bluuebunny/crossref_metadata_2025 huggingface dataset:
Experimental and Theoretical Characterization of the Aromatization, Epimerization, and Fragmentation Reactions of Bi‐2H‐azirin‐2‐yls Prepared from 1,4‐Diazidobuta‐1,3‐dienes
Authors: Banert Klaus , Köhler Frank , Melzer Antje , Scharf Ingolf , Rheinwald Gerd , Rüffer Tobias , Lang Heinrich , Herges Rainer , Heß Kirsten , Ghavtadze Nugzar , Würthwein Ernst‐Ulrich
Published: July 2011

1,4‐Diazidobuta‐1,3‐dienes (Z,Z)‐10, 17, and 21 were photolyzed and thermolyzed to yield the pyridazines 13, 20, and 23, respectively. To explain these aromatic final products, the generation of highly strained bi‐2H‐azirin‐2‐yls 12, 19, and 22 and their valence isomerization were postulated. In the case of meso‐ and rac‐22, nearly quantitative formation from diazide 21, isolation as stable solids, and complete characterization were possible. On the thermolysis of 22, aromatization to 23 was only a side reaction, whereas equilibration of meso‐ and rac‐22 and fragmentation, which led to alkyne 24 and acetonitrile, dominated. Prolonged irradiation of 22 gave mainly the pyrimidine 25. The change of the configuration at C‐2 of the 2H‐azirine unit was observed not only in the case of bi‐2H‐azirin‐2‐yls 22 but also for simple spirocyclic 2H‐azirines 29 at a relatively low temperature (75 °C). The fragmentation of rac‐22 to give alkyne 24 and two molecules of acetonitrile was also studied by high‐level quantum chemical calculations. For a related model system 30 (methyl instead of phenyl groups), two transition states TS‐30–31 of comparable energy with multiconfigurational electronic states could be localized on the energy hypersurface for this one‐step conversion. The symmetrical transition state complies with the definition of a coarctate mechanism.
Characterizing projective spaces on deformations of Hilbert schemes of K3 surfaces
Authors: Harvey David , Hassett Brendan , Tschinkel Yuri
Published: July 2011

We seek to characterize homology classes of Lagrangian projective spaces embedded in irreducible holomorphic‐symplectic manifolds, up to the action of the monodromy group. This paper addresses the case of manifolds deformation‐equivalent to the Hilbert scheme of length‐3 subschemes of a K3 surface. The class of the projective space in the cohomology ring has prescribed intersection properties, which translate into Diophantine equations. Possible homology classes correspond to integral points on an explicit elliptic curve; our proof entails showing that the only such point is two‐torsion. © 2011 Wiley Periodicals, Inc.
Static noise margin enhancement by flex‐pass‐gate SRAM
Authors: O'uchi Shin‐Ichi , Masahara Meishoku , Sakamoto Kunihiro , Endo Kazuhiko , Liu Yungxun , Matsukawa Takashi , Sekigawa Toshihiro , Koike Hanpei , Suzuki Eiichi
Published: July 2011

A flex‐pass‐gate SRAM, that is, a fin‐type field effect transistor‐based (FinFET‐based) SRAM, is proposed to enhance the noise margin during both read and write operations. In its cell, the flip‐flop is composed of the usual three‐terminal (3T) FinFETs and the pass gates are composed of four‐terminal (4T) FinFETs. The 4T‐FinFETs enable the adoption of dynamic threshold‐voltage (Vt) control in the pass gates. During a write operation, the Vt of the pass gates is lowered to enhance the writing speed and stability. During the read operation, on the other hand, the Vt is raised to enhance the static noise margin. An asymmetric‐oxide 4T‐FinFET is helpful in managing the leakage current through the pass gate. In this paper, a design strategy for the pass gate with an asymmetric gate oxide is considered, and a TCAD‐based Monte Carlo simulation reveals that the flex‐pass‐gate SRAM based on that design strategy may be expected to be effective in the half‐pitch 32‐nm technology for low standby power (LSTP) applications, even taking into account the variability in device performance. © 2011 Wiley Periodicals, Inc. Electron Comm Jpn, 94(8): 57–64, 2011; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.10217
Piezo‐driven vibrating insertion device for microelectrode array
Authors: Noda Takahiro , Kanzaki Ryohei , Takahashi Hirokazu
Published: July 2011

Microelectrode arrays are commonly used to measure neural activities in the brain, and arrays with some 100 electrodes are commercially available to date. However, insertion of a dense grid array deforms the brain, resulting in deterioration of the measurements. In order to overcome this problem, we propose a piezo‐driven vibrating insertion device to reduce the insertion‐induced deformation of the brain. We attempted under various conditions to insert the array into an agarose substrate, whose hardness was adjusted to that of the cerebral cortex of rats. Our experiments demonstrated that inverse‐sawtooth vibration reduced the insertion‐induced deformation of the substrate in proportion to the logarithm of the upstroke velocity when the velocity was higher than 10 mm/s, and vibrating insertion of the maximum velocity at 36.7 mm/s reduced the deformation by up to 40% as compared to insertion without vibration. In addition, we tested the vibrating insertion device in an electrophysiological experiment in the rat auditory cortex in vivo, and successfully measured tone‐evoked neural activities. © 2011 Wiley Periodicals, Inc. Electron Comm Jpn, 94(8): 37–46, 2011; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.10327
Surface hardness improvement of PMMA by low‐energy ion irradiation and electron irradiation
Authors: Sakurabayashi Yuya , Masaki Takahiro , Iwao Toru , Yumoto Motoshiga
Published: July 2011

Surface modification of PMMA (polymethylmethacrylate) was carried out by irradiation with low‐energy ions and/or electrons, which is expected to improve the surface hardness by introduction of a thin modified layer. The surface hardness was measured by the nanoindentation test. To clarify the structure and the properties of the modified layer, depth profiles of composition and chemical bonds were analyzed using XPS (X‐ray photoelectron spectroscopy). The formation of a cross‐linking structure that contributed to the surface hardness was analyzed by the dyeing method. From these results, it was confirmed that the surface hardness increased and a cross‐linking structure was present. It is suggested that the existence of the structure brought about the increase of surface hardness, showing the utility of the dyeing method. © 2011 Wiley Periodicals, Inc. Electron Comm Jpn, 94(8): 19–26, 2011; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.10350
Stabilizing predictive visual feedback control for fixed camera systems
Authors: Murao Toshiyuki , Kawai Hiroyuki , Fujita Masayuki
Published: July 2011

This paper investigates vision‐based robot control via a receding horizon control strategy for fixed camera systems, as stabilizing predictive visual feedback control. First, a visual motion robot error system with a fixed camera configuration is reconstructed in order to improve estimation performance. Next, stabilizing receding horizon control for three‐dimensional visual feedback systems, which are highly nonlinear and relatively fast systems, is proposed. The stability of the receding horizon control scheme is guaranteed by using a terminal cost derived from an energy function of the visual motion robot error system. Furthermore, simulation and actual nonlinear experimental results are assessed with respect to stability and performance. © 2011 Wiley Periodicals, Inc. Electron Comm Jpn, 94(8): 1–11, 2011; Published online in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/ecj.10357
Palladium‐Catalyzed Coupling of 3‐Halo‐Substituted Coumarins, Chromenes, and Quinolones with Various Nitrogen‐Containing Nucleophiles
Authors: Soussi Mohamed Ali , Audisio Davide , Messaoudi Samir , Provot Olivier , Brion Jean‐Daniel , Alami Mouâd
Published: July 2011

An efficient and general palladium‐catalyzed coupling reaction between 3‐bromocoumarins, 3‐bromoquinolin‐2(1H)‐ones, and 3‐iodo‐2H‐chromenes with a variety of nitrogen‐containing nucleophiles (azole, amide, lactam, sulfonamide, aniline, amine, and urea) is described. The reaction proceeded rapidly and cleanly in dioxane providing the coupling products in good to excellent yields. The chemoselectivity of this reaction was also studied with polyhalocoumarins. Under optimized conditions, these underwent a site‐selective amination at the C‐3–Br bond, regardless of the nature of the nitrogen‐containing nucleophile used.
Synthesis of Activated Cyclopropanes by an MIRC Strategy: An Enantioselective Organocatalytic Approach to Spirocyclopropanes
Authors: Russo Alessio , Meninno Sara , Tedesco Consiglia , Lattanzi Alessandra
Published: July 2011

An efficient cyclopropanation, by a Michael‐initiated ring‐closing (MIRC) reaction of 2‐arylidene‐1,3‐indandiones and 2‐arylidene malononitriles, has been developed by using different α‐monohalogenated methylene active compounds with triethylamine. The first enantioselective cyclopropanation to spirocyclopropanes derived by the reaction of 2‐arylidene‐1,3‐indandiones and dimethyl bromomalonate with a commercially available α,α‐L‐diarylprolinol as the organocatalyst and K2CO3 as the additive has been accomplished. The spirocyclopropanes were isolated in high yield and up to 85 % ee. Notably, the asymmetric one‐pot sequential approach to spirocyclopropanes proved to be a feasible process.
Key aroma compounds in Shanxi aged tartary buckwheat vinegar and changes during its thermal processing
Authors: Aili Wang , Huanlu Song , Changzhong Ren , Zaigui Li
Published: July 2011

Shanxi aged vinegar differs from other vinegars in that it has a unique thermal process (6 days at 85°C) which creates its typical flavour. The present study evaluated the key aroma compounds of Shanxi aged vinegar made from tartary buckwheat by gas chromatography–olfactometry–mass spectrometry. The influence of the thermal processing on product aroma was also studied. A total of 45 compounds were detected of which 24 were correctly identified while 21 were tentatively identified: the remaining six were unknown. Of the 45 compounds detected, 13 have not been previously reported in other vinegars. Most of the aroma compounds identified increased significantly during the first 3 days of heating and then decreased from the fourth day onwards. The changes may have resulted from different kinds of reactions, such as the Maillard reaction and other associated reactions such as degradation of amino acids. This study would be helpful for optimizing the processing conditions of Shanxi aged vinegar. Copyright © 2011 John Wiley & Sons, Ltd.
Liver cancer and non‐Hodgkin lymphoma in hepatitis C virus‐infected patients: Results from the DANVIR cohort study
Authors: Omland Lars Haukali , Jepsen Peter , Krarup Henrik , Christensen Peer Brehm , Weis Nina , Nielsen Lars , Obel Niels , Sørensen Henrik Toft , Stuver Sherri Oliver ,   on behalf of the DANVIR cohort study
Published: July 2011

Hepatitis C virus (HCV)‐infection can cause hepatocellular carcinoma (HCC) and most likely non‐Hodgkin lymphoma (NHL). No studies have compared the risk of these cancers between patients with chronic and cleared HCV‐infection. The aim of this study was to estimate the 10‐year risk of HCC and NHL in HCV‐infected patients and to compare the risk of these cancers between HCV‐infected patients and the general population in Denmark and between patients with chronic and cleared HCV‐infection. Nationwide cohorts were used: 11,975 HCV‐infected patients in the DANVIR cohort and 71,850 individuals from an age‐ and gender‐matched general population cohort. Within DANVIR, 4,158 patients with chronic HCV‐infection and 2,427 patients with cleared HCV‐infection were studied. The 10‐year risks of HCC and NHL in HCV‐infected patients were 1.0% [95% confidence interval (CI): 0.8–1.3%] and 0.1% (95% CI: 0.1–0.2%), respectively. Compared to the general population, HCV‐infected patients had a 62.91‐fold increased risk of HCC (95% CI: 28.99–136.52), a 29.97‐fold increased risk of NHL during the first year of follow‐up (95% CI: 6.08–147.84), and a 1.26‐fold increased risk of NHL after the first year (95% CI: 0.36–4.41). Chronic HCV‐infection was associated with a 4.71‐fold increased risk of HCC (95% CI: 1.67–13.32) compared to cleared HCV‐infection; 5 and 0 events of NHL occurred in patients with chronic and cleared HCV‐infection, respectively. HCC‐risk is increased substantially in HCV‐infected patients compared to the general population. Chronic as opposed to cleared HCV‐infection increases the risk of HCC and perhaps NHL.
P‐glycoprotein binds to ezrin at amino acid residues 149–242 in the FERM domain and plays a key role in the multidrug resistance of human osteosarcoma
Authors: Brambilla Daria , Zamboni Silvia , Federici Cristina , Lugini Luana , Lozupone Francesco , Milito Angelo De , Cecchetti Serena , Cianfriglia Maurizio , Fais Stefano
Published: July 2011

Overexpression of the mdr1 gene encoding P‐glycoprotein (Pgp) exerts a major role in reducing the effectiveness of cytotoxic therapy in osteosarcoma. The interaction between actin and Pgp has been shown to be instrumental in the establishment of multidrug resistance (MDR) in human tumor cells. The cytoskeleton linker ezrin exerts a pivotal role in maintaining the functional connection between actin and Pgp. We investigated the role of ezrin in a human multidrug‐resistant osteosarcoma cell line overexpressing Pgp and compared it to its counterpart that overexpresses an ezrin deletion mutant. The results showed that Pgp binds at amino acid residues 149–242 of the N‐terminal domain of ezrin. The interaction between ezrin and Pgp occurs in the plasma membrane of MDR cells, where they also co‐localize with the ganglioside GM1 located in lipid rafts. The overexpression of the ezrin deletion mutant entirely restored drug susceptibility of osteosarcoma cells, consistent with Pgp dislocation to cytoplasmic compartments and abrogation of GM1/Pgp co‐localization at the plasma membrane. Our study provides evidence that ezrin exerts a key role in MDR of human osteosarcoma cells through a Pgp‐ezrin‐actin connection that is instrumental for the permanence of Pgp into plasma membrane lipid rafts. We also show for the first time that Pgp‐binding site is localized to amino acid residues 149–242 of the ezrin Band 4.1, Ezrin/Radixin/Moesin (FERM) domain, thus proposing a specific target for future molecular therapy aimed at counteracting MDR in osteosarcoma patients.
Carbidopa abrogates L‐dopa decarboxylase coactivation of the androgen receptor and delays prostate tumor progression
Authors: Wafa Latif A. , Cheng Helen , Plaa Nathan , Ghaidi Fariba , Fukumoto Takahiro , Fazli Ladan , Gleave Martin E. , Cox Michael E. , Rennie Paul S.
Published: July 2011

The androgen receptor (AR) plays a central role in prostate cancer progression to the castration‐resistant (CR) lethal state. L‐Dopa decarboxylase (DDC) is an AR coactivator that increases in expression with disease progression and is coexpressed with the receptor in prostate adenocarcinoma cells, where it may enhance AR activity. Here, we hypothesize that the DDC enzymatic inhibitor, carbidopa, can suppress DDC‐coactivation of AR and retard prostate tumor growth. Treating LNCaP prostate cancer cells with carbidopa in transcriptional assays suppressed the enhanced AR transactivation seen with DDC overexpression and decreased prostate‐specific antigen (PSA) mRNA levels. Carbidopa dose‐dependently inhibited cell growth and decreased survival in LNCaP cell proliferation and apoptosis assays. The inhibitory effect of carbidopa on DDC‐coactivation of AR and cell growth/survival was also observed in PC3 prostate cancer cells (stably expressing AR). In vivo studies demonstrated that serum PSA velocity and tumor growth rates elevated ∼2‐fold in LNCaP xenografts, inducibly overexpressing DDC, were reverted to control levels with carbidopa administration. In castrated mice, treating LNCaP tumors, expressing endogenous DDC, with carbidopa delayed progression to the CR state from 6 to 10 weeks, while serum PSA and tumor growth decreased 4.3‐fold and 5.4‐fold, respectively. Our study is a first time demonstration that carbidopa can abrogate DDC‐coactivation of AR in prostate cancer cells and tumors, decrease serum PSA, reduce tumor growth and delay CR progression. Since carbidopa is clinically approved, it may be readily used as a novel therapeutic strategy to suppress aberrant AR activity and delay prostate cancer progression.
α1‐3/4 fucosylation at Asn 241 of β‐haptoglobin is a novel marker for colon cancer: A combinatorial approach for development of glycan biomarkers
Authors: Park Seung‐Yeol , Lee Sung‐Hyeon , Kawasaki Nana , Itoh Satsuki , Kang Keunsoo , Hee Ryu Soo , Hashii Noritaka , Kim Jin‐Man , Kim Ji‐Yeon , Hoe Kim Jung
Published: July 2011

Aberrant glycosylation has been observed in many types of cancer, but the mechanism of glycosylation change is still poorly understood. To elucidate relationships between glycosylation and colon cancer progression, we analyzed glycosylation status of β‐haptoglobin (β‐Hp) obtained from 46 cancer patients, 14 inflammatory bowel disease patients and 38 normal subjects. Aleuria aurantia lectin reactivity with cancer β‐Hp was much higher than in the other two study groups. These results were confirmed by lectin blotting and microarray assay using other lectins directed to fucosyl residues. Levels of such glycans were correlated with stage of colon cancer progression. Reactivity with fucosylated glycans was eliminated by treatment with α1‐3/4 fucosidase but not α1‐6 fucosidase, indicating that enhanced lectin reactivity with the fucose moiety of colon cancer β‐Hp is due to Fucα1‐3/4GlcNAc. Moreover, site‐specific glycan occupancy was determined by sequential LC/MS analysis. Mass spectrometric analysis showed that fucosylation of β‐Hp was higher in colon cancer patients than in other subjects. In particular, fucosylation at Asn 241 of β‐Hp in sera of colon cancer patients was clearly higher than in the other groups, and the ratio of fucosylated glycopeptides containing Asn 241 decreased greatly after treatment with α1‐3/4 fucosidase. In conclusion, the level of α1‐3/4 fucosyl epitope at Asn 241 of β‐Hp is potentially useful as a novel marker for colon cancer.
Proviral loads of human T‐lymphotropic virus Type 1 in asymptomatic carriers with different infection routes
Authors: Ueno Shiro , Umeki Kazumi , Takajo Ichiro , Nagatomo Yasuhiro , Kusumoto Norio , Umekita Kunihiko , Morishita Kazuhiro , Okayama Akihiko
Published: July 2011

High human T‐lymphotropic virus Type 1 (HTLV‐1) proviral DNA load (PVL) has been reported to be one risk factor for the development of adult T‐cell leukemia/lymphoma (ATL). ATL is also believed to develop in HTLV‐1 carriers who acquire infection perinatally. ATL cells have been reported to frequently harbor defective provirus. In our study, PVLs for three different regions of HTLV‐1 provirus (5′LTR‐gag, gag and pX) were measured in 309 asymptomatic carriers with different infection routes. PVLs for the pX region in 21 asymptomatic carriers with maternal infection was significantly higher than in 24 carriers with spousal infection. Among 161 carriers with relatively high pX PVLs (equal to or greater than 1 copy per 100 peripheral blood mononuclear cells), 26 carriers (16%) had low gag PVL/pX PVL (less than 0.5) and four (2%) had low 5′LTR‐gag PVL/pX PVL (less than 0.5). Low gag PVL/pX PVL ratio, which reflects deficiency and/or polymorphism of HTLV‐1 proviral DNA sequences for the gag region, was also associated with maternal infection. These data suggest that HTLV‐1 carriers with maternal infection tend to have high PVLs, which may be related to provirus with deficiency and/or the polymorphism of proviral DNA sequences. In addition, there is a possibility that this ratio may be used as a tool to differentiate the infection routes of asymptomatic HTLV‐1 carriers, which supports the need for a large scale study.
High expression of Galectin‐1 in pancreatic stellate cells plays a role in the development and maintenance of an immunosuppressive microenvironment in pancreatic cancer
Authors: Tang Dong , Yuan Zhongxu , Xue Xiaofeng , Lu Zipeng , Zhang Ye , Wang Hui , Chen Minyong , An Yong , Wei Jishu , Zhu Yi , Miao Yi , Jiang Kuirong
Published: July 2011

Galectin‐1 is implicated in making tumor cells immune privileged, in part by regulating the survival of infiltrating T cells. Galectin‐1 is strongly expressed in activated pancreatic stellate cells (PSCs); however, whether this is linked to tumor cell immune escape in pancreatic cancer is unknown. Galectin‐1 was knocked down in PSCs isolated from pancreatic tissues using small interfering RNA (siRNA), or overexpressed using recombinant lentiviruses, and the PSCs were cocultured with T cells. CD3+, CD4+ and CD8+ T cell apoptosis was detected by flow cytometry; T cell IL‐2, IL‐4, IL‐5 and INF‐γ production levels were quantified using ELISA. Immunohistochemical analysis showed increased numbers of PSCs expressed Galectin‐1 (p < 0.01) and pancreatic cancers had increased CD3+ T cell densities (p < 0.01) compared to normal pancreas or chronic pancreatitis samples. In coculture experiments, PSCs that overexpressed Galectin‐1 induced apoptosis of CD4+ T cells (p < 0.01) and CD8+ T cells (p < 0.05) significantly, compared to normal PSCs. Knockdown of Galectin‐1 in PSCs increased CD4+ T cell (p < 0.01) and CD8+ T cell viability (p < 0.05). Supernatants from T cells cocultured with PSCs that overexpressed Galectin‐1 contained significantly increased levels of Th2 cytokines (IL‐4 and IL‐5, p < 0.01) and decreased Th1 cytokines (IL‐2 and INF‐γ, p < 0.01). However, the knockdown of PSC Galectin‐1 had the opposite effect on Th1 and Th2 cytokine secretion. Our study suggests that the overexpression of Galectin‐1 in PSCs induced T cell apoptosis and Th2 cytokine secretion, which may regulate PSC‐dependent immunoprivilege in the pancreatic cancer microenvironment. Galectin‐1 may provide a novel candidate target for pancreatic cancer immunotherapy.
Expression of the nuclear bile acid receptor/farnesoid X receptor is reduced in human colon carcinoma compared to nonneoplastic mucosa independent from site and may be associated with adverse prognosis
Authors: Lax Sigurd , Schauer Georg , Prein Kurt , Kapitan Magdalena , Silbert Dagmar , Berghold Andrea , Berger Anton , Trauner Michael
Published: July 2011

The nuclear bile acid receptor/farnesoid X receptor (FXR; NR1H4) is involved in bile acid homeostasis, cell proliferation and apoptosis and has been linked to intestinal carcinogenesis in mice. Aim of this study was to analyze FXR expression in human normal intestinal mucosa and colon carcinoma. We achieved systematic mapping of FXR expression of human intestinal mucosa and analysis of 75 human colon carcinomas using FXR immunohistochemistry on formalin‐fixed, paraffin‐embedded tissue. FXR expression gradually decreased from terminal ileum to the sigmoid colon with strongest expression in the terminal ileum (p < 0.001). FXR expression in carcinomas was reduced compared to peritumoral nonneoplastic mucosa (p < 0.000). Loss of FXR expression was significantly correlated with grading in tumors of the right colon (p = 0.008). FXR expression in tumor and normal tissue showed an inverse correlation with stage. FXR expression in tumor was inversely correlated with clinical outcome. No association was found with patients' age and sex. In nonneoplastic mucosa FXR expression concurred with low expression of Ki‐67. In carcinomas, no association was found between FXR expression and Ki‐67 and cyclin D1, respectively. Development of colon carcinoma in humans is associated with reduced FXR expression independent of site and may reflect an impaired defense against potentially carcinogenic bile acids along their intestinal gradient. In contrast to normal colon mucosa, FXR expression in carcinomas is not associated with low proliferation. Colon carcinomas with FXR expression seem to be associated with lower stage and a more favourable outcome compared to FXR negative carcinomas.
Long‐lasting immunoprotective and therapeutic effects of a hyperstable E7 oligomer based vaccine in a murine human papillomavirus tumor model
Authors: Cerutti María L. , Alonso Leonardo G. , Tatti Silvio , de Prat‐Gay Gonzalo
Published: July 2011

Cervical cancer and many other anogenital and oropharyngeal carcinomas are strongly associated with high‐risk human papillomavirus (HPV) persistent infections. HPV E7 oncoprotein is the major viral transforming factor, emerging as a natural candidate for immunotherapy, since it is constitutively expressed in HPV‐induced cancer cells. We have previously shown that E7 can self‐assemble into soluble and homogeneous spherical oligomers, named E7 soluble oligomers (E7SOs). These are highly resistant to thermal denaturation, providing an additional advantage given the demand for highly stable vaccine formulations. Here, we present a new chemically stabilized form of the E7SOs (E7SOx) and analyzed its effect in a murine HPV‐tumor model. Vaccination of female mice with low doses of E7SOx combined with a CpG‐rich oligonucleotide (ODN) as adjuvant elicits a strong long‐lasting protection against E7‐expressing tumor cells, preventing tumor outgrowth after rechallenge 90‐days later. Therapeutic experiments showed that E7SOx/ODN vaccination significantly delays tumor growth and extends the time of survival of the treated mice in a dose‐dependent manner. These proof‐of‐principle preclinical experiments denote the potential applicability of our E7SOx‐based vaccine to the treatment of cervical cancer and other mucosal HPV‐related neoplastic lesions. In addition to thermal, chemical and proteolysis stability, the combined recombinant and chemical modification nature of the E7SOx vaccine candidate, results in low‐cost, of particular interest in developing countries, where most of the cervical cancer cases occur and the most affected population is at reproductive age.
Prognostic significance of osteopontin in hepatocellular carcinoma: A meta‐analysis
Authors: Zhang Chuan‐Hai , Xu Ge‐Liang , Jia Wei‐Dong , Ge Yong‐Sheng , Li Jian‐Sheng , Ma Jin‐Liang , Ren Wei‐Hua
Published: July 2011

Osteopontin (OPN) has been implicated in tumor development and progression for several years. However, the prognostic value of OPN overexpression in patients with hepatocellular carcinoma (HCC) remains controversial. We performed a meta‐analysis to assess the relationship between OPN overexpression and clinical outcome of HCC. A meta‐analysis of seven studies (1,158 patients) was carried out to evaluate the association between OPN and overall survival (OS) and disease‐free survival (DFS) in HCC patients. The correlation between OPN and tumor vascular invasion or other invasion‐related parameters was also assessed. Data were synthesized with random effect model of DerSimonian and Laird, hazard ratio (HR) or odds ratio (OR) with its 95% confidence interval (CI) was used as the effect size estimate. Our analysis results indicated that high OPN expression predicted poor OS (HR: 1.37, 95% CI: 1.21–1.55) and DFS (HR: 1.62, 95% CI: 1.24‐2.11) of HCC. OPN overexpression tended to be associated with the presence of tumor vascular invasion (OR: 1.93, 95% CI: 0.97–3.84) and advanced tumor grade (OR: 1.74, 95% CI: 0.95–3.18). By this study, we conclude that OPN overexpression indicates a poor prognosis for patients with HCC, it may also have predictive potential for HCC invasion and metastasis.
Novel insights in basic and applied stem cell therapy
Authors: Siniscalco Dario , Giordano Antonio , Galderisi Umberto
Published: July 2011

The achievement of novel findings in stem cell research were the subject of the meeting organized by Stem Cell Research Italy (SCR Italy) and by the International Society for Cellular Therapy‐Europe (ISCT). Stem cell therapy represents great promise for the future of molecular and regenerative medicine. The use of several types of stem cells is a real opportunity to provide a valid approach to curing several untreatable human diseases. Before it is suitable for clinical applications, stem cell biology needs to be investigated further and in greater detail. Basic stem cell research could provide exact knowledge regarding stem cell action mechanisms, and pre‐clinical research on stem cells on an in vivo model of disease provides scientific evidence for future human applications. Applied stem cell research is a promising new approach to handling several diseases. Along with tissue engineering, it offers a new and promising discipline that can help to manage human pathologies through stem cell therapy. All of these themes were discussed in this meeting, covering stem cell subtypes with their newest basic and applied research. J. Cell. Physiol. 227: 2283–2286, 2012. © 2011 Wiley Periodicals, Inc.
Enhanced delivery of mda‐7/IL‐24 using a serotype chimeric adenovirus (Ad.5/3) in combination with the apogossypol derivative BI‐97C1 (Sabutoclax) improves therapeutic efficacy in low CAR colorectal cancer cells
Authors: Azab Belal , Dash Rupesh , Das Swadesh K. , Bhutia Sujit K. , Shen Xue‐Ning , Quinn Bridget A. , Sarkar Siddik , Wang Xiang‐Yang , Hedvat Michael , Dmitriev Igor P. , Curiel David T. , Grant Steven , Dent Paul , Reed John C. , Pellecchia Maurizio , Sarkar Devanand , Fisher Paul B.
Published: July 2011

Adenovirus (Ad)‐based gene therapy represents a potentially viable strategy for treating colorectal cancer. The infectivity of serotype 5 adenovirus (Ad.5), routinely used as a transgene delivery vector, is dependent on Coxsackie‐adenovirus receptors (CAR). CAR expression is downregulated in many cancers thus preventing optimum therapeutic efficiency of Ad.5‐based therapies. To overcome the low CAR problem, a serotype chimerism approach was used to generate a recombinant Ad (Ad.5/3) that is capable of infecting cancer cells via Ad.3 receptors in a CAR‐independent manner. We evaluated the improved transgene delivery and efficacy of Ad.5/3 recombinant virus expressing melanoma differentiation associated gene‐7/interleukin‐24 (mda‐7/IL‐24), an effective wide‐spectrum cancer‐selective therapeutic. In low CAR human colorectal cancer cells RKO, wild‐type Ad.5 virus expressing mda‐7/IL‐24 (Ad.5‐mda‐7) failed to infect efficiently resulting in lack of expression of MDA‐7/IL‐24 or induction of apoptosis. However, a recombinant Ad.5/3 virus expressing mda‐7/IL‐24 (Ad.5/3‐mda‐7) efficiently infected RKO cells resulting in higher MDA‐7/IL‐24 expression and inhibition of cell growth both in vitro and in nude mice xenograft models. Addition of the novel Bcl‐2 family pharmacological inhibitor Apogossypol derivative BI‐97C1 (Sabutoclax) significantly augmented the efficacy of Ad.5/3‐mda‐7. A combination regimen of suboptimal doses of Ad.5/3‐mda‐7 and BI‐97C1 profoundly enhanced cytotoxicity in RKO cells both in vitro and in vivo. Considering the fact that Ad.5‐mda‐7 has demonstrated significant objective responses in a Phase I clinical trial for advanced solid tumors, Ad.5/3‐mda‐7 alone or in combination with BI‐97C1 would be predicted to exert significantly improved therapeutic efficacy in colorectal cancer patients. J. Cell. Physiol. 227: 2145–2153, 2012. © 2011 Wiley Periodicals, Inc.
Characterization of a versatile rhizospheric organism from cucumber identified as Ochrobactrum haematophilum
Authors: Zhao Lei , Teng Songshan , Liu Yanping
Published: July 2011

Several rhizobacteria play a vital role in promoting plant growth and protecting plants against fungal diseases and degrading pesticides in the environment. In this study, a bacterial strain, designated H10, was isolated from the rhizosphere at Laixi in Shandong Province, China, and was identified as Ochrobactrum haematophilum based on API 20 NE tests and 16S rRNA gene sequence analysis. The plant growth‐promoting characteristics of the strain were further characterized, and the results showed that strain H10 produces siderophore, indol‐3‐acetic (IAA) and solubilized phosphate but lacks 1‐aminocyclopropane‐1‐carboxylate (ACC) deaminase activity. Inoculation with the strain was found to significantly increase (p < 0.05) the growth of cucumber in pot experiments. Strain H10 was assessed in vitro for antagonism against several pathogenic fungi and showed high antifungal activity. The cell‐free culture filtrates, which had high extracellular chitinase, β;‐1,3‐glucanase and protease activities, could inhibit the growth of all pathogenic fungi tested, indicating that growth suppression was partly due to extracellular antifungal metabolites present in the culture filtrates. Changes in hyphal morphology were observed in phytopathogenic fungi after treatment with the culture filtrates. Additionally, strain H10 was able to degrade 80%, 85% and 58% of the pesticides chlorpyrifos, β;‐cypermethrin and imidacloprid, respectively, within 60 h in liquid culture. The inoculation of strain H10 into soil treated with 100 mg kg–1of the three pesticides accordingly resulted in a higher degradation rate than in noninoculated soils. These results highlight the potential of this bacterium for use as a biofertilizer and biopesticide and suggest that it may provide an alternative to the use of chemical fertilizers and pesticides in agriculture. Additionally, it may represent a bioremediation agent that can remove contaminating chemical pesticide residues from the environment. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Surveillance of single‐cell behavior in different subpopulations of Ralstonia pickettii AR1 during growth and polyhydroxybutyrate production phases by flow cytometry
Authors: Shakeri Shahryar , Roghanian Rasoul , Emtiazi Giti
Published: July 2011

Most bacterial strains accumulate intracellular polyhydroxybutyrate (PHB) granules as an energy reservoir, in response to fluctuations in their microenvironment. Flow cytometry was applied for the analysis of single cells of Ralstonia pickettii AR1 in response to changes in the culture conditions. Two parameters, the PHB production‐related FL2 and side scatter (SSC) parameters, were used to monitor, distinguish and characterize different subpopulations in the growth and PHB production phases. A high SSC level was observed in the mid‐log exponential growth phase. When the nitrogen source reached a limiting level, the SSC started to decrease, in contrast to the intracellular PHB granules‐related FL2 parameter. The results show that ammonium limitation is a critical and important factor for the accumulation of reserve compounds. Four subpopulations were observed and distinguished upon entrance of the cells into the exponential growth phase. When the cells entered the late exponential growth or early stationary phase, two subpopulations had disappeared and only two, different subpopulations were dominant. One of the subpopulations with changed SSC and PHB production activity switched to another subpopulation that was only active in PHB production in the stationary phase. The whole cells of R. pickettii AR1 tended to form a homogeneous population at the end of the stationary phase. In fact, the changes in the subpopulations of a single strain are related to different physiological states of the cells. The observation of different subpopulations suggests that each subpopulation shows a specific response to changes in the surrounding microenvironment, nutrients and limiting factors. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
GC content‐independent amino acid patterns in Bacteria and Archaea
Authors: Schmidt Andre , Rzanny Michael , Schmidt Astrid , Hagen Matthias , Schütze Eileen , Kothe Erika
Published: July 2011

Every organism can be characterized by the amino acid composition of its proteome. So far it was assumed that these compositions are determined by the GC content of the DNA or, in some cases, by extreme lifestyles, like thermophily or halophily. Here, we focussed our analysis on eight amino acids, each of which is encoded by both, GC and AT rich codons, to identify finer amino acid patterns beyond the GC dominance. We investigated the conceptually translated proteomes of 1029 bacterial and archaeal strains with sequenced genomes for amino acid composition. Using correspondence analysis, we found that phylogenetic groups within bacteria and archaea generally can be discriminated from other groups due to their amino acid composition. In some cases, single organisms, e.g. Treponema pallidum strains or Mycoplasma penetrans, are characterized by extreme amino acid compositions. We assume that our data could provide a basis for a new approach to analyze evolution of bacterial and archaeal groups. Furthermore, for single organisms, the detailed knowledge of the amino acid composition of the entire proteome encoded in the genome could lead to a better understanding, important for pharmaceutical or biotechnological applications. We recommend that information about amino acid compositions should be provided in databases, comparable to the GC content of genomes. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
40‐O ‐[2‐Hydroxyethyl]rapamycin modulates human dendritic cell function during exposure to Aspergillus fumigatus
Authors: Bauer Ruth , Mezger Markus , Blockhaus Christian , Schmitt Anna‐Lena , Kurzai Oliver , Einsele Hermann , Loeffler Juergen
Published: July 2011

40‐O ‐[2‐Hydroxyethyl]rapamycin (RAD), a novel derivative of the immunosuppressive drug rapamycin, was analyzed for its immunomodulatory influence during the interaction of human monocyte‐derived dendritic cells (moDC) with Aspergillus fumigatus. RAD is clinically used to prevent graft‐versus ‐host disease as well as solid organ and bone marrow transplant rejection. However, it may constitute a risk factor for the development of opportunistic infections, such as invasive aspergillosis which is mainly caused by the most common airborne fungal pathogen A. fumigatus. moDC were generated in the presence or absence of RAD. In this setting, RAD had various modulating effects on the immune function of DC. A decrease of pro‐ and anti‐inflammatory cytokines (IL‐12, TNF‐α, CCL20, IL‐10) was observed. Furthermore, RAD reduced the expression of innate immunity receptors (TLR2, TLR4, dectin‐1), impaired the maturation capacity of moDC observed through the reduction of costimulatory factors (CD40, CD80, CD83, CD86), and impaired their capacity to phagocytose and damage A. fumigatus. These data demonstrate that RAD influences the differentiation of DC. RAD modulates the cytokine response of DC to A. fumigatus and reduces their ability to kill germ tubes. Thus, RAD treatment might affect the risk of invasive aspergillosis independently of its capacity of blocking T cell activation. (© 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Isolation and characterization of Klebsiella oxytoca strain degrading crude oil from a Tunisian off‐shore oil field
Authors: Chamkha Mohamed , Trabelsi Yosra , Mnif Sami , Sayadi Sami
Published: July 2011

A facultatively anaerobic, Gram‐negative, mesophilic, moderately halotolerant, non‐motile, and non‐sporulated bacterium, designated strain BSC5 was isolated from an off‐shore “Sercina” oil field, located near the Kerkennah island, Tunisia. Yeast extract was not required for growth. Phenotypic characteristics and phylogenetic analysis of the 16S rRNA gene sequence of strain BSC5 revealed that it was related to members of the genus Klebsiella, being most closely related to the type strain of K. oxytoca (99% sequence similarity). Strain BSC5 was capable of using aerobically the crude oil as substrate growth. The growth of strain BSC5 on crude oil was followed by measuring the OD600 nm and by enumeration of viable cells at different culture's time. GC‐MS analysis showed that strain BSC5 was capable of degrading a wide range of aliphatic hydrocarbons from C13 to C30. The biodegradation rate for n ‐alkanes reached 44% and 75%, after 20 and 45 days of incubation, respectively. Addition of the synthetic surfactant, Tween 80, accelerated the crude oil degradation. The biodegradation rate for n ‐alkanes reached 61% and 98%, after 20 and 45 days of incubation, respectively. Moreover, three aromatic compounds, p ‐hydroxybenzoate, protocatechuate and gentisate, were metabolized completely by strain BSC5 after 24 h, under aerobic conditions. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Development of Loop‐mediated isothermal amplification method for rapid detection of Streptococcus iniae, the causative agent of streptococcicosis in fish
Authors: Cai Shuang‐Hu , Wang Bei , Lu Yi‐Shan , Jian Ji‐Chang , Wu Zao‐He
Published: July 2011

Streptococcus iniae is a major pathogen that causes sever economic losses in tilapia aquaculture. A set of four specific primers was designed by targeting lctO gene. With Bst DNA polymerase, the target DNA can be clearly amplified for 60 min at 64 °C in a simple water bath. The sensitivity of the LAMP assay for the detection of S. iniae is about 12.4 cells per reaction in both of pure cultures and added fish tissues cultures. LAMP products could be judged with agar gel or naked eye after addition of SYBR Green I. There were no cross‐reactions with other bacterial strains indicating high specificity of the LAMP. The LAMP method was also applied to detect S. iniae ‐infected tilapia tissues effectively. The LAMP assay reported here indicates the potential usefulness of the technique as a valuable simple, rapid alternative procedure for the detection of S. iniae during streptococcicosis monitoring of cultured fish. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Identification and biocontrol efficacy of Streptomyces miharaensis producing filipin III against Fusarium wilt
Authors: Kim Jeong Do , Han Jae Woo , Hwang In Cheon , Lee Dongho , Kim Beom Seok
Published: July 2011

A number of bacterial strains were isolated from the internal tissue of Trapa japonica. Of these, strain KPE62302H, which had a 16S rDNA sequence identical to that of Streptomyces miharaensis showed antifungal activity against several plant pathogens. Treatment of seeds with strain KPE62302H induced a significant reduction in the incidence of Fusarium wilt in tomato plants compared with untreated controls. An antifungal substance (FP‐1) was purified from the culture extract of strain KPE62302H using C18 flash and Sephadex LH‐20 column chromatography and reverse phase HPLC. Extensive spectrometric analysis using MS and NMR identified this as filipin III. FP‐1 inhibited the mycelial growth of plant pathogenic fungi such as Alternaria mali, Aspergillus niger, Colletotrichum gloeosporioides, C. orbiculare, Cylindrocarpon destructans, Diaporthe citiri, Fusarium oxysporum at 1–10 μg ml–1 and also markedly inhibited the development of Fusarium wilt caused by F. oxysporum f.sp. lycopersici in tomato plants by treatment with 10 μg ml–1 under greenhouse conditions. The efficacy of FP‐1 against Fusarium wilt was comparable to that of the synthetic fungicide benomyl. An egfp ‐tagged strain of KPE62302H confirmed its ability to colonize tomato plants. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Multidrug resistance reversal effect of DMC derived from buds of Cleistocalyx operculatus in human hepatocellular tumor xenograft model
Authors: Huang Hai‐ya , Niu Jiang‐long , Lu Yan‐hua
Published: July 2011

BACKGROUND: Multidrug resistance (MDR) is a major obstacle in the chemotherapeutic treatment of many human cancers. 2′,4′‐Dihydroxy‐6′‐methoxy‐3′,5′‐dimethylchalcone (DMC) isolated from the buds of Cleistocalyx operculatus (Roxb.) Merr. et Perry (Myrtaceae), was investigated for its reversal effects on cancer cell MDR.

RESULTS: A human hepatocellular tumor xenograft model was established with the BEL‐7402/5‐FU cell line. Combined 5‐fluorouracil (5‐FU) and DMC (40 mg kg−1) treatment significantly elevated tumor inhibition rate to 72.2%. DMC could also increase 5‐FU concentrations in tumor tissues and increase caspase‐3 activity. Also, combined therapy resulted in enhanced tumor apoptotic and reduced proliferative activities relative to 5‐FU alone. Examining body weight and other signs of unwanted toxicity of the different treatment groups revealed no significant signs of adverse effects.

CONCLUSION: All results suggested that DMC reverses 5‐FU resistance, with a benign side effects profile. Copyright © 2011 Society of Chemical Industry
Polyunsaturated fatty acid levels in rat tissues after chronic treatment with dietetic oils
Authors: Rustichelli Cecilia , Avallone Rossella , Campioli Enrico , Braghiroli Daniela , Baraldi Mario
Published: July 2011

BACKGROUND: The essential fatty acids can be helpful in the prevention of several pathologies. The purpose of this study was to quantify the major n‐3 and n‐6 fatty acids in tissues of rats fed with flaxseed oil and with a dietetic oil in order to evaluate how their chronic supplementation could influence the correspondent in vivo levels and to study the effectiveness of the dietetic oil compared to flaxseed oil.

RESULTS: Fatty acids were successfully extracted from biological samples, subjected to derivatization procedure and analysed by high‐performance liquid chromatography with UV detection under gradient elution mode. The developed method showed good linearity, precision and accuracy, with recoveries ranging from 89% to 92%. Animals treated with flaxseed and dietetic oils showed enhanced levels of n‐3 fatty acids compared to control groups, with significantly higher levels of eicosapentaenoic acid and docosahexaenoic acid in the brain and in the adipose tissue of the dietetic group compared to the flaxseed group.

CONCLUSION: The obtained data underline that the tested oils can effectively enhance the tissue levels of n‐3 fatty acids and therefore they could be successfully used in the dietetic treatment of lipid‐related diseases. Copyright © 2011 Society of Chemical Industry
Affinity capillary electrophoresis and quantum mechanical calculations applied to the investigation of hexaarylbenzene‐based receptor binding with lithium ion
Authors: Ehala Sille , Toman Petr , Rathore Rajendra , Makrlík Emanuel , Kašička Václav
Published: July 2011

In this study, two complementary approaches, affinity capillary electrophoresis (ACE) and quantum mechanical density functional theory (DFT) calculations, have been employed for quantitative characterization and structure elucidation of the complex between hexaarylbenzene (HAB)‐based receptor R and lithium ion Li+. First, by means of ACE, the apparent binding constant of LiR+ complex (K) in methanol was determined from the dependence of the effective electrophoretic mobilities of LiR+ complex on the concentration of lithium ions in the 25 mM Tris/50 mM chloroacetate background electrolyte (BGE) using non‐linear regression analysis. Prior to regression analysis, the effective electrophoretic mobilities of the LiR+ complex were corrected to reference temperature 25°C and constant ionic strength 25 mM. The apparent binding constant of the LiR+ complex in the above methanolic BGE was evaluated as logK = 1.15±0.09. Second, the most probable structures of nonhydrated LiR+ and hydrated LiR+·3H2O complexes were derived by DFT calculations. The optimized structure of the hydrated LiR+·3H2O complex was found to be more realistic than the nonhydrated LiR+ complex because of the considerably higher binding energy of LiR+·3H2O complex (500.4 kJ/mol) as compared with LiR+ complex (427.5 kJ/mol).
One‐step isolation of γ‐oryzanol from rice bran oil by non‐aqueous hydrostatic countercurrent chromatography
Authors: Angelis Apostolis , Urbain Aurélie , Halabalaki Maria , Aligiannis Nektarios , Skaltsounis Alexios‐Leandros
Published: July 2011

The value‐added γ‐oryzanol was purified in one step from crude rice bran oil (RBO) using a preparative hydrostatic countercurrent chromatography (hydrostatic CCC) method, operating in the dual mode. The fractionation was performed using a non‐aqueous biphasic solvent system consisting of heptane–acetonitrile–butanol (1.8:1.4:0.7, v/v/v), leading rapidly to the target compounds. Transfer of the analytical CCC method to large‐scale isolation was also carried out yielding a high quantity‐high purity fraction of γ‐oryzanol. In addition, a fraction of hydroxylated triterpene alcohol ferulates (polar γ‐oryzanol) was clearly separated and obtained. Furthermore, a fast HPLC‐APCI(±)‐HRMS method was developed and applied for the identification of γ‐oryzanol as well as the polar γ‐oryzanol in RBO and the resulting fractions. The purity of γ‐oryzanol fraction was estimated as 97% based on HPLC‐APCI‐HRMS analysis.