bluuebunny/crossref_metadata_2025 huggingface dataset:
Phanerochaete chrysosporium soluble proteome as a prelude for the analysis of heavy metal stress response
Authors: Özcan Servet , Yıldırım Volkan , Kaya Levent , Albrecht Dirk , Becher Dörte , Hecker Michael , Ozcengiz Gulay
Published: March 2007

A 2‐D reference map in pI range 3–10 was constructed for the soluble protein fraction of Phanerochaete chrysosporium growing vegetatively under standard conditions. Functional annotation could be made for 517 spots out of 720 that were subjected to MALDI‐TOF‐MS analysis, according to the specific accession numbers from the P. chrysosporium genomic database. Further analysis of the data revealed 314 distinct ORFs, 118 of which yielded multiple spots on the master gel. Functional classification of the proteins was made according to the eukaryote orthologous groups defined in the organism's genome website. The functional class of PTMs, protein turnover and chaperones was represented with the highest number (63) of the identified ORFs. Six proteins were assigned to the hypothetical proteins and 29 were predicted to have a signal peptide sequence. Subcellular localization predictions were also made for the identified proteins. Of the protein spots detected on the master gel, 380 were found to be probably phosphorylated and 96 of these matched to the identified proteins. The reference map was efficiently used in the identification of the proteins differentially expressed under cadmium and copper stress. Three new ribosomal proteins as well as zinc‐containing alcohol dehydrogenase, glucose‐6‐phosphate isomerase, flavonol/cinnamoyl‐CoA reductase, H+‐transporting two‐sector ATPase, ribosomal protein S7, ribosomal protein S21e, elongation factor EF‐1 alpha subunit were demonstrated as the most strongly induced.
2‐DE analysis of a new human cell line EM‐G3 derived from breast cancer progenitor cells and comparison with normal mammary epithelial cells
Authors: Selicharová Irena , Smutná Kateřina , Šanda Miloslav , Ubik Karel , Matoušková Eva , Buršíková Eva , Brožová Markéta , Vydra Jan , Jiráček Jiří
Published: March 2007

We performed a 2‐DE analysis of proteins of the newly established spontaneously immortalized clonal cell line EM‐G3 derived from a primary lesion of infiltrating ductal breast carcinoma. EM‐G3 cells may represent progenitors of the mammary epithelial cells spontaneously immortalized in early phase of cancerogenesis. We compared the protein profile of EM‐G3 line with proteins from populations of normal mammary epithelial cells (NME), and determined the phenotype of both types of cells. NME cells are a mixture of both main cell types in breast epithelia, myoepithelial and luminal cells. The EM‐G3 breast cancer cell line has a unique basal‐like phenotype. We identified proteins that are differently expressed in these cells. Cytokeratin 16, cytokeratin 19, squamous cell carcinoma antigen 1, caphepsin B and caspase 14 were predominantly expressed by NME cells. Cytokeratin 13, isoelectric variant of annexin 5, isoelectric variant of chloride intracellular channel protein 1, glyoxalase 1 and glutamine synthetase were predominantly expressed by EM‐G3 cells. The proteins up‐regulated in EM‐G3 cells may represent potential protein markers of mammary epithelial cells progenitors and may be important in early phase of carcinogenesis.
Proteome changes associated with hippocampal MRI abnormalities in the lithium pilocarpine‐induced model of convulsive status epilepticus
Authors: Greene Nicholas D. E. , Bamidele Abiodun , Choy ManKin , de Castro Sandra C. P. , Wait Robin , Leung Kit‐Yi , Begum Shajna , Gadian David G. , Scott Rod C. , Lythgoe Mark F.
Published: March 2007

Convulsive status epilepticus is associated with subsequent hippocampal damage and development of mesial temporal sclerosis in a subset of individuals. The lithium pilocarpine model of status epilepticus (SE) in the rat provides a model in which to investigate the molecular and pathogenic process leading to hippocampal damage. In this study, a 2‐DE‐based approach was used to detect proteome changes in the hippocampus, at an early stage (2 days) after SE, when increased T2 values were detectable by magnetic resonance imaging. Gel image analysis was followed by LC‐MS/MS identification of protein species that differed in abundance between pilocarpine‐treated and control rats. The most significantly up‐regulated species in the experimental animals was identified as heat shock 27‐kDa protein, in line with findings in humans and in other experimental models of epilepsy. Additional up‐regulated species included dihydropyrimidinase‐related protein‐2, cytoskeletal proteins (α‐tubulin and ezrin) and dihydropteridine reductase. In summary, the hippocampus of rats subject to pilocarpine‐induced SE exhibits specific changes in protein abundance, which likely relate to pathogenic, neuroprotective and neurogenic responses.
Novel heterocyclic poly(arylene ether ketone)s: Synthesis and polymerization of 4‐(4′‐hydroxyaryl)(2H)phthalazin‐1‐ones with methyl groups
Authors: Cheng Lin , Ying Lei , Feng Juan , Wang Chen Yi , Li Jian Ling , Xu Zhen
Published: March 2007

Based on green chemistry, a simple and efficient direct synthesis of 4‐(4′‐hydroxyaryl)(2H)phthalazin‐1‐ones (2a–2f) was developed in a two‐step reaction, in which the Friedel–Crafts acylation reaction of six phenols with phthalic anhydride was initially carried out and then followed by cyclization with hydrazine hydrate in good to excellent yields with high regioselectivity. A number of novel heterocyclic poly(arylene ether ketone)s were prepared conveniently from several unsymmetrical, twist, and noncoplanar phthalazinone‐containing monomers (2a–2f) and an activated difluoro monomer via a NC coupling reaction. It was very interesting that the obtained monomers and polymers exhibited diverse properties with the variation of the number and location of the substituted methyl groups. All these polymers had a high molecular weight with Mn and ηinh in the range of 44,960–169,000 Da and 0.38–0.79 dL/g, respectively. Actually, the obtained polymers displayed excellent thermal properties with Tg's ranging from 222 to 248 °C and 5% weight loss temperatures in nitrogen higher than 430 °C. Moreover, these polymers were readily soluble in common organic solvents, such as N‐methyl‐2‐pyrrolidone, chloroform, pyridine, and m‐cresol, and could be cast into flexible and colorless or nearly colorless films by spin‐coating or casting processes. © 2006 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 1525–1535, 2007
Thermally curable benzoxazine monomer with a photodimerizable coumarin group
Authors: Kiskan Baris , Yagci Yusuf
Published: March 2007

A new monomer, 4‐methyl‐9‐p‐tolyl‐9,10‐dihydrochromeno[8,7‐e][1,3]oxazin‐2(8H)‐one, possessing both benzoxazine and coumarin rings in its structure was synthesized by the reaction of 4‐methyl‐7‐hydroxycoumarin, paraformaldehyde, and p‐toluidine in methanol at 40 °C and characterized with spectral analysis. Upon photolysis around 300 nm, this monomer underwent dimerization via the [2πs+2πs] cycloaddition reaction. Photodimerization reactions were investigated with UV and 1H NMR spectroscopy measurements. The thermal ring‐opening reaction of the benzoxazine ring was demonstrated with differential scanning calorimetry measurements. The thermal behavior of the cured product was also investigated with thermogravimetric analysis. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 1670–1676, 2007
Helical poly(phenylacetylene) derived from L‐tyrosine: A promising candidate for functional helical polymers carrying transformable N‐ and C‐termini
Authors: Zhao Haichao , Sanda Fumio , Masuda Toshio
Published: March 2007

A novel optically active phenylacetylene derivative, N‐(tert‐butoxycarbonyl)‐4‐ethynyl‐L‐phenylalanine methyl ester (1), was synthesized from L‐tyrosine and polymerized with a rhodium catalyst. The corresponding polymer [poly(1)] with a moderate molecular weight was obtained in a high yield. The alkaline hydrolysis of poly(1) gave poly[N‐(tert‐butoxycarbonyl)‐4‐ethynyl‐L‐phenylalanine] [poly(2)] carrying free carboxy groups. Polarimetric, CD, and UV–vis spectroscopy analyses revealed that poly(1) took a predominantly one‐handed helical structure in MeOH and toluene, and poly(2) took a helical structure in MeOH. The secondary structures of poly(1) and poly(2) could be tuned with heat and solvents. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 45: 1691–1698, 2007
Integrated scenarios for assessing biodiversity risks
Authors: Spangenberg Joachim H.
Published: March 2007

The rapid loss of biodiversity (however measured) constitutes an urgent need to develop policy strategies that reduce the anthropogenic pressures on biodiversity. To go beyond short‐term curative measures, such strategies must address the driving forces causing the pressures in an integrated fashion, covering a wide range of policy domains. The development of scenarios and their illustration by modelling are essential tools to study the aggregate human impacts on biodiversity, and to derive well founded policy options to preserve it. However, so far socio‐economic, climate and biodiversity models exhibit a wide range of assumptions concerning population development, economic growth and the resulting pressures on biodiversity. This paper summarizes the efforts undertaken in the framework of the ALARM project by an interdisciplinary team of economists, climatologists, land use experts and modellers to identify pressures and drivers, and to derive effective policy strategies. It describes the challenges of such a kind of work, bringing together different world views necessarily inherent to the different fields of investigation, presents preliminary results, indicates necessary policy priorities and suggests urgent issues for future research. Copyright © 2007 John Wiley & Sons, Ltd and ERP Environment.
Protective effects of insulin on polychlorinated biphenyls‐induced disruption of actin cytoskeleton in hippocampal neurons
Authors: Tang Feige , Yan Chonghuai , Li Fei , Wu Shenghu , Yu Yongguo , Gao Yu , Jin Xingming , Tian Ying , Shen Xiaoming
Published: March 2007

Insulin receptors are widely distributed in the brain, and insulin improves learning and memory in some brain injury. Insulin elevates LIM kinase 1 (LIMK‐1) activity and induces actin polymerization in some cells, while actin cytoskeleton dynamics mediated via LIMK‐1/cofilin signal pathway is considered important to learning and memory formation. Our previous studies have shown that polychlorinated biphenyls (PCBs) disrupt the actin cytoskeleton by inhibiting LIMK‐1/cofilin signaling pathway in the cultured hippocampal neurons. To determine potential neuronal protective effects by insulin, we administered insulin to the cultured hippocampal neurons after exposure to PCBs mixture Aroclor 1254 (A 1254). We found that insulin antagonized a loss of filamentous actin and the cytotoxicity induced by A 1254. Similarly, insulin restored the decrease of LIMK‐1 and cofilin phosphorylation induced by A 1254. We concluded that insulin could protect neurons, probably partly by ameliorating filamentous actin cytoskeleton disruption mediated via the activation of LIMK‐1/cofilin signal pathway in cultured hippocampal neurons after exposure to A 1254. The above protective effects in hippocampal neuron may have important implications in the treatment of PCBs‐induced neurotoxicity and the mechanism by which insulin improves learning and memory. © 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 152–158, 2007.
Sampling and analysis of microcystins: Implications for the development of standardized methods
Authors: Tillmanns Angeline R. , Pick Frances R. , Aranda‐Rodriguez Rocio
Published: March 2007

Microcystins (MC), a group of cyanotoxins, have been found in lakes and rivers worldwide. One goal of MC research is to develop models which predict MC concentrations, but these efforts have been hampered by a lack of standardized methods necessary for comparing data across studies. Here, we investigate the effect of chemical analysis (HPLC‐PDA and ELISA), sample collection (whole water, plankton tow and surface scum), and choice of normalizing parameter (volume, dry weight, and chlorophyll a) on reported MC concentrations. Samples were collected over three years from a temperate mesotrophic, shallow lake with episodic blooms of cyanobacteria. We found that microcystins were up to four times higher in lake samples when analyzed by ELISA relative to HPLC‐PDA and that MC concentration measured by HPLC explained less than half of the variation in MC concentrations measured by ELISA. Also, samples collected by plankton tow gave consistently higher concentrations than whole water samples. An additional HPLC analysis of two chlorophyte cultures revealed the presence of compounds with a similar UV absorbance spectrum to MC‐LR, suggesting that identifying MC based solely on UV absorbance is not valid. Our results document the discrepancy in MC concentrations that can arise by using different methods throughout all stages of sampling, analysis, and reporting of MC concentrations. © 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 132–143, 2007.
Effect of dibutyltin on ATP levels in human natural killer cells
Authors: Dudimah Fred D. , Gibson Constance , Whalen Margaret M.
Published: March 2007

This study investigates the role that decreased ATP levels may play in dibutyltin (DBT)‐induced decreases in the tumor‐cell‐lysing (lytic) function of natural killer (NK) cells. NK cells are a subset of lymphocytes capable of killing tumor cells, virally infected cells, and antibody coated cells. DBT is used as stabilizer in PVC plastics and has also been used as a deworming product in poultry. NK cells were exposed to various concentrations of DBT for 1 h, 24 h, 48 h, and 6 days before determining ATP levels and lytic function. ATP levels and lytic function were also determined in NK cells that were exposed to DBT for 1 h followed by 24 h, 48 h, and 6 days in DBT‐free media. The results indicated that exposure of NK cells to 10 μM DBT for 1 h did not cause any significant decrease in NK cell ATP levels but did cause a very significant loss in lytic function. NK cells exposed to 500 nM DBT for 24 h showed significant loss of lytic function but showed no decrease in ATP levels. However, 48 h and 6 days exposures to those concentrations of DBT that caused decreases in tumor lysing function also caused significant decreases in ATP levels. Exposures of NK cells to varying DBT concentrations for 1 h followed by 24 h, 48 h, and 6 days in DBT free media produced effects on lytic function and ATP levels that were similar to those seen with continuous DBT exposures. The results indicate that DBT exposures decrease ATP levels in NK cells but that tumor lysing function can be reduced independent of any decreases in ATP levels. Additionally the results show that the effects of a range of DBT concentrations on ATP levels and tumor lysing function are irreversible. © 2007 Wiley Periodicals, Inc. Environ Toxicol 22: 117–123, 2007.
Pleural and pericardial effusion: a potential ultrasonographic marker for the prenatal differential diagnosis between congenital diaphragmatic eventration and congenital diaphragmatic hernia
Authors: Jeanty C. , Nien J. K. , Espinoza J. , Kusanovic J. P. , Gonçalves L. F. , Qureshi F. , Jacques S. , Lee W. , Romero R.
Published: March 2007

Objectives

To determine whether or not the presence of pleural and/or pericardial effusion can be used prenatally as an ultrasonographic marker for the differential diagnosis between diaphragmatic eventration and diaphragmatic hernia.

Methods

We present two case reports of non‐isolated diaphragmatic eventration associated with pleural and/or pericardial effusion. Additionally, we reviewed the literature for all cases of congenital diaphragmatic hernia (CDH) and diaphragmatic eventration that met the following criteria: (1) prenatal diagnosis of a diaphragmatic defect and (2) definitive diagnosis by autopsy or surgery. The frequencies of pleural effusion, pericardial effusion and hydrops were compared between the two conditions using Fisher's exact test. A subanalysis was conducted of cases with isolated diaphragmatic defects (i.e. diaphragmatic defects not associated with hydrops and other major structural or chromosomal anomalies).

Results

A higher proportion of fetuses with diaphragmatic eventration had associated pleural and pericardial effusions compared with fetuses with diaphragmatic hernia (58% (7/12) vs. 3.7% (14/382), respectively, P < 0.001). This observation remained true when only cases of diaphragmatic defects not associated with hydrops and other major structural or chromosomal anomalies were compared (29% (2/7) with eventration vs. 2.2% (4/178) with CDH, P < 0.02).

Conclusions

The presence of pleural and/or pericardial effusion in patients with diaphragmatic defects should raise the possibility of a congenital diaphragmatic eventration. This information is clinically important for management and counseling because the prognosis and treatment for CDH and congenital diaphragmatic eventration are different. Copyright © 2007 ISUOG. Published by John Wiley & Sons, Ltd.
A new putative Zygosaccharomyces yeast species isolated from traditional balsamic vinegar
Authors: Solieri Lisa , Cassanelli Stefano , Giudici Paolo
Published: March 2007

The taxonomic status and species number of the genus Zygosaccharomyces have rapidly changed in the last years. In this study, two new osmotolerant Zygosaccharomyces strains isolated from traditional balsamic vinegar, viz. ABT301 and ABT601, were investigated to elucidate their taxonomic relationships with Zygosaccharomyces rouxii species. A multi‐gene sequence approach was employed, including regions of the rDNA repeat [5.8S, two internal transcribed spacers (ITS) and the 26S D1/D2 domain], COX2 mitochondrial gene and two nuclear genes (SOD2 and HIS3). Cloning and sequence analysis of 5.8S‐ITS rDNA revealed that these strains bear an unusual polymorphism for this region. Three highly divergent 5.8S‐ITS sequences were detected, one identical to Z. rouxii, the other two showing some relatedness to Z. mellis. Sequence and gene number polymorphism was also observed for the protein‐encoding nuclear genes SOD2 and HIS3, as two copies for each gene different from those found in Z. rouxii were detected. Analysis of the D1/D2 26S domain showed that ABT301 and ABT601 have only one type of D1/D2 sequence statistically different from that of Z. rouxii. The findings obtained in this work suggest that the genomic background of strains ABT301 and ABT601 is different from the other Zygosaccharomyces species. We speculated that they could belong to a new putative species related to Z. rouxii. Copyright © 2007 John Wiley & Sons, Ltd.
Genetic and reproductive consequences of forest fragmentation for populations of Magnolia obovata
Authors: Isagi Yuji , Tateno Ryunosuke , Matsuki Yu , Hirao Akira , Watanabe Sonoko , Shibata Mitsue
Published: March 2007

In order to evaluate the consequences of forest fragmentation on populations of Magnolia obovata, we compared genetic diversity and reproductive characteristics at two nearby sites, one conserved and one fragmented. The genetic diversity between adults trees of the different sites was not significantly different. However, saplings in the conserved site showed a significantly higher genetic diversity than both adult trees in the conserved site and saplings in the fragmented sites; this was found to be the result of the larger gene flow into the conserved site. The density of the adult trees was significantly related to all of the reproductive traits analyzed (fertilization of ovules, insect attack to seeds, ovules that developed into seeds and outcrossing at the stage of seeds) at both sites. At both sites, fertilization of ovules and insect attack on seeds were positively correlated to adult tree density while outcrossing rate was negatively correlated to adult tree density. The fertilization of ovules and outcrossing were more dependent on adult tree density in the fragmented site than in the conserved site. The probability of ovules developing into outcrossed seeds showed a negative correlation with adult tree density at both sites, indicating the advantage of low density for this species and possibly implying a resilience to habitat fragmentation. A two‐generation‐analysis did not identify significant differences between sites in terms of the structure of the pollen pool and the number of pollen donors. Although fragmentation affected reproductive characteristics, the effect on seedling establishment and subsequent survival remains to be determined. Proposals for future studies that will assist in the development of management strategies for forests suffering fragmentation are made.
Europe combating cancer: The European Union's commitment to cancer research in the 6th Framework Programme
Authors: Jungbluth Stefan , Kelm Olaf , van de Loo Jan-Willem , Manoussaki Elengo , Vidal Maria , Hallen Manuel , Trias Octavi Quintana
Published: March 2007

As one of the major health issues in Europe, cancer was a research priority in the 6th Framework Programme (2002–2006). About €485 million were devoted to this theme, which allowed funding of 108 multidisciplinary transnational projects. A significant part of them was large‐scale initiatives addressing complex issues through a broad combination of competences. All major types of cancer were covered, as well as the three dimensions such as prevention, diagnostic and treatment, with a particular emphasis on translational research aiming at bringing basic knowledge on medical practice. This approach will be continued in the 7th Framework Programme (2007–2013), together with a strengthened effort to improve the coordination of European cancer research, which is fragmented and in which EU action represents only a small part. EU cancer research will also be addressed within the reinforced efforts in the areas of pharmaceutical and technological developments as well as common aetiological mechanisms of diseases that the 7th Framework Programme will undertake.
Cancer therapeutic antibodies come of age: Targeting minimal residual disease
Authors: Ben-Kasus Tsipi , Schechter Bilha , Sela Michael , Yarden Yosef
Published: March 2007

Ten years after the first clinical application of Rituximab, an anti‐CD20 recombinant monoclonal antibody, immunotherapy has become common practice in oncology wards. Thanks to the great diversity of the immune system and the powerful methodology of genetic engineering, the pharmacologic potential of antibody‐based therapy is far from exhaustion. The recent application of Trastuzumab, an antibody to a receptor tyrosine kinase, in adjuvant breast cancer therapy marks the beginning of a new phase in cancer treatment. Here we discuss molecular mechanisms of antibody‐based therapy, the emerging ability to target minimal disease and the therapeutic potential of combining antibodies with other modalities.
Functional role of Meox2 during the epithelial cytostatic response to TGF‐β
Authors: Valcourt Ulrich , Thuault Sylvie , Pardali Katerina , Heldin Carl-Henrik , Moustakas Aristidis
Published: March 2007

Transforming growth factor β (TGF‐β) suppresses epithelial cell growth. We have identified a new target gene of the TGF‐β/Smad pathway, Meox2, encoding the homeodomain transcription factor that is known to regulate endothelial cell proliferation and muscle development. Knockdown of endogenous Meox2 by RNA interference prevented the TGF‐β1‐induced cytostatic response. Moreover, ectopic Meox2 suppressed epithelial cell proliferation in cooperation with TGF‐β1, and mediated induction of the cell cycle inhibitor gene p21. Transcriptional induction of p21 by Meox2 required a distal region of the p21 promoter that spans the p53‐binding site. We show that Meox2 can form protein complexes with Smads leading to cooperative regulation of p21 gene expression. Finally, we found that in cell models that undergo both cell cycle arrest and epithelial–mesenchymal transition (EMT), ectopic Meox2 failed to induce EMT and inhibited the proper EMT response to TGF‐β. Thus, Meox2 is primarily involved in the TGF‐β tumor suppressor pathway.
The morphologies of breast cancer cell lines in three‐dimensional assays correlate with their profiles of gene expression
Authors: Kenny Paraic A. , Lee Genee Y. , Myers Connie A. , Neve Richard M. , Semeiks Jeremy R. , Spellman Paul T. , Lorenz Katrin , Lee Eva H. , Barcellos-Hoff Mary Helen , Petersen Ole W. , Gray Joe W. , Bissell Mina J.
Published: March 2007

3D cell cultures are rapidly becoming the method of choice for the physiologically relevant modeling of many aspects of non‐malignant and malignant cell behavior ex vivo. Nevertheless, only a limited number of distinct cell types have been evaluated in this assay to date. Here we report the first large scale comparison of the transcriptional profiles and 3D cell culture phenotypes of a substantial panel of human breast cancer cell lines. Each cell line adopts a colony morphology of one of four main classes in 3D culture. These morphologies reflect, at least in part, the underlying gene expression profile and protein expression patterns of the cell lines, and distinct morphologies were also associated with tumor cell invasiveness and with cell lines originating from metastases. We further demonstrate that consistent differences in genes encoding signal transduction proteins emerge when even tumor cells are cultured in 3D microenvironments.
Characterization of breast precancerous lesions and myoepithelial hyperplasia in sclerosing adenosis with apocrine metaplasia
Authors: Celis Julio E. , Moreira José M.A. , Gromova Irina , Cabezón Teresa , Gromov Pavel , Shen Tao , Timmermans Vera , Rank Fritz
Published: March 2007

The identification as well as the molecular characterization of breast precancerous lesions in terms of increased risk of progression and/or recurrence is becoming a critical issue today as improved non‐surgical procedures are detecting cancer at an earlier stage. The strategy we have been pursuing to identify early apocrine breast lesions is based on the postulate that invasive apocrine carcinomas evolve from epithelial cells in terminal duct lobular units (TDLUs) in a stepwise manner that involves apocrine metaplasia of normal breast epithelia, hyperplasia, atypia, and apocrine carcinoma in situ. First, we identify specific protein biomarkers for benign apocrine metaplasia and thereafter we search for biomarkers that are highly overexpressed by pure invasive apocrine carcinomas. Here we present studies in which we have used antibodies against components of a benign apocrine signature that includes 15‐prostaglandin dehydrogenase (15‐PGDH), a protein that is expressed by all benign apocrine lesions, and markers that are highly overexpressed by pure invasive apocrine carcinomas such as MRP14 (S100A9), psoriasin (S100A7), and p53 to identify precancerous lesions in sclerosing adenosis (SA) with apocrine metaplasia. The latter is a benign proliferative lesion of the breast that exhibits an increase in the size of the TDLUs and characterized by retained two‐cell lining, and myoepithelial (ME) and stromal hyperplasia. SA with apocrine metaplasia, i.e. apocrine adenosis (AA), presents with a higher degree of atypical apocrine hyperplasia, and these lesions are believed to be precursors of apocrine carcinoma, in situ and invasive. Analysis of 24 selected SA samples with apocrine metaplasia revealed non‐obligate putative apocrine precancerous lesions that displayed some, or in same cases all the three markers associated with pure invasive apocrine carcinomas. These studies also revealed p53 positive, non‐apocrine putative precancerous lesions as well as novel phenotypes for ME and some luminal cells characterized by the expression of cytokeratin 15.
Mice thrive without Cdk4 and Cdk2
Authors: Barrière Cédric , Santamaría David , Cerqueira Antonio , Galán Javier , Martín Alberto , Ortega Sagrario , Malumbres Marcos , Dubus Pierre , Barbacid Mariano
Published: March 2007

Mammalian cell division is thought to be driven by sequential activation of several Cyclin‐dependent kinases (Cdk), mainly Cdk4, Cdk6, Cdk2 and Cdk1. Since mice lacking Cdk4, Cdk6 or Cdk2 are viable, it has been proposed that they play compensatory roles. We report here that mice lacking Cdk4 and Cdk2 complete embryonic development to die shortly thereafter presumably due to heart failure. However, conditional ablation of Cdk2 in adult mice lacking Cdk4 does not result in obvious abnormalities. Moreover, these double mutant mice recover normally after partial hepatectomy. In culture, Cdk4−/−;Cdk2−/−embryonic fibroblasts become immortal, display robust pRb phosphorylation and have normal S phase kinetics. These observations indicate that Cdk4 and Cdk2 are dispensable for the mammalian cell cycle and for adult homeostasis.
The influence of inulin on the absorption of nitrogen and the production of metabolites of protein fermentation in the colon
Authors: Geboes Karen P. , De Hertogh Gert , De Preter Vicky , Luypaerts Anja , Bammens Bert , Evenepoel Pieter , Ghoos Yvo , Geboes Karel , Rutgeerts Paul , Verbeke Kristin
Published: March 2007

In the present study, the production and fate of bacterial metabolites in the colon were investigated in a direct way using two substrates labelled with stable isotopes: lactose [15N,15N]ureide as a source of labelled ammonia and egg proteins intrinsically labelled with [2H4]tyrosine as a precursor of [2H4]p-cresol. Both ammonia and phenolic compounds are believed to be carcinogenic. Stimulation of carbohydrate fermentation in order to prevent accumulation of these toxic metabolites was induced by inclusion of inulin in a test meal or by addition of inulin to the daily diet, allowing us to distinguish between changes induced by the actual presence of a fermentable carbohydrate and effects caused by a long-term dietary intervention. When a single dose of inulin was administered together with the labelled substrates, a significant increase in faecal15N excretion, accompanied by a proportional decrease in urinary15N excretion was observed, probably reflecting an enhanced uptake of ammonia for bacterial biosynthesis, since an increased concentration of labelled N in bacterial pellets was found. A statistically significant reduction of urinary [2H4]p-cresol excretion was also noted. Upon supplementation of inulin to the daily diet during 4 weeks, however, only a tendency towards decreased urinary excretion of both labelled and unlabelledp-cresol was noted. Further studies are warranted to confirm these results in a larger cohort.
Palmitic acid increase levels of pancreatic duodenal homeobox-1 and p38/stress-activated protein kinase in islets from rats maintained on a low protein diet
Authors: Arantes Vanessa C. , Reis Marise A. B. , Latorraca Márcia Q. , Ferreira Fabiano , Stoppiglia Luiz Fabrízio , Carneiro Everardo M. , Boschero Antonio C.
Published: March 2007

A severe reduction in insulin release in response to glucose is consistently noticed in protein-deprived rats and is attributed partly to the chronic exposure to elevated levels of NEFA. Since the pancreatic and duodenal transcription factor homeobox 1 (PDX-1) is important for the maintenance of β-cell physiology, and since PDX-1 expression is altered in the islets of rats fed a low protein (LP) diet and that rats show high NEFA levels, we assessed PDX-1 and insulin mRNA expression, as well as PDX-1 and p38/stress activated protein kinase 2 (SAPK2) protein expression, in islets from young rats fed low (6 %) or normal (17 %; control) protein diets and maintained for 48 h in culture medium containing 5·6 mmol/l glucose, with or without 0·6 mmol/l palmitic acid. We also measured glucose-induced insulin secretion and glucose metabolism. Insulin secretion by isolated islets in response to 16·7 mmol/l glucose was reduced in LP compared with control rats. In the presence of NEFA, there was an increase in insulin secretion in both groups. At 2·8 mmol/l glucose, the metabolism of this sugar was reduced in LP islets, regardless of the presence of this fatty acid. However, when challenged with 16·7 mmol/l glucose, LP and control islets showed a severe reduction in glucose oxidation in the presence of NEFA. The PDX-1 and insulin mRNA were significantly higher when NEFA was added to the culture medium in both groups of islets. The effect of palmitic acid on PDX-1 and p38/SAPK2 protein levels was similar in LP and control islets, but the increase was much more evident in LP islets. These results demonstrate the complex interrelationship between nutrients in the control of insulin release and support the view that fatty acids play an important role in glucose homeostasis by affecting molecular mechanisms and stimulus/secretion coupling pathways.
Zn-limited diet modifies the expression of the rate-regulatory enzymes involved in phosphatidylcholine and cholesterol synthesis
Authors: Gomez Nidia Noemí , Biaggio Verónica Silvina , Rozzen Esteban Javier , Alvarez Silvina Mónica , Gimenez María Sofía
Published: March 2007

Suboptimal intake of Zn is one of the most common nutritional worldwide problems. Previously, we showed that Zn deficiency produces alterations in lung lipid metabolism in rats. We studied the effect of a Zn-limited (ZL) diet on the expression of the enzymes involved in phosphatidylcholine and cholesterol synthesis. After 2 months of treatment with a ZL diet we found important variations in the lipid content of Wistar male rats: triacylglycerol (TG) decreased 60 % (P < 0·001) while esterified cholesterol (EC), free cholesterol and phospholipids (PL) increased 66 %, 24 % and 25 % respectively. We also observed a decrease of 40 % in the amount of3H incorporated into TG and an increase of 47 % and 28 % in the3H incorporated to PL and EC respectively. Fatty acid synthase and glucose-6-phosphate dehydrogenase activity was increased (P < 0·01 andP < 0·05 respectively). Glycerol-3-phosphate acyltransferase, lipoprotein lipase, diacyl glycerol acyl transferase and 3-hydroxy-3-methylglutaryl CoA reductase expression decreased (P < 0·01 in all cases), while acetyl CoA carboxylase and cholinephosphate cytidylyltransferase increased (P < 0·01 andP < 0·005 respectively). These results suggest that ZL alters the expression of enzymes involved in phosphatidylcholine and cholesterol synthesis, which could lead to increased PL and cholesterol and decreased TG. This study suggests that major changes in the lipid composition of lung are induced by a ZL condition. Therefore, Zn deficiency must be taken into account in order to design therapies and public health interventions, such as Zn supplementation for high-risk subjects or certain diseases, such as asthma.
The fatty acid pattern of dietary fat influences the oral bioavailability of the flavonol quercetin in pigs
Authors: Lesser Stephanie , Cermak Rainer , Wolffram Siegfried
Published: March 2007

We have shown recently that dietary fat content influences the bioavailability of the flavonol quercetin. In the present study, the influence of the fatty acid pattern of dietary fats on the oral bioavailability of quercetin was investigated. Quercetin (30 μmol/kg body weight) was administered to growing pigs (n 6) in test meals consisting either of 200 g of a standard pig diet (2 % crude fat) or of the same diet supplemented with 15 g fat/100 g diet using either medium-chain (MCT) or long-chain fatty acid triacylglycerols (LCT). Blood samples were drawn repeatedly over a period of 24 h and analysed by HPLC. In addition, the influence of the different diets on gastric emptying was investigated in rats. In pigs, the bioavailability of quercetin was measured by quantifying its plasma metabolites with an intact flavonol structure. Bioavailability was enhanced by 38 % (P < 0·05) and 12 % (P>0·05) after intake with the MCT and LCT diets, respectively, compared to the standard diet. Maximum plasma concentrations of quercetin were reached significantly later with the MCT diet than with the LCT or the standard diet (P < 0·05). No differences in dry matter of the gastric content were observed 60 min after intake of the experimental diets in rats. Thus, administration of quercetin together with a diet containing MCT fat enhances the bioavailability of the flavonol. Absorption of quercetin was delayed significantly with this diet. However, this was probably not due to slower gastric emptying of the MCT diet.
Levels of serum transferrin receptor and its response to Fe-supplement in Fe-deficient children
Authors: Lin Xiao-Ming , Ji Cheng-Ye , Liu Wen-Jing , Long Zhu , Shen Xiao-Yi
Published: March 2007

The object of the present study was to investigate the levels of serum transferrin receptor (sTfR) and its response to Fe supplementation in Fe-deficient children and the role of sTfR in detecting Fe deficiency and assessing the efficacy of Fe supplementation. According to the diagnostic standard, 1006 children, aged 6–14 years in Fangshan district, Beijing, Peoples Republic of China, were divided into four groups: normal; Fe store depletion (IDs); Fe deficiency erythropoiesis (IDE); Fe deficiency anaemia (IDA). sTfR was determined and transferrin receptor-ferritin (TfR-F) index was calculated in 238 children, sixty-four normal and 174 Fe deficient. Children were administered a NaFeEDTA capsule containing 60 mg Fe once per week for the IDs and IDE groups and three times per week for the IDA group for nine consecutive weeks. The parameters reflecting Fe status and sTfR were determined before and after Fe supplementation. The levels of sTfR and TfR-F index in Fe-deficient children were significantly higher than those in the normal group. The receiver operating characteristic curve showed that sTfR has proper diagnostic efficacy for functional Fe deficiency. After Fe supplementation, the level of sTfR was significantly decreased in children with IDs, but not in children with IDE and IDA, while TfR-F index was significantly decreased in Fe-deficient children. sTfR is a reliable indicator for detecting functional Fe deficiency, and TfR-F index is a sensitive parameter for assessing the efficacy of Fe supplementation.
Overweight induced by high-fat diet delays rat cutaneous wound healing
Authors: Nascimento Adriana P. , Costa Andréa M. A.
Published: March 2007

Prolonged wound healing is a complication that contributes to morbidity and mortality. Overweight people regularly undergo surgery and trauma, and often develop chronic wounds, but the effects of the adipose tissue excess on cutaneous wound healing are not well understood. This study tested the hypothesis that overweight induced by a high-fat diet impairs rat cutaneous wound healing. Male Wistar rats were fed with either a high-fat or a standard (control) diet. After 15 weeks, an excisional lesion was done and the animals were killed 21 d later. Wound contraction and re-epithelialization, blood pressure, glucose and retroperitoneal fat were evaluated. After killing, lesion and adjacent normal skin were formol-fixed and paraffin-embedded. Inflammatory infiltrate, myofibroblasts, collagen fibres and cellular proliferation were analysed and blood vessels were evaluated using stereological methods. There was no difference in blood pressure and glucose, but retroperitoneal fat increased in the high-fat diet group. Animals fed with the high-fat diet presented delayed wound contraction and re-epithelialization. It was found that 21 d after wounding, overweight induced by a high-fat diet increased the inflammatory infiltrate and delayed myofibroblastic differentiation, collagen deposition, epithelial and connective tissue cell proliferation, and angiogenesis. These findings support the hypothesis that a high-fat diet exerts negative effects on rat cutaneous wound healing, due mainly to the prolongation of the inflammatory phase.
Serum free testosterone, leptin and soluble leptin receptor changes in a 6-week strength-training programme
Authors: Ara I. , Perez-Gomez J. , Vicente-Rodriguez G. , Chavarren J. , Dorado C. , Calbet J. A. L.
Published: March 2007

Strength training is usually associated with a reduction in fat mass and with muscle hypertrophy. The aim of the present study was to examine whether the serum free leptin index (FLI), measured by the molar excess of soluble leptin receptor (sOB-R) over leptin, is increased by 6 weeks of strength training. Eighteen male, physical education students were randomly assigned to two groups: a strength-training (n12) and a control group (n6). Body composition (lean body mass and body fat) determined by dual-energy X-ray absorptiometry (DXA), muscle performance and leptin, sOB-R, total testosterone and free testosterone concentrations were determined before and after training. Fat mass was reduced by 1 kg with strength training (P < 0·05). Lean body mass of trained extremities was increased by 3 % (P < 0·05), while the concentration of free testosterone in serum was reduced by 17 % (P < 0·05) after training. However, despite the reduction in fat mass and free testosterone, serum leptin concentration was not significantly affected by strength training, even after accounting for the differences in body fat. By contrast, for a given fat mass, the sOB-R was increased by 13 % (P < 0·05) at the end of the strength-training programme, although the molar excess of sOB-R over leptin remained unchanged. Therefore, the quantity of free leptin available to bind to the target tissues was not significantly affected by the short strength-training programme, which elicited a 7 % reduction in fat mass.
No effect of increased water intake on blood viscosity and cardiovascular risk factors
Authors: Tonstad Serena , Klemsdal Tor Ole , Landaas Sverre , Høieggen Aud
Published: March 2007

Observational data have suggested that increased water intake decreases the risk of CHD. A postulated mechanism is that increased water ingestion reduces blood viscosity. The aim of the present study was to assess the effect of increased fluid intake on blood viscosity. Men (n 67) and postmenopausal women (n 27) with one or more risk factors for CVD who reported intake of ≤ 0·5 litres water daily were randomised to a control group (n 31), an intervention group (n 32) that increased their daily water intake by 1 litre/d and an intervention group (n 31) that ingested 1 litre blueberry juice/d. All were encouraged to continue their usual diet and lifestyle. Whole-blood viscosity and blood and urine chemistries were measured by standard techniques after 2 and 4 weeks. Urine volume increased (by a median of 872 and 725 ml in the water and blueberry juice groups, respectively, v. 10 ml in the control group; P ≤ 0·002), confirming the subjects' adherence to the protocol. Urine osmolality and urinary levels of Na, K and creatinine decreased in the water and blueberry juice groups v. the controls (P < 0·05). No change was seen in whole-blood viscosity or in levels of fibrinogen, total protein, lipids, glucose, insulin, C-peptide or other chemistry and haematology variables. In conclusion, a postulated protective effect of increased water or fluid intake is not explained by a change in blood viscosity and increased fluid intake does not influence CVD risk factors in the short term.
Effect of diet composition on pregnancy outcome in overnourished rapidly growing adolescent sheep
Authors: Wallace Jacqueline M. , Milne John S. , Redmer Dale A. , Aitken Raymond P.
Published: March 2007

When pregnant adolescent sheep are overnourished to promote maternal growth during pregnancy, growth of the placenta is impaired and results in the premature delivery of low birth weight lambs relative to control-fed adolescents of equivalent age. These effects have been achieved by feeding two levels of the same complete diet. The present study evaluated the role of protein in pregnancy outcome in our adolescent sheep paradigm. Adolescent ewes were implanted with single embryos on day 4 post-oestrus. Thereafter ewes were offered ad libitum an isoenergetic diet (11·4 MJ metabolisable energy/kg DM) containing either 12 % (basic, B) or 17 % (extra, E) crude protein. At day 75 of gestation, half the pregnant ewes on each protein level were switched to yield four groups, BB, EE, BE and EB protein. A further optimally nourished control group received a moderate quantity of a ration (14 % crude protein) designed to provide 100 % of the estimated energy and protein requirement of the adolescent sheep according to stage of pregnancy. Pregnancy outcome was determined at term. Feed intakes were independent of protein level in the four groups of ewes fed ad libitum and were higher (P < 0·001) than in the control group throughout. Maternal plasma urea concentrations reflected the current crude protein content of the diet offered and were elevated in the 17 % compared with 12 % protein groups (P < 0·001). Within groups fed ad libitum, maternal plasma insulin, glucose, NEFA and homocysteine concentrations were largely independent of protein level. Gestation length, placental weight, lamb birth weight and initial colostrum yield were reduced (P < 0·05) in all groups fed ad libitum relative to the optimally nourished control group. Similarly, total colostrum IgG, butterfat, lactose and crude protein content at parturition were attenuated in the ad libitum compared with the control groups. However, within ad libitum groups pregnancy outcome parameters were largely unaffected by level or timing of exposure to high protein intakes. The data imply that it is high-energy intakes that are the primary cause of impaired placental development and adverse pregnancy outcome in rapidly growing adolescent sheep.
The Westphalian–Stephanian macrofloral record from the South Wales Coalfield, UK
Authors: CLEAL CHRISTOPHER J.
Published: March 2007

The South Wales Coalfield has the most complete Westphalian macrofloral record anywhere on the Variscan Foreland or adjacent basins, with 135 biodiversity-meaningful morphospecies having been recognized. All of the standard macrofloral biozones of the Westphalian Stage have been recognized, although a detailed comparison with the Central Pennines Coalfields has indicated some discrepancies in the relative positions of the biozonal boundaries. Total Species Richness progressively increases through the Langsettian Substage, and then remains relatively stable through most of the Duckmantian and Bolsovian substages. There is a distinct reduction in Total Species Richness towards the top of the Bolsovian Substage, but this partially recovers in the middle Asturian Substage with the appearance of a range of marattialean ferns, and medullosalean and callistophytaleans pteridosperms. There is no evidence of any significant drop in Total Species Richness towards the top of the succession, indicating that conditions at this time were relatively stable. The change from coastal floodplain to alluvial braidplain conditions in middle Bolsovian times correlates with a marked increase in the proportion of medullosalean remains being preserved in the adpression record, reflecting an expansion of the clastic-substrate habitats.
DESIRED FAMILY SIZE, FAMILY PLANNING AND FERTILITY IN ETHIOPIA
Authors: BHARGAVA ALOK
Published: March 2007

This paper models the proximate determinants of children born to over 13,000 Ethiopian women and of the women’s stated preferences for additional births using the data from the Ethiopian Demographic and Health Survey 2000. Empirical models for the number of children born to women were estimated using Poisson and ordinal regressions. The results show the importance of variables such as maternal education for smaller family size, and that variables reflecting desired family size are strong predictors of the numbers of children born to women. Secondly, binary logistic models for dichotomous variables for women not wanting more children and if getting pregnant would be a ‘big problem’ showed non-linear effects of the surviving and ‘ideal’ number of children. Moreover, the results indicated a desire on the part of women to limit family size, especially as the number of surviving children increased. Probit models were estimated to address potential endogeneity of certain variables. Overall, the results indicated that counselling couples about small family size and increasing the utilization of health care services can lower fertility in Ethiopia.