bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased expression of fatty acid synthase and progesterone receptor in early steps of human mammary carcinogenesis
Authors: Esslimani‐Sahla Majida , Thezenas Simon , Simony‐Lafontaine Joelle , Kramar Andrew , Lavaill Roselyne , Chalbos Dany , Rochefort Henri
Published: October 2006

Progestins increase the risk of breast cancer in the hormone therapy of menopause, and progesterone receptor‐induced fatty acid synthase (FAS) is a potential therapeutical target of breast cancer. In a first attempt to specify in which lesions at risk of breast cancer progestins might be acting, we have compared the progesterone receptor (PR) and FAS expression in preinvasive breast lesions and in adjacent “normal” mammary glands. We used archive paraffin‐embedded tissues from 116 patients, with 164 lesions of increasing histological risk from nonproliferative “benign” breast disease (BBD) to in situ breast carcinomas. Immunostaining using our FAS antibody and a PR antibody from Dako was quantified as continuous variables by computer‐assisted image analysis. FAS level increased (p < 10−3 by the Kruskall–Wallis test) in all lesions, starting from nonproliferative BBD, and was maximal in in situ carcinoma. The % of PR‐positive cells increased from nonproliferative BBD and was higher in proliferative atypia (p < 10−3). It was very low in high‐grade DCIS corresponding to a likely different carcinogenesis pathway. There was a trend for a positive correlation between FAS and PR in normal glands. However, the 2 markers increased independently in BBD and were negatively correlated in in situ carcinomas. FAS and PR were positively correlated with Ki67 in BBD. The increased PR level in premalignant steps of mammary carcinogenesis suggests an early increased responsiveness to progestins. The increased FAS expression, in lesions parallel to their increased breast cancer risk, suggests further studies to develop new markers of high‐risk lesions and to prevent breast cancer. © 2006 Wiley‐Liss, Inc.
Synergism between GM–CSF and IFNγ: Enhanced immunotherapy in mice with glioma
Authors: Smith Karin E. , Janelidze Shorena , Visse Edward , Badn Wiaam , Salford Leif , Siesjö Peter , Darabi Anna
Published: October 2006

Glioblastoma multiforme is the most common malignant primary brain tumor and also one of the most therapy‐resistant tumors. Because of the dismal prognosis, various therapies modulating the immune system have been developed in experimental models. Previously, we have shown a 37–70% cure in a rat glioma model where rats were peripherally immunized with tumor cells producing IFNγ. On the basis of these results, we wanted to investigate whether a combination of GM–CSF and IFNγ could improve the therapeutic effect in a mouse glioma model, GL261 (GL‐wt). Three biweekly intraperitoneal (i.p.) immunizations with irradiated GM–CSF‐transduced GL261 cells (GL‐GM) induced a 44% survival in mice with intracranial glioma. While treatment of GL‐wt and GL‐GM with IFNγ in vitro induced upregulation of MHC I and MHC II on the tumor cells, it could not enhance survival after immunization. However, immunizations with GL‐GM combined with recombinant IFNγ at the immunization site synergistically enhanced survival with a cure rate of 88%. Tumors from mice receiving only 1 immunization on Day 10 after tumor inoculation were sectioned on Day 20 for analysis of leukocyte infiltration. Tumor volume was reduced and the infiltration of macrophages was denser in mice immunized with GL‐GM combined with IFNγ compared with that of both wildtype and nonimmunized mice. To our knowledge, this is the first study to demonstrate a synergy between GM–CSF and IFNγ in experimental immunotherapy of tumors, by substantially increasing survival as well as inducing a potent anti‐tumor response after only 1 postponed immunization. © 2006 Wiley‐Liss, Inc.
Broad tumor spectrum in a mouse model of multiple endocrine neoplasia type 1
Authors: Loffler Kelly A. , Biondi Christine A. , Gartside Michael , Waring Paul , Stark Mitchell , Serewko‐Auret Magdalena M. , Muller H. Konrad , Hayward Nicholas K. , Kay Graham F.
Published: October 2006

Multiple endocrine neoplasia type 1 (MEN1) is an inherited cancer predisposition syndrome typified by development of tumors in parathyroid, pituitary and endocrine pancreas, as well as less common sites including both endocrine and nonendocrine organs. Deletion or mutation of the tumor suppressor gene MEN1 on chromosome 11 has been identified in many cases of MEN1 as well as in sporadic tumors. The molecular biology of menin, the protein encoded by MEN1, remains poorly understood. Here we describe a mouse model of MEN1 in which tumors were seen in pancreatic islets, pituitary, thyroid and parathyroid, adrenal glands, testes and ovaries. The observed tumor spectrum therefore includes types commonly seen in MEN1 patients and additional types. Pancreatic pathology was most common, evident in over 80% of animals, while other tumor types developed with lower frequency and generally later onset. Tumors of multiple endocrine organs were observed frequently, but progression to carcinoma and metastasis were not evident. Tumors in all sites showed loss of heterozygosity at the Men1 locus, though the frequency in testicular tumors was only 36%, indicating that a different molecular mechanism of tumorigenesis occurs in those Leydig tumors that do not show loss of the normal Men1 allele. Menin expression was below the level of detection in ovary, thyroid and testis, but loss of nuclear menin immunoreactivity was observed uniformly in all pancreatic islet adenomas and in some hyperplastic islet cells, suggesting that complete loss of Men1 is a critical point in islet tumor progression in this model. © 2006 Wiley‐Liss, Inc.
Effects of copper on gill structure and transport function in the rainbow trout, oncorhynchus mykiss
Authors: Sola François , Isaia Jacques , Masoni Armand
Published: October 2006

Effects of copper were studied in freshwater adapted rainbow trout using the perfused head preparation. In its monovalent chemical form, copper at millimolar concentrations had no significant effects on Na+ and water transport. By contrast, the divalent form produced an increase in gill perfusion pressure, a significant reduction in Na+ influx and water fluxes and reversed Na+ net flux. Observations by light microscopy showed important cell damage (oedema, mucus production, cellular desquamation). By electron microscopy there was smoothing of apical membranes, swelling of the tubular system and destruction of mitochondria. The Na, K‐ATPase activity was totally suppressed and residual ATPase activity largely inhibited by 1 mM Cu2+. There was inhibition of the Na, K‐ATPase activity with an IC50 of ± 10 μM of total copper (free and bound cupric fractions). As active sodium transport is located on the secondary lamellae, our results show that its entry mechanism is inhibited at that level by cupric ions only. Results are discussed in relation to hydromineral balance of the trout.
Oxidative modifications produced in HL‐60 cells on exposure to benzene metabolites
Authors: Rao Nimmagudda R. , Snyder Robert
Published: October 2006

We have studied the effects of the benzene metabolites hydroquinone, p‐benzoquinone or 1,2,4‐benzenetriol on cytotoxicity, active oxygen formation, hydrogen peroxide (i.e. hydroperoxide) production and nitric oxide formation in HL‐60 cells. We also examined the effects of these compounds on antioxidant enzymes and intracellular antioxidants in these cells. The cytotoxicity of benzene metabolites to HL‐60 cells was found to be of the order of p‐benzoquinone>hydroquinone>benzenetriol. No appreciable changes in the basal levels of either superoxide anion production or nitric oxide formation were observed following exposures to the benzene metabolites, but significant increases in superoxide were seen on stimulation with TPA for each metabolite, whereas hydroquinone and p‐benzoquinone, but not 1,2,4‐benzenetriol, increased nitric oxide production under these conditions. Following exposure to the benzene metabolites, HL‐60 cells showed significant rises in hydrogen peroxide formation compared to controls. The study of antioxidant enzymes and intracellular antioxidants suggested that the benzene metabolites inhibit or reduce the levels of different antioxidant mechanisms and, thereby, cause the accumulation of free radicals in these cells predisposing them for oxidative damage.
Distinctive gene expression of prostatic stromal cells cultured from diseased versus normal tissues
Authors: Zhao Hongjuan , Ramos Cristiane F. , Brooks James D. , Peehl Donna M.
Published: October 2006

To obtain a comprehensive view of the transcriptional programs in prostatic stromal cells of different histological/pathological origin, we profiled 18 adult human stromal cell cultures from normal transition zone (TZ), normal peripheral zone (PZ), benign prostatic hyperplasia (BPH), and prostate cancer (CA) using cDNA microarrays. A hierarchical clustering analysis of 714 named unique genes whose expression varied at least threefold from the overall mean abundance in at least three samples in all 18 samples demonstrated that cells of different origin displayed distinct gene expression profiles. Many of the differentially expressed genes are involved in biological processes known to be important in the development of prostatic diseases including cell proliferation and apoptosis, cell adhesion, and immune response. Significance Analysis of Microarrays (SAM) analysis identified genes that showed differential expression with statistical significance including 24 genes between cells from TZ versus BPH, 34 between BPH versus CA, and 101 between PZ versus CA. S100A4 and SULF1, the most up‐ and downregulated genes in BPH versus TZ, respectively, showed expression at the protein level consistent with microarray analysis. In addition, sulfatase assay showed that BPH cells have lower SULF1 activity compared to TZ cells. Quantitative real‐time polymerase chain reaction (qRT‐PCR) analysis confirmed differential expression of ENPP2/autotoxin and six other genes between PZ versus CA, as well as differential expression of six genes between BPH versus CA. Our results support the hypothesis that prostatic stromal cells of different origin have unique transcriptional programs and point towards genes involved in actions of stromal cells in BPH and CA. J. Cell. Physiol. 210: 111–121, 2007. © 2006 Wiley‐Liss, Inc.
Bone anabolic effects of parathyroid hormone are blunted by deletion of the Wnt antagonist secreted frizzled‐related protein‐1
Authors: Bodine Peter V.N. , Seestaller‐Wehr Laura , Kharode Yogendra P. , Bex Frederick J. , Komm Barry S.
Published: October 2006

Secreted frizzled‐related protein (sFRP)‐1 is a Wnt antagonist that when deleted in mice leads to increased trabecular bone formation in adult animals after 13 weeks of age. Treatment of mice with parathyroid hormone (PTH) also increases trabecular bone formation, and some of the anabolic actions of this hormone may result from altered expression of Wnt pathway components. To test this hypothesis, we treated +/+ and −/− female sFRP‐1 mice with PTH 1–34 for 30 days and measured distal femur trabecular bone parameters by peripheral quantitative computed tomography (pQCT) and high‐resolution micro‐computed tomography. During the course of the 32‐week study, volumetric bone mineral density (vBMD) declined 41% in vehicle‐treated +/+ mice, but increased 24% in vehicle‐treated −/− animals. At 8 weeks of age when vBMD was not altered by deletion of sFRP‐1, treatment of +/+ and −/− mice with PTH increased vBMD by 147 and 163%, respectively. In contrast, at 24 weeks of age when vBMD was 75% higher in −/− mice than in +/+ controls, treatment with PTH increased vBMD 164% in +/+ animals, but only 58% in −/− mice. Furthermore, at 36 weeks of age when vBMD was 117% higher in −/− mice than in +/+ controls, treatment with PTH increased vBMD 74% in +/+ animals, while no increase was observed in −/− mice. At each of these time points, PTH treatment increased vBMD to a similar level in +/+ and −/− mice, and this level declined with age. In addition, at 36 weeks of age, the vBMD level reached by PTH treatment of +/+ mice was the same as that achieved solely by deletion of sFRP‐1. These results indicate that loss of sFRP‐1 and PTH treatment increase vBMD to a similar extent. Moreover, as the effects of sFRP‐1 deletion on vBMD increase, the ability of PTH to enhance vBMD declines suggesting that there are overlapping mechanisms of action. J. Cell. Physiol. 210: 352–357, 2007. © 2006 Wiley‐Liss, Inc.
AMP‐activated protein kinase mediates preconditioning in cardiomyocytes by regulating activity and trafficking of sarcolemmal ATP‐sensitive K+ channels
Authors: Sukhodub Andrey , Jovanović Sofija , Du Qingyou , Budas Grant , Clelland Allyson K. , Shen Mei , Sakamoto Kei , Tian Rong , Jovanović Aleksandar
Published: October 2006

Brief periods of ischemia and reperfusion that precede sustained ischemia lead to a reduction in myocardial infarct size. This phenomenon, known as ischemic preconditioning, is mediated by signaling pathway(s) that is complex and yet to be fully defined. AMP‐activated kinase (AMPK) is activated in cells under conditions associated with ATP depletion and increased AMP/ATP ratio. In the present study, we have taken advantage of a cardiac phenotype overexpressing a dominant negative form of the α2 subunit of AMPK to analyze the role, if any, that AMPK plays in preconditioning the heart. We have found that myocardial preconditioning activates AMPK in wild type, but not transgenic mice. Cardiac cells from transgenic mice could not be preconditioned, as opposed to cells from the wild type. The cytoprotective effect of AMPK was not related to the effect that preconditioning has on mitochondrial membrane potential as revealed by JC‐1, a mitochondrial membrane potential‐sensitive dye, and laser confocal microscopy. In contrast, experiments with di‐8‐ANEPPS, a sarcolemmal‐potential sensitive dye, has demonstrated that intact AMPK activity is required for preconditioning‐induced shortening of the action membrane potential. The preconditioning‐induced activation of sarcolemmal KATP channels was observed in wild type, but not in transgenic mice. HMR 1098, a selective inhibitor of sarcolemmal KATP channels opening, inhibited preconditioning‐induced shortening of action membrane potential as well as cardioprotection afforded by AMPK. Immunoprecipitation followed by Western blotting has shown that AMPK is essential for preconditioning‐induced recruitment of sarcolemmal KATP channels. Based on the obtained results, we conclude that AMPK mediates preconditioning in cardiac cells by regulating the activity and recruitment of sarcolemmal KATP channels without being a part of signaling pathway that regulates mitochondrial membrane potential. J. Cell. Physiol. 210: 224–236, 2007. © 2006 Wiley‐Liss, Inc.
Constitutive internalization of murine MHC class I molecules
Authors: Mahmutefendić Hana , Blagojević Gordana , Kučić Natalia , Lučin Pero
Published: October 2006

The total number of cell surface glycoprotein molecules at the plasma membrane results from a balance between their constitutive internalization and their egress to the cell surface from intracellular pools and/or biosynthetic pathway. Constitutive internalization is net result of constitutive endocytosis and endocytic recycling. In this study we have compared spontaneous internalization of murine major histocompatibility complex (MHC) class I molecules (Kd, Dd, full Ld, and empty Ld) after depletion of their egress to the cell surface (Cycloheximide [CHX], brefeldin A [BFA]) and internalization after external binding of monoclonal antibody (mAb). MHC class I alleles differ regarding their cell surface stability, kinetics, and in the way of internalization and degradation. Kd and Dd molecules are more stable at the cell surface than Ld molecules and, thus, constitutively internalized more slowly. Although the binding of mAbs to cell surface MHC class I molecules results in faster internalization than depletion of their egress, it is still slow and, thereby, can serve as a model for tracking of MHC class I endocytosis. Internalization of fully conformed MHC class I molecules (Kd, Dd, and Ld) was neither inhibited by chlorpromazine (CP) (inhibitor of clathrin endocytosis), nor with filipin (inhibitor of lipid raft dependent endocytosis), indicating that fully conformed MHC class I molecules are internalized via the bulk pathway. In contrast, internalization of empty Ld molecules was inhibited by filipin, indicating that non‐conformed MHC class I molecules require intact cholesterol‐rich membrane microdomains for their constitutive internalization. Thus, conformed and non‐conformed MHC class I molecules use different endocytic pathways for constitutive internalization. J. Cell. Physiol. 210: 445–455, 2007. © 2006 Wiley‐Liss, Inc.
Effects of TGF‐β and glucocorticoids on map kinase phosphorylation, IL‐6/IL‐11 secretion and cell proliferation in primary cultures of human lung fibroblasts
Authors: Pelaia Girolamo , Gallelli Luca , D'Agostino Bruno , Vatrella Alessandro , Cuda Giovanni , Fratto Donatella , Renda Teresa , Galderisi Umberto , Piegari Elena , Crimi Nunzio , Rossi Francesco , Caputi Mario , Costanzo Francesco S. , Vancheri Carlo , Maselli Rosario , Marsico Serafino A.
Published: October 2006

Transforming growth factor‐β1 (TGF‐β1) is crucially involved in the fibrotic events characterizing interstitial lung diseases (ILDs), as well as in the airway remodeling process typical of asthma. Within such a context, the aim of our study was to investigate, in primary cultures of normal and fibrotic human lung fibroblasts (HLFs), the effects of TGF‐β1 on mitogen‐activated protein kinase (MAPK) phosphorylation, cell proliferation, and production of interleukins 6 (IL‐6) and 11 (IL‐11), in the presence or absence of a pretreatment with budesonide (BUD). MAPK phosphorylation was detected by Western blotting, cell viability and proliferation were evaluated using Trypan blue staining and [3H]‐thymidine incorporation assay, respectively, and the release of IL‐6 and IL‐11 into cell culture supernatants was assessed by ELISA. TGF‐β1 (10 ng/ml) significantly stimulated MAPK phosphorylation (P < 0.01), and also enhanced cell proliferation as well as the secretion of both IL‐6 and IL‐11, which reached the highest increases at the 72nd h of cell exposure to this growth factor. All such effects were prevented by BUD (10−8 M) and, with the exception of IL‐6 release, also by a mixture of MAPK inhibitors. Therefore, our findings suggest that the fibrotic action exerted by TGF‐β1 in the lung is mediated at least in part by MAPK activation and by an increased synthesis of the profibrogenic cytokines IL‐6 and IL‐11; all these effects appear to be prevented by corticosteroids via inhibition of MAPK phosphorylation. J. Cell. Physiol. 210: 489–497, 2007. © 2006 Wiley‐Liss, Inc.
Alterations of hHrd1 expression are related to hyperphosphorylated tau in the hippocampus in Alzheimer's disease
Authors: Hou Hai‐Long , Shen Yu‐Xian , Zhu Hong‐Yan , Sun Hao , Yan Xue‐Bo , Fang Hui , Zhou Jiang‐Ning
Published: October 2006

The degradation of aberrantly phosphorylated tau in neurons plays an important role in the pathogenesis of Alzheimer's disease (AD). hHrd1 is a newly identified ubiquitin ligase involved in the endoplasmic reticulum (ER)‐associated protein degradation. The expression and function of hHrd1 in AD brain remains elusive. In the present study, the expression of hHrd1 in AD hippocampus and the morphological relations between hHrd1 expression and pretangle formation were studied by using immunohistochemical single‐ and double‐labeling methods. The results showed that hHrd1 was expressed in neurons and reactive astrocytes, especially in the CA2–CA4 hippocampal subfields. The ratio of hHrd1‐positive neurons/astrocytes to total neurons/astrocytes was increased in the CA1 subfield in AD hippocampus compared with the age‐matched controls (P < 0.05). Most Alz‐50 labeled pretangles were colocalized with hHrd1, and the expression levels showed an inversed change, implied that hHrd1 might be associated with the degradation of hyperphosphorylated tau. © 2006 Wiley‐Liss, Inc.
Contribution of valine 7′ of TMD2 to gating of neuronal α3 receptor subtypes
Authors: Nieves‐Cintrón Madeline , Caballero‐Rivera Daniel , Navedo Manuel F. , Lasalde‐Dominicci José A.
Published: October 2006

The second transmembrane domain (TMD2) of the Cys‐loop family of ligand‐gated ion channels forms the channel pore. The functional role of the amino acid residues contributing to the channel pore in neuronal nicotinic α3 receptors is not well understood. We characterized the contribution of TMD2 position V7′ to channel gating in neuronal nicotinic α3 receptors. Site‐directed mutagenesis was used to substitute position α3 (V7′) with four different amino acids (A, F, S, or Y) and coexpressed each mutant subunit with wild‐type (WT) β2 or β4 subunits inXenopusoocytes. Whole‐cell voltage clamp experiments show that substitution for an alanine, serine, or phenylalanine decreased by 2.3–6.2‐fold the ACh‐EC50for α3β2 and α3β4 receptor subtypes. Interestingly, mutation V7′Y did not produce a significant change in ACh‐EC50when coexpressed with the β2 subunit but showed a significant approximately two‐fold increase with β4. Similar responses were obtained with nicotine as the agonist. The antagonist sensitivity of the mutant channels was assessed by using dihydro‐β‐erythroidine (DHβE) and methyllycaconitine (MLA). The apparent potency of DHβE as an antagonist increased by ∼3.7‐ and 11‐fold for the α3β2 V7′S and V7′F mutants, respectively, whereas no evident changes in antagonist potency were observed for the V7′A and V7′Y mutants. The V7′S and V7′F mutations increase MLA antagonist potency for the α3β4 receptor by ∼6.2‐ and ∼9.3‐fold, respectively. The V7′A mutation selectively increases the MLA antagonist potency for the α3β4 receptor by ∼18.7‐fold. These results indicate that position V7′ contributes to channel gating kinetics and pharmacology of the neuronal nicotinic α3 receptors. © 2006 Wiley‐Liss, Inc.
A method for measuring bending stresses based on electromagnetic actuation and Raman spectroscopy
Authors: Ren Yongfeng , Xu Wenqiang , Xue Chenyang , Zhang Wendong , Zheng Lina
Published: October 2006

Bending stress and fracture strength of micromachined structures are the parameters determining the function and life span of microelectromechanical systems (MEMS) and are also indispensable data for MEMS design. This paper reports the results of a study in which a bending stress measurement method based on electromagnetic actuation and Raman spectroscopy has been examined. In this study, an eight‐cantilever mass structure is chosen as the experimental object. The results have indicated that it can accurately measure the bending stresses and fracture strength. Copyright © 2006 John Wiley & Sons, Ltd.
Isolation of eriocitrin (eriodictyol 7‐O‐rutinoside) as an arachidonate lipoxygenase inhibitor from Lumie fruit (Citrus lumia) and its distribution in Citrus species
Authors: Nogata Yoichi , Ohta Hideaki , Ishii Toshinao , Sekiya Keizo
Published: October 2006

An inhibitory compound acting against rat platelet 12‐lipoxygenase was isolated from the peel of Lumie fruit (Citrus lumia) by activity‐guided separation. It was identified as eriocitrin (eriodictyol 7‐O‐rutinoside) by spectroscopic analyses. Eriocitrin inhibited 5‐lipoxygenase (IC5029.1 µmol L−1) from rat peritoneal polymorphonuclear leukocytes in addition to 12‐lipoxygenase (IC5022.3 µmol L−1). Its aglycone, eriodictyol (5,7,3′, 4′‐tetrahydroxyflavanone), was a much more potent inhibitor of both 12‐lipoxygenase (IC500.07 µmol L−1) and 5‐lipoxygenase (IC500.20 µmol L−1). It also inhibited the production of leukotriene B4 in intact peritoneal polymorphonuclear leukocytes stimulated with calcium ionophore A23187 (IC5012.7 µmol L−1). The distribution of eriocitrin in 39 citrus fruits was investigated by high‐performance liquid chromatography analysis. Lumie, eureka lemon (Citrus limon), Sambokan (Citrus sulcata), Sudachi (Citrus sudachi) and Koji (Citrus leiocarpa) fruits were found to contain high levels of eriocitrin in both peel and juice vesicles. Copyright © 2006 Society of Chemical Industry
Evaluation of antioxidant capacity of some fruit and vegetable foods: efficiency of extraction of a sequence of solvents
Authors: Pellegrini Nicoletta , Colombi Barbara , Salvatore Sara , Brenna Oreste V , Galaverna Gianni , Del Rio Daniele , Bianchi Marta , Bennett Richard N , Brighenti Furio
Published: October 2006

All methods for assessing the total antioxidant capacity (TAC) of food samples are strongly affected by the solvents used during extraction. In recent years a sequential solvent extraction procedure utilising water and acetone has been widely used for TAC measurements of foods. To better understand the efficiency of this procedure in terms of the amount of extracted antioxidants and the subsequent measurement of TAC, two vegetables (onion and spinach) and two fruits (orange and tomato) were sequentially extracted with water, acetone and chloroform. Each extract fraction was analysed separately for its content of known antioxidant compounds by specific procedures and for its TAC by the Trolox equivalent antioxidant capacity assay. The results showed that the compounds in the water and acetone extracts were the main contributors to TAC. The chloroform extracts did not contribute to TAC, with the exception of the spinach extract, owing to the presence of low levels of carotenoids. In conclusion, the analysed extraction procedure was more effective for foods rich in water‐soluble antioxidants than for those rich in lipid‐soluble antioxidants. Copyright © 2006 Society of Chemical Industry
Effectiveness of chitosan‐based coating in improving shelf‐life of eggs
Authors: Caner Cengiz , Cansiz Ozge
Published: October 2006

Despite eggs having a natural packaging—shell—they are perishable and can lose their quality during storage. Chitosan‐based coatings were applied to shell eggs to examine potential effects on egg quality properties (weight loss, Haugh unit, yolk index) during 4 weeks of storage. Mineral amounts in yolks were also evaluated after 4 weeks of storage. Three chitosan‐based coatings produced with organic acids (acetic‐(C‐AA), lactic‐(C‐LA), and propionic (C‐PA)) were evaluated on shelf‐life enhancements of fresh egg quality. All chitosan‐coated eggs showed greater interior quality than the non‐coated eggs. The coatings significantly maintained weight loss compared to the control specimen (4.96%). Lower weight loss (3.45% for C‐PA, 3.53% for C‐LA) was observed in the coated eggs. Eggshell chitosan coat containing lactic and propionic acids maintained higher Haugh unit and yolk index than eggs coated with acetic acid. Uncoated (UC) eggs changed from grade ‘A’ to ‘B’ after 1 week of storage. Chitosan‐based coating containing lactic and propionic acids maintained eggs in grade ‘A’ for 4 weeks. Haugh unit showed that C‐LA and C‐PA effectively maintained eggs at grade ‘A’ quality for at least 3 weeks more than control and 1 week more than C‐AA. Results also indicated that the chitosan coating maintained mineral amounts at nutritional values (especially calcium, iron and magnesium concentration) in yolks after 4 weeks storage. Copyright © 2006 Society of Chemical Industry
Impact of European Union funding on tourism in the Grampian Region
Authors: Nash Robert , Koyabe Doris , Stansbie Paul
Published: October 2006

The European Union (EU) has no specific tourism policy. Even though there are divergent views on its involvement in regional development, there is a consensus on the fact that the EU is involved by its principle role of ‘Subsidiarity’. There is also continued participation of the EU in activities related to tourism development in the community. Undoubtedly, the EU is having an impact on regional development of members states. Research carried out shows the contribution of the EU in regional development and tourism. The EU funding has tended to focus on peripheral areas and where traditional industries are declining. Tourism related projects (such as visitor attractions) have benefited from such funding programmes, on the assumption that these projects will create employment and increase visitor numbers, as well as promote the economies of these regions. Consequently, this paper considers the contributions of EU funding of tourism attractions in the Aberdeen and Grampian region of Scotland and issues affecting the effectiveness of the attractions themselves. Copyright © 2006 John Wiley & Sons, Ltd.
G2019S LRRK2 mutation in familial and sporadic Parkinson's disease in Russia
Authors: Pchelina Sofya N. , Yakimovskii Andrei F. , Ivanova Olga N. , Emelianov Anton K. , Zakharchuk Andrei H. , Schwarzman Alexander L.
Published: October 2006

Among mutations associated with autosomal dominant and sporadic Parkinson's disease (PD) the G2019S substitution in the leucine‐rich repeat kinase 2 (LRRK2) gene is the most frequently identified. To estimate its frequency in Russia, we analyzed 208 patients with PD from the Northwestern region of Russia. Of these, 51 patients were probands from families with PD compatible with autosomal dominant inheritance. The control group represented 161 subjects without neurological disorders settled in the same region. The frequency of the G2019S mutation was greater in familial PD (2 [3.9%] of 51) than in sporadic PD (1 [0.6%] of 157). In addition, this mutation was found in the proband's father, who also had PD, in 1 PD family, and in 1 carrier without signs of PD at age 40 in another PD family. All carriers were heterozygous for the G2019S mutation and reported the Ashkenazi Jewish origin. The mutation was not found in the control group. © 2006 Movement Disorder Society
Parkinson's disease in Africa: A systematic review of epidemiologic and genetic studies
Authors: Okubadejo Njideka U. , Bower James H. , Rocca Walter A. , Maraganore Demetrius M.
Published: October 2006

Parkinson's disease (PD) occurs worldwide, but little is known about PD in Africa. We systematically reviewed publications on PD in Africa, with emphasis on epidemiologic and genetic studies. Articles published between 1944 and December 2004 were identified using several strategies. The studies emanated from 13 African countries (Kenya, Uganda, Tanzania, Ethiopia, Nigeria, Senegal, Ghana, Togo, Libya, Tunisia, Algeria, Zimbabwe, and South Africa). The publications fell into four categories: clinical series (n = 17), prevalence studies (n = 7), incidence studies (n = 1), and genetic studies (n = 3). The clinical series documented the occurrence of PD in Africa and described its clinical characteristics. The prevalence studies suggested some intracontinental geographic variation in PD prevalence. Overall, the prevalence figures and the incidence rates of PD in Africa appeared lower than those reported for European and North American populations. Few genetic studies of PD have been reported from Africa, and none in blacks. There are no case–control or cohort studies of PD reported from Africa. This review provides a summary of PD research in Africa over the past 60 years and highlights the information gaps and potential areas for future research. © 2006 Movement Disorder Society
Existence and exponential stability in Lr‐spaces of stationary Navier–Stokes flows with prescribed flux in infinite cylindrical domains
Authors: Ri Myong‐Hwan , Farwig Reinhard
Published: October 2006

We prove existence, uniqueness and exponential stability of stationary Navier–Stokes flows with prescribed flux in an unbounded cylinder of ℝn,n⩾3, with several exits to infinity provided the total flux and external force are sufficiently small. The proofs are based on analytic semigroup theory, perturbation theory and Lr − Lq‐estimates of a perturbation of the Stokes operator in Lq‐spaces. Copyright © 2006 John Wiley & Sons, Ltd.
Structural elucidation of nitro‐substituted five‐membered aromatic heterocycles utilizing GIAO DFT calculations
Authors: Katritzky Alan R. , Akhmedov Novruz G. , Doskocz Jacek , Hall C. Dennis , Akhmedova Rena G. , Majumder Suman
Published: October 2006

The GIAO (Gauge Including Atomic Orbitals) DFT (Density Functional Theory) method is applied at the B3LYP/6–31+G(d,p)//B3LYP/6–31+G(d), B3LYP/6–311++G(d,p)//B3LYP/6–31+G(d), B3LYP/6–311+G (2d,p)//B3LYP/6–31+G(d) and B3LYP/6–311++G(d,p)//B3LYP/6–311++G(d,p) levels of theory for the calculation of proton and carbon chemicals shifts and coupling constants for 25 nitro‐substituted five‐membered heterocycles. Difference (1D NOE) spectra in combination with long‐range gHMBC experiments were used as tools for the structural elucidation of nitro‐substituted five‐membered heterocycles. The assigned NMR data (chemical shifts and coupling constants) for all compounds were found to be in good agreement with theoretical calculations using the GIAO DFT method. The magnitudes of one‐bond (1JCH) and long‐range (nJCH, n > 1) coupling constants were utilized for unambiguous differentiation between regioisomers of nitro‐substituted five‐membered heterocycles. Copyright © 2006 John Wiley & Sons, Ltd.
Hybrid approximation algorithm with Kriging and quadratic polynomial‐based approach for approximate optimization
Authors: Sakata S. , Ashida F. , Zako M.
Published: October 2006

This paper describes a new hybrid algorithm that uses a Kriging and quadratic polynomial‐based approach for approximate optimization. The Kriging method is used for generating a global approximation model, and the polynomial‐based approximation method is used for generating a local approximation model.

The Kriging system is only used to construct a polynomial‐based locally approximate model by estimating some function values and Hessian components of an estimated surface. The number of Kriging estimations can be reduced in comparison with direct Kriging‐based optimization, and a local optimum solution on an approximated surface can be clearly estimated without use of an optimization procedure based on a local appropriate quadratic polynomial model.

Numerical examples of engineering optimization using the proposed method illustrate validity and effectiveness of the proposed method. Copyright © 2006 John Wiley & Sons, Ltd.
A preconditioned conjugate gradient approach to structural reanalysis for general layout modifications
Authors: Li Zhengguang , Wu Baisheng
Published: October 2006

This paper presents a preconditioned conjugate gradient approach to structural static reanalysis for general layout modifications. It is suitable for all types of layout modifications, including the general case in which some original members and nodes are deleted and other new members and nodes are added concurrently. The approach is based on the preconditioned conjugate gradient technique. The preconditioner is constructed, and an efficient implementation for applying the preconditioner is presented, which requires the factorization of the stiffness matrix corresponding to the newly added degrees of freedom only. In particular, the approach can adaptively monitor the accuracy of approximate solutions. Numerical examples show that the condition number of the preconditioned matrix is remarkably reduced. Therefore, the fast convergence and accurate results can be achieved by the approach. Copyright © 2006 John Wiley & Sons, Ltd.
Optimization of structural dynamic behaviour based on effective modal parameters
Authors: Besset S. , Jézéquel L.
Published: October 2006

Optimization of complex structures often leads to high calculation costs. Indeed, the structure has to be frequently reanalysed in order to update the optimization criteria. We propose an optimization method based on effective modal parameters. These parameters are close to the modal matrices used for the modal analysis of a structure. Thus, once the structure has been analysed, it becomes very easy to calculate optimization criteria. First, we will explain the modal analysis that we will use in this paper. A modal model will be used to analyse the hollow parts of the structure. The modal analysis of the whole structure will be performed using substructuring and ‘double modal synthesis’ proposed by Jezequel. Secondly, we will explain how to obtain effective modal parameters and their use for optimization. Finally, we will show the efficiency of these parameters through the optimization of a complex structure, using two types of optimization methods. Copyright © 2006 John Wiley & Sons, Ltd.
Sexual dimorphism in limb bones of ibex (Capra ibex L.): mixture analysis applied to modern and fossil data
Authors: Fernández H. , Monchot H.
Published: October 2006

Estimating sex ratios of fossil bone assemblages is an important step in the determination of demographic profiles, which are essential for understanding the palaeobiology and palaeoethology of any particular species, as well as its exploitation patterns by humans. This is especially true for ibex (Capra ibex), which was a main source of food for hominids during Pleistocene times. Classical methods for determining sexual dimorphism and sex ratio, such as analyses using uni‐ and bivariate plots, are based on an arbitrary fixing of limits between sexes. Here we use a more robust statistical method termed mixture analysis (MA) to determine the sex of postcranial remains (long bones, metapodials and tarsals) from ibex. For the first time, we apply MA to both a modern and a fossil sample of one species, by using metric data taken from (i) a collection of present‐day ibex skeletons and (ii) a Palaeolithic sample of the same species. Our results clearly show that the forelimb (humerus and radius) is more dimorphic than the hindlimb (femur and tibia) and is therefore better suited for sexing ibex. It also appears that metapodials should be used carefully for estimating sex ratios. On the basis of these results, we propose a classification of bone measurements that are more or less reliable for sexing ibex. The results of MA applied to the ibex fossil bones from the Upper Palaeolithic site of the Observatoire (Monaco) lead us to the conclusion that this assemblage consists of a majority of males. The quantitative estimations calculated by the MA make it possible to compare the size of Pleistocene and modern ibex for the whole set of variables used in this study. Copyright © 2006 John Wiley & Sons, Ltd.
Integrating forward and reverse proteomics to unravel protein function
Authors: Palcy Sandrine , Chevet Eric
Published: October 2006

To date, proteomics approaches have aimed to either identify novel proteins or change in protein expression/modification in various organisms under normal or disease conditions. One major aspect of functional proteomics is to identify protein biological properties in a given context, however, forward proteomics approaches alone cannot complete this goal. Indeed, with the increasing successes of such proteomics‐based research strategies and the subsequent increasing amounts of proteins identified with unknown molecular functions, approaches allowing for systematic analyses of protein functions are desired. In this review, we propose to depict the complementarities of forward and reverse proteomics approaches in the definite understanding of protein functions. This dual strategy requires a data integration loop which allows for systematic characterization of protein function(s). The details of the integrative process combining both in silico and experimental resources and tools are presented. Altogether, we believe that the integration of forward and reverse proteomics approaches supported by bioinformatics will provide an efficient path towards systems biology.
Crystal structure of human TMDP, a testis‐specific dual specificity protein phosphatase: Implications for substrate specificity
Authors: Kim Seung Jun , Jeong Dae‐Gwin , Yoon Tae‐Sung , Son Jeong‐Hee , Cho Somi Kim , Ryu Seong Eon , Kim Jae‐Hoon
Published: October 2006

The testis‐ and skeletal‐muscle‐specific dual‐specificity phosphatase (TMDP) is a member of the dual‐specificity phosphatase (DSP) subgroup of protein tyrosine phosphatases. TMDP has similar activities toward both tyrosine and threonine phosphorylated substrates, and is supposed to be involved in spermatogenesis. Here, we report the crystal structure of human TMDP at a resolution of 2.4 Å. In spite of high sequence similarity with other DSPs, the crystal structure of TMDP shows distinct structural motifs and surface properties. In TMDP, the α1–β1 loop, a substrate recognition motif is located further away from the active site loop in comparison to prototype DSP Vaccinia H1 related phophatase (VHR), which preferentially dephosphorylates tyrosine phosphorylated substrates and down‐regulates MAP kinase signaling. Residues in the active site residues of TMDP are smaller in size and more hydrophobic than those of VHR. In addition, TMDP cannot be aligned with VHR in loop β3–α4. These differences in the active site of TMDP result in a flat and wide pocket structure, allowing equal binding of phosphotyrosine and phosphothreonine substrates. Proteins 2007. © 2006 Wiley‐Liss, Inc.
Preparation and conversion electron Mössbauer study of Fe3O4/α‐Fe2O3 composite films
Authors: Niu Z. P. , Wang Y. , Li F. S.
Published: October 2006

Fe3O4 and Fe3O4/α‐Fe2O3 composite films have been prepared from a sintered Fe2O3 target, using an rf magnetron sputtering apparatus with various argon pressures. The phase and magnetic properties of the samples have been characterized by X‐ray‐diffraction (XRD) measurements, conversion electron Mössbauer spectroscopy (CEMS), and a vibrating sample magnetometer (VSM). With the increasing argon pressure during sputtering, the increase of the average center shift of Mössbauer lines indicates the decreasing valency state of Fe ions induced by the enhancement of presumed oxygen loss. An enhanced magnetoresistance (MR) has been found in the Fe3O4/α‐Fe2O3 composite film that was prepared under an Ar pressure of 1.0 Pa. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Liquid chromatography/electrospray ionization mass spectrometric characterization of Harpagophytum in equine urine and plasma
Authors: Colas Cyril , Garcia Patrice , Popot Marie‐Agnès , Bonnaire Yves , Bouchonnet Stéphane
Published: October 2006

A method has been developed for the analysis and characterization in equine urine and plasma of iridoid glycosides: harpagide, harpagoside and 8‐para‐coumaroyl harpagide, which are the main active principles of Harpagophytum, a plant with antiinflammatory properties. The method involves liquid chromatography coupled with positive electrospray ionization mass spectrometry. The addition of sodium or lithium chloride instead of formic acid in the eluting solvent has been studied in order to enhance the signal and to modify the ion's internal energy. Fragmentation pathways and associated patterns are proposed for each analyte. A comparison of three types of mass spectrometer: a 3D ion trap, a triple quadrupole and a linear ion trap, has been conducted. The 3D ion trap was selected for drug screening analysis whereas the linear ion trap was retained for identification and quantitation analysis. Copyright © 2006 John Wiley & Sons, Ltd.
Determination of the immunomodulator laquinimod in human plasma by liquid chromatography/tandem mass spectrometry; development, validation and application of two methods in clinical pharmacokinetic profiling
Authors: Sennbro Carl Johan , Olin Magnus , Edman Karin , Hansson Gunnar , Gunnarsson Per Olov , Svensson Leif D
Published: October 2006

Laquinimod (ABR‐215062) is a synthetic compound currently undergoing clinical development for oral treatment of multiple sclerosis. The present paper describes the development, validation and clinical application of two rapid and sensitive methods for the determination of ABR‐215062 in human plasma. Both methods use liquid chromatography/tandem mass spectrometry (LC/MS/MS) with electrospray ionization in positive mode and a stable isotope (13C6)‐labeled ABR‐215062 as internal standard for calibration. The selected reaction monitoring was based on the transitions m/z 357.1 → 236.1 for ABR‐215062, and either m/z 363.2 → 236.1 or 365.2 → 238.1 for 13C6‐ABR‐215062. Method 1, aimed and validated for low level determinations (0.4–100 nmol/L) of ABR‐215062, utilizes solid‐phase extraction followed by isocratic elution. Method 2, aimed and validated for wide range determinations (0.75–15000 nmol/L) of ABR‐215062, utilizes protein precipitation followed by fast gradient elution. The methods were validated with respect to selectivity, lower limit of quantification (LLOQ), dynamic range, precision, accuracy, extraction recovery and ruggedness. Furthermore, the stability of low level ABR‐215062 in plasma was investigated. The methods were also applied in clinical trials. The LLOQs were 0.4 and 0.75 nmol/L for methods 1 and 2, respectively. The intra‐ and inter‐day precision for the methods were 1.6–3.5% and 2.1–5.7%. The extraction recoveries were 90–97% for both methods. ABR‐215062 was stable in plasma for at least 3 months when stored at −20°C. In conclusion, our developed methods were found to be selective, sensitive, robust and suitable for applications in clinical pharmacokinetic profiling. Copyright © 2006 John Wiley & Sons, Ltd.
Nickel quantification in serum by a validated sector‐field inductively coupled plasma mass spectrometry method: assessment of tentative reference values for an Italian population
Authors: Bocca Beatrice , Forte Giovanni , Ronchi Anna , Gaggeri Raffaella , Alimonti Alessandro , Minoia Claudio
Published: October 2006

The daily exposure to Ni from food, industrial processes, jewellery and coins makes the determination of Ni in human serum an important way to monitor the health status in non‐occupationally exposed subjects. To this end, a method based on sector‐field inductively coupled plasma mass spectrometry was developed and validated. The limits of detection (LoD) and quantification (LoQ), sensitivity, linearity range, trueness, repeatability, within‐laboratory reproducibility and robustness were the considered issues of the validation process. The uncertainty associated with the measurements was also calculated, according to the Eurachem/Citac Guide. The method LoD and LoQ were 0.03 and 0.09 ng mL−1, linearity was over two order of magnitude, trueness was −3.57%, and the repeatability and reproducibility showed relative standard deviations equal to 4.56% and 6.52%, respectively. The relative expanded uncertainty was 21.8% at the Ni levels found in the general population. The tentative reference value for serum Ni was 0.466 ± 0.160 ng mL−1 with a related interval between 0.226 and 1.026 ng mL−1. Copyright © 2006 John Wiley & Sons, Ltd.
An objective means of diagnosing anorexia nervosa and bulimia nervosa using 15N/14N and 13C/12C ratios in hair
Authors: Hatch Kent A. , Crawford Morgan A. , Kunz Amanda W. , Thomsen Steven R. , Eggett Dennis L. , Nelson Stephen T. , Roeder Beverly L.
Published: October 2006

An objective means based on the carbon and nitrogen stable isotope analysis of five hairs per individual is presented for distinguishing between individuals with anorexia nervosa and/or bulimia nervosa from non‐clinical individuals (i.e. clinically normal controls). Using discriminant analysis, an algorithm has been developed that provides both sensitivity and specificity of 80% in making diagnoses of individuals with these eating disorders. With further refinements, the results suggest that it may be also possible to distinguish between individuals with anorexia or bulimia. Finally, the study shows the value of conducting blind tests and using larger sample sizes of both control and treatment groups. Both groups are needed to validate the diagnostic value of a method and to provide measures of sensitivity and specificity of any diagnostic test. Copyright © 2006 John Wiley & Sons, Ltd.