bluuebunny/crossref_metadata_2025 huggingface dataset:
The 14-3-3 proteins in the teleost fish rainbow trout (Oncorhynchus mykiss)
Authors: Koskinen Heikki , Krasnov Aleksei , Rexroad Caird , Gorodilov Yuri , Afanasyev Sergey , Mölsä Hannu
Published: August 2004

We report studies of 14-3-3 genes in rainbow trout, adding to current understanding of the molecular evolution of this multigene family and its functional importance in fish. Ten genes were identified that are apparent duplicates of five ancestors. The duplicated 14-3-3 genes diverged rapidly and their cladogram is markedly different from the phylogenetic tree. The mean rate of nonsynonymous divergence of trout 14-3-3 genes is one order of magnitude greater than that of mammalian genes. An evolutionarily recent genome duplication in salmonid fish relaxed functional constraints, and selection favored establishment of novel isoforms. Differences in tissue distribution of 14-3-3 genes were minor; however, results of 31 microarray experiments showed divergence of expression profiles, which was related to structural divergence of the duplicates. We observed remarkable coordination of expression of all isoforms in our study of stress response in the brain. Profiles of the 14-3-3 genes correlated with a large group of chaperones and genes involved in cell communication and signal transduction. We studied embryonic expression of 14-3-3 genes and found abundant transcripts in the rapidly growing and differentiating parts of embryos such as eyes, tail bud and yolk syncytium during somitogenesis and in gills and pectoral fins after completion of somitogenesis. Consistent expression was observed in the neural crest, which is known to have high morphogenetic potential.
Arachidonic acid reduces the stress response of gilthead seabream Sparus aurata L.
Authors: Van Anholt R. D. , Spanings F. A. T. , Koven W. M. , Nixon O. , Bonga S. E. Wendelaar
Published: August 2004

In this study the influence of the dietary level of the fatty acid arachidonic acid (ArA, 20:4n-6) was determined on the acute stress response and osmoregulation of adult gilthead seabream Sparus aurata L. Seabream were fed a diet containing either 0.9% or 2.4% of total fatty acids as ArA for 18 days before being subjected to a 5 min period of net confinement. Prior to this stressor, a subgroup of fish from both dietary treatment groups was treated with acetylsalicylic acid (ASA), an irreversible blocker of cyclooxygenase (COX). This would indicate whether any effects were caused by an enhanced synthesis of prostaglandins derived from ArA. The highest ArA levels were found in the kidneys, and these were further enhanced by dietary ArA-supplementation. In gill tissues, there were significant changes in all selected fatty acid classes 24 h after confinement, except for the docosahexaenoic acid (DHA, 22:6n-3): eicosapentaenoic acid (EPA, 20:5n-3)ratio. ArA feeding strongly reduced the cortisol response to confinement,which was partially counteracted by ASA treatment. ArA also attenuated the stress-associated increase in plasma osmolality and, in combination with ASA,enhanced the osmolality and plasma chloride levels, but reduced plasma sodium levels after confinement. Furthermore, ArA enhanced the branchial Na+, K+-ATPase activity both before and after confinement, whereas feeding ASA diminished this effect. It appeared that the effects of ArA-supplementation could not always be ascribed to an increase in prostaglandin synthesis. It is advisable to determine the long-term effects of replacing fish oils in commercial diets with vegetable oils that contain no long-chain fatty acids, particularly in carnivorous/marine species with low fatty acid elongation and desaturation activities. The effects of a low dietary intake of ArA (and other polyunsaturated fatty acids) should be studied over a longer term, taking into account any consequences for the health of the fish.
Bioconvective pattern formation ofTetrahymenaunder altered gravity
Authors: Mogami Yoshihiro , Yamane Akiko , Gino Atsuko , Baba Shoji A.
Published: August 2004

Bioconvection is a result of the negative gravitactic behavior of microorganisms. When the top-heavy density gradient generated by gravitaxis grows sufficiently large, an overturning convection occurs leading to a formation of characteristic patterns, which involve highly concentrated aggregation of cells into extended two-dimensional structures. Although gravity is a crucial factor, few experiments have been done with reference to gravity as an experimental variable. In order to gain an insight into the hydrodynamic as well as biological dependence of the convective motion on gravity, we investigated changes in bioconvective patterns of Tetrahymena under altered gravity acceleration generated by a long-arm centrifuge. Bioconvective patterns recorded of three different cell strains (T. pyriformis, T. thermophila and its behavioral mutant,TNR) were analyzed quantitatively using space-time plot and Fourier analysis. For example, under subcritical conditions, when T. pyriformis(1.0×106 cells ml-1) was placed in a 2 mm-deep chamber, no spatial pattern was observed at 1 g. When the suspension was centrifuged, however, patterns began to appear as acceleration increased over a critical value (1.5 g), and then remained steady. The formation was reversible, i.e. the patterns disappeared again as acceleration decreased. Under supracritical conditions, i.e. when a suspension of the same density was placed in a 4 mm-deep chamber, a steady state pattern was formed at 1 g. The pattern spacing in the steady state was observed to decrease stepwise in response to step increases in acceleration. Fourier analysis demonstrated that for TNR the mean wave number changed almost simultaneously with step changes in acceleration, whereas the responses were less sharp in the wild-type strains. This may suggest that the locomotor phenotype of the cell, such as its avoiding response ability, has a crucial role in bioconvective pattern formation. These findings are discussed in relation to former theoretical studies.
Renal function in Palestine sunbirds: elimination of excess water does not constrain energy intake
Authors: McWhorter Todd J. , del Rio Carlos Martínez , Pinshow Berry , Roxburgh Lizanne
Published: August 2004

Although the renal responses of birds to dehydration have received significant attention, the consequences of ingesting and processing large quantities of water have been less studied. Nectar-feeding birds must often deal with exceptionally high water intake rates in order to meet their high mass-specific energy demands. Birds that ingest large volumes of water may either eliminate excess water in the kidney or regulate the volume of water absorbed in the gastrointestinal tract. Because water absorption in the gastrointestinal tract of Palestine sunbirds (Nectarinia osea)decreases with increasing water ingestion rate, we predicted that glomerular filtration rate (GFR) in these birds would not be unusually high in spite of large ingested water loads. When feeding on dilute sucrose solutions, sunbirds ingested between 4 and 6 times their body mass in nectar per day, yet they were able to compensate for varying nectar energy density and increased thermoregulatory energy demands with no apparent difficulty. GFR was lower than predicted (1976.22±91.95 μl h-1), and was not exceptionally sensitive to water loading. Plasma glucose concentrations were high, and varied 1.8-fold between fasted (16.08± 0.75 mmol l-1) and fed (28.18±0.68 mmol l-1) sunbirds, but because GFR was low, glucose filtered load also remained relatively low. Essentially the entire glucose filtered load (98%) was recovered by the kidney. Renal fractional water reabsorption (FWR) decreased from 0.98 to 0.64 with increasing water intake. The ability of Palestine sunbirds to reduce the absorption of ingested water in the gastrointestinal tract may resolve the potential conflict between filtering a large excess of absorbed water in the kidney and simultaneously retaining filtered metabolites.
Electrodepositing Redox Polymer on Sandwich Complex for the Improvement of Sensitivity in Sandwich Enzyme-linked Immunoassay
Authors: Gao Qiang , Qi Bin , Sha Yufang , Yang Xiurong
Published: August 2004

Improvement of the sensitivity of electrochemical sandwich enzyme immunoassay has been achieved by electrodepositing redox polymer on screen-printed carbon electrode surface, on which the sandwich complex was formed.
Preparation of a Nanocrystalline TiO2 Photocatalyst Using a Dry-process with Acetylene Black
Authors: Tang Weiping , Chen Zaihua , Katoh Shunsaku
Published: August 2004

Well-dispersed anatase-type TiO2 nanocrystals showing good photocatalytic activity were prepared for the first time by a dry-process involving only the heating of a mixture of titanium tetraisopropoxide and acetylene black.
Artificial Assembly of Myoglobin and Flavodoxin Reductase Using Designed Coiled-coil Peptides
Authors: Sakamoto Seiji , Itoh Atsushi , Kudo Kazuaki
Published: August 2004

A unique and artificial assembly of myoglobin and flavodoxin reductase was constructed using a heterodimeric coiled-coil formation of de novo designed polypeptides in order to demonstrate a novel method for the artitecture of the active-site directed fusion protein.
Biomimetic Hydrolysis of p-Nitrophenyl Alkanoates with Functionalized Mesoporous Silicas
Authors: Chang Jeong Ho , Kim Kyung Ja , Shin Young-Kook , Liu Jun
Published: August 2004

A biomimetic catalysis of p-nitrophenyl alkanoate esters as a function of their alkyl chain lengths through multifunctionalization of molecular self-assembly onto ordered silica nanopores has been investigated in Tris-HCl buffer, and their kinetic parameters were calculated including a Michaelis–Menten kinetics.
4-Difluoromethylated Quinoline Synthesis via Intramolecular SN2′ Reaction of α-Trifluoromethylstyrenes Bearing Imine Moieties
Authors: Mori Takashi , Ichikawa Junji
Published: August 2004

Intramolecular cyclization of o-methyleneamino-substituted α-trifluoromethylstyrenes is promoted by DBU and a catalytic amount of KCN to provide 4-(difluoromethyl)quinolines. The reaction proceeds via (i) the generation of carbon nucleophiles from the imine moieties, (ii) their intramolecular SN2′ reaction with loss of a fluoride ion, and (iii) subsequent aromatization by alkene isomerization.
A Novel BEDT-TTF-based Organic Conducting Salt with a Ferrocene-containing Dianion, α-(BEDT-TTF)4(Fe(Cp–CONHCH2SO3)2)·4H2O
Authors: Furuta Keigo , Akutsu Hiroki , Yamada Jun-ichi , Nakatsuji Shin’ichi
Published: August 2004

We have prepared a new organic dianion containing the ferrocene derivative, Fe(C5H4–CONHCH2SO3−)2, as a PPh4 salt. Cyclic voltammetry indicates its oxidation potential is 0.14 V higher than that of BEDT-TTF. Electrocrystallization with BEDT-TTF gave the title compound. Its crystal structure and physical properties are reported.
Photophysical and Photocatalytic Properties of Molybdates and Tungstates with a Scheelite Structure
Authors: Kato Hideki , Matsudo Naoko , Kudo Akihiko
Published: August 2004

Photophysical and photocatalytic properties of molybdates and tungstates with a scheelite structure were investigated. The band gaps of scheelite compounds narrowed when the compounds were composed of Ag+ and Bi3+ ions. (NaBi)0.5MoO4 (BG: 3.1 eV) and (AgBi)0.5MoO4 (BG: 3.0 eV) showed the photocatalytic activities for O2 evolution from an aqueous solution containing an electron acceptor. On the other hand, (NaBi)0.5WO4 (BG: 3.5 eV) produced H2 from an aqueous solution containing an electron donor.
Hydrogen Adsorption/Desorption Property of Activated Carbon Loaded with Platinum
Authors: Takagi Hideyuki , Hatori Hiroaki , Yamada Yoshio
Published: August 2004

The hydrogen adsorption isotherms of activated carbon fiber loaded with platinum (Pt–ACF) in the 303–673 K range indicated that a temperature above 573 K was required to give rise to an increase in the amount of adsorbed hydrogen due to the spillover phenomenon. Two peaks at 800 and 1200 K in the D2 desorption profile of Pt–ACF clarified that two kinds of sites for adsorption or bonding of spiltover deuterium exist on the carbon surface.
Failure of a trial evaluating the effect of venous surgery on healing and recurrence rates in venous ulcers? The USABLE trial: rationale, design and methodology, and reasons for failure
Authors: Davies A H , Hawdon A J , Greenhalgh R M , Thompson S
Published: August 2004

Objective: This Medical Research Council funded trial was set up to investigate whether venous surgery provided any additional benefit in the healing and recurrence rates of venous ulcers over compression bandaging alone.

Methods: Ulcer surgery as adjuvant to compression bandaging for leg ulcers (USABLE) was a multicentre, randomized controlled trial, which planned to recruit 1000 venous ulcer patients to receive either compression bandaging alone or compression bandaging plus venous surgery. Follow up recorded ulcer healing, recurrence and patient quality of life.

Results: Recruitment was slow, and involved screening a large number of patients (759) with only 75 randomized over an 18-month recruitment period.

Conclusions: The results would infer that surgery is unlikely to be a management option for the majority of patients presenting to a hospital with a gaiter area leg ulcer.
Causal attributions in delusional thinking: An investigation using qualitative methods
Authors: Beese A. G. , Stratton P.
Published: August 2004

Objectives: To investigate the attributions offered spontaneously during interview by patients who have delusional beliefs. Specifically, to assess whether people with delusional beliefs reason in a way that is analysable. If so, to extend the findings from previous research to provide a detailed picture of the patients' accounts that could inform cognitive therapies for psychosis.

Method: A group of 31 patients with delusional beliefs were interviewed using a semi‐structured open‐ended interview. Their causal attributions were identified and analysed in terms of standard definitions of attributional theory using the Leeds Attributional Coding System.

Results: Patients made an average of 96 attributions each. These showed a high degree of recognizable coherence in reasoning, including when directly discussing delusional beliefs. The patterns of attributing were partly consistent with previous research findings, with external attributions being offered for negative events. Several other aspects emerged, indicating a pervasive negativity, with non‐delusional aspects of the illness being a major concern. When delusions were described as a cause of events, the dominant pattern of attributing was stable, internal, personal and uncontrollable, with negative outcomes. When delusions were reported as an outcome, the patient was more in control.

Conclusions: The reasoning processes of patients with delusional beliefs are in many aspects normal and include patterns not identified by the ASQ. Attributing in relation to delusions differs from talk about other subjects. The qualitative attributional approach provided a detailed and rich picture of the world of these patients. Interpretable tendencies are described that should provide information useful to those practising and evaluating cognitive therapies.
Life events, depression and social support in dementia
Authors: Waite Allyson , Bebbington Paul , Skelton‐Robinson Martin , Orrell Martin
Published: August 2004

Objectives: Recent research has highlighted the influence of psychosocial factors on the course of dementia, and previous studies have demonstrated an association between the experience of life events and depression. This study aimed to investigate the interrelationships between life events, depression and social support in people with dementia.

Design: Participants were 72 people with dementia and their carers. Data were collected on severity of dementia, cognitive impairment, depression, life events and social support.

Results: The results showed that 62% of people with dementia who had experienced a severely threatening and independent life event in the preceding 3 months were depressed, compared with only 28% of those people with dementia without such events (p = .005). No relationships were found between social support and depression in people with dementia.

Conclusions: This study builds on previous research indicating that people with dementia who experience stressful life events have a higher risk of depression.
An evaluation of gender differences on the Belbin Team Role Self‐Perception Inventory
Authors: Anderson Neil , Sleap Sarah
Published: August 2004

This paper presents results into gender differences and potential adverse impact on Belbin's original BTRSPI. Based on a sample of 311 sales and customer service representatives (208 females, 103 males), a series of descriptive statistics, parametric and non‐parametric tests are reported. Findings suggest significant differences between genders in response patterns favouring males on the team roles of Chair, Shaper, and Plant. Conversely, females scored significantly higher on the role of Team Worker. Implications for potential adverse impact, Four‐Fifths Rule differences, and the use of Belbin's questionnaire for selection, particularly into team leader positions, are raised.
The effects of psychological contract breach on union commitment
Authors: Turnley William H. , Bolino Mark C. , Lester Scott W. , Bloodgood James M.
Published: August 2004

This research investigates the relationship between psychological contract breach and union commitment among 109 union employees. As expected, psychological contract breach is positively related to commitment to the union. The results also suggest that union instrumentality moderates the relationship between psychological contract breach and union commitment. Specifically, the findings indicate that the relationship between psychological contract breach and union commitment is stronger when individuals perceive that their union is highly instrumental in protecting their rights and benefits. Implications of this study are discussed and recommendations for future research are offered.
The relationship between disposition and career success: A British study
Authors: Bozionelos Nikos
Published: August 2004

The relationship of the Five‐Factor Model of personality and general mental ability with extrinsic and intrinsic career success was investigated in a British sample of 308 white‐collar workers. Extrinsic career success was associated with both experiential variables and dispositional variables, while intrinsic career success was almost exclusively associated with personality traits. General mental ability contributed to eventual, but not to organization‐specific extrinsic career success; and neuroticism reduced both extrinsic career prospects and intrinsic career evaluations, while agreeableness reduced extrinsic career prospects but enhanced intrinsic career evaluations. However, in sharp contrast to hypotheses, conscientiousness primarily and extraversion secondarily were negatively associated with extrinsic career success. The findings are discussed with respect to extant knowledge and the British national cultural context.
Confirmatory factor analysis and invariance of an organizational citizenship behaviour measure across samples in a Dutch‐speaking context
Authors: Lievens Filip , Anseel Frederik
Published: August 2004

Although Organizational Citizenship Behaviour (OCB) has been studied extensively over the years in the US, the measurement of OCB has received relatively limited attention in other international contexts. This study investigates the dimensionality of a specific OCB measure in a Dutch‐speaking context (Flemish part of Belgium). In addition, we examine the invariance of this measure across two different samples: supervisor ratings of 259 subordinates and peer ratings of 215 employees. Generally, we found clear support for the discriminant validity of five OCB factors but convergent validity was only established for three of the five factors. This measurement structure found was relatively invariant across the two different samples These results show that the forms of OCB that are predominantly studied in the US seem to hold relatively well in another international context, although there are some differences.
Measurement of exhaled hydrogen peroxide from rabbit lungs
Authors: Weissmann N. , Vogels H. , Schermuly R. T. , A. Ghofrani H. , Hänze J. , Fink L. , Rose F. , Seeger W. , Grimminger F.
Published: August 2004

Exhaled H2O2 is considered an indicator of lung inflammatory and oxidative stress. Moreover, H2O2 may be involved in signal transduction processes. It is not fully elucidated to what extent (i) H2O2 escapes from the intravascular compartment, and (ii) pulmonary H2O2 generation and nasopharyngeal H2O2 generation contribute to exhaled H2O2. We investigated H2O2 concentrations in breath condensate from isolated bufferperfused and ventilated rabbit lungs, and from both intubated and spontaneously breathing rabbits with a horseradish peroxidase/ 2,7dichlorofluorescin assay. For the perfused lungs, a H2O2 concentration of 58±19 nM was found. Addition of H2O2 to the buffer fluid resulted in only minute appearance in the exhaled air (<0.001%). Levels of exhaled H2O2 in intubated rabbits and perfused lungs were virtually identical. Nearly tenfold higher levels were detected in spontaneously breathing rabbits. Decreasing the inspired oxygen concentration from 21% to 1% resulted in a tendency toward decreased H2O2 exhalation in perfused lungs. In contrast, phorbol-12-myristate-13- acetate (PMA) prompted a ~4-fold increase in H2O2 exhalation. We conclude that the horseradish peroxidase/ 2',7'dichlorofluorescin assay is a feasible technique to measure H2O2 in exhaled breath condensate in rabbits. When collecting exhaled air via the tracheal tube, the signal represents pulmonary H2O2 generation with the contribution of the remaining body being negligible.
Simultaneous exposure of rats to dioxin and carbon monoxide reduces the xenobiotic but not the hypoxic response
Authors: Hofer T. , Pohjanvirta R. , Spielmann P. , Viluksela M. , Buchmann D. P. , Wenger R. H. , Gassmann M.
Published: August 2004

Aryl hydrocarbon receptor (AhR) and hypoxiainducible factor-1α (HIF-1α) are conditionally regulated transcription factor subunits that form heterodimeric complexes with their common partner, AhR nuclear translocator (ARNT/HIF-1β). Whereas the environmentally toxic compound 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD) initiates the trans-activation activity of AhR:ARNT/HIF-1β, hypoxic exposure stabilizes HIF-1α and functionally activates the HIF-1α:ARNT/HIF-1β complex. To analyze a possible crosstalk between these two pathwaysin vivo, rats were given dioxin orally and/or were exposed to carbon monoxide (CO), causing functional anemia. We found that exposure to CO inhibited the xenobiotic response while dioxin application had no significant negative impact on hypoxia-mediated gene transcription.
Structure and expression of two kininogen genes in mice
Authors: Cardoso C. C. , Garrett T. , Cayla C. , Meneton P. , Pesquero J. B. , Bader M.
Published: August 2004

Kininogens serve dual functions by forming a scaffold for the assembly of the protein complex initiating the surface-activated blood coagulation cascade and as precursors for the kinin hormones. While rats have three kininogen genes, for mice, cattle, and humans only one gene has been described. Here, we present sequence and expression data of a second mouse kininogen gene. The two genes, kininogen-I and kininogen-II, are located in close proximity on chromosome 16 in a headtohead arrangement. In liver and kidney, both genes are expressed and for each gene three alternative splice variants are synthesized. Two of them are the expected high and low molecular weight isoforms known from all mammalian kininogens. However, for both genes also a third, hitherto unknown splice variant was detected which lacks part of the high molecular weight mRNA due to splicing from the low molecular weight donor site to alternative splice acceptor sites in exon 10. The physiological functions of the six kininogen isoforms predicted by these findings need to be determined.
Structure of the Q237W mutant of HhaI DNA methyltransferase: an insight into protein-protein interactions
Authors: Dong A. , Zhou L. , Zhang X. , Stickel S. , Roberts R.J. , Cheng X.
Published: August 2004

We have determined the structure of a mutant (Q237W) of HhaIDNA methyltransferase, complexed with the methyl-donor product AdoHcy. The Q237W mutant proteins were crystallized in the monoclinic space group C2 with two molecules in the crystallographic asymmetric unit. Protein-protein interface calculations in the crystal lattices suggest that the dimer interface has the specific characteristics for homodimer protein-protein interactions, while the two active sites are spatially independent on the outer surface of the dimer. The solution behavior suggests the formation of HhaIdimers as well. The same HhaIdimer interface is also observed in the previously characterized binary (M.HhaIAdoMet) and ternary (M.HhaIDNAAdoHcy) complex structures, crystallized in different space groups. The dimer is characterized either by a noncrystallographic two-fold symmetry or a crystallographic symmetry. The dimer interface involves three segments: the aminoterminal residues 28, the carboxyterminal residues 313327, and the linker (amino acids 179184) between the two functional domains the catalytic methylation domain and the DNA target recognition domain. Both the amino- and carboxy-terminal segments are part of the methylation domain. We also examined proteinprotein interactions of other structurally characterized DNA MTases, which are often found as a 2-fold related dimer with the largest dimer interface area for the group-β MTases. A possible evolutionary link between the Type I and Type II restriction-modification systems is discussed.
Activation of NF-κB/Rel transcription factors in human primary peripheral blood mononuclear cells by interleukin 7
Authors: Lamberti A. , Petrella A. , Pascale M. , Romano M.F. , Bisogni R. , Vita N. , Venuta S. , Turco M.C.
Published: August 2004

Pathways that regulate the activation of NF-κB/Rel transcription factors are known to include signaling through a number of cytokine receptors. Interleukin 7 (IL-7), produced by bone marrow and other stromal cells, is a key factor for differentiation and survival in the lymphoid and other compartments. We found that human recombinant IL-7 induced NF-κB/Rel activation, analyzed by electrophoretic mobility shift assay (EMSA), in human peripheral blood T lymphocytes from healthy donors. Induced complexes included p65 and p50 NF-κB/Rel subunits. These results demonstrate for the first time that IL-7 can participate in signaling leading to NF-κB/Rel activation.
Searching for the most effective screening system to identify cell-active inhibitors of β-secretase
Authors: Middendorp O. , Lüthi U. , Hausch F. , Barberis A.
Published: August 2004

The β-secretase BACE1 is an attractive drug target for reducing the level of the Alzheimers disease-promoting Aβ peptide in the brain. Whereas potent peptidomimetic in vitro inhibitors of BACE1 have been designed, screening approaches to identify cell-permeable small molecule inhibitors have had limited success so far. In the present minireview we summarize existing screening methods, discuss their scope of application in the drug discovery process and compare them to a novel cell-based screening system to identify BACE1 inhibitors by a positive yeast growth selection.
Nutrient-gene interactions: a single nutrient and hundreds of target genes
Authors: Daniel H. , Dieck H. tom
Published: August 2004

Based on the effects of a selective experimental zinc deficiency in a rodent model we explore the use of transcriptome profiling for assessing nutrient-gene interactions in the liver at the molecular and cellular levels. Zinc deficiency caused pleiotropic alterations in mRNA/protein levels of hundreds of genes. In the context of observed metabolic alterations in hepatic metabolism, possible mechanisms are discussed for how a low zinc status may be sensed and transmitted into changes in various metabolic pathways. However, it also becomes obvious that analysis of such complex nutrient-gene interactions beyond the descriptional level is a real challenge for systems biology.
Linking nutrition to genomics
Authors: Bauer M. , Hamm A. , Pankratz M.J.
Published: August 2004

The new scientific field of nutrigenomics utilizes genomic tools, like microarrays, to analyze metabolic adaptations induced by variations in nutritional status. Here we describe how transcriptional regulation patterns caused by nutritional changes can be identified using gene expression profiling. This includes technical remarks on microarray analysis and data processing, as well as giving biological meaning to statistically solid data. We highlight our recent findings of transcriptional regulation of genes representing specific signaling and metabolic pathways in mouse liver under starvation. The results show strong correlations to previously identified responses to caloric restriction, which can be linked to lifespan extension.
Metal-responsive transcription factor (MTF-1) and heavy metal stress response in Drosophila and mammalian cells: a functional comparison
Authors: Balamurugan K. , Egli D. , Selvaraj A. , Zhang B. , Georgiev O. , Schaffner W.
Published: August 2004

The zinc finger transcription factor MTF-1 (metalresponsive transcription factor-1) is conserved from insects to vertebrates. Its major role in both organisms is to control the transcription of genes involved in the homeostasis and detoxification of heavy metal ions such as Cu[2+], Zn[2+] and Cd [2+]. In mammals, MTF-1 serves at least two additional roles. First, targeted disruption of the MTF-1 gene results in death at embryonic day 14 due to liver degeneration, revealing a stagespecific developmental role. Second, under hypoxicanoxic stress, MTF-1 helps to activate the transcription of the gene placental growth factor (PlGF), an angiogenic protein. Recently we characterized dMTF-1, theDrosophilahomolog of mammalian MTF-1. Here we present a series of studies to compare the metal response in mammals and insects, which reveal common features but also differences. A human MTF-1 transgene can restore to a large extent metal tolerance to flies lacking their own MTF-1 gene, both at low and high copper concentrations. Likewise,DrosophilaMTF-1 can substitute for human MTF-1 in mammalian cell culture, although both the basal and the metalinduced transcript levels are lower. Finally, a clear difference was revealed in the response to mercury, a highly toxic heavy metal: metallothioneintype promoters respond poorly, if at all, to Hg[2+] in mammalian cells but strongly inDrosophila, and this response is completely dependent on dMTF-1.
All-trans retinoic acid-responsive genes identified in the human SH-SY5Y neuroblastoma cell line and their regulated expression in the nervous system of early embryos
Authors: Merrill R.A. , Ahrens J.M. , Kaiser M.E. , Federhart K.S. , Poon V.Y. , Clagett-Dame M.
Published: August 2004

The vitamin A metabolite, all-trans retinoic acid (atRA), is required for embryonic development. atRA binds to the nuclear retinoic acid receptors and regulates the transcription of specific target genes. In order to identify atRA-induced genes that play a role in neural development, a subtractive library was created from SH-SY5Y neuroblastoma cells, a human cell line that exhibits changes in cell adhesion and neurite outgrowth after exposure to the vitamin A acid. We report here the identification of 14 genes that are rapidly induced by atRA (retinoic acid induced in neurobIlastoma or RAINB), eight of which were previously not known to be atRA responsive (BTBD11, calmin, cyclin M2, ephrin B2, HOXD10, NEDD9, RAINB6 and tenascin R). mRNA regulation by atRA was confirmed in SHSY5Y cells by Northern blotting, and gene regulation was studied in additional human cell lines using the quantitative polymerase chain reaction. The majority of the atRA-responsive clones revealed in this screen are highly expressed in the nervous system of developing rat embryos. Further, the expression of several of these genes is perturbed in developing rat embryos exposed to excess atRA or conversely, deprived of sufficient retinoid during early development. We propose that a subset of these genes lie downstream of atRA and its receptors in the regulation of neurite outgrowth and cell adhesion in both neural and nonneural tissues within the developing embryo.
GAPDH enhances group II intron splicing in vitro
Authors: Böck-Taferner P. , Wank H.
Published: August 2004

Group II introns are autocatalytic RNAs which selfsplice
Metal-responsive transcription factor-1 (MTF-1) selects different types of metal response elements at low vs. high zinc concentration
Authors: Wang Y. , Lorenzi I. , Georgiev O. , Schaffner W.
Published: August 2004

Metalresponsive transcription factor-1 (MTF-1) is a zinc finger protein with a central role in heavy metal homeostasis/ detoxification. MTF-1 binds to DNA sequence motifs known as metal response elements (MREs) with a core consensus TGCRCNC. Since MTF-1 is also involved in other stress responses, we tested whether it is able to recognize different types of DNA sequence motifs. To this end we selected MTF-1-binding oligonucleotides from a collection of random sequences. Since MTF-1 binds to known target sequences at relatively high zinc concentrations, oligonucleotide selection was performed in a mammalian cell nuclear extract both at high and low zinc concentrations. Irrespective of zinc concentration, we find a robust representation of MRE consensus sequences, however with specific features. Selection was most efficient at 100 M zinc, yielding many oligonucleotides with two MRE motifs in divergent orientation of the sequence
Inactivation of protein disulfide isomerase by alkylators including α,β-unsaturated aldehydes at low physiological pHs
Authors: Liu X.-W. , Sok D.-E.
Published: August 2004

Protein disulfide isomerase (PDI) is known to contain the thioredoxin box motif with a low pKa cysteine residue. To investigate the reactivity of PDI with thiol modifiers at low physiological pHs, either the reduced (PDI[red]) or oxidized form (PDI[oxid]) of PDI was exposed to various alkylating ragents. When PDI was incubated with iodoacetamide at pH 6.3 for 30 min at 38C, a remarkable inactivation (>90%) of PDI[red] was caused by iodoacetamide (IC[50]=8 M). However, PDI[oxid] was only slightly inactivated (approximately 18%) by iodoacetamide. Similarly, PDI[red] was significantly inactivated by Nethylmaleimide (NEM), but PDI[oxid] was not. When the inactivation by these alkylators was analyzed by pseudofirst order kinetics, NEM (k[3]=1.7510[-2] s[-1]; K[i]=124 M) was observed to be more potent than iodoacetamide (k[3]=9.110[-3] s[-1]; K=311 M). Interestingly, the inactivation of PDI[red] by iodoacetamide was greater at pH 6.3 than pH 7.0, in contrast to a similar inactivation potency of NEM at both pHs. Moreover, the maximal inactivation of PDI[red] or PDI[oxid] by iodoacetamide was mainly observed around pH 6.0. In addition, PDI[red] was found to be inactivated by acrolein (IC[50]=10 M) at pH 6.3, and this inactivation was also greater at pH 6.3 than at pH 7. Based on these results, we suggest that PDI[red] is susceptible to inactivation by alkylators including endogenous α,β-unsaturated aldehydes at low physiological pHs.
SMAD-signaling in chronic obstructive pulmonary disease: transcriptional down-regulation of inhibitory SMAD 6 and 7 by cigarette smoke
Authors: Springer J. , Scholz F.R. , Peiser C. , Groneberg D.A. , Fischer A.
Published: August 2004

Transforming growth factor-β1 is a potent mediator of fibrosis stimulating the secretion of extracellular matrix proteins and is involved in airway remodeling in chronic obstructive pulmonary disease (COPD). Signals from the TGF superfamily are mediated by the SMAD group of transcription factors. Here, the expression of the regulatory SMAD2, 3, the co-SMAD4 and the inhibitory SMAD6 and 7 was assessed in bronchial biopsies of COPD patients and controls by quantitative RTPCR. While SMAD2 was not expressed and SMAD3 and 4 displayed no change, the inhibitory SMAD6 and 7 were significantly downregulated in COPD. To reveal the molecular basis of tobacco smoke-induced airway remodeling and to test whether it may interfere with intracellular SMAD signaling, the airway epithelial cell line A549 was incubated with cigarette smoke extract (1% and 10%) for 48 hours, which led to down-regulation of SMAD6 and 7 at both concentrations tested. It can be concluded that TGF-β-mediated effects in COPD are influenced by a disturbed intracellular feedback mechanism of inhibitory SMADs. Also, the effects of non-volatile components in tobacco smoke may partly be regulated via a smoke-induced down-regulation of inhibitory SMADs.
Exogenously added GPI-anchored tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) displays enhanced and novel biological activities
Authors: Djafarzadeh R. , Mojaat A. , Vicente A. Belen , Lüttichau I. von , Nelson P.J.
Published: August 2004

The family of tissue inhibitors of metalloproteinases (TIMPs) exhibits diverse physiological/biological functions including the inhibition of active matrix metalloproteinases, regulation of proMMP activation, cell growth, and the modulation of angiogenesis. TIMP-1 is a secreted protein that can be detected on the cell surface through its interaction with surface proteins. The diverse biological functions of TIMP-1 are thought to lie, in part, in the kinetics of TIMP-1/MMP/surface protein interactions. Proteins anchored by glycoinositol phospholipids (GPIs), when purified and added to cells
Collagen, ageing and nutrition
Authors: Frey Jacques
Published: August 2004

The relationship between ageing and nutrition is considered with collagen as the intermediate target. Some data showed that diet restriction resulted in decreased collagen accumulation and collagen ageing. Conversely, being overweight reduced the lifespan and increased collagen ageing.Collagen ageing, which includes low turnover and glycoxidation, involves an increase in both stiffness and weakness. Their consequences concern all tissues including those with vital importance such as cartilage, heart ventricle or arterial wall.
Evaluation of a new efficient procedure for single-nucleotide polymorphism genotyping: tetra-primer amplification refractory mutation system-polymerase chain reaction
Authors: Okayama Naoko , Fujimura Kozue , Nakamura Junji , Suehiro Yutaka , Hamanaka Yuichiro , Hinoda Yuji
Published: August 2004

Tetra-primer amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) is a new efficient method for single-nucleotide polymorphism (SNP) genotyping. To determine the optimal conditions for ARMS-PCR we attempted to genotype ten SNPs. DNA was extracted from the peripheral blood of 168 unrelated healthy Japanese volunteers. Two problems inhibited uniform efficiency of the amplification of three bands. The first problem was the lower amplification efficiency of the shorter and allele-specific products compared with the largest product. This phenomenon was overcome by increasing the relative concentration of the inner primers. The second problem was non-specific amplification of the shorter products. To reduce the amplification of these nonspecific bands, adjusting any one of the following PCR conditions was effective: i) reducing the ratio of the inner primer concentration relative to that of the outer primers; ii) increasing the annealing temperature for the initial 5–10 cycles; iii) hot start PCR. With these procedures all ten of the SNPs were successfully genotyped. Our present data may be useful in the further application of tetra-primer ARMS-PCR to SNP genotyping.
Limited usefulness of the PFA-100™ for the monitoring of ADP receptor antagonists–in vitro experience
Authors: Golanski Jacek , Pluta Justyna , Baraniak Janina , Watala Cezary
Published: August 2004

We have evaluated the usefulness of the PFA-100™ system (collagen/ADP and collagen/epinephrine cartridges) to assess the in vitro effects of a few platelet function inhibitors: Aspisol
Isolation of plasma small-dense low-density lipoprotein using a simple air-driven ultracentrifuge and quantification using immunassay of apolipoprotein B
Authors: Menys Valentine C. , Liu Yifen , Mackness Michael I. , Kwok See , Caslake Muriel J. , Stewart Grace , Durrington Paul N.
Published: August 2004

Small-dense low-density lipoprotein (SD-LDL) is associated with coronary heart disease risk. Current methods for its quantification are expensive, complex and time-consuming. Plasma was adjusted to a density (D) of 1.044 g/ml in a volume of 0.18 ml and centrifuged in a Beckman Airfuge at 160 000×
Analytical performance and diagnostic accuracy of a fully-automated electrochemiluminescent assay for the N-terminal fragment of the pro-peptide of brain natriuretic peptide in patients with cardiomyopathy: comparison with immunoradiometric assay methods for brain natriuretic peptide and atrial natriuretic peptide
Authors: Prontera Concetta , Emdin Michele , CarloZucchelli Gian , Ripoli Andrea , Passino Claudio , Clerico Aldo
Published: August 2004

We evaluated the analytical performance of a fully-automated electrochemiluminescence “sandwich” immunoassay method for the N-terminal fragment of the pro-peptide of brain natriuretic peptide (BNP). We then compared the diagnostic accuracy of this method in discriminating between normal subjects and patients with cardiomyopathy with that found with two previously described immunoradiometric assay methods for the assay of atrial natriuretic peptide (ANP) and BNP. We studied 193 consecutive patients (mean age 64.4±12.3 years, range 20–89 years, including 56 women and 137 men) with chronic cardiomyopathy and a group of 85 healthy subjects (mean age 52.3±12.0 years, 42 women and 43 men, range 20–79 years). N-terminal fragment of proBNP
Calcium oxalate crystallization in untreated urine, centrifuged and filtered urine and ultrafiltered urine
Authors: Guerra Angela , Meschi Tiziana , Allegri Franca , Schianchi Tania , Adorni Giuditta , Novarini Almerico , Borghi Loris
Published: August 2004

Centrifuged and filtered urine is often used to evaluate in vitro the crystallization processes of calcium oxalate (CaOx), but even such simple manipulations can alter the composition of the urine, as regards its protein and lipid concentrations. In urine samples taken from 17 normal male adults, we evaluated CaOx crystallization by simultaneously using three different types of urine: untreated (U), centrifuged at 2000 rpm (800
Matrix effects in clinical immunoassays and the effect of preheating and cooling analytical samples
Authors: Yoshida Hiroshi , Imafuku Yuji , Nagai Toshihiko
Published: August 2004

Immunological reactions are influenced by various factors including antigens, antibodies and other variables. We focused on two items: i) matrix effects, especially of detergents and ii) temperature effects: preheating sera, especially effects on rheumatoid factor (RF) measurement and false-positive reactions in ELISAs, and cold storage of sera, especially effects on complement. Among various additives, detergents affected the agglutination reaction for fecal hemoglobin and hepatitis B surface (HBs) antigen. Some of the detergents examined abolished these antigenicities, however, polyethylenglycols enhanced the reactions. Heat-inactivation of sera at 56°C for 30 min was employed in serological testing. However, in RF measurement, 10 min of preheating was sufficient to abolish C1q (subcomponent of C1), which could participate in the agglutination reaction. In ELISA for antibodies, false-positive reactions were caused by preheating sera. By the analyses of assays for antibodies to hepatitis C virus (HCV) and cardiolipin, it was found that they were induced by immunoglobulin G (IgG) modified by preheating. Cold storage induced activation of complement (cold activation) in anti-HCV antibody positive sera. CH50 titers in the sera were lowered by one cycle of freezing at −20°C and thawing, and the decrease was affected by the containers.
Association between the CYP2C9 polymorphism and the drug metabolism phenotype
Authors: Topic Elizabeta , Štefanovic Mario , Samardžija Marina
Published: August 2004

CYP2C9, an isoform of the cytochrome P450 enzyme, is involved in the metabolism of most of the drugs of choice for the treatment of thromboembolic disorders. Functional polymorphism is associated with two variant alleles (alleles
Analytical goals for coagulation tests based on biological variation
Authors: Wada Yui , Kurihara Masako , Toyofuku Mitsuko , Kawamura Minako , Iida Hiroko , Kayamori Yuzou , Kinoshita Sachiko , Hamasaki Naotaka
Published: August 2004

Allowable imprecision and bias reference limits for laboratory data can be calculated based on measurements of biological variation. Although biological variation of clinical chemical data has been reported from many laboratories, there have been few reports of biological variation in coagulation tests. In this study, we calculated the biological variation of 13 coagulation tests in the clinical laboratory of Kyushu University Hospital and determined allowable imprecision and bias limits of variation. The participating subjects were 17 healthy individuals: three males and two females in their 20s, two males and two females in their 30s, one male and four females in their 40s, and two males and one female in their 50s. Monthly measurements were performed before breakfast 12 times from June 2001 to May 2002 and allowable imprecision and bias limits were calculated. Taken together with coefficient of variation of control plasma used in daily laboratory work at the hospital, the allowable imprecision limits of intra-laboratory variation determined in this study appear to be in attainable ranges.