bluuebunny/crossref_metadata_2025 huggingface dataset:
Stable isotope evidence for the consumption of millet and other plants in Bronze Age Italy
Authors: Tafuri Mary Anne , Craig Oliver E. , Canci Alessandro
Published: December 2008

Stable carbon and nitrogen isotope analysis was carried out on human and animal bones from four inland Early and Middle Bronze Age sites in Northern and Southern Italy. The main aims of the investigation were to explore the contribution of plant foods to the human diet and to examine any dietary differences between and within each of the sites. At two of the sites in Northern Italy, human and animal bones were significantly enriched in 13C. This finding was attributed to the consumption of domestic millets (Panicum miliaceum and/or Setaria italica), which are C4 pathway plants. Conversely, individuals from the two Bronze Age sites in Southern Italy were significantly depleted in 13C compared to those from the north. Here, millet was absent from the diet, and protein from C3 plants made a much greater dietary contribution than animal protein. This finding highlights the importance of cereal cultivation, most likely of wheat and barley, in the south of Italy during the Bronze Age. Overall, our results support the idea that the widespread cultivation of millet first occurred in Northern Italy, following its introduction from across the Alps in Central Europe. Finally, we found no significant differences in the stable isotope values between individuals at each site, when grouped by their sex or presence of grave goods. This leads to the conclusion that any status difference that may have existed is not reflected in the long‐term dietary record, or at least not as measurable by stable isotope analysis. Am J Phys Anthropol, 2009. © 2008 Wiley‐Liss, Inc.
Topography characterization and initial cellular interaction of plasma‐based Ar+ beam‐treated PDMS surfaces
Authors: Keranov I. , Vladkova T. G. , Minchev M. , Kostadinova A. , Altankov G. , Dineff P.
Published: December 2008

Assuming that the existence of an ion‐flow in the plasma volume could strength the surface modifying effect, including its durability, a parallel plate reactor in reactive ion etching mode was employed to obtain surface modified PDMS with improved cellular interaction. The discharge power was varied at 100, 1200, and 2500 W to ensure varied ion‐flow density. The changes in the surface topography were observed by SEM and AFM, and the surface roughness was characterized by both: mean roughness, Ra, and root‐mean‐square, Rq. Time dependent water contact angle measurements were performed to control the durability of the hydrophilizing effect. Anisotropic etching, accompanied with decrease of the PDMS surface roughness, was observed up to discharge power of 1200 W that turns in intense isotropic one, accompanied with a sharp increase of the surface roughness over 1200 W, most probably because of arise of reverse sputtered neutrals diffracting the main plasma Ar+ flow. Human fibroblasts were applied as an in vitro model to learn more about the initial cellular interaction of the modified surfaces and to identify the optimal treatment conditions. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009
Polyurethane/multiwalled carbon nanotube nanowebs prepared by an electrospinning process
Authors: Kimmer Dušan , Slobodian Petr , Petráš David , Zatloukal Martin , Olejník Robert , Sáha Petr
Published: December 2008

Polyurethane (PU) and PU/multiwalled carbon nanotube (MWCNT) nanocomposite nanofibers, both with diameters of 350 nm, were prepared by an electrospinning process from PU dimethylformamide solutions. The appearance of nanowebs in PU/MWCNT nanofiber structures containing PU fibers with diameters of 20–40 nm was observed. The existence of these structures could have been based on the occurrence of strong secondary electric fields, which were created between individual conducting MWCNTs (distributed in the PU/MWCNT nanocomposites), which started to behave as the local moving nanoelectrodes promoting the creation of additional very fine nanowebs during the electrospinning processes. To our knowledge, this is the first report describing nanowebs from synthetic polymers prepared by an electrospinning process. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009
Immobilized metal ions cellophane–PGMA‐grafted membranes for affinity separation of β‐galactosidase enzyme. I. Preparation and characterization
Authors: Mohy Eldin M. S. , Soliman E. A. , Hassan E. A. , Abu‐Saied M. A.
Published: December 2008

Immobilized Cu2+ ions affinity cellophane–poly(glycidyl methacrylate) (PGMA)‐grafted membranes have been prepared through three steps. The first step was introducing of epoxy groups to its chemical structure through grafting process with PGMA. Factors affecting the grafting process have been studied and grafting percentage (GP) up to 233% has been obtained. The second step was converting the introduced epoxy groups to sulfonic ones. It was found that maximum amount of sulfonic groups (2.7 mmol/g) was obtained with minimum GP (46.08%). The third and last step was the immobilization of Cu2+ ions into sulfonated grafted membranes obtained from the previous step. Maximum amount of immobilized Cu2+ ions was found to be 60.9 ppm per gram of polymer. The verification of the grafting and sulfonation steps has been performed through characterization of the obtained membranes using FTIR, TGA, and EDAX analysis. Finally, Cu2+‐immobilized membranes have been evaluated in separation of β‐galactosidase (β‐Gal) enzyme from its mixture with bovine serum albumin (BSA) in different pH medium. Maximum protein adsorption, for both proteins, has been obtained at pH range 4–4.5; as 90 and 45% for β‐Gal and BSA, respectively. The results showed high affinity toward β‐Gal separation although BSA concentration (0.5%) is 20‐folds of β‐Gal (0.025%). © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009
Characterization and Classification of Zebrafish Brain Morphology Mutants
Authors: Lowery Laura Anne , De Rienzo Gianluca , Gutzman Jennifer H. , Sive Hazel
Published: December 2008

The mechanisms by which the vertebrate brain achieves its three‐dimensional structure are clearly complex, requiring the functions of many genes. Using the zebrafish as a model, we have begun to define genes required for brain morphogenesis, including brain ventricle formation, by studying 16 mutants previously identified as having embryonic brain morphology defects. We report the phenotypic characterization of these mutants at several timepoints, using brain ventricle dye injection, imaging, and immunohistochemistry with neuronal markers. Most of these mutants display early phenotypes, affecting initial brain shaping, whereas others show later phenotypes, affecting brain ventricle expansion. In the early phenotype group, we further define four phenotypic classes and corresponding functions required for brain morphogenesis. Although we did not use known genotypes for this classification, basing it solely on phenotypes, many mutants with defects in functionally related genes clustered in a single class. In particular, Class 1 mutants show midline separation defects, corresponding to epithelial junction defects; Class 2 mutants show reduced brain ventricle size; Class 3 mutants show midbrain–hindbrain abnormalities, corresponding to basement membrane defects; and Class 4 mutants show absence of ventricle lumen inflation, corresponding to defective ion pumping. Later brain ventricle expansion requires the extracellular matrix, cardiovascular circulation, and transcription/splicing‐dependent events. We suggest that these mutants define processes likely to be used during brain morphogenesis throughout the vertebrates. Anat Rec, 2009. © 2008 Wiley‐Liss, Inc.
Ontogenetic Patterning of Cortical Bone Microstructure and Geometry at the Human Mid‐Shaft Femur
Authors: Goldman H.M. , Mcfarlin S.C. , Cooper D.M.L. , Thomas C.D.L. , Clement J.G.
Published: December 2008

The bone growth process has long‐lasting effects on adult bone structure and mechanical adaptation, yet the tissue level dynamics of growth are poorly studied. The specific aims of this study were to (1) quantify changes in bone size and shape through ontogeny, (2) describe the distribution of tissue types and patterns of cortical drift and expansion through ontogeny, and (3) explore relationships between cortical drift and ontogenetic variation geometric size and shape. The study utilized 14 juvenile (ages 2–19) mid‐shaft femur blocks removed at autopsy from individuals who died suddenly. Eighty‐μm‐thick sections were imaged using polarized and brightfield microscopy. For descriptive purposes the sample was divided into five age groups. Features of collagen fiber matrix orientation, vascularity (e.g., pore orientation and density), and osteocyte lacunar density and shape were used to classify primary and secondary tissue types in LM images. This information, combined with evaluation of resorptive versus depositional bone surfaces, was used to identify cortical drift direction. A pattern of posterior and medial drift was identified at the mid‐shaft femur in the toddler years. The drift pattern shifts antero‐laterally in late childhood, predating the appearance of a more adult‐like geometry. On the basis of the presence of transitional fibrolamellar bone complex, growth is more rapid during the toddler years and peri‐puberty, and slower in early to late childhood and in later adolescence. Extensive variability in histological and geometric organization typifies the sample, particularly beginning in late childhood. The potential implications of this variability for adult fracture risk warrant further study. Anat Rec, 2009. © 2008 Wiley‐Liss, Inc.
Gene Expression Profiling in TWIST‐Depleted Gastric Cancer Cells
Authors: Feng Mei‐Yan , Wang Kuan , Shi Qing‐Tao , YU Xiu‐Wen , Geng Jing‐Shu
Published: December 2008

TWIST is an important transcription factor during embryonic development and has recently been found to promote the epithelial‐mesenchymal transition (EMT) phenomenon seen during the initial steps of tumor metastasis. To further investigate the potential targets and interacting genes of TWIST in human gastric cancer, we performed microarray analysis to compare the gene expression profiles in HGC‐27 cells, with or without small interfering RNA (siRNA)‐mediated depletion of TWIST. Our results showed that NF1, RAP1A, SRPX, RBL2, PFDN4, ILK, F2R, ERBB3, and MYB were up‐regulated, whereas AKR1C2, FOS, GDF15, NR2F1, ATM, and CTPS were down‐regulated after TWIST depletion. Moreover, TWIST‐depleted HGC‐27 cells showed a reversal of the morphologic and molecular changes associated with EMT. These results provide evidence that TWIST regulates the expression of several genes involved in the differentiation, adhesion, and proliferation of gastric cancer cells. The role of TWIST in the development of certain types of gastric cancer is discussed. Anat Rec, 2009. © 2008 Wiley‐Liss, Inc.
Pancreatitis‐Associated Protein Is Related Closely to Neoplastic Proliferative Activity in Patients with Colorectal Carcinoma
Authors: Cao Guang , Ma Jing , Zhang Yafang , Liu Baoquan , Li Fei
Published: December 2008

Pancreatitis‐associated protein (PAP) is a secretory protein that is not only expressed during acute pancreatitis but also in pancreatic adenocarcinoma, gastric carcinoma, hepatocellular carcinoma, and colorectal carcinoma. Expression in carcinoma might be another characteristic of PAP. The aim of our study was to assess, in 27 patients undergoing surgery for colorectal carcinoma, the expression of the PAP mRNA and to evaluate its association with DNA ploidy and proliferative activity (S‐phase fraction, SPF) by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and flow cytometric analysis (FCM). PAP mRNA was expressed in 29.6% (8 of 27) of the patients with colorectal carcinoma. DNA aneuploid and high SPF were found in 87.5% (7 of 8) of patients with PAP mRNA positive colorectal carcinoma. The serum PAP level was significantly elevated in patients with colorectal carcinoma when compared with the healthy subjects. Twelve of the 27 patients with colorectal carcinoma had high serum PAP concentrations (>25 ng/mL) and the mean SPF was 17.82% ± 8.02%, which was significantly higher compared with the normal colorectal tissue group (7.33% ± 3.18%, P < 0.05). The mean serum PAP concentration of DNA aneuploidy colorectal carcinomas was 46.67 ± 17.58 ng/mL, which was significantly different when compared with the DNA diploidy group (19.18 ± 8.89 ng/mL, P < 0.05). PAP mRNA expression and serum PAP levels are closely related to neoplastic proliferative activity in patients with colorectal carcinoma. No significant differences are observed between PAP mRNA expression and clinicopathologic parameters (P > 0.05). Anat Rec, 2009. © 2008 Wiley‐Liss, Inc.
Lung Development of Monotremes: Evidence for the Mammalian Morphotype
Authors: Ferner Kirsten , Zeller Ulrich , Renfree Marilyn B.
Published: December 2008

The reproductive strategies and the extent of development of neonates differ markedly between the three extant mammalian groups: the Monotremata, Marsupialia, and Eutheria. Monotremes and marsupials produce highly altricial offspring whereas the neonates of eutherian mammals range from altricial to precocial. The ability of the newborn mammal to leave the environment in which it developed depends highly on the degree of maturation of the cardio‐respiratory system at the time of birth. The lung structure is thus a reflection of the metabolic capacity of neonates. The lung development in monotremes (Ornithorhynchus anatinus, Tachyglossus aculeatus), in one marsupial (Monodelphis domestica), and one altricial eutherian (Suncus murinus) species was examined. The results and additional data from the literature were integrated into a morphotype reconstruction of the lung structure of the mammalian neonate. The lung parenchyma of monotremes and marsupials was at the early terminal air sac stage at birth, with large terminal air sacs. The lung developed slowly. In contrast, altricial eutherian neonates had more advanced lungs at the late terminal air sac stage and postnatally, lung maturation proceeded rapidly. The mammalian lung is highly conserved in many respects between monotreme, marsupial, and eutherian species and the structural differences in the neonatal lungs can be explained mainly by different developmental rates. The lung structure of newborn marsupials and monotremes thus resembles the ancestral condition of the mammalian lung at birth, whereas the eutherian newborns have a more mature lung structure. Anat Rec, 2009. © 2008 Wiley‐Liss, Inc.
Synthesis and Pharmacological Evaluation ofN‐(Dimethylamino)ethyl Derivatives of Benzo‐ and Pyridopyridazinones
Authors: Pakulska Wanda , Malinowski Zbigniew , Szczesniak Aleksandra K. , Czarnecka Elzbieta , Epsztajn Jan
Published: December 2008

NewN‐(dimethylamino)ethyl derivatives of phthalazinones and pyridopyridazinones7,9were synthesized and assayed as potential analgesic agents in the hot‐plate, tail‐flick, and writhing tests. Pharmacological assay demonstrated that eight (in ten) of the newly synthesized compounds showed antinociceptive activity. Especially, 2‐[2‐(dimethylamino)ethyl]‐4‐phenyl‐2H‐phthalazin‐1‐one7ashowed remarkably higher antinociceptive activity in all tests. This is connected with influence on supraspinal, spinal, and peripheral structures. The decreased sensitivity to the pain stimulus in the hot‐plate was higher than that of metamizole.
Synthesis and Antimicrobial Activity of Some Novel 2‐[4‐(Substituted Piperazin‐/Piperidin‐1‐ylcarbonyl)phenyl]‐1H‐benzimidazole Derivatives
Authors: Kus Canan , Sözüdönmez Fatma , Altanlar Nurten
Published: December 2008

In this study, we report the synthesis and antimicrobial evaluation of several new 4‐(1H‐benzimidazol‐2‐yl)benzamides (11–30) and 5‐chloro‐1‐(p‐fluorobenzyl)‐2‐{4‐[(4‐methylpiperazin‐1‐yl)carbonyl]phenyl}‐1H‐benzimidazole (33). Compound 20 exhibited the best antibacterial activity with MIC value of 6.25 μg/mL against Staphylococcus aureus and methicillin‐resistant Staphylococcus aureus (MRSA). Significant antifungal activities were obtained with the compounds 13, 14, 18, 19, and 33 with MIC values of 3.12 μg/mL which are close to fluconazole.
Influence of the dye presence on the conformational preferences of CREKA, a tumor homing linear pentapeptide
Authors: Zanuy David , Curcó David , Nussinov Ruth , Alemán Carlos
Published: December 2008

The conformational properties of the Cys‐Arg‐Glu‐Lys‐Ala (CREKA) peptide sequence labeled with fluorescein, a fluorescent dye attached to the Cys through a flexible linker have been examined using molecular dynamics simulations. The CREKA sequence has been identified as a tumor‐homing peptide that effectively binds to clotted plasma proteins. Before conformational exploration, the molecular geometry, basicity and spectroscopic properties of this dye, which is essential for the imaging the peptide activity, have been examined using quantum mechanical calculations, with the results also allowing determination of the force‐field parameters required for classical simulations. Minimum energy conformations derived from the conformational search have been classified using clustering analyses with criteria based on both the existence of interactions and backbone geometric similarity. The results have been compared with those reported for isolated CREKA (peptide without dye). We found that the fluorescein affects the energy distribution of the minimum energy conformations, with the repulsive steric interactions induced by the dye producing shifting the distribution towards higher energy values. Interestingly, although the structural characteristics of the bioactive conformation identified for CREKA are not perturbed by the dye, it is less stable when the peptide is attached to the dye than in other chemical environments previously studied (isolated peptide, peptide attached to the surface of a protein, and peptide inserted in a phage display protein). © 2008 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 92: 83–93, 2009.

This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com
Constructing a legitimate history: crisis, legitimacy, and the atlantic schools of Business Conferences
Authors: Long Brad , Pyper Rhonda , Rostis Adam
Published: December 2008

The Atlantic Schools of Business (ASB) conference is one of the longest running academic business conferences in Canada. It continued without the structure of an organizing committee for 34 years. In its 35th year, however, a formal structure was adopted. The adoption of formal structure provides an opportunity to look critically at the change process from various perspectives. This paper takes three interrelated approaches: reviews of archival material to provide context; surveys of ASB membership to ascertain individual perceptions of the organization's purpose; and interviews with key individuals involved in the change process to examine the decision to formalize. Our analysis shows that perceived organizational crisis and a search for enhanced legitimacy can precipitate isomorphism and exclude alternative avenues for change. Copyright © 2008 ASAC. Published by John Wiley & Sons, Ltd.
Strategic ambidexterity and performance in international new ventures
Authors: Han Mary , Celly Nikhil
Published: December 2008

We drew on the literature on ambidextrous organizations to propose that international new ventures (INVs) that concurrently pursue paradoxical pairs of strategies—known as the capability of strategic ambidexterity—can achieve superior performance. These paradoxical strategies are: (a) few investments and many countries, and (b) standardization and innovation. Our hypotheses were tested using data from 70 Canadian INVs. This is the first study that attempts to theoretically and empirically link ambidexterity and INV performance. Our findings suggest that INVs that are capable of pursuing and implementing paradoxical strategies achieve superior performance over those lacking such capability. Copyright © 2008 ASAC. Published by John Wiley & Sons, Ltd.
Thirty years of international business scholarship in Canada
Authors: Rugman Alan M.
Published: December 2008

Upon the invitation of the Hemant Merchant (Editor, Strategic Management and International Business Division) I have reviewed the development of international business (IB) scholarship in Canada and related this to my own research conducted while I was a faculty member at the University of Winnipeg 1970–79, Concordia University 1979–80, Dalhousie University 1980–87, and the University of Toronto 1987–98. I have developed a retrospective analysis of four key thematic areas of research that are specifically relevant in a Canadian context. Copyright © 2008 ASAC. Published by John Wiley & Sons, Ltd.
Connexin36, an essential element in the rod pathway, is highly expressed in the essentially rodless retina of Gallus gallus
Authors: Kihara A.H. , Paschon V. , Cardoso C.M. , Higa G.S.V. , Castro L.M. , Hamassaki D.E. , Britto L.R.G.
Published: December 2008

Electrical coupling provided by connexins (Cx) in gap junctions (GJ) plays important roles in both the developing and the mature retina. In mammalian nocturnal species, Cx36 is an essential component in the rod pathway, the retinal circuit specialized for night, scotopic vision. Here, we report the expression of Cx36 in a species (Gallus gallus) that phylogenetic development endows with an essentially rodless retina. Cx36 gene is very highly expressed in comparison with other Cxs previously described in the adult retina, such as Cx43, Cx45, and Cx50. Moreover, real‐time PCR, Western blot, and immunofluorescence all revealed that Cx36 expression massively increased over time during development. We thoroughly examined Cx36 in the inner and outer plexiform layers, where this protein was particularly abundant. Cx36 was observed mainly in the off sublamina of the inner plexiform layer rather than in the on sublamina previously described in the mammalian retina. In addition, Cx36 colocalized with specific cell markers, revealing the expression of this protein in distinct amacrine cells. To investigate further the involvement of Cx36 in visual processing, we examined its functional regulation in retinas from dark‐adapted animals. Light deprivation markedly up‐regulates Cx36 gene expression in the retina, resulting in an increased accumulation of the protein within and between cone synaptic terminals. In summary, the developmental regulation of Cx36 expression results in particular circuitry‐related roles in the chick retina. Moreover, this study demonstrated that Cx36 onto‐ and phylogenesis in the vertebrate retina simultaneously exhibit similarities and particularities. J. Comp. Neurol. 512:651–663, 2009. © 2008 Wiley‐Liss, Inc.
Age‐related changes in hypothalamic androgen receptor and estrogen receptor α in male rats
Authors: Wu Di , Lin Grace , Gore Andrea C.
Published: December 2008

The control of reproductive function involves actions of sex steroids upon their nuclear receptors in the hypothalamus and preoptic area (POA). Whether hypothalamic hormone receptors change their expression in aging male mammals has not been extensively pursued, although such changes may underlie functional losses in reproductive physiology occurring with aging. We performed a stereologic analysis of immunoreactive androgen receptor (AR) and estrogen receptor alpha (ERα) cells in three POA nuclei of male Sprague‐Dawley rats (anteroventral periventricular nucleus [AVPV], median preoptic area [MePO], and medial preoptic nucleus [MPN]), at young (3 months), middle‐aged (12 months), and old (20 months) ages. Serum testosterone and estradiol levels were assayed. Testosterone concentrations decreased significantly and progressively with aging. Estradiol concentrations were significantly higher in middle‐aged than either young or old rats. Stereologic analyses of the POA demonstrated that AR‐immunoreactive cell numbers and density in the AVPV, MePO, and MPN were significantly higher in old compared with young or middle‐aged rats. No change in the total number or density of ERα‐immunoreactive cells was detected with age, although when cells were subdivided by intensity of immunolabeling, the most heavily labeled ERα cells increased in number with aging in the AVPV and MePO, and in density in the AVPV. There are several interpretations to our finding of substantially increased AR cell numbers during aging, including a potential compensatory upregulation of the AR under diminished testosterone concentrations. These results provide further information about how the neural targets of steroid hormones change with advancing age. J. Comp. Neurol. 512:688–701, 2009. © 2008 Wiley‐Liss, Inc.
Stem cells of the adult cornea: From cytometric markers to therapeutic applications
Authors: Takács Lili , Tóth Enikö , Berta András , Vereb György
Published: December 2008

The cornea is a major protective shield of the interior of the eye and represents two thirds of its refractive power. It is made up of three tissue layers that have different developmental origins: the outer, epithelial layer develops from the ectoderm overlying the lens vesicle, whereas the stroma and the endothelium have mesenchymal origin. In the adult organism, the outermost corneal epithelium is the most exposed to environmental damage, and its constant renewal is assured by the epithelial stem cells that reside in the limbus, the circular border of the cornea. Cell turnover in the stromal layer is very slow and the endothelial cells probably do not reproduce in the adult organism. However, recent experimental evidence indicates that stem cells may be found in these layers. Damage to any of the corneal layers leads to loss of transparency and low vision. Corneal limbal stem cell deficiency results in severe ocular surface disease and its treatment by transplantating ex vivo expanded limbal epithelial cells is becoming widely accepted today. Stromal and endothelial stem cells are potential tools of tissue engineering and regenerative therapies of corneal ulcers and endothelial cell loss. In the past few years, intensive research has focused on corneal stem cells aiming to improve the outcomes of the current corneal stem cell transplantation techniques. This review summarizes the current state of knowledge on corneal epithelial, stromal and endothelial stem cells. Special emphasis is placed on the molecular markers that may help to identify these cells, and the recently revealed mechanisms that could maintain their “stemness” or drive their differentiation. The techniques for isolating and culturing/expanding these cells are also described. © 2008 International Society for Advancement of Cytometry
Suitability of various membrane lipophilic probes for the detection of trogocytosis by flow cytometry
Authors: Daubeuf Sandrine , Bordier Christine , Hudrisier Denis , Joly Etienne
Published: December 2008

Trogocytosis is a recently discovered phenomenon whereby lymphocytes capture fragments of the plasma membrane from antigen presenting cells (APCs). Using APCs labeled with widely used fluorescent lipophilic probes, we previously described a trogocytosis analysis protocol (TRAP) useful to understand the mechanisms and biological consequences of this process and to identify lymphocytes reacting specifically with antigen‐bearing APCs. We have compared the suitability of 22 different fluorescent lipophilic probes for use in TRAP assays with cytotoxic T lymphocytes (CTL). The criteria we used were: simple and efficient incorporation in APC membranes, minimal passive diffusion among cells but efficient transfer onto T cells during trogocytosis. Sphingosin‐based probes were found to incorporate inefficiently into cells. For others with unsaturated lipid chains, we found a tendency for extensive passive diffusion. In the end, about a third of the probes tested were found to be suitable in TRAP assays, which all carry either C16 or C18 saturated carbon chains, including some that can be excited with a red laser. Moreover, we found it possible to combine TRAP assays based on lipophilic probes with intracellular cytokine detection. We have identified a set of new lipophilic fluorescent probes suitable for TRAP assays in combination with intracellular staining. © 2008 International Society for Advancement of Cytometry
Single cell analysis reveals unexpected growth phenotype ofS. cerevisiae
Authors: Kortmann Hendrik , Blank Lars M. , Schmid Andreas
Published: December 2008

Single cell analysis is mainly limited to single time‐point measurements, without the possibility to track behavioral changes of a single cell or its descendents. Here, the integration of a spatiotemporal single cell lab‐on‐a‐chip system with an automated cultivation device allows single cell analysis under defined growth conditions and, especially, semiautomated cell retrieval and growth kinetic analysis of the single cell descendants. Performance of the new platform was evaluated using the yeastSaccharomyces cerevisiae. The yeast was singularized in the lab‐on‐a‐chip (Envirostat), which combines the possibility of cell cultivation with cell analysis. Singularized cells were collected in a microtiter plate and cultivated in a semiautomated cultivation device (Bioscreen C).S. cerevisiaeshowed highly reproducible and glucose concentration independent growth kinetics in population experiments. Yet, growth kinetics in cultures inoculated with only one or few cells exhibited strong variations, because of an unexpected growth phenotype: colony formation in submerse cultures. Interestingly, the colony‐like structures grew for more then 60 h and were stable for at least 82 h, despite rigorous shaking. Cell agglomeration due to pseudohyphal growth could be excluded, suggesting changes in cell wall properties of yeast populations starting from a single yeast. Single cell analysis still exhibits unexpected obstacles. Nevertheless, this new single cell analysis platform can be used for studying cellular dynamics of single cells and expanded cell populations thereof. © 2008 International Society for Advancement of Cytometry
Diabetes reduces bone marrow and circulating porcine endothelial progenitor cells, an effect ameliorated by atorvastatin and independent of cholesterol
Authors: Mohler Emile R. , Shi Yuquan , Moore Jonni , Bantly Andrew , Hamamdzic Damir , Yoder Mervin , Rader Daniel J. , Putt Mary , Zhang Lifeng , Parmacek Michael , Wilensky Robert L.
Published: December 2008

Bone marrow derived endothelial progenitor cells (EPCs) are early precursors of mature endothelial cells which replenish aging and damaged endothelial cells. The authors studied a diabetic swine model to determine if induction of DM adversely affects either bone marrow or circulating EPCs and whether a HMG‐CoA reductase inhibitor (statin) improves development and recruitment of EPCs in the absence of cholesterol lowering. Streptozotocin was administered to Yorkshire pigs to induce DM. One month after induction, diabetic pigs were treated with atorvastatin (statin, n = 10), ezetimibe (n = 10) or untreated (n = 10) and evaluated for number of bone marrow and circulating EPCs and femoral artery endothelial function. There was no effect of either medication on cholesterol level. One month after induction of DM prior to administration of drugs, the number of bone marrow and circulating EPCs significantly decreased (P < 0.0001) compared to baseline. Three months after DM induction, the mean proportion of circulating EPCs significantly increased in the atorvastatin group, but not in the control or ezetimibe groups. The control group showed progressive reduction in percentage of flow mediated vasodilatation (no dilatation at 3 months) whereas the atorvastatin group and ezetimibe exhibited vasodilatation, 6% and 4% respectively. DM results in significant impairment of bone marrow and circulating EPCs as well as endothelial function. The effect is ameliorated, in part, by atorvastatin independent of its cholesterol lowering effect. These data suggest a model wherein accelerated atherosclerosis seen with DM may, in part, result from reduction in EPCs which may be ameliorated by treatment with a statin. © 2008 International Society for Advancement of Cytometry
Rational Design of a Novel Chiral Palladacycle: Synthesis, Optical Resolution, and Stereochemical Evaluation
Authors: Ding Yi , Li Yongxin , Pullarkat Sumod A. , Leng Yap See , Leung Pak‐Hing
Published: December 2008

A novel palladacycle was designed and prepared by direct cyclopalladation of the ligand 1‐(3,6‐dimethylnaphthalen‐1‐yl)‐N,N‐dimethylethanamine, which was synthesized by a multistep sequence starting from 2,7‐dimethylnaphthalene. The palladacycle structure and ring conformations of its triphenylphosphane derivative was investigated by X‐ray structural analysis in the solid state and 2D 1H–1H ROESY NMR spectroscopy in solution. The ortho‐palladated ring is stereochemically rigid, and the five‐membered ring conformation is locked by repulsive interactions between the methyl group on the prochiral carbon atom and its neighboring naphthylene proton. The racemic cyclopalladated complex could be efficiently resolved through the formation of its (S)‐prolinato derivatives, and efficient separation of the resulting diastereomeric complexes was achieved by simple silica‐gel chromatography. The structures and absolute configurations of the two optically resolved palladium complexes were determined by X‐ray crystallography. Both the (R,R)‐ and (S,S)‐di‐μ‐chloride dimeric palladium complexes could be obtained chemoselectively by treating the corresponding prolinato derivatives with 1 M hydrochloric acid. Despite the severe interchelate steric constraints within this new organopalladium complex, the bulky monodentate ligand 3,4‐dimethyl‐1‐phenylphosphole (dmpp) was coordinated regiospecifically to the ortho‐palladated amine unit trans to the NMe2 group. In comparison to its naphthylamine analogue, the new palladacycle showed a much higher stereoselectivity in the chiral template‐promoted asymmetric Diels–Alder reaction of coordinated dmpp and N,N‐dimethylacrylamide.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Chlorophyll Catabolites – Chemical and Structural Footprints of a Fascinating Biological Phenomenon
Authors: Moser Simone , Müller Thomas , Oberhuber Michael , Kräutler Bernhard
Published: December 2008

Twenty years ago, the molecular basis for the seasonal disappearance of chlorophyll was still enigmatic. In the meantime, our knowledge on chlorophyll breakdown has grown considerably. As outlined here, it has been possible to decipher the basic transformations involved in natural chlorophyll breakdown by identification of chlorophyll catabolites in higher plants, and with the help of the synthesis of (putative) catabolic intermediates. In vascular plants, chlorophyll breakdown typically converts the green plant pigments efficiently into colorless and non‐fluorescent tetrapyrroles. It involves colored intermediates only fleetingly and in an (elusive) enzyme‐bound form. The non‐fluorescent chlorophyll catabolites accumulate in the vacuoles of degreened leaves and are considered the products, primarily, of a detoxification process. However, they are effective antioxidants, and may thus also have physiologically beneficial chemical properties.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Synthesis of Optically Active Monoacid Side‐Chains of Cephalotaxus Alkaloids
Authors: Berhal Farouk , Tardy Sébastien , Pérard‐Viret Joëlle , Royer Jacques
Published: December 2008

The general preparation of enantiopure monoacid side‐chains of several esters of cephalotaxine is described. The strategy, similar to Weinreb's approach to the synthesis of deoxyharringtonine, used as key intermediate the chiral nonracemic epoxide 11a prepared from the commercially available monomethyl itaconate (8). The key step of the strategy was the ring‐opening of the epoxide 11a by using different organocuprate nucleophiles. Hydrogenolysis as the final step gave the monacid side‐chains of the corresponding esters of cephalotaxine in moderate to good overall yields from epoxide 11a.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Incremental dynamic analysis of woodframe buildings
Authors: Christovasilis Ioannis P. , Filiatrault Andre , Constantinou Michael C. , Wanitkorkul Assawin
Published: December 2008

The collapse of wood buildings was one of the main contributors to the heavy death toll and economic losses during the 1995 Hyogo‐ken Nanbu (Kobe) earthquake in Japan. In California, half of the property loss from the 1994 Northridge earthquake was attributed to wood construction. Based on damage observed in recent earthquakes, the seismic vulnerability of existing wood buildings under maximum credible seismic events is uncertain. The main objective of this study is to quantify the seismic collapse fragilities and collapse mechanisms of a two‐story townhouse and three‐story woodframe apartment building through numerical analyses. Three construction quality variants (poor, typical and superior) were considered for each building in order to assess the effects of construction qualities on seismic collapse fragilities. The buildings were also re‐designed according to the 2006 edition of the International Building Code to quantify the seismic fragilities of modern woodframe construction. The results obtained suggest that the construction quality, excitation direction and wall finish materials can influence significantly the collapse fragilities of woodframe buildings. Copyright © 2008 John Wiley & Sons, Ltd.
Hybrid mesh generation with embedded surfaces using a multiple marching direction approach
Authors: Ito Yasushi , Shih Alan M. , Soni Bharat K.
Published: December 2008

This paper proposes a method for the creation of hybrid meshes with embedded surfaces for viscous flow simulations as an extension of the multiple marching direction approach (AIAA J. 2007; 45(1):162–167). The multiple marching direction approach enables to place semi‐structured elements around singular points, where valid semi‐structured elements cannot be placed using conventional hybrid mesh generation methods. This feature is discussed first with a couple of examples. Elements sometimes need to be clustered inside a computational domain to obtain more accurate results. For example, solution features, such as shocks, vortex cores and wake regions, can be extracted during the process of adaptive mesh generation. These features can be represented as surface meshes embedded in a computational domain. Semi‐structured elements can be placed around the embedded surface meshes using the multiple marching direction approach with a pretreatment method. Tetrahedral elements can be placed easily instead. A couple of results are presented to demonstrate the capability of the mesh generation method. Copyright © 2008 John Wiley & Sons, Ltd.
Are household subjective forecasts of personal finances accurate and useful? A directional analysis of the British Household Panel Survey
Authors: Easaw Joshy , Heravi Saeed
Published: December 2008

The purpose of the paper is to analyse the accuracy and usefulness of household subjective forecasts of personal finance. We use non‐parametric directional analysis to assess the subjective forecasts which are based on qualitative judgments. Using the British Household Panel Survey (BHPS) we are able to analyse a large number of individuals over a number of years. We also take into account individual characteristics such as gender, age, education and employment status when assessing their subjective forecasts. The paper extends the existing literature in two ways: the accuracy and usefulness of subjective forecasts, based on directional analysis, are assessed at the household level for the first time. Secondly, we adapt and extend the methods of directional analysis, which are applied to the household panel or longitudinal survey. Copyright © 2008 John Wiley & Sons, Ltd.
Long‐term accumulation of microglia with proneurogenic phenotype concomitant with persistent neurogenesis in adult subventricular zone after stroke
Authors: Thored Pär , Heldmann Ursula , Gomes‐Leal Walace , Gisler Ramiro , Darsalia Vladimer , Taneera Jalal , Nygren Jens M. , Jacobsen Sten‐Eirik W. , Ekdahl Christine T. , Kokaia Zaal , Lindvall Olle
Published: December 2008

Neural stem cells (NSCs) in the adult rat subventricular zone (SVZ) generate new striatal neurons during several months after ischemic stroke. Whether the microglial response associated with ischemic injury extends into SVZ and influences neuroblast production is unknown. Here, we demonstrate increased numbers of activated microglia in ipsilateral SVZ concomitant with neuroblast migration into the striatum at 2, 6, and 16 weeks, with maximum at 6 weeks, following 2 h middle cerebral artery occlusion in rats. In the peri‐infarct striatum, numbers of activated microglia peaked already at 2 weeks and declined thereafter. Microglia in SVZ were resident or originated from bone marrow, with maximum proliferation during the first 2 weeks postinsult. In SVZ, microglia exhibited ramified or intermediate morphology, signifying a downregulated inflammatory profile, whereas amoeboid or round phagocytic microglia were frequent in the peri‐infarct striatum. Numbers of microglia expressing markers of antigen‐presenting cells (MHC‐II, CD86) increased in SVZ but very few lymphocytes were detected. Using quantitative PCR, strong short‐ and long‐term increase (at 1 and 6 weeks postinfarct) of insulin‐like growth factor‐1 (IGF‐1) gene expression was detected in SVZ tissue. Elevated numbers of IGF‐1‐expressing microglia were found in SVZ at 2, 6, and 16 weeks after stroke. At 16 weeks, 5% of microglia but no other cells in SVZ expressed the IGF‐1 protein, which mitigates apoptosis and promotes proliferation and differentiation of NSCs. The long‐term accumulation of microglia with proneurogenic phenotype in the SVZ implies a supportive role of these cells for the continuous neurogenesis after stroke. © 2008 Wiley‐Liss, Inc.
Microglial cell migration stimulated by ATP and C5a involve distinct molecular mechanisms: Quantification of migration by a novel near‐infrared method
Authors: Miller Aaron M. , Stella Nephi
Published: December 2008

Microglial cells, the macrophages of the brain, play an essential role in the propagation of neuroinflammation. Increased microglial cell migration in response to specific chemoattractants has been documented, but less is known about the differences between these stimuli and the signal transduction pathways that mediate their effects. Current methods to measure cell migration are often labor‐intensive and rely on the manual counting of cell number, so more efficient and objective methods are needed. Here we present an improved and higher‐throughput Boyden chamber technique that measures microglial cell migration by using DRAQ5, a nuclear dye that emits in the near‐infrared. Out of a panel of chemoattractants tested, we found that ATP and C5a potently stimulate the migration of mouse primary microglial cells. The stimulatory effects of ATP and C5a displayed significant additivity, suggesting that each chemoattractant stimulated migration through independent molecular mechanisms. Accordingly, we found key differences in these responses: ATP stimulated a combination of both chemokinesis and chemotaxis, and this response was mediated by the ROCK signaling pathway; whereas C5a stimulated only chemotaxis and this response was mediated by the Rac1 signaling pathway. Finally, we found that functional PI3‐kinase is only required for random basal microglial cell migration. Thus, our results show that distinct nonoverlapping signal transduction pathways control different modes of microglial cell migration and suggest that the targeting of these distinct molecular mechanisms should modulate different aspects of neuroinflammation propagation. © 2008 Wiley‐Liss, Inc.
Effects of progesterone on oligodendrocyte progenitors, oligodendrocyte transcription factors, and myelin proteins following spinal cord injury
Authors: Labombarda Florencia , González Susana L. , Lima Analia , Roig Paulina , Guennoun Rachida , Schumacher Michael , de Nicola Alejandro F.
Published: December 2008

Progesterone is emerging as a myelinizing factor for central nervous system injury. Successful remyelination requires proliferation and differentiation of oligodendrocyte precursor cells (OPC) into myelinating oligodendrocytes, but this process is incomplete following injury. To study progesterone actions on remyelination, we administered progesterone (16 mg/kg/day) to rats with complete spinal cord injury. Rats were euthanized 3 or 21 days after steroid treatment. Short progesterone treatment (a) increased the number of OPC without effect on the injury‐induced reduction of mature oligodendrocytes, (b) increased mRNA and protein expression for the myelin basic protein (MBP) without effects on proteolipid protein (PLP) or myelin oligodendrocyte glycoprotein (MOG), and (c) increased the mRNA for Olig2 and Nkx2.2 transcription factors involved in specification and differentiation of the oligodendrocyte lineage. Furthermore, long progesterone treatment (a) reduced OPC with a concomitant increase of oligodendrocytes; (b) promoted differentiation of cells that incorporated bromodeoxyuridine, early after injury, into mature oligodendrocytes; (c) increased mRNA and protein expression of PLP without effects on MBP or MOG; and (d) increased mRNA for the Olig1 transcription factor involved in myelin repair. These results suggest that early progesterone treatment enhanced the density of OPC and induced their differentiation into mature oligodendrocytes by increasing the expression of Olig2 and Nkx2.2. Twenty‐one days after injury, progesterone favors remyelination by increasing Olig1 (involved in repair of demyelinated lesions), PLP expression, and enhancing oligodendrocytes maturation. Thus, progesterone effects on oligodendrogenesis and myelin proteins may constitute fundamental steps for repairing traumatic injury inflicted to the spinal cord. © 2008 Wiley‐Liss, Inc.
Non‐antagonistic relationship between mitogenic factors and cAMP in adult Schwann cell re‐differentiation
Authors: Monje Paula V. , Rendon Sayuri , Athauda Gagani , Bates Margaret , Wood Patrick M. , Bunge Mary Bartlett
Published: December 2008

The expression of myelination‐associated genes (MGs) can be induced by cyclic adenosine monophosphate (cAMP) elevation in isolated Schwann cells (SCs). To further understand the effect of known SC mitogens in the regulation of SC differentiation, we studied the response of SCs isolated from adult nerves to combined cAMP, growth factors, including neuregulin, and serum. In adult SCs, the induction of MGs by cAMP coincided with the loss of genes expressed in non‐myelin‐forming SCs and with a change in cell morphology from a bipolar to an expanded epithelial‐like shape. Prolonged treatment with high doses of cAMP‐stimulating agents, as well as low cell density, was required for the induction of SC differentiation. Stimulation with serum, neuregulin alone, or other growth factors including PDGF, IGF and FGF, increased SC proliferation but did not induce the expression of MGs or the associated morphological change. Most importantly, when these factors were administered in combination with cAMP‐stimulating agents, SC proliferation was synergistically increased without reducing the differentiating activity of cAMP. Even though the initiation of DNA synthesis and the induction of differentiation were mostly incompatible events in individual cells, SCs were able to differentiate under conditions that also supported active proliferation. Overall, the results indicate that in the absence of neurons, cAMP can trigger SC re‐differentiation concurrently with, but independently of, growth factor signaling. © 2008 Wiley‐Liss, Inc.