐1A antigen gene regulation in nonexpressing A549 and A431 cells, as compared to expressing pancreatic carcinoma (CAPAN 2) cells, reveal a complex mechanism of repression of this gene.
Authors: Siemieniako Barbara , Wiland Ewa , Trzeciak Wiesław H.
Published: October 2002

Elements controlling high expression of the 17‐1A antigen gene in pancreatic carcinoma cells (Capan 2) reside within the two regions: proximal (−193 to +3) and distal (−877 to −518). We demonstrate here that some factors present in nuclear extracts from nonexpressing cells bind specifically to the control elements, important for gene expression. Our results suggest that nonexpressing cells may either lack at least one of the factors necessary for activation or may contain their modified forms. A major difference between expressing and nonexpressing cells was found in the region containing core enhancer sequence. Moreover, nonexpressing cells display a complex pattern of DNA‐protein interactions in this region, suggesting that these cells contain factors acting negatively mainly on the enhancer sequence. Our results however, indicate that the mechanism of repression is much more complicated than expected.
Transforming growth factor β1 increases the phosphoenolpyruvate carboxykinase mRNA level in cultured rat hepatocytes
Authors: Thoresen G. Hege , Christoffersen Thoralf
Published: October 2002

Transforming growth factor β1 (TGFβ1) elevated the phosphoenolpyruvate carboxykinase (PEPCK) mRNA abundance in primary cultures of rat hepatocytes. Although this increase was not as large as the rise in PEPCK gene expression induced by the cAMP‐elevating agents glucagon or isoproterenol, the effect of TGFβ1 was several‐fold and concentration‐dependent, with ED50 at about 2.5 pM, which is in the same concentration range as the previously found growth‐inhibitory effect of TGFβ. The data show that the level of mRNA for PEPCK, an enzyme typically expressed in the liver, can be regulated in the same direction by TGFβ1 and cAMP.
Some aspects of zinc transport across eel (Anguilla anguilla) red blood cell membranes.
Authors: Mandolfino M. , Cimino G. , Scuteri A. , Cordì R. , Romano L.
Published: October 2002

Zinc movement across eel and human red blood cell membranes was measured by atomic absorption spectrophotometry. It was observed that:

In human red blood cells zinc uptake is twice as rapid as in fish red blood cells over a temperature range of 10‐40°C. The low rate of zinc uptake in eel red blood cell may be simply the side effect of different surface area to volume ratios for the differences in cell size or, it may be due to the low permeability of bicarbonate through the red blood cell membranes.

Zinc uptake measured in eel and human red blood cells treated and untreated with external trypsin shows different features. The zinc uptake was reduced by about 40% in treated eel red blood cells with respect to the total uptake of untreated red blood cells. Human red blood cells treated and untreated with trypsin do not show any differences in the amount of zinc transported.

In fish red blood cells, zinc uptake in NANO3 medium is markedly reduced, compared with that measured in NaCl medium. The [Zn2+i slightly increases in the presence of bicarbonate. In human red blood cells in NANO3 medium the zinc uptake is strongly reduced and the presence of bicarbonate marginally increases the zinc influx.

In eel red blood cells there seem to be two independent pathways for zinc uptake: one DIDS‐sensitive and the other DIDS‐insensitive. DIDS 10 μM inhibits only 64% of the total zinc transported. Iincreasing the DIDS concentration did not give more inhibition. In human red blood cells only one DIDS‐sensitive pathway for zinc transported seems to exist, because, 2,5 μM DIDS inhibits 97% of zinc uptake.