bluuebunny/crossref_metadata_2025 huggingface dataset:
The diagnostic value of endobronchial ultrasound‐guided needle biopsy in lung cancer and mediastinal adenopathy
Authors: Sun Wei , Song Kunchang , Zervos Michael , Pass Harvey , Cangiarella Joan , Bizekis Costas , Crawford Bernard , Wang Beverly Y.
Published: November 2009

Endobronchial ultrasonography (EBUS) has emerged as a new diagnostic tool that allows the bronchoscopist to see beyond the airway, including pulmonary and mediastinal lesion. The real time EBUS‐guided transbronchial needle aspiration (TBNA) has advanced the diagnostic yield in primary lung pathology and mediastinal lymph node staging of lung carcinoma. Sixty‐four patients (36 males, 28 females, ages ranging from 16 to 86 years) with peribronchial lung lesions and mediastinal and/or hilar lymph node lesions underwent EBUS‐TBNA. All patients had intraoperative cytological assessment by smears on aspiration samples or touch preparation on needle core biopsies.

The cytological final diagnoses were categorized as negative, suspicious/positive, and non‐diagnostic. Forty‐nine samples were obtained from lymph node lesions and 15 samples were obtained from lung lesions. In cytology specimens, 32 patients had suspicious/positive diagnoses and 32 patients had negative diagnosis. In follow‐up histology specimens, 35 patients had malignant diagnoses, including 18 adenocarcinomas, 8 small cell carcinomas, 6 squamous cell carcinomas, 1 metastatic hepatocellular carcinoma, 1 metastatic melanoma, and 1 lymphoma. Twenty‐nine patients had negative diagnoses. Sensitivity and specificity were 88.9% and 96.4%, respectively. Positive and negative predictive values were 97.0% and 87.1%, respectively. Diagnostic accuracy was 92.2%. EBUS‐TBNA is an efficient and effective technique for diagnosis of intrapulmonary and mediastinal/hilar lymph nodes. It becomes significantly invaluable on clinical management for staging in those patients with lung cancer of other metastatic malignancies. This technique enables us to obtain tissue samples for quick diagnoses beyond central airway with minimal complications. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.
A rapid and efficient method of genotyping zebrafish mutants
Authors: Parant John M. , George Stephen A. , Pryor Rob , Wittwer Carl T. , Yost H. Joseph
Published: November 2009

In order to facilitate high throughput genotyping of zebrafish, we have developed a novel technique that uses High Resolution Melting Analysis (HRMA) to distinguish wild‐type, heterozygous mutants and homogyzous mutants. This one hour technique removes the need for restriction enzymes and agarose gels. The generated melting curve profiles are sensitive enough to detect non‐specific PCR products. We have been able to reliably genotype three classes of mutations in zebrafish, including point mutants, apchu745 (apcmcr), and p53zy7 (p53I166T), a small deletion mutant (bap28y75) and a retroviral insertion mutant (wdr43hi821a). This technique can genotype individual zebrafish embryos and adults (by tail‐clip) and is applicable to other model organisms. Developmental Dynamics 238:3168–3174, 2009. © 2009 Wiley‐Liss, Inc.
Micrographia and hypophonia in anorexia nervosa
Authors: Sekar Murali Krishnan , Arcelus Jon , Palmer Robert L.
Published: November 2009

Micrographia is a condition, in which the sufferers write with small handwriting. Hypophonia is described as low volume speech. Both conditions have been described in neurological conditions, such as lesions in the deep white matter of the brain, Parkinson's disease etc., In this case report, we are presenting a 22‐years‐old female patient with anorexia nervosa who suffered from both these conditions. The patient also suffered from epilepsy. The onset of these symptoms, progress, and current status provides scope for discussing both the possible biological and psychodynamic etiology for these symptoms in this young woman. © 2009 by Wiley Periodicals, Inc. Int J Eat Disord 2010; 43:762–765
Dinuclear Manganese and Cobalt Complexes with Cyclic Polyoxovanadate Ligands: Synthesis and Characterization of [Mn2V10O30]6– and [Co2(H2O)2V10O30]6–
Authors: Inami Shinnosuke , Nishio Masaki , Hayashi Yoshihito , Isobe Kiyoshi , Kameda Hiroyuki , Shimoda Tatsuya
Published: November 2009

An all‐inorganic complex, [Mn2{(VO3)5}2]6– (1), was synthesized, and the structure determination reveals a dinuclear manganese complex coordinated by two cyclic pentavanadate ligands. The cyclic pentavanadate units sandwich the edge‐sharing octahedral dimanganese core through coordination of the oxido group of the pentavanadate. A dinuclear cobalt complex with a cyclic decavanadate, [Co2(OH2)2(VO3)10]6– (2), was also synthesized. The structure analysis reveals a dinuclear cobalt complex with a macrocyclic decavanadate, which is composed of 10 VO4 units joined by the vertex sharings. The CoO6 octahedrons are edge‐shared, with each cobalt octahedron coordinated to five oxido groups from the decavanadate. The remaining site is occupied by water. The coordinated water molecules are supported with hydrogen bonds in two directions. Complex 2 in acetonitrile shows no reactivity with dioxygen even at low temperature, and the cyclic voltammogram of 2 shows no redox chemistry in acetonitrile. Complex 2 exhibits chromism by water exposure both in the solid state and in acetonitrile. Complex 2 is green–yellow in color, and the addition of water causes the complex to turn brown. After heating the sample, it returns to its original color in a reversible manner. The EXAFS data in acetonitrile is also reported and is consistent with the solid‐state structure. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Silver and Gold Catalysis for Cycloisomerization Reactions
Authors: Belmont Philippe , Parker Evelyne
Published: November 2009

We have focused our attention on cycloisomerization reactions involving silver‐ and gold‐based homogeneous catalysis. This topic is divided into two parts: one relating to cycloisomerization with an allene group and the other with an alkyne group. In each case we have then studied different partners such as carbonyl/carboxy groups, imines, amides or alkenes. Reactions involving allene and alkyne systems as partners are also briefly reported, along with miscellaneous transformations. Our purpose is also to stimulate some discussion by setting together the contributions of differentauthors to each type of cycloisomerization reaction. Meanwhile, we have outlined when necessary the impact of the choice of counterion or metal (silver vs. gold) in the course of the reaction. Finally, some mechanistic discussion and reaction intermediates are also mentioned if available.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Silylative Coupling of Olefins with Vinylsilanes in the Synthesis of π‐Conjugated Double Bond Systems
Authors: Pawluć Piotr , Prukała Wiesław , Marciniec Bogdan
Published: November 2009

The design and development of highly efficient and selective methods for the synthesis of π‐conjugated arylvinyl derivatives, based on sequential catalytic reactions of organometallic reagents, have been the subject of extensive study because of their versatile application in organic synthesis and materials science. The silylative coupling of olefins with vinyl‐substituted silicon compounds represents one of the most efficient and selective methods for the synthesis of alkenylsilanes, which are particularly attractive scaffolds for further transformations, including transition‐metal‐catalysed cross‐coupling with organic halides or substitution with organic and inorganic electrophiles. The microreview highlights recent developments in sequential synthetic strategies including ruthenium‐catalysed silylative coupling followed by desilylative cross‐coupling and halogenation, leading to stereodefined organic derivatives containing arylene‐vinylene units widely applied as fine chemicals, functional materials or unsaturated building blocks in organic synthesis.
Convenient Synthesis of N‐Terminal Tfm‐Dipeptides from Unprotected Enantiopure α‐Tfm‐Proline and α‐Tfm‐Alanine (Eur. J. Org. Chem. 33/2009)
Authors: Chaume Grégory , Lensen Nathalie , Caupène Caroline , Brigaud Thierry
Published: November 2009

The cover picture shows that enantiopure fluorinated amino acids and peptides are conveniently obtained from ethyl trifluoropyruvate and trifluoroacetone. Details are discussed in the article by T. Brigaud et. al. on p. 5717 ff. The background is a picture of the Zen garden of the Tôfuku‐ji temple in Kyoto.
FEM modelling of faults occurred in buried power cables due to the removal of tree roots
Authors: Klimenta D. , Kuč V. , Raičević V. , Radosavljević J. , Jevtić M. , Tasić D.
Published: November 2009

An engineering approach to numerical modelling of power cable heating due to a line‐to‐earth short‐circuit caused by previous mechanical damages of a cable sheath and insulation is proposed in this paper. Nonlinearity of the finite element model is based on the soil thermal conductivity and heat sources as functions of temperature and time. The nonlinear transient analysis is applied to an authentic example of line‐to‐earth short‐circuit caused by a mechanical damage of the cable sheath and insulation that has arisen during the previous removal of tree roots. The represented fault occurred on 14 November 2007 in a buried 0.4 kV cable line installed in the city of Bijelo Polje, Republic of Montenegro. Also, this paper outlines a proposition of a technical report prescribing procedures for the removal of tree roots from the soil surrounding cables and for laying power cables nearby high vegetation. The proposed engineering approach is applicable to power cables of all voltage levels laid in soils with changeable moisture content. Copyright © 2009 John Wiley & Sons, Ltd.
Open‐loop analysis and self‐noise performance of an NDA synchroniser for carrier‐independent symbol timing recovery at baud rate
Authors: Gappmair Wilfried
Published: November 2009

Feedback loops for symbol timing recovery are an attractive solution for continuous transmission of data, where rapid acquisition is not so important as in burst‐mode systems. In this context, carrier‐independent non‐data‐aided (NDA) timing error detectors, operated at only one sample per symbol, can be a most interesting option. AssumingM‐ary PSK signals and focusing on a linearised loop model for analytical work, such an algorithm has been discussed already in the technical literature. On the other hand, the linear approach does not apply for larger deviations from the stable equilibrium point as they are encountered during initial acquisition. In this case, the detector characteristic (S‐curve) must be known. Since not available from the open literature, it is derived in this paper not only for PSK but for linear modulation schemes in general. The slope in the stable equilibrium point is given in closed form such that the linearised tracker can be specified immediately. Furthermore, it turned out that the complexity of the detector, but also the computation of both S‐curve and slope, is most simple for a special value of the additional design parameter characterizing the algorithm. Based on this, the self‐noise variance is derived in closed form. ForM‐PSK schemes, it is proved that they experience no jitter floor due to pattern (data) noise. On the other hand, for non‐constant modulus signals, like QAM or APSK constellations, it is shown that the performance is proportional to the loop bandwidth. Finally, a detailed comparison with NDA maximum‐likelihood and Gardner algorithm, as standard method for carrier‐blind NDA tracking, is presented. Copyright © 2009 John Wiley & Sons, Ltd.
Volatile chemistry of a plant: mono‐sesquiterpenoid pattern in the growth cycle of Curcuma haritha
Authors: Baby Sabulal , Raj Gopan , Thaha Abdul Rasheed Muhammad , Dan Mathew
Published: November 2009

Volatile molecules, mostly terpenoids, play a significant role in plant defence. The growth pattern–terpenoid interrelationship of Curcuma haritha, a perennial herb in the ginger family, is tracked by probing the contents of individual mono/sesquiterpenoids throughout its growth cycle by GC‐based techniques (GC–FID, GC–MS, Co‐GC, LRI determination, database and literature search), supported by NMR/mass spectroscopy. Forty to sixty‐four volatile compounds were identified in various growth stages. Major monoterpenes in C. haritha volatile oils were camphor (35.3 ± 0.89%/15.3 ± 0.16%), 1,8‐cineole (13.9 ± 0.27%/7.1 ± 0.09%), isoborneol (9.4 ± 0.27%/4.6 ± 0.10%) and borneol (4.4 ± 0.12%/2.3 ± 0.07%). Total monoterpene (C10) content in C. haritha was highest (69.7–67.0%) in its early growth stages (sprouting/leaf development) and gradually declined towards underground dormancy (lowest 36.9%). Major sesquiterpenes were germacrane‐type compounds: curdione (19.8 ± 0.37%/11.1 ± 0.11%), furanogermenone (18.8 ± 0.16%/trace), neocurdione (6.2 ± 0.14%/2.6 ± 0.04%) and germacrone (5.7 ± 0.12%/1.5 ± 0.06%). Total sesquiterpene (C15) content showed an opposite trend, i.e. 29.9% in sprouting stage to a maximum of 60.9% in dormancy. Guaiane‐type sesquiterpene lactones zedoarondiol, isozedoarondiol, zedoarolide B and zedoalactone B were also isolated from the acetone extract/ethyl acetate fraction of C. haritha rhizomes. The reverse progression of mono‐ and sesquiterpene levels from sprouting/leaf development/leaf‐withering stage to dormancy reveals an energetically feasible biosynthetic/defence plan in C. haritha. Copyright © 2009 John Wiley & Sons, Ltd.
Performance and persistence of Commodity Trading Advisors: Further evidence
Authors: Gregoriou Greg N. , Hübner Georges , Kooli Maher
Published: November 2009

We re‐examine the performance of Commodity Trading Advisors (CTAs) over the January 1995 to October 2008 period. We compare abnormal performance based on a number of alternative existing models, as well as a category‐specific model introducing asset‐, option‐, and moments‐based factors. Taking more factors into account significantly raises the explanatory power, and 9 out of 12 CTA categories significantly outperform the market. We find that numerous CTAs show persistence over a horizon of at least three months and they are also more likely to be persistent over a longer period. Yet, most of the persistence fades away upon the “acid test” of considering only the top and bottom quartiles of CTAs. © 2009 Wiley Periodicals, Inc. Jrl Fut Mark 30:725–752, 2010
RBSP3 is frequently altered in premalignant cervical lesions: Clinical and prognostic significance
Authors: Mitra Sraboni , Mazumder Indra Dipanjana , Bhattacharya Nilanjana , Singh Ratnesh K. , Basu Partha S. , Mondal Ranajit K. , Roy Anup , Zabarovsky Eugene R. , Roychoudhury Susanta , Panda Chinmay K.
Published: November 2009

To understand the importance of frequent deletion of 3p22.3 in cervical carcinogenesis, alterations (deletion/methylation/expression) of the candidate genes STAC, MLH1, ITGA9, and RBSP3, located in the region, were analyzed in 24 cervical intraepithelial neoplasia (CIN) and 137 uterine cervical carcinoma (CACX) samples. In CIN, RBSP3 deletion (48%) and methylation (26%) were high compared with the other genes (4–9%). In CACX, alterations of these genes were as follows: deletion: STAC (54%) > MLH1 (46%) > RBSP3 (45%) > ITGA9 (41%), methylation: RBSP3 (25%) > ITGA9 (24%) > STAC (19%) > MLH1 (13%). Overall, alterations of RBSP3 showed association with CIN, whereas for STAC and MLH1, this frequency increased significantly from CIN → Stage I/II and for ITGA9 from CIN → Stage I/II and also from Stage I/II → Stage III/IV. Quantitative mRNA expression analysis showed differential reduced expression of these genes in CACX concordant to their molecular alterations. The more active RBSP3B splice variant was underexpressed in CACX. RB1 was infrequently deleted in CACX. Concordance was seen between (i) inactivation of RBSP3 and intense p‐RB1 nuclear immunostaining and (ii) low/absence of MLH1 expression and its molecular alterations in CACX. In normal cervical epithelium, p‐RB1 immunostaining was low in differentiated cells, whereas MLH1 staining was seen in both nucleus and cytoplasm irrespective of differentiation stage. Alterations of the genes were significantly associated with poor prognosis. High parity (≥5)/early sexual debut (≤19 years) coupled with RBSP3 alterations/RB1 deletion predicted worst prognosis. Thus, inactivation of RBSP3 might be one of the early events in cervical carcinogenesis. © 2009 Wiley‐Liss, Inc.
Influence of coherence between multiple cortical columns on alpha rhythm: A computational modeling study
Authors: Naruse Yasushi , Matani Ayumu , Miyawaki Yoichi , Okada Masato
Published: November 2009

In electroencephalographic (EEG) and magnetoencephalographic (MEG) signals, stimulus‐induced amplitude increase and decrease in the alpha rhythm, known as event‐related synchronization and desynchronization (ERS/ERD), emerge after a task onset. ERS/ERD is assumed to reflect neural processes relevant to cognitive tasks. Previous studies suggest that several sources of alpha rhythm, each of which can serve as an alpha rhythm generator, exist in the cortex. Since EEG/MEG signals represent spatially summed neural activities, ERS/ERD of the alpha rhythm may reflect the consequence of the interactions between multiple alpha rhythm generators. Two candidates modulate the magnitude of ERS/ERD: (1) coherence between the activities of the alpha rhythm generators and (2) mean amplitude of the activities of the alpha rhythm generators. In this study, we use a computational model of multiple alpha rhythm generators to determine the factor that dominantly causes ERS/ERD. Each alpha rhythm generator is modeled based on local column circuits in the primary visual cortex and made to interact with the neighboring generators through excitatory connections. We observe that the model consistently reproduces spontaneous alpha rhythms, event‐related potentials, phase‐locked alpha rhythms, and ERS/ERD in a specific range of connectivity coefficients. Independent analyses of the coherence and amplitude of multiple alpha rhythm generators reveal that the ERS/ERD in the simulated data is dominantly caused by stimulus‐induced changes in the coherence between multiple alpha rhythm generators. Nonlinear phenomena such as phase‐resetting and entrainment of the alpha rhythm are related to the neural mechanism underlying ERS/ERD. Hum Brain Mapp, 2010. © 2009 Wiley‐Liss, Inc.
Water and heat fluxes above a lowland dipterocarp forest in Peninsular Malaysia
Authors: Takanashi Satoru , Kosugi Yoshiko , Ohkubo Shinjiro , Matsuo Naoko , Tani Makoto , Nik Abdul Rahim
Published: November 2009

We measured the fluxes of sensible and latent heat between a low‐land dipterocarp forest in Peninsular Malaysia and the atmosphere. No clear seasonal or interannual changes in latent heat flux were found from 2003 to 2005, while sensible heat flux sometimes fluctuated depending on the fluctuation of incoming radiation between wet and dry seasons. The evapotranspiration rates averaged for the period between 2003 and 2005 were 2·77 and 3·61 mm day−1 using eddy covariance data without and with an energy balance correction, respectively. Average precipitation was 4·74 mm day−1. Midday surface conductance decreased with an increasing atmospheric water vapour pressure deficit and thus restricted the excess water loss on sunny days in the dry season. However, the relationship between the surface conductance and vapour pressure deficit did not significantly decline with an increase in volumetric soil water content even during a period of extremely low rainfall. Copyright © 2009 John Wiley & Sons, Ltd.
Observe, listen, wonder, and respond: A preliminary exploration of reflective function skills in early care providers
Authors: Tomlin Angela M. , Sturm Lynne , Koch Steven M.
Published: November 2009

Parents' behaviors demonstrating reflective capacity towards their children, parental reflective functioning, have been identified as central to both the formation of a secure attachment and therapeutic efforts to remediate attachment problems (Fonagy & Target, 2005; Sadler, Slade, & Mayes, 2006). We hypothesize that reflective skills in providers may be key elements in effecting change through the parent–professional relationship. As a first step, the present study examined early care and intervention providers' self‐report of the importance of reflective practice skills in their work with families of young children. We further examined whether provider characteristics such as professional affiliation and experience related to importance ratings. Lastly, we examined, in a preliminary fashion, whether provider valuing of reflective functioning skills is associated with reported practice using hypothetical vignettes drawn from common home‐visiting scenarios.
Perspectives on helping traumatized infants, young children, and their families
Authors: Osofsky Joy D.
Published: November 2009

Traumatized infants, toddlers, and young children can affect adults in different ways but most often pull extreme reactions ranging from empathy to anger. It is important for those who support, intervene, and provide therapeutic services for traumatized young children and their families to understand that various traumatization, compassion, fatigue, and burnout can be an integral part of the work. All interveners, including child welfare workers, clinicians, home visitors, teachers, and even nontraditional responders, such as those who supervise therapeutic visitation, must find their own ways to cope with the overwhelming feelings that may be aroused. Support or regularly scheduled reflective supervision as well as self‐care is crucial for those who work with trauma. This paper describes helpful ways to intervene and provide support for infant mental health therapists and others working with traumatized young children who may experience vicarious traumatization.
Thinner inhalation effects on oxidative stress and DNA repair in a rat model of abuse
Authors: Martínez‐Alfaro Minerva , Cárabez‐Trejo Alfonso , Gallegos‐Corona Marco‐Antonio , Pedraza‐Aboytes Gustavo , Hernández‐Chan Nancy Georgina , Leo‐Amador Guillermo Enrique
Published: November 2009

Humans can come into contact with thinner by occupational exposure or by intentional inhalation abuse. Numerous studies of workers for genotoxic effects of thinner exposure have yielded conflicting results, perhaps because co‐exposure to variable other compounds cannot be avoided in workplace exposure studies. In contrast, there is no data concerning the genotoxic effects of intentional inhalation abuse. The aim of this project was to examine the genotoxic effects of thinner inhalation in an animal model of thinner abuse (rats exposed to 3000 ppm toluene, a high solvent concentration over a very short, 15 min time period, twice a day for 6 weeks). The data presented here provides evidence that thinner inhalation in our experimental conditions is able to induce weight loss, lung abnormalities and oxidative stress. This oxidative stress induces oxidative DNA damage that is not a characteristic feature of genotoxic damage. No significant difference in DNA damage and DNA repair (biomarkers of genotoxicity) in lymphocytes from thinner‐treated and control rats was found. Lead treatment was used as a positive control in these assays. Finally, bone marrow was evaluated as a biomarker of cellular alteration associated with thinner inhalation. The observed absence of hemopoietic and genetic toxicity could be explained in part by the absence of benzene, the only carcinogenic component of thinner; however, benzene is no longer a common component of thinner. In conclusion, thinner did not cause genotoxic effects in an experimental model of intentional abuse despite the fact that thinner inhalation induces oxidative stress. Copyright © 2009 John Wiley & Sons, Ltd.
Bradykinin enhances cell migration in human chondrosarcoma cells through BK receptor signaling pathways
Authors: Yang Wei‐Hung , Chang Jung‐Tzu , Hsu Sheng‐Feng , Li Te‐Mao , Cho Der‐Yang , Huang Chun‐Yin , Fong Yi‐Chin , Tang Chih‐Hsin
Published: November 2009

Bradykinin (BK) is an inflammatory mediator, and shows elevated levels in regions of severe injury and inflammatory diseases. BK has recently been shown to be involved in carcinogenesis and cancer progression. In this study, we found that BK increased the migration and the expression of α2β1 integrin in human chondrosarcoma cells. We also found that human chondrosarcoma tissues had significantly higher expression of the B1 and B2 receptors comparing to normal cartilage. BK‐mediated migration and integrin up‐regulation was attenuated by B1 and B2 BK receptor siRNA or antagonist. Activations of phospholipase C (PLC), protein kinase Cδ (PKCδ), and NF‐κB pathways after BK treatment was demonstrated, and BK‐induced integrin expression and migration activity was inhibited by the specific inhibitor of PLC, PKCδ, and NF‐κB cascades. Taken together, our results indicated that BK enhances the migration of chondrosarcoma cells by increasing α2β1 integrin expression through the BK receptors/PLC/PKCδ/NF‐κB signal transduction pathway. J. Cell. Biochem. 109: 82–92, 2010. © 2009 Wiley‐Liss, Inc.
Identification of microRNAs regulated by activin A in human embryonic stem cells
Authors: Tsai Zong‐Yun , Singh Sher , Yu Sung‐Liang , Kao Li‐Pin , Chen Bo‐Zhi , Ho Bing‐Ching , Yang Pan‐Chyr , Li Steven Shoei‐Lung
Published: November 2009

Human embryonic stem (hES) cells have the capacities to propagate for extended periods and to differentiate into cell types from all three germ layers both in vitro and in vivo. These characteristics of self‐renewal and pluripotency enable hES cells having the potential to provide an unlimited supply of different cell types for tissue replacement, drug screening, and functional genomics studies. The hES‐T3 cells with normal female karyotype cultured on either mouse embryonic fibroblasts (MEF) in hES medium (containing 4 ng/ml bFGF) (T3MF) or feeder‐free Matrigel in MEF‐conditioned medium (supplemented with additional 4 ng/ml bFGF) (T3CM) were found to express very similar profiles of mRNAs and microRNAs, indicating that the unlimited self‐renewal and pluripotency of hES cells can be maintained by continuing culture on these two conditions. However, the expression profiles, especially microRNAs, of the hES‐T3 cells cultured on Matrigel in hES medium supplemented with 4 ng/ml bFGF and 5 ng/ml activin A (T3BA) were found to be different from those of T3MF and T3CM cells. In T3BA cells, four hES cell‐specific microRNAs miR‐372, miR‐302d, miR‐367, and miR‐200c, as well as three other microRNAs miR‐199a, miR‐19a, and miR‐217, were found to be up‐regulated, whereas five miRNAs miR‐19b, miR‐221, miR‐222, let‐7b, and let‐7c were down‐regulated by activin A. Thirteen abundantly differentially expressed mRNAs, including NR4A2, ERBB4, CXCR4, PCDH9, TMEFF2, CD24, and COX6A1 genes, targeted by seven over‐expressed miRNAs were identified by inverse expression levels of these seven microRNAs to their target mRNAs in T3BA and T3CM cells. The NR4A2, ERBB4, and CXCR4 target genes were further found to be regulated by EGF and/or TNF. The 50 abundantly differentially expressed genes targeted by five under‐expressed miRNAs were also identified. The abundantly expressed mRNAs in T3BA and T3CM cells were also analyzed for the network and signaling pathways, and roles of activin A in cell proliferation and differentiation were found. These findings will help elucidate the complex signaling network which maintains the self‐renewal and pluripotency of hES cells. J. Cell. Biochem. 109: 93–102, 2010. © 2009 Wiley‐Liss, Inc.
Inducible activation of IFI 16 results in suppression of telomerase activity, growth suppression and induction of cellular senescence
Authors: Clarke Christopher J.P. , Hii Linda L. , Bolden Jessica E. , Johnstone Ricky W.
Published: November 2009

Expression of the human HIN‐200 family member IFI 16 has been reported to suppress cell growth and contribute to the onset of cellular senescence. However the molecular events involved in this process have not been fully characterised. We fused IFI 16 to the estrogen receptor ligand‐binding domain to establish an inducible model for studying the molecular events that cause these phenomena. In cells induced to express the ER‐IFI 16 within the nucleus there was a decrease in cellular proliferation and concomitant growth arrest in the G1 phase of the cell cycle. Unlike previous reports, this did not appear to involve the p53‐p21WAF1/CIP1‐cdk2‐pRb pathway. Following nuclear expression of ER‐IFI 16 we noted senescence‐like morphological changes and expression of senescence‐associated β‐galactosidase in growth arrested cells. Importantly, we also found a marked reduction in telomerase activity in arrested cells compared to controls. Moreover, IFI 16 and hTERT co‐localised within the nucleus and these two proteins physically interacted in vivo and in vitro. Together, these data suggest that IFI 16 may act as an endogenous regulator of telomerase activity and, through its interaction with hTERT, contributes to the inhibition of proliferation and induces a senescence‐like state. J. Cell. Biochem. 109: 103–112, 2010. © 2009 Wiley‐Liss, Inc.
Restricted dead‐end elimination: Protein redesign with a bounded number of residue mutations
Authors: Safi Maria , Lilien Ryan H.
Published: November 2009

Dead‐end elimination (DEE) has emerged as a powerful structure‐based, conformational search technique enabling computational protein redesign. Given a protein withnmutable residues, the DEE criteria guide the search toward identifying the sequence of amino acids with the global minimum energy conformation (GMEC). This approach does not restrict the number of permitted mutations and allows the identified GMEC to differ from the original sequence in up tonresidues. In practice, redesigns containing a large number of mutations are often problematic when taken into the wet‐lab for creation via site‐directed mutagenesis. The large number of point mutations required for the redesigns makes the process difficult, and increases the risk of major unpredicted and undesirable conformational changes. Preselecting a limited subset of mutable residues is not a satisfactory solution because it is unclear how to select this set before the search has been performed. Therefore, the ideal approach is what we define as the κ‐restricted redesign problem in which any κ of thenresidues are allowed to mutate. We introduce restricted dead‐end elimination (rDEE) as a solution of choice to efficiently identify the GMEC of the restricted redesign (the κGMEC). Whereas existing approaches requiren‐choose‐κ individual runs to identify the κGMEC, the rDEE criteria can perform the redesign in a single search. We derive a number of extensions to rDEE and present a restricted form of the A* conformation search. We also demonstrate a 10‐fold speed‐up of rDEE over traditional DEE approaches on three different experimental systems. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010
Electronic structure and reactivity of guanylthiourea: A quantum chemical study
Authors: Mehdi Ahmed , Adane Legesse , Patel Dhilon S. , Bharatam Prasad V.
Published: November 2009

Electronic structure analysis of guanylthiourea (GTU) and its isomers has been carried out using quantum chemical methods. Two major tautomeric classes (thione and thiol) have been identified on the potential energy (PE) surface. In both the cases conjugation of pi‐electrons and intramolecular H‐bonds have been found to play a stabilizing role. Various isomers of GTU on its PE surface have been analyzed in two different groups (thione and thiol). The interconversion from the most stable thione conformer (GTU‐1) to the most stable thiol conformer (GTU‐t1) was found to take place via bimolecular process which involves protonation at sulfur atom of GTU‐1 followed by subsequent CN bond rotation and deprotonation. The detailed analysis of the protonation has been carried out in gas phase and aqueous phase (using CPMC model). Sulfur atom (S1) was found to be the preferred protonation site (over N4) in GTU‐1 in gas phase whereas N4 was found to be the preferred site of protonation in aqueous medium. The mechanism of S‐alkylation reaction in GTU has also been studied. The formation of alkylated analogs of thiol isomers (alkylated guanylthiourea) is believed to take place via bimolecular process which involves alkyl cation attack at S atom followed by CN bond rotation and deprotonation. The reactive intermediate RS(NH2)CNC(NH2)2+ belongs to the newly identified ⊕N(←L)2 class of species and provides the necessary dynamism for easy conversion of thione to thiol. © 2009 Wiley Periodicals, Inc. J Comput Chem, 2010
Molecular dissection of Otx1 functional domains in the zebrafish embryo
Authors: Bellipanni Gianfranco , Murakami Tohru , Weinberg Eric S.
Published: November 2009

Otx proteins are involved in the induction of neurectoderm patterning and morphogenetic movements, leading to the formation of the vertebrate central nervous system. Despite lack of homology of sequence outside the homeodomain, a large body of evidence has shown that the Otx/Otd class of proteins has similar functions in many animal phyla. Thus, characterization of functional domains in proteins of this family would help in understanding how this functional equivalence operates. Our previous analysis using the zebrafish embryo (Bellipanni et al., 2000, Dev Biol 223:339–353), has suggested that induction of cell aggregation is a morphoregulatory role of Otx/Otd factors in embryonic development. We now use the induction of cell aggregation as an in vivo assay to examine the functional requirement for particular domains of the zOtx1 protein. We demonstrate that zOtx1 induces cell aggregation by acting as a transcriptional activator through its C‐terminal region. Further, we show that a region of 37 amino acids in the C‐terminal third of zOtx1 is necessary but not sufficient for this activation potential. The effects of selective deletion of each of the three homeodomain α‐helices of zOtx1 on cell aggregation were also tested. Surprisingly, we find that helix 3, which is required for binding to DNA, is dispensable for stimulation of cell aggregation. Our results suggest that for transcriptional activation of at least one gene in the cell aggregation pathway, zOtx1 need not bind directly to DNA, but does require helix 1 and 2 of its homeodomain to interact with an as yet undefined DNA binding protein. J. Cell. Physiol. 222: 286–293, 2010. © 2009 Wiley‐Liss, Inc.
FEZ1/LZTS1 protein expression in ovarian cancer
Authors: Califano Daniela , Pignata Sandro , Pisano Carmela , Greggi Stefano , Laurelli Giuseppe , Losito Nunzia Simona , Ottaiano Alessandro , Gallipoli Adolfo , Pasquinelli Rosa , De Simone Veronica , Cirombella Roberto , Fusco Alfredo , Chiappetta Gennaro
Published: November 2009

The FEZ1/LZTS1 (FEZ1) gene maps to chromosome 8p22 and is frequently altered in human cancer. FEZ1 has been proposed as a candidate tumour suppressor gene and its loss may contribute to tumour progression. We have analysed the expression of FEZ1 protein in tissues from ovarian carcinomas in relation to clinico‐pathological variables, response to chemotherapy and disease‐free and overall survival. FEZ1 status was assessed by immunohistochemistry. Cytoplasmic staining for FEZ1 protein was absent or drastically reduced in 38% of tumours. FEZ1 protein expression was not related to tumour grade, histotype, disease‐free survival, or overall survival. On the contrary, it was significantly correlated with age and with FIGO stage of disease. This finding indicates that FEZ1 is involved in ovarian carcinogenesis. Moreover, loss of FEZ1 protein significantly predicted a complete treatment response in patients who received taxane‐based chemotherapy. In conclusion, the reduction or loss of FEZ1 protein could be an aid to the clinical management of patients affected by ovarian carcinoma. J. Cell. Physiol. 222: 382–386, 2010. © 2009 Wiley‐Liss, Inc.
Inhibition of PARP activity by PJ‐34 leads to growth impairment and cell death associated with aberrant mitotic pattern and nucleolar actin accumulation in M14 melanoma cell line
Authors: Chevanne Marta , Zampieri Michele , Caldini Riccardo , Rizzo Angela , Ciccarone Fabio , Catizone Angela , D'Angelo Carmen , Guastafierro Tiziana , Biroccio Anna , Reale Anna , Zupi Gabriella , Caiafa Paola
Published: November 2009

The capability of PARP activity inhibitors to prevent DNA damage recovery suggested the use of these drugs as chemo‐ and radio‐sensitisers for cancer therapy. Our research, carried out on cultured human M14 melanoma cells, was aimed to examine if PJ‐34, a potent PARP activity inhibitor of second generation, was per se able to affect the viability of these cancer cells without any DNA damaging agents. Using time‐lapse videomicroscopy, we evidenced that 10 µM PJ‐34 treatment induced severe mitotic defects leading to dramatic reduction of cell proliferation and to cell death. PJ‐34 cytotoxic effect was further confirmed by analysis of cell viability and clonogenic assay. Absence of canonic apoptosis markers allowed us to exclude this kind of cell death. No single and/or double stranded DNA damage was evidenced. Immunofluorescence analysis showed an aberrant mitotic scenario in several cells and subsequent multinucleation suggesting an atypical way for cells to die: the mitotic catastrophe. The detection of aberrant accumulation of polymerised actin inside the nucleolus was noteworthy. Taken toghether, our results demonstrate that, targeting PARP activity by PJ‐34, cancer cell survival is affected independently of DNA damage repair. Two findings are remarkable: (a) cisplatin concentration can be reduced by three quarters if it is followed by treatment with 10 µM PJ‐34 for 24 h to obtain the same citotoxic effect; (b) effects dependent on PJ‐34 treatment are reversible. Our data suggest that, to reduce the harm done to non‐tumour cells during chemotherapy with cisplatin, the latter could be coupled with PJ‐34 treatment. J. Cell. Physiol. 222: 401–410, 2010. © 2009 Wiley‐Liss, Inc.
Characterization of Bombyx embryo segmentation process: expression profiles of engrailed, even‐skipped, caudal, and wnt1/wingless homologues
Authors: Nakao Hajime
Published: November 2009

To gain insight into segmentation processes, the expression at embryonic stages of the silkmoth Bombyx mori homologues of even‐skipped (eve), engrailed (en), caudal (cad), and wnt1/wingless (wg) transcripts were examined by whole mount in situ hybridization. Pair‐rule eve stripes and segmental en and wnt1/wg stripes were generated sequentially from anterior to posterior, confirming the previous results that showed that Bombyx belongs to short‐germ insects. However, unlike in previously described short germ insects, the segmentation of Bombyx occurred without marked germ band elongation: the putative growth zone was expanded compared with previously described short germ insects. This may indicate that Bombyx represents an evolutionarily intermediate state in a transition from short to long germ type. The expressions of cad and wnt1/wg, which are known to be present in the growth zone in short germ insects, initially showed a large median expression domain that, as segmentation proceeded, later retracted to the posterior pole. This is also unique to this insect. Detailed analysis of their relative expressions indicated that wnt1/wg domain retracted faster than the cad domain, and double stain in situ hybridization suggested that the eve stripe appears from cells that have ceased to express wnt1/wg. Another unique aspect of Bombyx embryogenesis is that gastrulation began at later embryonic stage compared with other insects and proceeded slowly from anterior to posterior. On the basis of these results, conserved and divergent aspects of the evolution of insect segmentation mechanisms and germ cell formation are discussed. J. Exp. Zool. (Mol. Dev. Evol.) 314B:224–231, 2010. © 2009 Wiley‐Liss, Inc.
Synthesis and molecular recognition of novel multiimidazole cyclophanes
Authors: Xu Xiao‐Wei , Wang Xin‐Long , Wu Ai‐Ming , Zheng Zhi‐Ming , Yi Mei‐Gui , Xiao Rong
Published: November 2009

magnified image

Cyclophanes based on 2,2′‐biimidazole and 2,2′‐bibenzimidazole were synthesized as receptors. UV spectroscopic titration in chloroform at 25°C showed 1:1 complexes between the cyclophanes and the guests, and the binding constants (K) and Gibbs free energy changes (−ΔG0) were calculated according to the modified Benesi‐Hildebrand equation. J. Heterocyclic Chem., 00, 00 (2009).
111In‐labeled KCCYSL peptide as an imaging probe for ErbB‐2‐expressing ovarian carcinomas
Authors: Deutscher Susan L. , Figueroa Said D. , Kumar Senthil R.
Published: November 2009

Aberrant expression of ErbB‐2, a member of the epidermal growth factor family of receptors, has been implicated in the formation of various malignancies including ovarian cancer. The objective of this study was to determine whether the phage display‐selected ErbB‐2 targeting peptide, KCCYSL, once radiolabeled with 111In would serve as a tumor targeting and single photon emission computed tomography (SPECT/CT) imaging agent in a mouse model of human ovarian carcinoma expressing ErbB‐2. The KCCYSL peptide was synthesized with a chelator 1,4,7,10‐tetra‐azacyclododecane‐N,N′,N″,N′″‐tetraacetic acid (DOTA), and a Gly‐Ser‐Gly (GSG) spacer between DOTA and amino terminus of the peptide and radiolabeled with 111InC13. In vitro cell binding studies indicated that 111In‐DOTA‐GSG‐KCCYSL bound to cultured ovcar‐3 carcinoma cells. Bio‐distribution studies in scid mice bearing human ovcar‐3 tumor xenografts revealed a tumor uptake of 0.50±0.05 percent injected dose per gram (%ID/g) at 1 h, and 0.39±0.1 %ID/g at 2 h. Blocking studies with non‐radiolabeled counterpart indicated a partial inhibition (41%) (P=0.04) in tumor uptake of 111In‐DOTA‐GSG‐KCCYSL. In vivo tumor uptake of 111In‐DOTA‐GSG‐KCCYSL was clearly evident through SPECT/CT images after 2 h post injection. These studies suggest the potential of this peptide as a radiopharmaceutical for imaging of ErbB‐2‐expressing ovarian tumors. Copyright © 2009 John Wiley & Sons, Ltd.
Periaqueductal gray afferents synapse onto dopamine and GABA neurons in the rat ventral tegmental area
Authors: Omelchenko Natalia , Sesack Susan R.
Published: November 2009

The midbrain central gray (periaqueductal gray; PAG) mediates defensive behaviors and is implicated in the rewarding effects of opiate drugs. Projections from the PAG to the ventral tegmental area (VTA) suggest that this region might also regulate behaviors involving motivation and cognition. However, studies have not yet examined the morphological features of PAG axons in the VTA or whether they synapse onto dopamine (DA) or GABA neurons. In this study, we injected anterograde tracers into the rat PAG and used immunoperoxidase to visualize the projections to the VTA. Immunogold‐silver labeling for tyrosine hydroxylase (TH) or GABA was then used to identify the phenotype of innervated cells. Electron microscopic examination of the VTA revealed axons labeled anterogradely from the PAG, including myelinated and unmyelinated fibers and axon varicosities, some of which formed identifiable synapses. Approximately 55% of these synaptic contacts were of the symmetric (presumably inhibitory) type; the rest were asymmetric (presumably excitatory). These findings are consistent with the presence of both GABA and glutamate projection neurons in the PAG. Some PAG axons contained dense‐cored vesicles indicating the presence of neuropeptides in addition to classical neurotransmitters. PAG projections synapsed onto both DA and GABA cells with no obvious selectivity, providing the first anatomical evidence for these direct connections. The results suggest a diverse nature of PAG physiological actions on midbrain neurons. Moreover, as both the VTA and PAG are implicated in the reinforcing actions of opiates, our findings provide a potential substrate for some of the rewarding effects of these drugs. © 2009 Wiley‐Liss, Inc.
Oxidative stress up‐regulates presenilin 1 in lipid rafts in neuronal cells
Authors: Oda Akiko , Tamaoka Akira , Araki Wataru
Published: November 2009

Oxidative stress is associated with β‐amyloid peptide (Aβ) accumulation in the brains of Alzheimer's disease patients. Aβ is generated upon the sequential proteolytic cleavage of transmembrane amyloid precursor protein (APP) by two membrane‐bound proteases, β‐secretase (BACE1) and the γ‐secretase complex comprising presenilin 1 (PS1), nicastrin, APH‐1 and PEN‐2. Recent evidence suggests that significant amounts of BACE1 and γ‐secretase components localize in the cholesterol‐rich region of membranes known as lipid rafts, where Aβ production occurs preferentially. In this study, we investigated the effects of oxidative stress on the BACE1 and γ‐secretase components in lipid rafts using human neuroblastoma SH‐SY5Y cells exposed to ethacrynic acid (EA), a compound that induces cellular glutathione depletion. Following exposure of cells to EA, heme oxygenase‐1, a marker protein of oxidative stress, was strongly induced. Moreover, treatment with EA resulted in a significant increase in PS1 protein levels, but not those of nicastrin, APH‐1, PEN‐2 or BACE1, in both cell lysates and the lipid raft fraction. This increase in PS1 protein expression was prevented by co‐treatment with an antioxidant, N‐acetylcysteine (NAC). EA additionally induced a significant increase in PS1 mRNA expression, which was inhibited by NAC. Finally, EA treatment was found to promote Aβ secretion from cells expressing Swedish mutant APP. It appears that in our cell culture model, oxidative stress enhances PS1 protein levels in lipid rafts via up‐regulation of PS1 transcription, which may constitute the mechanism underlying the oxidative stress‐associated promotion of Aβ production. © 2009 Wiley‐Liss, Inc.
Protective role of COMP‐Ang1 in ischemic rat brain
Authors: Shin Hye Young , Lee Yoon Jin , Kim Hak Joo , Park Chul‐Kee , Kim Jong Hoon , Wang Kyu Chang , Kim Dong Gyu , Koh Gou Young , Paek Sun Ha
Published: November 2009

In cerebral ischemia, the induction of angiogenesis may represent a natural defense mechanism that enables the hypoxic brain to avoid progression into infarction. Angiopoietin‐1 (Ang1) is known to produce non‐leaky and stable blood vessel formation mainly by the Tie2 receptor. Therefore, we envisioned that the application of cartilage oligomeric matrix protein‐Ang1 (COMP‐Ang1), a soluble, stable, and potent form of Ang1, would promote angiogenesis and provide a protective effect following unilateral middle cerebral artery occlusion (MCAO) in rats. To this end, we employed a 2‐hour‐MCAO model, and treated rats with adenovirus encoding COMP‐Ang1 (Ade‐COMP‐Ang1) or control virus encoding β‐gal (Ade‐β‐gal). Time course magnetic resonance images (MRIs) revealed significantly reduced infarct volume in the rats treated with Ade‐COMP‐Ang1 with an improvement of post‐ischemic neurological deficits compared with rats treated with Ade‐β‐gal. Moreover, compared to the rats treated with Ade‐β‐gal, the rats treated with Ade‐COMP‐Ang1 showed an increase in blood vessels, especially in the border zone adjacent to the infarction, increased number of endogenous neuronal progenitor cells in the ischemic brain, and decreased number of TUNEL‐positive cells. Taken together, COMP‐Ang1 reduced infarct volume and consequently attenuated post‐ischemic neurological deficits through enhanced angiogenesis and increased viable cell mass of neuronal cells. © 2009 Wiley‐Liss, Inc.