bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of [5]Rotaxanes Containing Bi‐ and Tridentate Coordination Sites in the Axis
Authors: Collin Jean‐Paul , Durot Stéphanie , Keller Michel , Sauvage Jean‐Pierre , Trolez Yann , Cetina Mario , Rissanen Kari
Published: November 2010

A new example of a linear [5]rotaxane has been synthesized by using the traditional “gathering‐and‐threading” approach but based on an unusual axle incorporating a symmetrical bis(bidentate) chelating fragment built on a 4,7‐phenanthroline core. The stoppering reaction is particularly noteworthy since, instead of using a trivial bulky stopper as precursor to the blocking group, two semistoppered copper‐complexed [2]pseudorotaxanes (namely [2]semirotaxanes) are used, which leads to the desired [5]rotaxane in good yield. The efficiency of the method relies on the use of “click” chemistry, with its very mild conditions, and on the protection by a transition‐metal (copper(I)) of the various coordinating groups present in the fragments to be interconnected (terpy and bidentate chelating groups), thus inhibiting potential detrimental side reactions during the copper‐catalyzed stoppering reaction. Since the external fragments and the central core of the system contain tri‐ and bidentate chelating units, respectively, the axle of the final [5]rotaxane incorporates two types of coordinating units: two external terpy groups (terpy: 2,2′:6′,2′′‐terpyridine) and two central bidentate ligands. Such a situation enables the system to tidy two different metals centers, and to localize them in a priori well‐defined positions. This is what was observed when mixing the free ligand with a mixture of Zn2+ and Li+: the zinc(II) ions were unambiguously shown to occupy the external sites, whereas the Li+ cations were found in the central part of the [5]rotaxane. An X‐ray diffraction study carried out on a [3]pseudorotaxane, the axis of which is similar to the central part of the [5]rotaxane axle, demonstrates that Zn2+ is clearly five‐coordinate, the fifth ligand being a counterion, even when the coordination site of the pseudorotaxane is designed for four‐coordinate metals, which is in marked contrast with copper(I) or Li+.
Smooth Photocatalytic Preparation of 2‐Substituted 1,3‐Benzodioxoles
Authors: Ravelli Davide , Albini Angelo , Fagnoni Maurizio
Published: November 2010

A mild and general method for the synthesis of potentially bioactive 2‐substituted‐1,3‐benzodioxoles is presented. This is based on the photocatalyzed activation of methylene hydrogen atoms in the presence of tetrabutylammonium decatungstate (TBADT). The method gave yields ranging from 46–77 % with no interference by benzene ring substituents, such as OR, COOMe, Me, or CHO. The OH group interfered, but protection regenerated the reactivity. 5‐Chloro‐1,3‐benzodioxole was converted into a safrole derivative through a one‐pot process involving two consecutive irradiations, at 366 nm for the photocatalyzed alkylation at position 2 and at 310 nm for the alkylation at position 5.
Laparoscopic extraperitoneal para‐aortic lymphadenectomy in locally advanced cervical cancer1
Authors: Ramirez Pedro T. , Jhingran Anuja , Macapinlac Homer A. , Euscher Elizabeth D. , Munsell Mark F. , Coleman Robert L. , Soliman Pamela T. , Schmeler Kathleen M. , Frumovitz Michael , Ramondetta Lois M.
Published: November 2010

BACKGROUND:

Failure to detect metastasis to para‐aortic nodes in patients with locally advanced cervical cancer leads to suboptimal treatment. No previous studies have prospectively compared positron emission tomography (PET)/computed tomography (CT) with laparoscopic extraperitoneal staging in the evaluation of para‐aortic lymph nodes.

METHODS:

Sixty‐five patients were enrolled; 60 were available for analysis. Patients with stage IB2‐IVA cervical cancer without evidence of para‐aortic lymphadenopathy on preoperative CT or magnetic resonance imaging (MRI) were prospectively enrolled. All patients underwent preoperative PET/CT. Laparoscopic extraperitoneal lymphadenectomy was performed from the common iliac vessels to the left renal vein.

RESULTS:

The median age at diagnosis was 48 years (range, 23‐84). The median operative time was 140 minutes (range, 89‐252). The median blood loss was 22.5 mL (range, 5‐150). The median length of hospital stay was 1 day (range, 0‐4). The median number of lymph nodes retrieved was 11 (range, 1‐39). Fourteen (23%) patients had histopathologically positive para‐aortic nodes. Of the 26 patients with negative pelvic and para‐aortic nodes on PET/CT, 3 (12%) had histopathologically positive para‐aortic nodes. Of the 27 patients with positive pelvic but negative para‐aortic nodes on PET/CT, 6 (22%) had histopathologically positive para‐aortic nodes. The sensitivity and specificity of PET/CT in detecting positive para‐aortic nodes when nodes were negative on CT or MRI were 36% and 96%, respectively. Eleven (18.3%) patients had a treatment modification based on surgical findings.

CONCLUSIONS:

Laparoscopic extraperitoneal para‐aortic lymphadenectomy is safe and feasible. Surgical staging of patients with locally advanced cervical cancer should be considered before planned radiation and chemotherapy. Cancer 2011. © 2010 American Cancer Society.
Local control, survival, and late toxicities of locally advanced nasopharyngeal carcinoma treated by simultaneous modulated accelerated radiotherapy combined with cisplatin concurrent chemotherapy
Authors: Xiao Wei‐Wei , Huang Shao‐Min , Han Fei , Wu Shao‐Xiong , Lu Li‐Xia , Lin Cheng‐Guang , Deng Xiao‐Wu , Lu Tai‐Xiang , Cui Nian‐Ji , Zhao Chong
Published: November 2010

BACKGROUND:

The aim of this phase 2 study was to determine the long‐term local control, survival, and late toxicities among patients with locally advanced nasopharyngeal carcinoma (NPC) treated with intensity‐modulated radiotherapy (IMRT) with the simultaneous modulated accelerated radiation therapy (SMART) boost technique and concurrent chemotherapy.

METHODS:

Eighty‐one patients with pathologically diagnosed locally advanced NPC were enrolled in this study. IMRT was delivered with the SMART boost technique at prescribed doses of 68 grays (Gy)/30 fraction to the nasopharynx gross target volume. Concurrent cisplatin chemotherapy (80 mg/m2/d on Days 1 and 22) was administered.

RESULTS:

The mean actual physical dose delivered to the nasopharynx gross target volume was 73.8 Gy, and the mean biologically effective dose (BED) for the nasopharynx gross target volume was 84.8 Gy. With a median follow‐up of 54 months, 4 (4.9%) patients experienced local recurrence. The 5‐year local control rate was 94.9%. Eighteen patients died. Among them, 66.7% died of distant metastasis. The 5‐year disease‐free and overall survivals were 76.7% and 74.5%, respectively. The most common late toxicities among 68 patients with ≥4 years follow‐up were grade 1‐2 xerostomia, hearing loss, skin dystrophy, and subcutaneous fibrosis. No grade 4 late toxicities were noted.

CONCLUSIONS:

IMRT with SMART to enhance BED and concurrent chemotherapy is feasible in patients with locally advanced NPC. Long‐term results showed excellent local control with fewer late toxicities, although no further improvement was noted in overall survival, and the major cause of death was distant metastasis. Exploration of more effective combined chemoradiation strategies is warranted. Cancer 2011. © 2010 American Cancer Society.
Expression of glutamate and inhibitory amino acid vesicular transporters in the rodent auditory brainstem
Authors: Ito Tetsufumi , Bishop Deborah C. , Oliver Douglas L.
Published: November 2010

Glutamate is the main excitatory neurotransmitter in the auditory system, but associations between glutamatergic neuronal populations and the distribution of their synaptic terminations have been difficult. Different subsets of glutamatergic terminals employ one of three vesicular glutamate transporters (VGLUT) to load synaptic vesicles. Recently, VGLUT1 and VGLUT2 terminals were found to have different patterns of organization in the inferior colliculus, suggesting that there are different types of glutamatergic neurons in the brainstem auditory system with projections to the colliculus. To positively identify VGLUT‐expressing neurons as well as inhibitory neurons in the auditory brainstem, we used in situ hybridization to identify the mRNA for VGLUT1, VGLUT2, and VIAAT (the vesicular inhibitory amino acid transporter used by GABAergic and glycinergic terminals). Similar expression patterns were found in subsets of glutamatergic and inhibitory neurons in the auditory brainstem and thalamus of adult rats and mice. Four patterns of gene expression were seen in individual neurons. 1) VGLUT2 expressed alone was the prevalent pattern. 2) VGLUT1 coexpressed with VGLUT2 was seen in scattered neurons in most nuclei but was common in the medial geniculate body and ventral cochlear nucleus. 3) VGLUT1 expressed alone was found only in granule cells. 4) VIAAT expression was common in most nuclei but dominated in some. These data show that the expression of the VGLUT1/2 and VIAAT genes can identify different subsets of auditory neurons. This may facilitate the identification of different components in auditory circuits. J. Comp. Neurol. 519:316‐340, 2011. © 2010 Wiley‐Liss, Inc.
2D Measurement of Edge Plasma Dynamics by Using High‐Speed Camera Based on HeI Line Intensity Ratio Method
Authors: Ohno N. , Miyamoto K. , Kodama K. , Kajita S. , Kitagawa K.
Published: November 2010

In order to investigate dynamic plasma behaviors in scrape‐off layer (SOL) regions, such as ELMs and plasma blobs, we have developed a 4‐channel high‐speed camera equipped with four interference filters to obtain 2D successive images of spectral intensity of He I emission lines at 667.8 nm, 706.5 nm and 728.1 nm in addition to HeII at 656.0 nm. By using calculated results based on a Collisional Radiative (CR) model for He, the intensity ratios of the He I lines (728.1/706.5 and 667.8/706.5), provides 2D distributions of the electron temperature and density. The 2D profiles of electron density and temperature reconstructed from 2D images of He I emission lines taken in a small tokamak, HYBTOK‐II were compared with those measured with the triple probe in order to evaluate the accuracy of this diagnostics. Electron density and temperature measured with the 4‐channel high‐speed camera are in good agreement with those obtained with the triple probe (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Excitation Mechanisms and Sheath Dynamics in Capacitively Coupled Radio‐Frequency Oxygen Plasmas
Authors: Dittmann K. , Matyash K. , Nemschokmichal S. , Meichsner J. , Schneider R.
Published: November 2010

Phase resolved optical emission spectroscopy (PROES) was used to determine the spatio‐temporal behavior of excitation rates in rf sheaths of capacitively coupled plasmas at 13.56 MHz. The plasmas were ignited in pure oxygen at pressures from 20 to 100 Pa and rf powers from 10 to 100 W. The spatial and phase resolved excitation rates have shown four characteristic patterns, which differing in their spatial and temporal position.

PIC‐MCC simulations of the oxygen capacitively coupled radio‐frequency discharge were used to get a detailed microscopic description of the dynamic processes in rf plasmas. The PIC‐MCC simulations reproduced the excitation patterns observed in experiment quite well and allowed to identify the underlying physics.

Three excitation patterns appearing in front of the powered electrode were found tobe due to electron impact dissociative excitation of molecular Oxygen, whereas the fourth pattern very close to the powered electrode is attributed to collisions of the positive ions with the background gas (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Progress in the Theory of Dense Strongly Coupled Plasmas
Authors: Redmer R. , Röpke G.
Published: November 2010

We report the great progress in the physics of dense strongly coupled plasmas that has been achieved over the past decade. Driven by ambitious inertial confinement fusion programs worldwide, matter at unprecedented high energy densities is generated and studied at various research facilities. Furthermore, progress in computational capacity has allowed to apply ab initio methods to strongly coupled systems so that a better understanding of the complex interplay between quantum and correlation effects is possible. However, despite this great progress many problems in the theory of dense strongly coupled plasmas have not been solved yet and new challenges in the theoretical treatment of many‐particle systems have appeared. We discuss these problems and possible approaches. Since progress in analytical methods can still be accomplished, and computational methods allow to evaluate basic expressions for a broad parameter domain, a combination of both methods gives presently an optimal approach to thermodynamic and transport properties of dense strongly coupled plasmas. This will have a strong impact on technical applications such as the diagnostics of inertial confinement fusion plasmas via Thomson scattering and other spectroscopic methods, but is also of paramount importance in astrophysics when modeling the interior of solar and extrasolar giant planets as well as other compact objects (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Hydrocarbon Collision Cross Sections for Magnetic Fusion: The Methane, Ethane and Propane Families
Authors: Reiter D. , Janev R. K.
Published: November 2010

Kinetic analysis of the behaviour of hydrocarbons in fusion plasmas requires knowledge of cross sections for their most relevant collision processes. Upgrades and extensions of the commonly used HYDKIN (HYDride KINetics) online cross section database (based on compilations in 2002) and its data analysis toolbox for collisions of hydrocarbons with electrons and protons in relevant fusion plasma conditions are presented. Newly identified channels for dissociative recombination provide strongly reduced D/XB ratios for Gerd‐Band spectroscopy at low temperatures compared to earlier results, in accordance with experimental findings (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Proteomic identification and comparative analysis of asymmetrically arginine‐methylated proteins in immortalized, young and senescent cells
Authors: Lim Yongchul , Hong Eunyoung , Kwon Daeho , Lee Eunil
Published: November 2010

Protein‐arginine methylation is one of the modifications that yields mono and dimethyl (asymmetric or symmetric) arginine residues in proteins. Previously, we found that asymmetric arginine methylation is decreased proportionately with a decrease of cell proliferation potential of cells, and such arginine methylation is greatest in immortalized cells, followed by normal young cells, and lowest in replicatively senescent cells. Using an asymmetric dimethyl‐arginine‐specific antibody, we identified arginine‐methylated proteins in these cell types by immunoprecipitation and 2‐D immunoblotting followed by MS. As a result, arginine methylation of chaperone molecules and RNA‐binding proteins was differentially regulated between immortalized or young cells and senescent cells. Immortalized cells had significantly higher levels of methyl‐accepting proteins, such as cleavage stimulation factor 2 (CstF2) and heterogenous nuclear ribonucleoprotein (hnRNP) R, than young cells. However, senescent cells contained hypomethylaed CstF2, hnRNP K, and chaperone containing TCP1 subunit 7, as well as decreased hnRNP R level. Further, significant reduction of arginine modification in CstF2 and chaperone containing TCP1 subunit 7 was observed in prematurely senescent fibroblasts, induced by treatment with adenosine dialdehyde, a transmethylation inhibitor, or subcytotoxic concentration of H2O2. These results suggest that asymmetric modification of RNA‐binding proteins and molecular chaperones plays an essential role in maintaining cell proliferation capability.
Mapping spatial aggregation from counts data: a penalized likelihood approach
Authors: Grevstad Nels
Published: November 2010

A model for counts data on a spatial domain is presented. The counts are assumed to follow the negative binomial distribution, with both the mean and dispersion parameter allowed to vary spatially. The ratio of the mean to the dispersion parameter is estimated via non‐parametric penalized likelihood regression, and is equivalent to David and Moore's ecological index of aggregation. The dispersion parameter is assumed to follow a parametric model whose parameters are estimated via minimization of a variant of the well‐known generalized approximate cross‐validation (GACV) score, which is simultaneously used to estimate the smoothing parameter associated with estimation of the mean to dispersion parameter ratio. The increased flexibility of the model permitted by allowing the dispersion parameter to vary spatially is illustrated through spatial maps of David and Moore's and other indices of aggregation, such as Lloyd's indices of crowding and patchiness, using real and simulated data. Copyright © 2010 John Wiley & Sons, Ltd.
The spatial variation of weathering and soil depth across a Triassic sandstone outcrop
Authors: Tye A. M. , Lawley R. L. , Ellis M. A. , Rawlins B. G.
Published: November 2010

In this study, we used an archive of borehole logs from the British Geological Survey to collect information on the spatial structure of weathering that extends from the surface to competent bedrock across the Triassic Sherwood Sandstone Group outcrop (750 km2), in the East Midlands, UK. The borehole logs were used to estimate the thickness of the soil (n = 280) and soil and saprolite (S&S) to competent rock (n = 500). The weathering profile of the sandstone consisted of soil (median thickness ∼ 1·5 m) overlying a transition zone of compacted and weakly cemented weathered sandstone saprolite over bedrock. Topographic analysis using a NEXTMAP 5 m × 5 m digital elevation model (DEM) revealed no significant relationships between slope properties (relief, flow length, flow accumulation or slope angle) and soil or S&S thickness. A weak, but statistically significant correlation was found between the thickness of the soil and S&S (rs = 0·25, p < 0·001, n = 192). The variation in soil thickness may be related to changes in current and historic and land‐use, variation in sandstone properties and the influence of glacial/peri‐glacial processes. The thickness of the saprolite was more variable towards the southern part of the study area, where it increased to a maximum 40 m. We hypothesize and provide evidence that the greater weathering thickness is related to the occurrence of increased faulting in this part of the study region, allowing increased access to meteoric waters. A possible source of increased water supply is meltwater from Quaternary ice sheets; the overburden of ice may have increased sub‐glacial pore water pressure, with the fractures and faults acting as a drainage system for the removal of dissolved weathering products. British Geological Survey © NERC 2010
Regulatory consideration of bioavailability for metals: Simplification of input parameters for the chronic copper biotic ligand model
Authors: Peters Adam , Merrington Graham , de Schamphelaere Karel , Delbeke Katrien
Published: November 2010

The chronic Cu biotic ligand model (CuBLM) provides a means by which the bioavailability of Cu can be taken into account in assessing the potential chronic risks posed by Cu at specific freshwater locations. One of the barriers to the widespread regulatory application of the CuBLM is the perceived complexity of the approach when compared to the current systems that are in place in many regulatory organizations. The CuBLM requires 10 measured input parameters, although some of these have a relatively limited influence on the predicted no-effect concentration (PNEC) for Cu. Simplification of the input requirements of the CuBLM is proposed by estimating the concentrations of the major ions Mg2+, Na+, K+, , Cl−, and alkalinity from Ca concentrations. A series of relationships between log10 (Ca, mg l−1) and log10 (major ion, mg l−1) was established from surface water monitoring data for Europe, and applied in the prediction of Cu PNEC values for some UK freshwater monitoring data. The use of default values for major ion concentrations was also considered, and both approaches were compared to the use of measured major ion concentrations. Both the use of fixed default major ion concentrations, and major ion concentrations estimated from Ca concentrations, provided Cu PNEC predictions which were in good agreement with the results of calculations using measured data. There is a slight loss of accuracy when using estimates of major ion concentrations compared to using measured concentration data, although to a lesser extent than when fixed default values are applied. The simplifications proposed provide a practical evidence-based methodology to facilitate the regulatory implementation of the CuBLM. Integr Environ Assess Manag 2011;7:437–444. © 2011 SETAC
Alcohol consumption and risk of glioblastoma; evidence from the Melbourne collaborative cohort study
Authors: Baglietto Laura , Giles Graham G. , English Dallas R. , Karahalios Amalia , Hopper John L. , Severi Gianluca
Published: November 2010

Despite the brain being highly susceptible to the action of alcohol and, therefore, potentially susceptible to its carcinogenic effects, it is not clear whether alcohol consumption is associated with risk of glioblastoma. We analyzed data from 39,766 participants of the Melbourne Collaborative Cohort Study recruited in 1990–1994 and followed to the end of 2008 for an average of 15 years. Incidence of glioblastoma of the brain was ascertained via linkage to the Victorian and other State cancer registries in Australia. During a structured face‐to‐face interview at baseline we elicited each participant's history of alcoholic beverage consumption during the current decade at baseline. We used Cox regression models with age as the time metric, adjusted for country of birth, sex, total energy intake, educational attainment and coffee consumption to estimate hazard ratios (HR) and corresponding 95% confidence intervals (CI). A total of 67 glioblastomas was diagnosed in the cohort during follow‐up. The HRs associated with each additional 10 grams per day of alcohol intake was 1.16 (95% CI, 1.05 to 1.29; p for linear trend = 0.007). Compared to lifetime abstainer, the HR for glioblastoma associated with alcohol consumption were 1.07 (0.55 to 2.10) for 1 to 19 g/day, 1.79 (0.81 to 3.95) for 20 to 39 g/day, 3.07 (1.26 to 7.47) for 40 to 59 g/day and 2.54 (0.92 to 7.00) for 60 or more g/day. Alcohol consumption at baseline was associated with the risk of glioblastoma in a dose‐response relationship.
Autophagy‐mediated chemosensitization by cysteamine in cancer cells
Authors: Wan Xiao‐Mei , Zheng Fang , Zhang Li , Miao Yan‐Yan , Man Na , Wen Long‐Ping
Published: November 2010

Cysteamine (CS) has many biomedical and clinical applications because of its excellent water solubility, low cytotoxicity and good biocompatibility. A previous study by Brawer et al. reported the occurrence of many Gomori inclusion bodies in CS‐treated astrocytes, which would suggest the induction of autophagy. Here we provided a comprehensive line of evidence demonstrating that CS caused autophagosome accumulation in cancer cells. CS exerted a biphasic effect on the autophagy process, increasing the formation of autophagosomes in the early phase and blocking the autophagic degradation in a later phase. Furthermore, we showed that CS sensitized doxorubicin‐elicited chemotherapeutic killing in HeLa, B16 melanoma and doxorubicin‐resistant MCF‐7 cells and also enhanced chemotherapeutic efficacy of doxorubicin in a mouse melanoma model. Finally, we demonstrated that the chemosensitizing effect of CS was at least partly dependent on its ability to modulate autophagy. Our results revealed a novel biological function for CS in enhancing the chemotherapeutic effect of doxorubicin through autophagy modulation and pointed to the potential use of CS in adjunct cancer chemotherapy.
UV irradiation of immunized mice induces type 1 regulatory T cells that suppress tumor antigen specific cytotoxic T lymphocyte responses
Authors: Toda Masaaki , Wang Linan , Ogura Suguru , Torii Mie , Kurachi Makoto , Kakimi Kazuhiro , Nishikawa Hiroyoshi , Matsushima Kouji , Shiku Hiroshi , Kuribayashi Kagemasa , Kato Takuma
Published: November 2010

We previously showed that exposure to UV radiation after immunization suppresses Th1 and Th2 immune responses, leading to impaired Ab and allo‐immune responses, but the impact of UV radiation after immunization on anti‐tumor immune responses mediated by tumor‐specific CD8+ T cell responses remains less clear. Furthermore, the exact phenotypic and functional characteristics of regulatory T cell population responsible for the UV‐induced immunosuppression still remain elusive. Using the MBL‐2 lymphoma cell line engineered to express OVA as a surrogate tumor Ag, here we demonstrate that UV irradiation after tumor Ag‐immunization suppresses the anti‐tumor immune response in a manner dependent on the immunizing Ag. This suppression was mediated by interleukin (IL)‐10 released from CD4+CD25+ T cells, by which impaired the induction of cytotoxic T lymphocytes (CTL) able to kill Ag‐expressing tumor cells. In addition, we generated a panel of T cell clones from UV‐irradiated and non‐irradiated mice, and all of the clones derived from UV‐irradiated mice had a Tr1‐type regulatory T cell phenotype with expression of IL‐10 and c‐Maf, but not Foxp3. These Tr1‐type regulatory T cell clones suppressed tumor rejection in vivo as well as Th cell activation in vitro in an IL‐10 dependent manner. Given that suppression of Ag‐specific CTL responses can be induced in Ag‐sensitized mice by UV irradiation, our results may imply that exposure to UV radiation during premalignant stage induces tumor‐Ag specific Tr1 cells that mediate tumor‐Ag specific immune suppression resulting in the promotion of tumor progression.
MAGE‐A10 is a nuclear protein frequently expressed in high percentages of tumor cells in lung, skin and urothelial malignancies
Authors: Schultz‐Thater Elke , Piscuoglio Salvatore , Iezzi Giandomenica , Le Magnen Clémentine , Zajac Paul , Carafa Vincenza , Terracciano Luigi , Tornillo Luigi , Spagnoli Giulio C.
Published: November 2010

MAGE‐A10 is a highly immunogenic member of the MAGE‐A family of cancer/testis tumor‐associated antigens (C/T TAAs). Studies performed with broadly reactive antibodies have helped to initially characterize this TAA. However, no specific reagents have been developed so far, thus preventing a thorough analysis of its expression in healthy and tumoral tissues. We have produced MAGE‐A10 gene product in soluble recombinant form, and we have used it to generate specific monoclonal antibodies (mAbs). One of these reagents, recognizing an epitope located at the COOH terminus of the MAGE‐A10 gene product, was used to stain a multitumor tissue microarray comprising more than 2,500 paraffin‐embedded specimens including healthy tissues, benign tumors and malignancies of different histological origin. MAGE‐A10 protein was identified as an intranuclear protein of an apparent molecular weight of 70 kDa, expressed in normal spermatogonia and spermatocytes but in no other healthy tissue. Most importantly, this C/T TAA appears to be expressed in high (>50%) percentages of cancer cells from a number of malignancies, including lung, skin and urothelial tumors. Unexpectedly, high expression of MAGE‐A10 TAA at the protein level was also detectable in gynecological malignancies and stomach and gall bladder cancers. The characterization of MAGE‐A10‐specific reagents might set the stage for the development of targeted active immunotherapy by clarifying potential indications and by allowing the selection of patients eligible for treatment and the monitoring of its effectiveness.
Direct visualization of repair of oxidative damage by OGG1 in the nuclei of live cells
Authors: Zielinska Agnieszka , Davies Owain T. , Meldrum Rosalind A. , Hodges Nikolas J.
Published: November 2010

Oxidative DNA damage caused by intracellular reactive oxygen species (ROS) is widely considered to be important in the pathology of a range of human diseases including cancer as well as in the aging process. A frequently occurring mutagenic base lesion produced by ROS is 8‐oxo deoxyguanine (8‐oxo dG) and the major enzyme for repair of 8‐oxo dG is 8‐oxoguanine‐DNA glycosylase 1 (OGG1). There is now substantial evidence from bulk biochemical studies that a common human polymorphic variant of OGG1 (Ser326Cys) is repair deficient, and this has been linked to individual risk of pathologies related to oxidative stress. In the current study, we have used the technique of multiphoton microscopy to induce highly localized oxidative DNA damage in discrete regions of the nucleus of live cells. Cells transfected with GFP‐tagged OGG1 proteins demonstrated rapid (<2 min) accumulation of OGG1 at sites of laser‐induced damage as indicated by accumulation of GFP‐fluorescence. This was followed by repair as evidenced by loss of the localized fluorescence over time. Quantification of the rate of repair confirmed that the Cys326 variant of OGG1 is repair deficient and that the initial repair rate of damage by Cys326 OGG1 was 3 to 4 fold slower than that observed for Ser326 OGG1. These values are in good agreement with kinetic data comparing the Ser326 and Cys326 proteins obtained by biochemical studies. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 25:1–7 2011; View this article online at wileyonlinelibrary.com. DOI 10.1002/jbt.20346
Effects of the crotalus durissus terrificus snake venom on hepatic metabolism and oxidative stress
Authors: Silva Jonas Golart da , Soley Bruna da Silva , Gris Vanessa , Pires Amanda do Rocio Andrade , Caderia Silvia Maria Suter Correia , Eler Gabrielle Jackin , Hermoso Aparecida Pinto Munhos , Bracht Adelar , Dalsenter Paulo Roberto , Acco Alexandra
Published: November 2010

Snake venoms present different action mechanisms because of their complex composition, represented mainly by toxins and enzymes. This work aimed to investigate the effects of the Crotalus durissus terrificus(Cdt) venom in the liver. Wistar rats were inoculated intraperitoneally with saline (control) or Cdt venom. After 3, 4, or 6 h, the following parameters were analyzed: (a) hepatic function, (b) oxidative stress parameters, and (c) the metabolism of alanine in the isolated perfused liver. Plasma activities of alanine aminotransferase and aspartate aminotransferase and hepatic glutathione S‐transferase and catalase presented significant elevation in rats inoculated with 300 μg ⋅ kg−1 Cdt venom. Liver lipoperoxidation was enormously increased by venom doses of 100, 200, and 300 μg ⋅kg−1, whereas glutathione S‐transferase was not changed. Perfused livers from rats inoculated with 1500 μg ⋅kg−1 venom showed increased production of lactate, pyruvate, and ammonia when alanine was the metabolic substrate. These results demonstrate that the Cdt venom can produce several changes in hepatocytes. The causes of the changes are possibly related to the disequilibrium in the redox homeostasis but also to specific needs of the poisoned organism, for example, an increased supply of lactate and pyruvate in response to an increased activity of the Cori cycle. © 2010 Wiley Periodicals, Inc. J Biochem Mol Toxicol 25:195–203, 2011; View this article online at wileyonlinelibrary.com. DOI 10.1002/jbt.20376
The differential impact of PDE4 subtypes in human lung fibroblasts on cytokine‐induced proliferation and myofibroblast conversion
Authors: Selige Jens , Hatzelmann Armin , Dunkern Torsten
Published: November 2010

Lung fibroblast proliferation and differentiation into myofibroblasts are pathological key events during development of lung fibrosis. Cyclic nucleotide signaling is described as a negative modulator of these cellular processes, and cyclic nucleotide degrading type 4 phosphodiesterases (PDE4) are important regulators of these pathways. In this study, we elucidated expression and the role of individual subtypes of PDE4 in primary normal human lung fibroblast (NHLF) in controlling cytokines‐induced proliferation and conversion to myofibroblasts by short‐interfering RNAs (siRNAs) induced knockdown. We verified the expression of PDE4A, B, and D, while PDE4C was only minor or even not expressed in NHLF. An efficient liposome‐mediated transfection method for mRNA silencing and a knockdown of the expressed PDE4 subtypes was achieved in these cells. This knockdown was further validated by PDE4 protein expression analysis and PDE4 activity measurements. Functionally, the knockdown of PDE4A and PDE4B inhibited proliferation induced by the cytokine combination of bFGF and IL‐1β, whereas knockdown of PDE4D was ineffective. In contrast, TGF‐β induced differentiation into myofibroblasts was affected by knockdown of PDE4B and PDE4D, but not by PDE4A knockdown. In summary, our data allow to assign different PDE4 subtypes to distinct functions of human lung fibroblasts and highlight the predominant role of PDE4B in controlling pathophysiological processes of human lung fibroblasts. This provides a scientific rationale for focused therapeutic targeting of PDE4B to treat respiratory diseases with fibrotic lesions in the lung. J. Cell. Physiol. 226: 1970–1980, 2011. © 2010 Wiley‐Liss, Inc.
Analysis of reactive oxygen species in the guard cell of wheat stoma with confocal microscope
Authors: Liu Dongwu , Chen Zhiwei , Shi Peiguo , Wang Xue , Cai Weiwei
Published: November 2010

Recently, the laser‐scanning confocal microscope has become a routine technique and indispensable tool for cell biological studies. Previous studies indicated that reactive oxygen species (ROS) were generated in tobacco epidermal cells with confocal microscope. In the present studies, the probe 2′,7′‐dichlorof luorescein diacetate (H2DCF‐DA) was used to research the change of ROS in the guard cell of wheat stoma, and catalase (CAT) was used to demonstrate that ROS had been labeled. The laser‐scanning mode of confocal microscope was XYT, and the time interval between two sections was 1.6351 s. Sixty optical sections were acquired with the laser‐scanning confocal microscope, and CAT (60,000 U mg−1) was added after four optical sections were scanned. Furthermore, the region of interest (ROI) was circled and the fluorescence intensity of ROS was quantified with Leica Confocal Software. The quantitative data were exported and the trend chart was made with software Excell. The results indicated that ROS were produced intracellularly in stomatal guard cells, and the quantified fluorescence intensity of ROS was declined with CAT added. It is a good method to research the instantaneous change of ROS in plant cells with confocal microscope and fluorescence probe H2DCF‐DA. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.
A morphological study of the vertebral venous plexus and its connections in the Cape dune mole‐rat, Bathyergus suillus (Bathyergidae)
Authors: Kotzé S.H. , Boonzaier J. , Vorster W. , Hoogland P.V.J.M.
Published: November 2010

Bathyergus suillus are subterranean rodents found in the Western Cape of South Africa, where they inhabit sandy, humid burrows. Vertebral venous plexuses around the vertebral column have been implicated in aiding the maintenance of a constant central nervous system temperature via its connections with muscles and interscapular brown adipose tissue. The morphology of the vertebral venous plexuses and its connections in B.suillus were investigated. Frozen (n = 10) animals were defrosted; the venous system injected with latex and the vertebral venous plexuses, azygos‐ and intercostal veins dissected along the dorsal and ventral aspects of the vertebral column. Specimens (n = 4) were used for histological serial cross sections of the thoracic vertebrae. Veins drained from the interscapular brown adipose tissue to the external vertebral venous plexus, via a dorsal vein at the spinous process of T2 which might represent the “vein of Sulzer” described in rats. The intercostal veins cranial to the level of T8 drained directly into the ventral external vertebral venous plexus instead of into the azygos vein as seen in rats. The azygos vein was situated ventrally on the thoracic vertebral bodies in the median plane as opposed to most rodents that have a left sided azygos vein. The internal vertebral venous plexus consisted of two ventrolateraly placed longitudinal veins in the spinal epidural space. Veins from the forelimbs entered the internal vertebral venous plexus directly at the levels of C7 and T1 and have not been described in other rodents. Serial histological sections, revealed no regulatory valves in vessels leading toward the internal vertebral venous plexus, allowing blood to presumably move in both directions within the vertebral venous plexus. The vertebral venous plexus of B. suillus shows similarities to that of the rat but the vessels from the forelimbs draining directly into to the internal vertebral venous plexus and the position of the azygos vein and the intercostal veins draining into the external vertebral venous plexus are notable exceptions. J. Morphol., 2011. © 2010 Wiley‐Liss, Inc.
Antibody‐mediated sialidase activity in blood serum of patients with multiple myeloma
Authors: Bilyy Rostyslav , Tomin Andriy , Mahorivska Iryna , Shalay Olga , Lohinskyy Volodymyr , Stoika Rostyslav , Kit Yuriy
Published: November 2010

Cell surface sialylation is known to be tightly connected with tumorigenicity, invasiveness, metastatic potential, clearance of aged cells, while the sialylation of IgG molecules determines their anti‐inflammatory properties. Four sialidases – hydrolytic enzymes responsible for cleavage of sialic residues – were described in different cellular compartments. However, sialidases activity in body fluids, and specifically in blood serum, remains poorly studied. Here, we characterize first known IgG antibodies possessing sialidase‐like activity in blood serum of multiple myeloma (MM) patients.

Ig fractions were precipitated with ammonium sulfate (50% of saturation) from blood serum of 12 healthy donors and 14 MM patients, and screened for the presence of sialidase activity by using 4‐MUNA (2′‐(4‐methylumbelliferyl)‐α‐D‐N‐acetylneuraminic acid) as substrate. High level of sialidase activity was detected in the MM patients, but not in healthy donors. Subsequent antibody purification by protein‐G affinity chromatography and HPLC size exclusion chromatography at acidic conditions demonstrated that sialidase activity was attributable to IgG molecules. Sialidase activity was also specific for (Fab)2 fragment of IgG and blocked by sialidase inhibitor DANA. Sialidase activity of IgG molecule was also confirmed by in gel assay for cleavage of sialidase substrate. Kinetic parameters of the catalysis reaction were described by Michaelis–Menten equation with Km = 44.4–108 µM and kcat = 2.7–23.1 min−1. The action of IgG possessing sialidase‐like activity towards human red blood cells resulted in a subsequent increase in their agglutination by the peanut agglutinin, that confirms their desialylation by the studied IgG. This is the first demonstration of the intrinsic sialidase activity of IgG isolated from blood serum of MM patients. Copyright © 2010 John Wiley & Sons, Ltd.
Binding dynamics and energetic insight into the molecular forces driving nucleotide binding by guanylate kinase
Authors: Kandeel Mahmoud , Kitade Yukio
Published: November 2010

Plasmodium deoxyguanylate pathways are an attractive area of investigation for future metabolic and drug discovery studies due to their unique substrate specificities. We investigated the energetic contribution to guanylate kinase substrate binding and the forces underlying ligand recognition. In the range from 20 to 35°C, the thermodynamic profiles displayed marked decrease in binding enthalpy, while the free energy of binding showed little changes. GMP produced a large binding heat capacity change of −356 cal mol−1 K−1, indicating considerable conformational changes upon ligand binding. Interestingly, the calculated ΔCp was −32 cal mol−1 K−1, indicating that the accessible surface area is not the central change in substrate binding, and that other entropic forces, including conformational changes, are more predominant. The thermodynamic signature for GMP is inconsistent with rigid‐body association, while dGMP showed more or less rigid‐body association. These binding profiles explain the poor catalytic efficiency and low affinity for dGMP compared with GMP. At low temperature, the ligands bind to the receptor site under the effect of hydrophobic forces. Interestingly, by increasing the temperature, the entropic forces gradually vanish and proceed to a nonfavorable contribution, and the interaction occurs mainly through bonding, electrostatic forces, and van der Waals interactions. Copyright © 2010 John Wiley & Sons, Ltd.
Fully automated determination of N‐nitrosamines in environmental waters by headspace solid‐phase microextraction followed by GC–MS–MS
Authors: Llop Anna , Borrull Francesc , Pocurull Eva
Published: November 2010

A fully automated method for determining nine Environmental Protection Agency N‐nitrosamines in several types of environmental waters at ng/L levels is presented. The method is based on a headspace solid‐phase microextraction followed by GC–MS–MS using chemical ionization. Three different fibers (carboxen/PDMS, divinylbenzene/carboxen/PDMS, and PEG) were tested. Solid‐phase microextraction conditions were best when a divinylbenzene/carboxen/PDMS fiber was exposed for 60 min in the headspace of 10 mL water samples at pH 7 containing 360 g/L of NaCl, at 45°C. All compounds were analyzed by GC–MS–MS within 18 min. The method was validated using effluent from an urban wastewater treatment plant and the LODs ranged from 1 to 5 ng/L. The method was then applied to determine the N‐nitrosamines in samples of different complexities, such as tap water and several influent and effluent wastewater samples from urban and industrial wastewater treatment plants and a potable water treatment plant. Although the analysis of influent industrial wastewater revealed high concentrations of some compounds (N‐nitrosomorpholine and N‐nitrosodimethylamine at μg/L levels), in industrial effluents and other samples, the concentrations were substantially lower (ng/L levels). The new method is suitable for the simple and reliable determination of N‐nitrosamines in highly complex water samples in a completely automated procedure.
A simple and reliable stir bar sorptive extraction‐liquid chromatography procedure for the determination of chlorpromazine and trifluoperazine in human serum using experimental design methodology
Authors: Bazhdanzadeh Shaghayegh , Talebpour Zahra , Adib Noushin , Aboul‐Enein Hassan Y.
Published: November 2010

A sensitive and reproducible stir bar sorptive extraction and HPLC‐UV detection method was used for the therapeutic drug monitoring of chlorpromazine and trifluoperazine in human serum. The separation was achieved using a C18 column. The mobile phase consisted of methanol/sodium acetate buffer (pH 4.1; 0.1 M) (95:5, v/v) including 0.5% triethylamine. This miniaturized method can result in faster analysis, lower solvent consumption and less workload per sample while maintaining or even improving sensitivity. In the second part, stir bar sorptive extraction/HPLC‐UV method was optimized by a chemometrics approach. An experimental design was therefore used to evaluate the statistically influential and/or interacting factors, among those described in the literature, and to find the best extraction and desorption conditions. Optimal sample volume of 1 mL, extraction time of 24 min at 31°C with pH 8.1 were obtained in a screening 25 half fractional factorial design followed by a Box–Behnken design. For the desorption conditions, a Box–Behnken design showed that the best conditions were 150 μL mobile phase for 20 min at 50°C. The optimized method was repeatable (CV<10%, linear (LOQ‐500 ng/mL)), with the LOQs equal to 0.7 and 1.5 ng/mL for chlorpromazine and trifluoperazine, respectively.
Robotic‐assisted transoral removal of a submandibular megalith
Authors: Walvekar Rohan R. , Tyler Patrick D. , Tammareddi Neelima , Peters Geoffrey
Published: November 2010

The majority of salivary stones are less than 8 mm in size and most frequently occur in the submandibular gland. Traditional management of larger stones involves gland resection. Sialendoscopy combined with an external or a transoral sialolithotomy, also called the combined approach technique, permits stone removal and gland preservation. A 31‐year‐old male presented to our service with a 20‐mm megalith in the left submandibular gland. Here we report the first description of a combined approach using the da Vinci Si Surgical System to facilitate transoral stone removal and salivary duct repair. Laryngoscope, 2011
Endoplasmic reticulum stress is a mediator of posttransplant injury in severely steatotic liver allografts
Authors: Anderson Christopher D. , Upadhya Gundumi , Conzen Kendra D. , Jia Jianlou , Brunt Elizabeth M. , Tiriveedhi Venkataswarup , Xie Yan , Ramachandran Sabarinathan , Mohanakumar Thalachallour , Davidson Nicholas O. , Chapman William C.
Published: November 2010

Hepatic steatosis continues to present a major challenge in liver transplantation. These organs have been shown to have increased susceptibility to cold ischemia/reperfusion (CIR) injury in comparison with otherwise comparable lean livers; the mechanisms governing this increased susceptibility to CIR injury are not fully understood. Endoplasmic reticulum (ER) stress is an important link between hepatic steatosis, insulin resistance, and metabolic syndrome. In this study, we investigated ER stress signaling and blockade in the mediation of CIR injury in severely steatotic rodent allografts. Steatotic allografts from genetically leptin-resistant rodents had increased ER stress responses and increased markers of hepatocellular injury after liver transplantation into strain-matched lean recipients. ER stress response components were reduced by the chemical chaperone taurine-conjugated ursodeoxycholic acid (TUDCA), and this resulted in an improvement in the allograft injury. TUDCA treatment decreased nuclear factor kappa B activation and the proinflammatory cytokines interleukin-6 and interleukin-1β. However, the predominant response was decreased expression of the ER stress cell death mediator [CCAAT/enhancer-binding protein homologous protein (CHOP)]. Furthermore, activation of inflammation-associated caspase-11 was decreased, and this linked ER stress/CHOP to proinflammatory cytokine production after steatotic liver transplantation. These data confirm ER stress in steatotic allografts and implicate this as a mediating mechanism of inflammation and hepatocyte death in the steatotic liver allograft. Liver Transpl 17:189–200, 2011. © 2011 AASLD