bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis and Biological Evaluation of N‐Acylhydrazones as Inhibitors of MurC and MurD Ligases
Authors: Šink Roman , Kovač Andreja , Tomašić Tihomir , Rupnik Veronika , Boniface Audrey , Bostock Julieanne , Chopra Ian , Blanot Didier , Mašič Lucija Peterlin , Gobec Stanislav , Zega Anamarija
Published: July 2008

The Mur ligases have an essential role in the intracellular biosynthesis of bacterial peptidoglycan, and they represent attractive targets for the design of novel antibacterials. A series of compounds with an N‐acylhydrazone scaffold were synthesized and screened for inhibition of the MurC and MurD enzymes from Escherichia coli. Compounds with micromolar inhibitory activities against both MurC and MurD were identified, and some of them also showed antibacterial activity.
Integrating Structure‐ and Ligand‐Based Virtual Screening: Comparison of Individual, Parallel, and Fused Molecular Docking and Similarity Search Calculations on Multiple Targets
Authors: Tan Lu , Geppert Hanna , Sisay Mihiret T. , Gütschow Michael , Bajorath Jürgen
Published: July 2008

Similarity searching is often used to preselect compounds for docking, thereby decreasing the size of screening databases. However, integrated structure‐ and ligand‐based screening schemes are rare at present. Docking and similarity search calculations using 2D fingerprints were carried out in a comparative manner on nine target enzymes, for which significant numbers of diverse inhibitors could be obtained. In the absence of knowledge‐based docking constraints and target‐directed parameter optimisation, fingerprint searching displayed a clear preference over docking calculations. Alternative combinations of docking and similarity search results were investigated and found to further increase compound recall of individual methods in a number of instances. When the results of similarity searching and docking were combined, parallel selection of candidate compounds from individual rankings was generally superior to rank fusion. We suggest that complementary results from docking and similarity searching can be captured by integrated compound selection schemes.
A 0.6 V, 2.11 MHz, 62 dB SFDR active‐RC filter in 0.13µm CMOS process
Authors: Seok Ji‐Hwan , Jo Jun‐Gi , Yoo Changsik
Published: July 2008

Active‐RC biquad is proposed, which allows the DC level of the input of operational amplifier (op‐amp) to be different from that of the op‐amp output, enabling the low‐voltage operation. The proposed biquad realizes a second‐order transfer function with only one op‐amp, rendering even lower power consumption. By cascading two biquads, a 0.6 V fourth‐order filter is realized in a 0.13µm CMOS technology. While dissipating only 0.42 mW, the filter shows 2.11 MHz cut‐off frequency and 62 dB spurious‐free dynamic range. Copyright © 2008 John Wiley & Sons, Ltd.
Homocysteine enhances cardiac neural crest cell attachment in vitro by increasing intracellular calcium levels
Authors: Heidenreich David J. , Reedy Mark V. , Brauer Philip R.
Published: July 2008

Elevated homocysteine (Hcys) increases the risk of neurocristopathies. Previous studies show Hcys inhibits neural crest (NC) cell migration in vivo. However, the mechanisms responsible for this effect are unknown. Here, we evaluated the effect of Hcys on NC cell attachment in vitro and determined if any of the effects were due to altered Ca2+ signaling. We found Hcys enhanced NC cell attachment in a dose and substrate‐dependent manner. Ionomycin mimicked the effect of Hcys while BAPTA‐AM and 2‐APB blocked the effect of Hcys on NC attachment. In contrast, inhibitors of plasma membrane Ca2+ channels had no effect on NC attachment. Hcys also increased the emission of the intracellular Ca2+‐sensitive probe, Fluo‐4. These results show Hcys alters NC attachment by triggering an increase in intracellular Ca2+ possibly by generating inositol triphosphate. Hence, the teratogenic effect ascribed to Hcys may be due to perturbation of intracellular Ca2+ signaling. Developmental Dynamics 237:2117–2128, 2008. © 2008 Wiley‐Liss, Inc.
1,25‐dihydroxyvitamin D3 promotes IL‐10 production in human B cells
Authors: Heine Guido , Niesner Uwe , Chang Hyun‐Dong , Steinmeyer Andreas , Zügel Ulrich , Zuberbier Torsten , Radbruch Andreas , Worm Margitta
Published: July 2008

1,25‐dihydroxyvitamin D3 (calcitriol) regulates immune responses, e.g., inhibits expression of IgE by B cells and enhances expression of IL‐10 by dendritic cells and T cells. We report here that activation of human B cells by B cell receptor, CD40 and IL‐4 signals induces expression of the gene for 25‐hydroxyvitamin‐D3–1α‐hydroxylase (CYP1α). Accordingly, these B cells generate and secrete significant amounts of calcitriol. In activated B cells calcitriol induces expression of the genes Cyp24, encoding a vitamin D hydroxylase, and Trpv6, encoding a calcium selective channel protein. Calcitriol enhances IL‐10 expression of activated B cells more than threefold, both by recruiting the vitamin D receptor to the promoter of Il‐10, and to lesser extent by modulation of calcium‐dependent signaling. The molecular link in activated B cells between vitamin D signaling, expression of IgE and IL‐10, and their ability to produce calcitriol from its precursor, suggest that pro‐vitamin D (25‐hydroxyvitamin D3) can be used as a modulator of allergic immune responses.

Supporting Information for this article is available at http://www.wiley‐vch.de/contents/jc_2040/2008/38216_s.pdf
CD11c provides an effective immunotarget for the generation of both CD4 and CD8 T cell responses
Authors: Castro Fernanda V. V. , Tutt Alison L. , White Ann L. , Teeling Jessica L. , James Sonya , French Ruth R. , Glennie Martin J.
Published: July 2008

The magnitude and quality of T cell responses generated when Ag is targeted to receptors on DC is influenced by both the specific receptor targeted and its distribution among DC subsets. Here we examine the targeting of the model Ag OVA to potential DC targets, including CD11c, CD205, MHC class II, CD40, TLR2 and FcγRII/III, using a panel of (Fab' × OVA) conjugates. In vitro studies identified CD11c, CD205 and MHC class II as superior and comparably effective immunotargets for the delivery of OVA to APC for presentation to T cells. In vivo studies, however, showed a marked advantage of targeting Ag to CD11c for both CD4 (OT‐II) and CD8 (OT‐I) responses, with robust stimulation after a single, low dose (equivalent to 0.5 μg OVA); in contrast, (anti‐CD205 × OVA) and (anti‐MHC class II × OVA) resulted in markedly less proliferation of both OT‐I and OT‐II cells. Biodistribution and immunohistochemical studies suggest that the exceptional ability of CD11c to capture Ag in lymphoid tissues may, at least partially, explain its ability to promote T cell responses. These results suggest that targeting antigen via CD11c offers a previously unappreciated strategy for vaccine development which, unlike most targets, delivers robust responses of both CD4 and CD8 T cells.

See accompanying article: http://dx.doi.org/10.1002/eji.200838645

Supporting Information for this article is available at http://www.wiley‐vch.de/contents/jc_2040/2008/38302_s.pdf
CD11c: Not merely a murine DC marker, but also a useful vaccination target
Authors: Kurts Christian
Published: July 2008

The induction of robust CD4+ and CD8+ T cell responses is a central aim in antiviral and anticancer vaccination. To this end, dendritic cells (DC) need to capture the vaccine, process and present it on MHC class I and II molecules. The mechanisms by which DC acquire antigen predetermine the quantity and quality of the ensuing T cell activation. In this issue of the European Journal of Immunology, it is demonstrated that targeting antigen towards CD11c, an integrin expressed preferentially by murine DC, facilitates efficient CD4+ and CD8+ T cell activation. The underlying mechanisms involve efficient antigen delivery into the marginal zone and T/DC zone of the spleen. This commentary discusses these findings in the context of current knowledge on antigen presentation.

See accompanying article http://dx.doi.org/10.1002/eji.200838302
Alum adjuvanticity: Unraveling a century old mystery
Authors: De Gregorio Ennio , Tritto Elaine , Rappuoli Rino
Published: July 2008

The development of vaccine adjuvants for human use has been one of the slowest processes in the history of medicine. For almost one century, aluminium hydroxide (alum) has been the only vaccine adjuvant approved worldwide. Only in the past decade have two oil‐in‐water emulsions and one TLR agonist been approved by the European authorities as new vaccine adjuvants. Despite the fact that alum has been injected into billions of people, its mechanism of action is not fully understood. Recently, several reports have greatly increased our knowledge of the molecular and cellular events triggered by alum; however, the contribution of each of these processes to alum adjuvanticity is still unclear. A study published in this issue of the European Journal of Immunology, together with two recent publications, have demonstrated that the NOD‐like receptor, pyrin domain containing 3 (Nlrp3)‐inflammasome is the molecular target of alum immunostimulatory activity in vitro. Surprisingly, these three studies reported conflicting results on the requirement of the Nlrp3 inflammasome complex for alum adjuvant effects in vivo. This commentary attempts to resolve some of these discrepancies.

See accompanying article http://dx.doi.org/10.1002/eji.200838549
Insights into a Nontoxic and High‐Yielding Synthesis of Methyltrioxorhenium (MTO)
Authors: Mitterpleininger Josef K. M. , Szesni Normen , Sturm Stefanie , Fischer Richard W. , Kühn Fritz E.
Published: July 2008

The versatile catalyst methyltrioxorhenium(VII) (MTO) is now available in high yields by utilizing easily accessible nontoxic starting materials. The new synthetic pathway allows an inexpensive, large‐scale production of MTO, paving the way for industrial applications. A variety of starting materials is compared with respect to their applicability, availability and ease of handling. The reaction times and the byproducts formed are compared under different reaction conditions. It was seen that silver perrhenate in combination with methylzinc acetate, which was derived from trimethylaluminum and zinc acetate, are the best starting materials for a high‐yielding large‐scale synthesis of MTO. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Deaggregation of Trimethylsilylmethyllithium
Authors: Tatic Tanja , Ott Holger , Stalke Dietmar
Published: July 2008

Polyamine donor bases such as N,N,N′,N′‐tetramethylethylenediamine (TMEDA), (–)‐sparteine and N,N,N′,N″,N″‐pentamethyldiethylentriamine (PMDETA) have been employed to deaggregate parent trimethylsilylmethyllithium (1). The crystal structures of the dimers [(TMEDA)·LiCH2SiMe3]2 (2) and [{(–)‐sparteine}·LiCH2SiMe3]2 (3) were determined and reveal a four‐membered ring as the central structural motif. The two lithium atoms are each coordinated by the chelating ligands and the carbanions. The Li–C contacts show alternating bond lengths. The monomer [(PMDETA)·LiCH2SiMe3] (4) has the shortest Li–C contact of the aggregates discussed, and the Me3Si group points toward the central nitrogen atom of the PMDETA ligand, even if this alignment seems unfavourable for steric reasons. The bond lengths and angles of compounds 2–4 are discussed in relation to the orientation of the lone pair at the carbanion. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
The Interaction of Iodide Film with Platinum Microparticles on Different Electrode Materials for Various Electrocatalytic Reactions
Authors: Su Buo‐Wei , Thiagarajan Soundappan , Chen Shen‐Ming
Published: July 2008

The electrochemical depositions of Pt microparticles and KI film were successfully carried out on glassy carbon electrodes (GCE), gold electrodes (GE), and indium tin oxide electrodes (ITO). The electrochemical studies of Pt micro/KI film on GCE show that the film was stable, active at pH 1.5 electrolyte solutions. The Pt microparticle/KI film modified ITO electrodes were examined by using scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques. The SEM and AFM results show that the Pt particle sizes were in the range of 120 nm–1.4 μm, respectively. The proposed film on GCE shows efficient electrocatalysis for oxygen, Cr2O$\rm{ {_{7}^{2-}}}$ reduction by using cyclic voltammetry. Further the electrochemical oxidation of sodium meta‐arsenite (As(III)), H2O2 were successfully carried out and the detection of H2O2 in real samples has been validated.
Performance characterization of an insulator‐based dielectrophoretic microdevice
Authors: Ozuna‐Chacón Sandra , Lapizco‐Encinas Blanca H. , Rito‐Palomares Marco , Martínez‐Chapa Sergio O. , Reyes‐Betanzo Claudia
Published: July 2008

Dielectrophoresis (DEP), the motion of particles in nonuniform electric fields, is a nondestructive electrokinetic (EK) transport mechanism can be used to concentrate and separate bioparticles. Traditionally, DEP has been performed employing microelectrodes, an approach that is expensive due to the cost of microelectrode fabrication. An alternative is insulator‐based DEP (iDEP), an inexpensive method where nonuniform electric fields are created with arrays of insulating structures. This study presents the effects of operating conditions on the dielectrophoretic behavior of polystyrene microparticles under iDEP. Experiments were performed employing microchannels containing insulating structures that worked as insulators. The parameters varied were pH (8–9) and conductivity (25–100 μS/cm) of the bulk medium, and the magnitude of the applied field (200–850 V/cm). Optimal operating conditions in terms of pH and conductivity were obtained, and the microdevice performance was characterized in terms of concentration factor and minimum electric field required (minimum energy consumption). This is the first report on improving iDEP processes when EOF is present. DEP and EOF have been studied extensively, however, this study integrates the effect of suspending medium characteristics on both EK phenomena. These findings will allow improving the performance of iDEP microdevices achieving the highest concentration fold with the lowest energy consumption.
Capillary electrophoresis immunoassay using magnetic beads
Authors: Chen Hong‐Xu , Busnel Jean‐Marc , Gassner Anne‐Laure , Peltre Gabriel , Zhang Xin‐Xiang , Girault Hubert H.
Published: July 2008

Protein A‐coated magnetic beads (0.3 μm) have been trapped in a small portion of a neutrally coated capillary (50 μm id). Anti‐β‐lactoglobulin (β‐LG) antibodies have then been immobilized on the beads through strong affinity with protein A to subsequently capture β‐LG from model or real samples. Once the immunocomplexes formed at physiological pH, a discontinuous buffer system has been used to release the partners and preconcentrate them by transient ITP. The antigens and antibodies have finally been separated by CZE and detected by UV absorbance. An LOQ of 55 nM has been achieved. This methodology has been applied to quantify native β‐LG in pasteurized and ultra‐high‐temperature‐treated bovine milk. All the described procedures, including immunosorbent preparation, sample extraction, cleanup, preconcentration, and separation are completely automated on a commercial CE instrument. As this CE immunoassay method is simple, rapid, selective, and sensitive, it should be a practical and attractive technology for the analysis of complicated biological samples.
Current awareness in geriatric psychiatry
Authors:   John Wiley & Sons, Ltd.
Published: July 2008

In order to keep subscribers up‐to‐date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of geriatric psychiatry. Each bibliography is divided into 9 sections: 1 Reviews; 2 General; 3 Assessment; 4 Epidemiology; 5 Therapy; 6 Care; 7 Dementia; 8 Depression; 9 Psychology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted
Switch from type II to I Fas/CD95 death signaling on in vitro culturing of primary hepatocytes
Authors: Walter Dorothée , Schmich Kathrin , Vogel Sandra , Pick Robert , Kaufmann Thomas , Hochmuth Florian Christoph , Haber Angelika , Neubert Karin , McNelly Sabine , von Weizsäcker Fritz , Merfort Irmgard , Maurer Ulrich , Strasser Andreas , Borner Christoph
Published: July 2008

Fas/CD95-induced apoptosis of hepatocytes

in vivo

proceeds through the so-called type II pathway, requiring the proapoptotic BH3-only Bcl-2 family member Bid for mitochondrial death signaling. Consequently, Bid-deficient mice are protected from anti-Fas antibody injection induced fatal hepatitis. We report the unexpected finding that freshly isolated mouse hepatocytes, cultured on collagen or Matrigel, become independent of Bid for Fas-induced apoptosis, thereby switching death signaling from type II to type I. In such

in vitro

cultures, Fas ligand (FasL) activates caspase-3 without Bid cleavage, Bax/Bak activation or cytochrome c release, and neither Bid ablation nor Bcl-2 overexpression is protective. The type II to type I switch depends on extracellular matrix adhesion, as primary hepatocytes in suspension die in a Bid-dependent manner. Moreover, the switch is specific for FasL-induced apoptosis as collagen-plated Bid-deficient hepatocytes are protected from tumor necrosis factor alpha/actinomycin D (TNFα/ActD)-induced apoptosis.

Conclusion:

Our data suggest a selective crosstalk between extracellular matrix and Fas-mediated signaling that favors mitochondria-independent type I apoptosis induction. (Hepatology 2008;48:1942-1953.)
Modulation of neurotoxicant‐induced increases in intracellular calcium by phytoestrogens differ for amyloid beta peptide (Aβ) and 1‐methyl‐4‐phenyl‐pyridine (MPP+)
Authors: Sørensen Izel Fourie , Purup Stig , Ehrich Marion
Published: July 2008

Neurotoxicant‐induced elevation of intracellular calcium (Ca2+) and modulation by phystoestrogens were examinedin vitrousing human neuroblastoma SH‐SY5Y cells cultured with amyloid beta‐peptide (Aβ) and 1‐methyl‐4‐phenyl‐pyridine (MPP+). Although Aβitself did not increase Ca2+, it exacerbated the effects of carbachol. The elevation of Ca2+caused by the agents in combination could be reduced by pretreatment with the phytoestrogens equol and genistein, as well as by the L‐type Ca2+channel blocker nifedipine. MPP+ exposure also elevated Ca2+, an effect blocked by nifedipine but not by the phytoestrogens. As opposed to phytoestrogens, nifedipine was also able to significantly reduce cell death caused by higher concentrations of MPP+in the LDH viability assay. The results suggest that phytoestrogens are unlikely to serve as general cellular protectants for neurotoxicants with different mechanisms of action. The concentrations of Aβand MPP+affecting Ca2+release did not inhibit cell viability as measured with the LDH release assay. This indicates that mechanisms involved with toxicity can be studied at doses that are not lethal. Copyright © 2008 John Wiley & Sons, Ltd.
Optical investigations of the RNA polymerase molecular motor
Authors: Strick Terence R.
Published: July 2008

Molecular motors have been extensively studied over the past 25 years using a range of optical microscopy methods. The specific example of RNA polymerase, responsible for transcribing the information encoded in DNA into RNA, illustrates the strength of the convergence between these recent approaches and the vast store of data collected over the past half century using the more “classical” approaches of genetics, biochemistry, and structure elucidation. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Surface functionalization of bioactive glasses
Authors: Verné E. , Vitale‐Brovarone C. , Bui E. , Bianchi C. L. , Boccaccini A. R.
Published: July 2008

Different cleaning and silanization methods have been applied to bioactive glasses with the aim of covalently bonding bone morphogenetic proteins (BMP‐2) to the surface. Several glasses, with different bioactivity index, were cleaned with acidic, basic, or neutral aqueous media to investigate the role of pH in the formation of silanols on glass surfaces of different reactivity. The cleaned glasses were then functionalized using 3‐aminopropyl‐triethoxysilane (APTS). After the optimization of the silanization procedure, proteins of different complexity were immobilized on the functionalized glasses. To optimize the protein immobilization, a model protein (carnosine) was first used, and the procedure was then used to bind human BMP‐2. The glass surfaces were characterized during each step of the treatment by water contact angles and X‐ray photoelectron analyses. The APTS functionalization was then used to immobilize bone morphogenetic protein on the bioactive glasses. This result suggested that such a treatment could be successfully used as an efficient alternative to systemic administration of transforming growth factors for the development of local delivery vehicle implants. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009
Influence of calcium phosphate crystal morphology on the adhesion, spreading, and growth of bone derived cells
Authors: Liu Y. , Shelton R. , Gbureck U. , Barralet J.
Published: July 2008

In this study, rat periosteal cells and bone marrow stromal cells (BMSC) were examined when cultured on octacalcium phosphate (OCP) crystals to determine the effects of crystal morphology on their adhesion, spreading, and growth. Three type of OCP crystals with different morphology were prepared by different precipitation methods. OCPI was prepared by homogeneous precipitation; while OCPII and III were prepared by heterogeneous precipitation dripping a calcium ion containing solution into a phosphate ion solution or vice versa, respectively. The effects of crystal morphology on the attachment and spreading of cultured cells on OCP crystals were observed using SEM. The effects of crystal morphology on the growth of periosteal cell or BMSC were evaluated using the MTT assay, while OCP crystal cytotoxicity was determined by measuring lactic dehydrogenase (LDH) released from 2.12 × 105 cells incubated with 60 mg of the different OCPs. BMSCs and periosteal cells attached to or between the crystals of OCPI and the cell number increased over the 17 days of culture. In contrast, cell numbers could not be measured for either periosteal cells or BMSC on OCPII and III. BMSC incubated with OCPII and III showed a higher rate of LDH release, compared with the cells incubated with OCPI. This study demonstrated that the morphology of OCP crystals influenced the attachment, spreading and growth of periosteal cells and BMSC. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009
Designing tailored biomaterial surfaces to direct keratinocyte morphology, attachment, and differentiation
Authors: Bush K. A. , Driscoll P. F. , Soto E. R. , Lambert C. R. , McGimpsey W. G. , Pins G. D.
Published: July 2008

Precisely engineering the surface chemistry of biomaterials to modulate the adsorption and functionality of biochemical signaling molecules that direct cellular functions is critical in the development of tissue engineered scaffolds. Specifically, this study describes the use of functionalized self‐assembled monolayers (SAMs) as a model system to assess the effects of biomaterial surface properties on controlling fibronectin (FN) conformation and concentration as well as keratinocyte function. By systematically analyzing FN adsorption at low and saturated surface densities, we distinguished between SAM‐dependent effects of FN concentration and conformation on presenting cellular binding domains that direct cellular functions. Quantitative image analyses of immunostained samples showed that modulating the availability of the FN synergy site directly correlated with changes in keratinocyte attachment, spreading, and differentiation, through integrin‐mediated signaling mechanisms. The results of this study will be used to elucidate design features that can be incorporated into dermal equivalents and percutaneous implants to enhance the rate of re‐epithelialization and tissue regeneration. Furthermore, these findings indicate that SAM‐based model systems are a valuable tool for designing and investigating the development of scaffolds that regulate the conformation of extracellular matrix cues and cellular functions that accelerate the rate of tissue regeneration. © 2008 Wiley Periodicals, Inc. J Biomed Mater Res, 2009
Characterization of the 5′‐flanking region and regulation of expression of human anion exchanger SLC26A6
Authors: Saksena Seema , Dwivedi Alka , Singla Amika , Gill Ravinder K. , Tyagi Sangeeta , Borthakur Alip , Alrefai Waddah A. , Ramaswamy Krishnamurthy , Dudeja Pradeep K.
Published: July 2008

SLC26A6 (putative anion transporter 1, PAT1) has been shown to play an important role in mediating the luminal Cl−/OH−(HCO) exchange process in the intestine. Very little is known about the molecular mechanisms involved in the transcriptional regulation of intestinal SLC26A6 gene expression in the intestine. Current studies were, therefore, designed to clone and characterize the 5′‐regulatory region of the human SLC26A6 gene and determine the mechanisms involved in its regulation. A 1,120 bp (p−964/+156) SLC26A6 promoter fragment cloned upstream to the luciferase reporter gene in pGL2‐basic exhibited high promoter activity when transfected in Caco2 cells. Progressive deletions of the 5′‐flanking region demonstrated that −214/−44 region of the promoter harborscis‐acting elements important for maximal SLC26A6 promoter activity. Since, diarrhea associated with inflammatory bowel diseases is attributed to increased secretion of pro‐inflammatory cytokines, we examined the effects of IFNγ (30 ng/ml, 24 h) on SLC26A6 function, expression and promoter activity. IFNγ decreased both SLC26A6 mRNA and function and repressed SLC26A6 promoter activity. Deletion analysis indicated that IFNγ response element is located between −414/−214 region and sequence analysis of this region revealed the presence of potential Interferon Stimulated Responsive Element (ISRE), a binding site (−318/−300 bp) for interferon regulatory factor‐1 transcription factor (IRF‐1). Mutations in the potential ISRE site abrogated the inhibitory effects of IFNγ. These studies provided novel evidence for the involvement of IRF‐1 in the regulation of SLC26A6 gene expression by IFNγ in the human intestine. J. Cell. Biochem. 105: 454–466, 2008. © 2008 Wiley‐Liss, Inc.
Inflammation as the primary aetiological agent of human prostate cancer: A stem cell connection?
Authors: Maitland Norman J. , Collins Anne T.
Published: July 2008

Inflammation has been implicated for some time as a potential aetiological agent in human prostate cancer. Viral and bacterial infections or even chemical carcinogens such as those found in cooked meat have been proposed as the inflammatory stimuli, but the mechanism of cancer induction is unknown. Recent information about gene expression patterns in normal and malignant epithelial stem cells from human prostate provides a new hypothesis for inflammation‐induced carcinogenesis. The hypothesis states that in the stem cells located in the basal cell compartment of the prostate, activated prostate epithelial stem cells acquire a survival advantage, by expressing one of more of the same cytokines such as IL6. The establishment of one or more autocrine signalling loops results in an expansion of these cells in the absence of inflammation, as a potential first stage in the development of the tumour. J. Cell. Biochem. 105: 931–939, 2008. © 2008 Wiley‐Liss, Inc.
Intricate gene regulatory networks of helix‐loop‐helix (HLH) proteins support regulation of bone‐tissue related genes during osteoblast differentiation
Authors: Zhang Ying , Hassan Mohammad Q. , Li Zhao‐Yong , Stein Janet L. , Lian Jane B. , van Wijnen Andre J. , Stein Gary S.
Published: July 2008

Helix‐loop‐helix (HLH) transcription factors are key regulators of neurogenesis, myogenesis and osteogenesis. Here the relative contributions of multiple classes of HLH factors to the expression of bone related genes during osteoblast maturation were compared. We examined the expression of a panel of HLH proteins (e.g., Twist1/2, USF1/2, c‐Myc, Id1 ∼ 4, E12/47, Stra13) and one Zn finger protein (Snail which recognizes a subset of E‐boxes), during osteoblast differentiation and their functional contributions to bone phenotypic gene regulation. While expression of Twist1, Stra13, E12/47 and Snail transcripts remains relatively constant, expression of Twist2 as well as the inhibitory factors Id1, Id2, Id3, and Id4 decreases and USF1 is up‐regulated during osteoblastic differentiation of MC3T3 cells. Forced expression of selected HLH transcription factors shows that Myc, Snail and USF factors increase expression of the bone markers osteocalcin (OC) and/or alkaline phosphatase (AP), while E12/47, Twist and Id factors decrease their expression. None of these factors affect Runx2 gene expression. Interestingly, Snail enhances expression of osteoblast markers, while Twist1 and Twist2 factors are cross‐regulated and inhibit bone specific gene expression and other HLH proteins (e.g., Id) indirectly. Thus, our data suggest that the integrated activities of negative and positive E‐box related regulatory factors control osteoblast differentiation. J. Cell. Biochem. 105: 487–496, 2008. © 2008 Wiley‐Liss, Inc.
Quercetin suppresses hypoxia‐induced accumulation of hypoxia‐inducible factor‐1α (HIF‐1α) through inhibiting protein synthesis
Authors: Lee Dae‐Hee , Lee Yong J.
Published: July 2008

Quercetin, a ubiquitous bioactive plant flavonoid, has been shown to inhibit the proliferation of cancer cells and induce the accumulation of hypoxia‐inducible factor‐1α (HIF‐1α) in normoxia. In this study, under hypoxic conditions (1% O2), we examined the effect of quercetin on the intracellular level of HIF‐1α and extracellular level of vascular endothelial growth factor (VEGF) in a variety of human cancer cell lines. Surprisingly, we observed that quercetin suppressed the HIF‐1α accumulation during hypoxia in human prostate cancer LNCaP, colon cancer CX‐1, and breast cancer SkBr3 cells. Quercetin treatment also significantly reduced hypoxia‐induced secretion of VEGF. Suppression of HIF‐1α accumulation during treatment with quercetin in hypoxia was not prevented by treatment with 26S proteasome inhibitor MG132 or PI3K inhibitor LY294002. Interestingly, hypoxia (1% O2) in the presence of 100 µM quercetin inhibited protein synthesis by 94% during incubation for 8 h. Significant quercetin concentration‐dependent inhibition of protein synthesis and suppression of HIF‐1α accumulation were observed under hypoxic conditions. Treatment with 100 µM cycloheximide, a protein synthesis inhibitor, replicated the effect of quercetin by inhibiting HIF‐1α accumulation during hypoxia. These results suggest that suppression of HIF‐1α accumulation during treatment with quercetin under hypoxic conditions is due to inhibition of protein synthesis. J. Cell. Biochem. 105: 546–553, 2008. © 2008 Wiley‐Liss, Inc.
EVI1 recruits the histone methyltransferase SUV39H1 for transcription repression
Authors: Cattaneo Francesca , Nucifora Giuseppina
Published: July 2008

EVI1 is an oncoprotein inappropriately expressed in acute myeloid leukemia and myelodysplastic syndrome cells. In vitro studies indicate that diverse biological properties can be attributed to this protein. Its role in leukemogenesis is still unclear but it is thought that overall EVI1 can act mostly as a transcription repressor through its interaction with a subset of histone deacetylases. Studies with histone deacetylase inhibitors have however indicated that EVI1‐mediated repression can be only partially rescued by deacetylase inhibitor drugs, suggesting that additional chromosomal modifications might occur to induce gene repression by EVI1. To investigate whether histone methylation contributes to the repressive potential of EVI1, we examined a potential association between EVI1, the histone methyltransferase (HMT) SUV39H1, and methyltransferase activity in vitro. We find that EVI1 directly interacts with SUV39H1 and that the proteins form an active complex with methyltransferase activity in vitro. Our data indicate that SUV39H1 enhances the transcription repressive potential of EVI1 in vivo. We suggest that EVI1 affects promoters' activity in two different pathways, by association with histone deacetylases and by recruiting chromatin‐modifying enzymes to impose a heterochromatin‐like structure establishing a lasting transcription repression. J. Cell. Biochem. 105: 344–352, 2008. © 2008 Wiley‐Liss, Inc.
Does clofazimine (B663) reach Mycobacterium leprae persisting in Schwann cells and endothelial cells of endoneurial blood vessels in peripheral nerves?
Authors: Kumar V.
Published: July 2008

Peripheral nerve biopsies from 10 Lepromatous leprosy (LL) patients who were on multidrug treatment (MDT) were investigated by light and electron microscopy. Clofazimine (CLF) has been included as an essential component of MDT, which is the standard WHO regimen for treatment of leprosy. The patients receiving continuous MDT for a long period had viable bacilli in Schwann cells (SCs) of peripheral nerves whereas they had disappeared from the skin. Our ultrastructural observations clearly indicated the presence of CLF crystals in SCs. The crystals were in the form of osmiophilic rods of various shapes and sizes. On the other hand, the blood nerve barrier was clearly noticed in endoneurial blood vessels (EBV), and the barrier seems to play an important role for penetration of antileprosy drugs especially CLF. Microsc. Res. Tech., 2008 © 2008 Wiley‐Liss, Inc.
David Shakow and schizophrenia research at Worcester State Hospital: The roots of the scientist‐practitioner model
Authors: Cautin Robin L.
Published: July 2008

As Chief Psychologist and Director of Psychological Research at Worcester State Hospital (WSH), David Shakow (1901–1981) made substantial contributions to the scientific study of schizophrenia and by extension to the study of psychopathology in general. His methodological innovations—particularly on issues of diagnosis and conditions of testing—set a new standard for experimental rigor in the field. Shakow helped to establish many of what are considered basic facts about schizophrenia. His empirical work at WSH—specifically on the crossover effect—provided the scientific foundation for his theory of schizophrenic cognition, known as segmental set. Moreover, Shakow's schizophrenia work informed his developing ideas on the synergy between clinical practice and research. © 2008 Wiley Periodicals, Inc.
Science and culture around the Montessori's first “Children's Houses” in Rome (1907–1915)
Authors: Foschi Renato
Published: July 2008

Between 1907 and 1908, Maria Montessori's (1870–1952) educational method was elaborated at the Children's Houses of the San Lorenzo district in Rome. This pioneering experience was the basis for the international fame that came to Montessori after the publication of her 1909 volume dedicated to her “Method.” The “Montessori Method” was considered by some to be scientific, liberal, and revolutionary. The present article focuses upon the complex contexts of the method's elaboration. It shows how the Children's Houses developed in relation to a particular scientific and cultural eclecticism. It describes the factors that both favored and hindered the method's elaboration, by paying attention to the complex network of social, institutional, and scientific relationships revolving around the figure of Maria Montessori. A number of “contradictory” dimensions of Montessori's experience are also examined with a view to helping to revise her myth and offering the image of a scholar who was a real early‐twentieth‐century prototype of a “multiple” behavioral scientist. © 2008 Wiley Periodicals, Inc.
Body mass estimation in xenarthra: A predictive equation suitable for all quadrupedal terrestrial placentals?
Authors: De Esteban‐Trivigno Soledad , Mendoza Manuel , De Renzi Miquel
Published: July 2008

The Magnorder Xenarthra includes strange extinct groups, like glyptodonts, similar to large armadillos, and ground sloths, terrestrial relatives of the extant tree sloths. They have created considerable paleobiological interest in the last decades; however, the ecology of most of these species is still controversial or unknown. The body mass estimation of extinct species has great importance for paleobiological reconstructions. The commonest way to estimate body mass from fossils is through linear regression. However, if the studied species does not have similar extant relatives, the allometric pattern described by the regression could differ from those shown by the extinct group. That is the case for glyptodonts and ground sloths. Thus, stepwise multiple regression were developed including extant xenarthrans (their taxonomic relatives) and ungulates (their size and ecological relatives). Cases were weighted to maximize the taxonomic evenness. Twenty‐eight equations were obtained. The distribution of the percent of prediction error (%PE) was analyzed between taxonomic groups (Perissodactyla, Artiodactyla, and Xenarthra) and size groups (0–20 kg, 20–300 kg, and more than 300 kg). To assess the predictive power of the functions, equations were applied to species not included in the regression development [test set cross validation, (TSCV)]. Only five equations had a homogeneous %PE between the aforementioned groups. These were applied to five extinct species. A mean body mass of 80 kg was estimated for Propalaehoplophorus australis (Cingulata: Glyptodontidae), 594 kg for Scelidotherium leptocephalum (Phyllophaga: Mylodontidae), and 3,550.7 kg for Lestodon armatus (Phyllophaga: Mylodontidae). The high scatter of the body mass estimations obtained for Catonyx tarijensis (Phyllophaga: Mylodontidae) and Thalassocnus natans (Phyllophaga: Megatheriidae), probably due to different specializations, prevented us from predicting its body mass. Surprisingly, although obtained from ungulates and xenarthrans, these five selected equations were also able to predict the body mass of species from groups as different as rodents, carnivores, hyracoideans, or tubulidentates. This result suggests the presence of a complex common allometric pattern for all quadrupedal placentals. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.
Phage display screening in low dielectric media
Authors: Olofsson Linus , Söderberg Pernilla , Ankarloo Jonas , Nicholls Ian A.
Published: July 2008

Here we report the first application of phage display screening in low dielectric media. Two series of phage clones with affinity for α‐chymotrypsin (CT) were selected from a Ph.D.TM‐C7C library, using either a buffer or acetonitrile in buffer (50%, v/v). The affinity of lysates, individual clones or selected cyclic peptides for the enzyme was studied by examining their influence on CT activity. Peptides displayed on phage selected in buffer provided significant protection from enzyme autolysis resulting in marked increase in CT activity (>100%). Phage selected in ACN provided some, albeit weak, protection from the detrimental influence on CT from ACN. In conclusion, the results demonstrate the potential for the application of phage display screening protocols to targets in media of low dielectricity. Copyright © 2008 John Wiley & Sons, Ltd.
Structural and dynamic roles of permanent water molecules in ligand molecular recognition by chicken liver bile acid binding protein
Authors: Ricchiuto Piero , Rocco Alessandro Guerini , Gianazza Elisabetta , Corrada Dario , Beringhelli Tiziana , Eberini Ivano
Published: July 2008

Chicken liver bile acid binding protein (cL‐BABP) crystallizes with water molecules in its binding site. To obtain insights on the role of internal water, we performed two 100 ns molecular dynamics (MD) simulations in explicit solvent for cL‐BABP, as apo form and as a complex with two molecules of cholic acid, and analyzed in detail the dynamics properties of all water molecules. The diffusion coefficients of the more persistent internal water molecules are significantly different from the bulk, but similar between the two protein forms. A different number of molecules and a different organization are observed for apo‐ and holo‐cL‐BABP. Most water molecules identified in the binding site of the apo‐crystal diffuse to the bulk during the simulation. In contrast, almost all the internal waters of the holo‐crystal maintain the same interactions with internal sidechains and ligands, which suggests they have a relevant role in protein‐ligand molecular recognition. Only in the presence of these water molecules we were able to reproduce, by a classical molecular docking approach, the structure of the complex cL‐BABP::cholic acid with a low ligand root mean square deviation (RMSD) with respect to its reference positioning. Literature data reported a conserved pattern of hydrogen bonds between a single water molecule and three amino acid residues of the binding site in a series of crystallized FABP. In cL‐BABP, the interactions between this conserved water molecule and the three residues are present in the crystal of both apo‐ and holo‐cL‐BABP but are lost immediately after the start of molecular dynamics. Copyright © 2008 John Wiley & Sons, Ltd.
Pain relief by gabapentin and pregabalin via supraspinal mechanisms after peripheral nerve injury
Authors: Tanabe Mitsuo , Takasu Keiko , Takeuchi Yuichi , Ono Hideki
Published: July 2008

The antihypersensitivity actions of gabapentin and pregabalin have been well characterized in a large number of studies, although the underlying mechanisms have yet to be defined. We have been focusing on the supraspinal structure as a possible site for their action and have demonstrated that intracerebroventricular (i.c.v.) administration of gabapentin and pregabalin indeed decreases thermal and mechanical hypersensitivity in a murine chronic pain model involving partial ligation of the sciatic nerve. This novel supraspinally mediated analgesic effect was markedly suppressed by either depletion of central noradrenaline (NA) or blockade of spinal α2‐adrenergic receptors. Moreover, i.c.v. injection of gabapentin and pregabalin increased spinal NA turnover in mice only after peripheral nerve injury. In locus coeruleus (LC) neurons in brainstem slices prepared from mice after peripheral nerve injury, gabapentin reduced the γ‐aminobutyric acid (GABA) type A receptor‐mediated inhibitory postsynaptic currents (IPSCs). Glutamate‐mediated excitatory synaptic transmission was hardly affected. Moreover, gabapentin did not reduce IPSCs in slices taken from mice given a sham operation. Although gabapentin altered neither the amplitude nor the frequency of miniature IPSCs, it reduced IPSCs together with an increase in the paired‐pulse ratio, suggesting that gabapentin acts on the presynaptic GABAergic nerve terminals in the LC. Together, the data suggest that gabapentin presynaptically reduces GABAergic synaptic transmission, thereby removing the inhibitory influence on LC neurons only in neuropathic pain states, leading to activation of the descending noradrenergic system. © 2008 Wiley‐Liss, Inc.
In vivo effects of phosphodiesterase inhibition on basal cyclic guanosine monophosphate levels in the prefrontal cortex, hippocampus and cerebellum of freely moving rats
Authors: Marte Antonella , Pepicelli Olimpia , Cavallero Anna , Raiteri Maurizio , Fedele Ernesto
Published: July 2008

We have characterized the various phosphodiesterases (PDE) that degrade cyclic GMP in the prefrontal cortex, hippocampus, and cerebellum using the microdialysis technique to measure in vivo extracellular cyclic GMP in awake rats. The following PDE blockers were used (100 and 1,000 μM): 8‐methoxymethyl‐IBMX (8‐MM‐IBMX), erythro‐9‐(2‐hydroxy‐3‐nonyl)adenine (EHNA), milrinone, rolipram, and zaprinast. For solubility reasons, sildenafil was tested only at 100 μM. All drugs were administered locally in the brain regions through the dialysis probe. At 100 μM, 8‐MM‐IBMX enhanced the cyclic nucleotide extracellular levels in the prefrontal cortex and hippocampus but not in the cerebellum; EHNA and milrinone were active only in the hippocampus; rolipram was devoid of any effect; zaprinast and sildenafil were effective in all three brain areas. At 1 mM, 8‐MM‐IBMX, milrinone, and zaprinast increased extracellular cyclic GMP in all the brain regions examined, EHNA became active also in the prefrontal cortex and rolipram showed a significant effect only in the cerebellum. This is the first in vivo functional study showing that, in cortex, PDE1, ‐2, and ‐5/9 degrade cGMP, with PDE9 probably playing a major role; in hippocampus, PDE5/9 and PDE1 are mainly involved and seem almost equally active, but PDE2 and ‐3 also contribute; in cerebellum, PDE5/9 are the main cGMP hydrolyzing enzymes, but also PDE1 and ‐4 significantly operate. © 2008 Wiley‐Liss, Inc.
Up‐regulation of brain‐derived neurotrophic factor by application of fibroblast growth factor‐2 to the cut optic nerve is important for long‐term survival of retinal ganglion cells
Authors: Blanco Rosa E. , Soto Ileana , Duprey‐Díaz Mildred , Blagburn Jonathan M.
Published: July 2008

Application of basic fibroblast growth factor (FGF‐2) to the optic nerve after axotomy promotes the survival of retinal ganglion cells (RGCs) in the frog Rana pipiens and results in a rapid up‐regulation of brain‐derived neurotrophic factor (BDNF) and TrkB synthesis by the RGCs. Here we investigate whether this up‐regulation is maintained over the long term and whether it is required for FGF‐2's survival effect. At 6 weeks after axotomy and FGF‐2 treatment, we found more RGCs immunopositive for BDNF protein and higher intensity of BDNF and TrkB immunostaining, accompanied by increases in BDNF and TrkB mRNA in RGCs. Application of fluorescently labeled siRNA targeted against BDNF to the cut RGC axons showed that it was transported to the cell bodies. Axonal siRNA treatment eliminated the increases in BDNF immunostaining and mRNA that were induced by FGF‐2 and had no effect on TrkB mRNA. This reduction in BDNF synthesis by siRNA greatly reduced the long‐term survival effect of FGF‐2 on RGCs. This, taken together with previous results, suggests that, although FGF‐2 may initially activate survival pathways via ERK signaling, its main long‐term survival effects are mediated via its up‐regulation of BDNF synthesis by the RGCs. © 2008 Wiley‐Liss, Inc.