bluuebunny/crossref_metadata_2025 huggingface dataset:
Econazole‐induced Ca2+ fluxes and apoptosis in human oral cancer cells
Authors: Kuo Daih‐Huang , Liu Li‐Min , Chen Hsin‐Wei , Chen Fu‐An , Jan Chung‐Ren
Published: March 2010

The effect of econazole on cytosolic free Ca2+ concentrations ([Ca2+]i) and viability was explored in human oral cancer cells (OC2), using the fluorescent dyes fura‐2 and WST‐1, respectively. Econazole at concentrations of >1 µM increased [Ca2+]i in a concentration‐dependent manner. The Ca2+ signal was reduced partly by removing extracellular Ca2+. The econazole‐induced Ca2+ influx was sensitive to blockade of aristolochic acid (phospholipase A2 inhibitor) and GF109203X (PKC inhibitor). In Ca2+‐free medium, after treatment with 1 µM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor), 30 µM econazole failed to induce a [Ca2+]i rise. Inhibition of phospholipase C with 2 µM U73122 substantially suppressed econazole‐induced [Ca2+]i rise. At concentrations of 5–70 µM econazole killed cells in a concentration‐dependent manner. The cytotoxic effect of 50 µM econazole was enhanced by prechelating cytosolic Ca2+ with 1,2‐bis(2‐aminophenoxy)ethane‐N,N,N′,N′‐tetraacetic acid (BAPTA). The ERK MAPK inhibitor, PD98059 (10 µM), also enhanced 20 µM econazole‐induced cell death. Propidium iodide staining data suggest that econazole induced apoptosis between concentrations of 10–70 µM. Collectively, in OC2 cells, econazole induced [Ca2+]i rises by causing Ca2+ release from the endoplasmic reticulum and Ca2+ influx from phospholipase A2/PKC‐regulated Ca2+ channels. Furthermore, econazole caused cell death appeared to be regulated by ERK MAPK. Drug Dev Res 71: 240–248, 2010. © 2010 Wiley‐Liss, Inc.
Unexpected roles for DEAD‐box protein 3 in viral RNA sensing pathways
Authors: Mulhern Orla , Bowie Andrew G.
Published: March 2010

Detection of viral nucleic acid within infected cells is essential to an effective anti‐viral response. The retinoic acid‐inducible gene‐I‐like receptors (RLR) form part of the virus detection repertoire and are critically important in sensing viral RNA in the cytoplasm. Efforts continue to define the signalling components downstream of RLR that are required to induce type I IFN (IFN‐α and promoter stimulator‐1) after viral infection. One surprising finding was that the Asp‐Glu‐Ala‐Asp box helicase DEAD/H Box 3 (DDX3), known for some time to have a number of roles in cellular RNA regulation in the nucleus, has a role in the RLR cytoplasmic signalling pathway involved in promoter stimulator‐1 induction. In this issue of theEuropean Journal of Immunology, an article reports an additional distinct positive role for DDX3 in the RLR RNA sensing pathway. This further emphasises the importance of DDX3 in anti‐viral immunity, and is consistent with the idea that viruses target DDX3 for immune evasion.
Bystander activation of CD4+ T cells
Authors: Boyman Onur
Published: March 2010

Bystander activation of T cells, i.e. the stimulation of unrelated (heterologous) T cells by cytokines during an Ag‐specific T‐cell response, has been best described for CD8+ T cells. In the CD8+ compartment, the release of IFN and IFN‐inducers leads to the production of IL‐15, which mediates the proliferation of CD8+ T cells, notably memory‐phenotype CD8+ T cells. CD4+ T cells also undergo bystander activation, however, the signals inducing this Ag‐nonspecific stimulation of CD4+ T cells are less well known. A study in this issue of the European Journal of Immunology sheds light on this aspect, suggesting that common γ‐chain cytokines including IL‐2 might be involved in bystander activation of CD4+ T cells.
Photoactivated Biological Activity of Transition‐Metal Complexes (Eur. J. Inorg. Chem. 10/2010)
Authors: Schatzschneider Ulrich
Published: March 2010

The cover picture shows examples of metal complexes with photoactivated biological activity, as symbolized by the lightning in the background. This includes liberation of small‐molecule messengers, such as carbon monoxide, nitric oxide, and neuroactive amines from the metal coordination sphere, as well as light‐triggered oxidative or covalent modification of DNA. More examples are presented in the Microreview by U. Schatzschneider on p. 1451 ff.
Study of elution behaviour with gradient voltage in CEC using methacrylate monolithic columns
Authors: Baeza‐Baeza Juan José , Bernabé‐Zafón Virginia , Herrero‐Martínez José Manuel , Simó‐Alfonso Ernesto F.
Published: March 2010

A theoretical study on the retention behaviour and chromatographic performance of neutral solutes using a lauryl methacrylate‐based monolithic column under voltage gradient mode in CEC was carried out. Through a flexible mathematical function based on a modified Gaussian model, the peak shape of compounds was firstly fitted under constant and gradient voltage. Using the peak shape parameters and retention time, the estimation of global chromatographic performance, efficiency and peak capacity under several voltage conditions was performed. The influence of voltage gradient on the separation efficiency is discussed and simple equations are presented to calculate retention and peak widths under voltage gradient conditions. A comparison in terms of chromatographic performance of a test mixture of neutral solutes under constant and gradient voltage modes was also carried out. The experiments carried out under gradient voltage showed better efficiencies (172 000 plates/m) and lower peak widths than those obtained under constant voltage (52 000 plates/m).
Glycogen: A novel branched polysaccharide chiral selector in CE
Authors: Chen Jiaquan , Du Yingxiang , Zhu Fenxia , Chen Bin
Published: March 2010

Various chiral selectors have been employed in CE and among them linear polysaccharides exhibited powerful enantioselective properties. Different from linear polysaccharides, the use of branched polysaccharides as chiral selectors in CE has not been reported previously. In this study glycogen belonging to the class of branched polysaccharides was used as a novel chiral selector for the enantiomeric separations for the first time. Since glycogen is electrically neutral, the method is applicable to ionic compounds. Eighteen chiral compounds including 12 basic drugs and six acidic drugs have been tested to demonstrate the potential of this chiral selector. BGE and selector concentrations and buffer pH were systematically optimized in order to obtain successful chiral separations. Among the tested compounds, the enantiomers of ibuprofen, which is an acidic drug, were successfully recognized by 3.0% w/v glycogen with 90 mM Tris‐H3PO4 buffer (pH 7.0). The enantiomers of basic drugs such as citalopram, cetirizine and nefopam were also baseline‐resolved with 50 mM Tris‐H3PO4 buffer (pH 3.0) containing 3.0% glycogen. Amlodipine belonging to basic compound only gave partial enantioseparation under the above‐mentioned condition.
Heart‐cutting 2D‐CE with on‐line preconcentration for the chiral analysis of native amino acids
Authors: Anouti Suzanne , Vandenabeele‐Trambouze Odile , Cottet Hervé
Published: March 2010

The use of transient moving chemical reaction boundary (tMCRB) was investigated for the on‐line preconcentration of native amino acids in heart‐cutting 2D‐CE with multiple detection points using contactless conductivity detection. The tMCRB focusing was obtained by using ammonium formate (pH 8.56) as sample matrix and acetic acid (pH 2.3) as a BGE in the first dimension of the heart‐cutting 2D‐CE. Different experimental parameters such as the injected volume and the concentration in ammonium formate were optimized for improving the sensitivity of detection. A stacked fraction from the first dimension was selected, isolated in the capillary, and then separated in the second dimension in the presence of a chiral selector ((+)‐(18‐crown‐6)‐2,3,11,12‐tetracarboxylic acid). This on‐line tMCRB preconcentration coupled with heart‐cutting 2D‐CE was applied with success to the chiral separation of D,L‐phenylalanine, and D,L‐threonine in a mixture of 22 native amino acids. The sample mixture was diluted in 0.8 M of ammonium formate, and injected at a concentration of 2.5 μM for each enantiomer with a volume corresponding to 10% of the total capillary volume. An LOD (S/N=3) of 2 μM was determined for L‐threonine.
Microchip electrophoresis of Alu elements for gender determination and inference of human ethnic origin
Authors: Njoroge Samuel K. , Witek Małgorzata A. , Hupert Mateusz L. , Soper Steven A.
Published: March 2010

We performed a series of multi‐locus PCRs followed by the rapid and efficient microchip electrophoretic sorting of Alu products with LIF detection. Five polymorphic human‐specific Alu insertions (RC5, A1, PV92, TPA and ACE) were used for inference of human ethnicity and two monomorphic Alu insertions for sex typing, one fixed on the X chromosome (AluSTXa) and the other on the Y chromosome (AluSTYa). These markers were used to generate unique DNA profiles for five different DNA samples. The PCR‐based assays used primers that flank the insertion point to determine genotypes based on the presence or absence of the Alu element. A1, RC5, PV92, TPA and ACE were used for ethnicity determinations and have two alleles, each indicating the presence (+) or absence (−) of the Alu element on the paired chromosomes, which results in three genotypes (+/+, +/− or −/−). RC5 and A1 did not show ethnic heterogeneity resulting in a homozygous (−/−) genotype, which correctly inferred that DNA samples originating from a Caucasian male and an Asian male were not of African ancestry. The results from the five Alu markers indicated that these Alu loci could assist in identifying the individual's ethnicity using microchip electrophoresis in under 15 min of separation time. Using microchip electrophoresis and mixed genotype ratios, male DNA‐to‐female DNA of 1:9, corresponding to a ratio of Y‐to‐X chromosomes of 1:19, was also detected for both AluSTXa and AluSTYa to provide gender identification without requiring separation of female from male cells prior to the assay.
Triblock copolymer matrix‐based capillary electrophoretic microdevice for high‐resolution multiplex pathogen detection
Authors: Kim Se Jin , Shin Gi Won , Choi Seok Jin , Hwang Hee Sung , Jung Gyoo Yeol , Seo Tae Seok
Published: March 2010

Rapid and simple analysis for the multiple target pathogens is critical for patient management. CE‐SSCP analysis on a microchip provides high speed, high sensitivity, and a portable genetic analysis platform in molecular diagnostic fields. The capability of separating ssDNA molecules in a capillary electrophoretic microchannel with high resolution is a critical issue to perform the precise interpretation in the electropherogram. In this study, we explored the potential of poly(ethyleneoxide)‐poly(propyleneoxide)‐poly(ethyleneoxide) (PEO‐PPO‐PEO) triblock copolymer as a sieving matrix for CE‐SSCP analysis on a microdevice. To demonstrate the superior resolving power of PEO‐PPO‐PEO copolymers, 255‐bp PCR amplicons obtained from 16S ribosomal RNA genes of four bacterial species, namely Proteus mirabilis, Haemophilus ducreyi, Pseudomonas aeruginosa, and Neisseria meningitidis, were analyzed in the PEO‐PPO‐PEO matrix in comparison with 5% linear polyacrylamide and commercial GeneScan™ gel. Due to enhanced dynamic coating and sieving ability, PEO‐PPO‐PEO copolymer displayed fourfold enhancement of resolving power in the CE‐SSCP to separate same‐sized DNA molecules. Fivefold input of genomic DNA of P. aeruginosa and/or N. meningitidis produced proportionally increased corresponding amplicon peaks, enabling correct quantitative analysis in the pathogen detection. Besides the high‐resolution sieving capability, a facile loading and replenishment of gel in the microchannel due to thermally reversible gelation property makes PEO‐PPO‐PEO triblock copolymer an excellent matrix in the CE‐SSCP analysis on the microdevice.
Simultaneous electrophoretic analysis of proteins of very high and low molecular mass using Tris‐acetate polyacrylamide gels
Authors: Cubillos‐Rojas Monica , Amair‐Pinedo Fabiola , Tato Irantzu , Bartrons Ramon , Ventura Francesc , Rosa Jose Luis
Published: March 2010

To separate and analyze giant and small proteins in the same electrophoresis gel, we have used a 3–15% polyacrylamide gradient gel containing 2.6% of the crosslinker bisacrylamide and 0.2 M of Tris‐acetate buffer (pH 7.0). Samples were prepared in a sample buffer containing lithium dodecyl sulphate and were run in the gel described above using Tris‐Tricine‐SDS‐sodium bisulfite buffer, pH 8.2, as electrophoresis buffer. Here, we show that this system can be successfully used for general applications of SDS‐PAGE such as CBB staining and immunoblot. Thus, by using Tris‐acetate 3–15% polyacrylamide gels, it is possible to simultaneously analyze proteins, in the mass range of 10–500 kDa, such as HERC1 (532 kDa), HERC2 (528 kDa), mTOR (289 kDa), Clathrin heavy chain (192 kDa), RSK (90 kDa), S6K (70 kDa), β‐actin (42 kDa), Ran (24 kDa) and LC3 (18 kDa). This system is highly sensitive since it allows detection from as low as 10 μg of total protein per lane. Moreover, it has a good resolution, low cost, high reproducibility and allows for analysis of proteins in a wide range of weights within a short period of time. All these features together with the use of a standard electrophoresis apparatus make the Tris‐acetate‐PAGE system a very helpful tool for protein analysis.
Simplified analysis of mid‐story seismically isolated buildings
Authors: Wang Shiang‐Jung , Chang Kuo‐Chun , Hwang Jenn‐Shin , Lee Bo‐Han
Published: March 2010

The mid‐story isolation design method is recently gaining popularity for the seismic protective design of buildings located in the areas of high population. In a mid‐story isolated building, the isolation system is incorporated into the mid‐story rather than the base of the building. In this paper, the dynamic characteristics and seismic responses of mid‐story isolated buildings are investigated using a simplified three‐lumped‐mass structural model for which equivalent linear properties are formulated. From the parametric study, it is found that the nominal frequencies of the superstructure and the substructure, respectively, above and below the isolation system have significant influences on the isolation frequency and equivalent damping ratio of a mid‐story isolated building. Moreover, the mass and stiffness of the substructure are of greater significance than the superstructure in affecting the dynamic characteristics of the isolated building. Besides, based on the response spectrum analysis, it is noted that the higher mode responses may contribute significantly to the story shear force of the substructure. Consequently, the equivalent lateral force procedure of design codes should carefully include the effects of higher modes. Copyright © 2010 John Wiley & Sons, Ltd.
A facile route to 1,5‐benzodiazepin‐2‐yl‐phosphonates via ytterbium chloride‐catalyzed three‐component reaction
Authors: Ji Xiaofeng , Cai Lijian , Yao Zhigang , Fan Xu , Qi Shen
Published: March 2010

1,5‐Benzodiazepin‐2‐yl‐phosphonates were facilely synthesized via a one‐pot three‐component condensation of o‐diaminobenzene, 1,3‐diketone and diethyl phosphite in the presence of a catalytic amount of ytterbium chloride under mild reaction conditions. © 2010 Wiley Periodicals, Inc. Heteroatom Chem 21:89–95, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20573
One‐pot synthesis of α‐aminophosphonates catalyzed by boric acid at room temperature
Authors: Karimi‐Jaberi Zahed , Amiri Mohammad
Published: March 2010

A simple, efficient, and general method has been developed for the one‐pot, three‐component synthesis of α‐aminophosphonates from a condensation reaction of trimethyl phosphite, aldehydes, and amines in the presence of catalytic amount of boric acid (10 mol%) under solvent‐free conditions. Thus α‐aminophosphonates were synthesized relatively quickly in good yields at room temperature. © 2010 Wiley Periodicals, Inc. Heteroatom Chem 21:96–98, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20577
A new family of platinum(II) complexes incorporating five‐ and six‐membered cyclic phosphine ligands
Authors: Kerényi Andrea , Kovács Viktória , Körtvélyesi Tamás , Ludányi Krisztina , Drahos László , Keglevich György
Published: March 2010

New platinum complexes of the type cis‐Pt(L)2Cl2 have been synthesized from five‐ and six‐membered cyclic phosphines, which were prepared after deoxygenating a series of phosphine oxides (3‐phospholene oxides, phospholane oxides, a 1,4‐dihydrophosphinine oxide, and a 1,2,3,6‐tetrahydrophosphinine oxide). The complexes were characterized by NMR and mass spectral data, and their stereostructures were elucidated by B3LYP/6‐31G(d)‐LANL2DZ ECP calculations. The phosphine intermediates were characterized as the corresponding phosphine‐boranes. © 2010 Wiley Periodicals, Inc. Heteroatom Chem 21:63–70, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20579
Synthesis of biologically active phenoxy derivatives of substituted benzothiazole organophosphates
Authors: Jhajharia Priyanka , Samota Mahesh Kumar , Seth Gita
Published: March 2010

The reaction of S‐(phenyl benzothiazolyl‐2)phosphorodichloridothioate/phosphorodichloridodithioate with 2 mol of phenol/4‐chlorophenol/4‐nitrophenol in the presence of stoichiometric amounts of triethylamine in dry THF/CH2Cl2 has afforded a series of the corresponding organophosphate phenoxy derivatives (1a, 1b, 2a, 2b, and 3a, 3b). Plausible structures have been proposed on the basis of elemental analysis, IR, 1H NMR, 31P NMR, and mass spectral studies. The antibacterial activity of these organophosphate phenoxy derivatives has been evaluated against pathogenic bacteria Staphylococcus aureus (+ve) and Escherichia coli (−ve). The antifungal activity of these organophosphate phenoxy derivatives has been evaluated against pathogenic fungi Aspergillus niger and Fusarium oxysporium. The results indicate that organophosphate phenoxy derivatives are found more active than the parent compounds. © 2010 Wiley Periodicals, Inc. Heteroatom Chem 21:84–88, 2010; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/hc.20582
Hydroxyapatite versus titanium implant: Comparison of the functional outcome after vocal fold medialization in unilateral recurrent nerve paralysis
Authors: Storck Claudio , Fischer Claude , Cecon Mikis , Schmid Stephan , Gambazzi Franco , Wolfensberger Markus , Brockmann Meike
Published: March 2010

Background.

Results of medialization thyroplasty for treatment of unilateral vocal fold paralysis are often unsatisfactory. This study compares glottal closure and voice quality after use of 2 different medialization implant types: VoCoM and TVFMI.

Methods.

In all, 26 patients with unilateral vocal fold paralysis following thoracic surgery underwent medialization thyroplasty. In 11 patients (group I), a hydroxyapatite implant (VoCoM) was used. In 15 patients (group II), a titanium implant (TVFMI) was used. Preoperative and postoperative glottal closure and voice function were assessed with videostroboscopy, perceptual and objective voice measures, and the Voice Dysfunction Index (VDI).

Results.

Group II showed a higher rate of complete glottal closure and greater improvement in perceived hoarseness, maximal phonation time, and maximal voice intensity than those in group I.

Conclusions.

With the individually adjustable titanium implant, better glottal closure and better functional outcome (phonation time and voice quality) were achieved. © 2010 Wiley Periodicals, Inc. Head Neck, 2010
Pseudoaneurysm of the lingual artery after concurrent intra‐arterial chemotherapy with radiotherapy for advanced tongue cancer
Authors: Murono Shigeyuki , Nakanishi Yosuke , Inoue Dai , Ozaki Kumi , Kondo Satoru , Wakisaka Naohiro , Matsui Osamu , Yoshizaki Tomokazu
Published: March 2010

Background.

Aneurysms of the external carotid artery and its branches are rare.

Methods.

We present an extremely rare case of pseudoaneurysm of the lingual artery secondary to concurrent intra‐arterial chemotherapy via the superficial temporal artery for advanced tongue cancer.

Results.

The patient developed sudden and significant oral bleeding from the left tongue base 10 weeks after the completion of treatment. Angiography provided a diagnosis of pseudoaneurysm of the lingual artery, and subsequent endovascular intervention was an effective means of controlling bleeding.

Conclusion.

Rupture of a pseudoaneurysm causes massive, life‐threatening bleeding, which requires adequate diagnosis and treatment. Pseudoaneurysm should be considered in cases showing sudden oral bleeding after intra‐arterial chemotherapy for advanced head and neck cancer. © 2010 Wiley Periodicals, Inc. Head Neck, 2011
Parathyroid carcinoma: A review
Authors: Owen Randall P. , Silver Carl E. , Pellitteri Phillip K. , Shaha Ashok R. , Devaney Kenneth O. , Werner Jochen A. , Rinaldo Alessandra , Ferlito Alfio
Published: March 2010

Background.

Parathyroid carcinoma is a rare entity, comprising fewer than 1% of cases of hyperparathyroidism. The disease generally presents with severe hyperparathyroidism and occasionally with vocal cord paralysis or a firm palpable cervical mass.

Methods.

The classic histopathologic features of trabecular pattern, mitotic figures, thick fibrous bands, and capsular and vascular invasion are not present in every case but are useful in assessing whether a particular hyperfunctional parathyroid lesion is a carcinoma. Nevertheless, recognition of malignancy at initial operation may be difficult. Management primarily involves complete surgical removal through en bloc resection, which may include adjacent central neck structures.

Results.

Adjuvant radiation therapy has not been proven to be uniformly effective, but several series show results suggestive of a possible survival advantage. Chemotherapy, genetic, and other biomodifying agents remain experimental. Long‐term outcome for this neoplasm remains problematic, and complications from intractable hypercalcemia constitute the foremost cause of death. Reported 5‐year survivals have ranged between 70% and 85%.

Conclusions.

Initial surgical success offers the greatest opportunity for cure. Reoperation for recurrence offers the potential for short‐ and intermediate‐term relief from the sequelae of hypercalcemia, but rarely results in cure. Severely elevated calcium levels should be controlled before initial surgery, and much of the treatment for recurrent or persistent disease, including reoperative surgery, is directed at control of hypercalcemia. © 2010 Wiley Periodicals, Inc. Head Neck, 2010
Novel germline SDHD deletion associated with an unusual sympathetic head and neck paraganglioma
Authors: Cadiñanos Juan , Llorente José L. , de la Rosa Jorge , Villameytide José A. , Illán Rafael , Durán Noelia S. , Murias Eduardo , Cabanillas Rubén
Published: March 2010

Background.

Paragangliomas (PGLs) are rare tumors arising either from sympathetic or parasympathetic‐associated chromaffin tissue. PGLs can occur either sporadically or as part of a hereditary syndrome. Sympathetic head and neck PGLs are extremely rare tumors and only a few cases have been reported to date.

Methods.

We report the pedigree of a patient with a head and neck PGL arising from the right sympathetic trunk. SDHD mutation analysis was performed using standard sequencing, multiplex ligation‐dependent probe amplification, chromosome 11‐specific comparative genome hybridization, and long‐range/short‐range polymerase chain reaction (PCR) approaches.

Results.

A previously unreported chromosome 11q deletion encompassing 5 annotated genes (SDHD, DLAT, PIH1D2, C11Orf57, and TIMM8B) was detected in the proband.

Conclusion.

PGL families considered “mutation‐negative” may be attributable to large gene deletions not detectable by standard sequencing methods. Therefore, deletion analysis should be offered to families or individuals at risk for hereditary PGLs. © 2010 Wiley Periodicals, Inc. Head Neck, 2011
Intratumoral injection of interferon‐α and systemic delivery of agonist anti‐CD137 monoclonal antibodies synergize for immunotherapy
Authors: Dubrot Juan , Palazón Asis , Alfaro Carlos , Azpilikueta Arantza , Ochoa María Carmen , Rouzaut Ana , Martinez‐Forero Iván , Teijeira Alvaro , Berraondo Pedro , Le Bon Agnes , Hervás‐Stubbs Sandra , Melero Ignacio
Published: March 2010

CD137 artificial costimulation results in complete tumor rejection in several mouse models. Type I interferons (IFN) exert antitumor effects through an array of molecular functions on malignant cells, tumor stroma and immune system cells. The fact that agonist anti‐CD137 mAb induce tumor regressions in mice deficient in the unique receptor for Type I IFNs (IFNAR−/−) indicated potential for treatment combinations. Indeed, combination of intratumor injections of mouse IFN‐α and intraperitoneal injections of anti‐CD137 mAb synergized as seen on subcutaneous lesions derived from the MC38 colon carcinoma, which is resistant to each treatment if given separately. Therapeutic activity was achieved both against lesions directly injected with IFN‐α and against distant concomitant tumors. Experiments in bone marrow chimeras prepared with IFNAR−/− and WT mice concluded that expression of the receptor for Type I interferons is mainly required on cells of the hematopoietic compartment. Synergistic effects correlated with a remarkable cellular hyperplasia of the tumor draining lymph nodes (TDLNs). Enlarged TDLNs contained more plasmacytoid and conventional dendritic cells (DC) that more readily cross‐presented. Importantly, numbers of both DC subtypes inversely correlated with the tumor size. Numbers of CD8 T cells specific for a dominant tumor antigen were increased at TDLNs by each separate treatment but only with slight augments due to the combination. Combined antitumor effects of the therapeutic strategy were also seen on subcutaneous TC‐1 tumors established for 24 days before treatment onset. The described strategy is realistic because (i) agents of each kind are clinically available and (ii) equivalent procedures in humans are feasible.
The leader peptide of a human rec. MnSOD as molecular carrier which delivers high amounts of Cisplatin into tumor cells inducing a fast apoptosis in vitro
Authors: Borrelli Antonella , Schiattarella Antonietta , Mancini Roberto , Morelli Franco , Capasso Clemente , De Luca Viviana , Gori Enrico , Mancini Aldo
Published: March 2010

The leader peptide of a recombinant MnSOD (rMnSOD‐Lp) constitutes the carrier that allows rMnSOD to penetrate tumor cells. A synthetic preparation of rMnSOD‐Lp was 68Ga labeled (rMnSOD‐Lp‐68Ga) and injected into animals bearing spontaneous mammary cancers, followed by PET examinations, which demonstrated unambiguously the tumor sites in all the animals, suggesting that if rMnSOD‐Lp was able to transport the radioisotope into tumor cells, it would also be able to deliver cytotoxic molecules. The rMnSOD‐Lp was, therefore, conjugated to cisplatin (rMnSOD‐Lp‐CC) and added to cultured tumor cells. Equal concentrations of cisplatin were used for the tests. After treating the ovarian cancer cells with 11.1 μg of cisplatin alone, analysis by atomic absorbance spectrophotometry was able to detect only 6 ng of platinum, whereas when the same cells were treated with the same amount of cisplatin conjugated to leader peptide rMnSOD, 387 ng of platinum were detected, i.e., an amount 80 times greater. Only the tumor cells died following treatment with rMnSOD‐Lp‐CC; molecular analysis revealed that its addition generated an increasing expression of Erk‐2 and Bax products, which could be inhibited only by a selective MAP/ERK kinase inhibitor (PD98059), revealing that rMnSOD‐Lp‐CC has an apoptotic function, exactly as occurs when using the cisplatin alone. Data are statistically significant and indicate that by using rMnSOD‐Lp‐CC, the cisplatin can be transformed from an agent with antireplicative activity into a specific and selective antitumor molecule, increasing its therapeutic index. We think that rMnSOD‐Lp‐CC deserves to be considered as a new antitumor agent.
Hsa‐let‐7g inhibits proliferation of hepatocellular carcinoma cells by downregulation of c‐Myc and upregulation of p16INK4A
Authors: Lan Fei‐Fei , Wang Hua , Chen Yang‐Chao , Chan Chu‐Yan , Ng Samuel S. , Li Kui , Xie Dan , He Ming‐Liang , Lin Marie C. , Kung Hsiang‐Fu
Published: March 2010

zMicroRNAs (miRNAs) are endogenously expressed small noncoding RNAs that regulate approximately one‐third of human genes at post‐transcription level. Previous studies have shown that miRNAs were implicated in many cellular processes and participated in the progress of various tumors including hepatocellular carcinoma (HCC). Among all miRNAs, the let‐7 family is well recognized to play pivotal roles in tumorigenesis by functioning as potential growth suppressor. In the present study, we aimed to investigate the role of let‐7 family, particularly the hsa‐let‐7g, in the molecular pathogenesis of HCC. By use of MTT, qPCR, Western blotting and 2‐dimensional electrophoresis (2‐DE), over‐expression of hsa‐let‐7g was found to inhibit the proliferation of HCC cell line via negative and positive regulations of c‐Myc and p16INK4A, respectively. The expression of hsa‐let‐7g was noted to be markedly lowered in the HepG2, Hep3B and Huh7 cells, yet higher in the Bel‐7404 HCC cell line. Proliferation of HCC cell line was significantly inhibited after the transfection of hsa‐let‐7g mimics, while hsa‐let‐7g inhibitor transfection exerted an opposite effect. Concurrently, the mRNA and protein levels of c‐Myc were found significantly decreased in HepG2 cells after transfection of hsa‐let‐7g mimics, but obviously increased in Bel‐7404 cells after transfection of hsa‐let‐7g inhibitor. As revealed by 2‐DE, a significant upregulation of p16INK4A was revealed after the gain‐of‐function study using hsa‐let‐7g. Therefore, we suggest that hsa‐let‐7g may act as a tumor suppressor gene that inhibits HCC cell proliferation by downregulating the oncogene, c‐Myc, and upregulating the tumor suppressor gene, p16INK4A.
ABO blood group and incidence of epithelial ovarian cancer
Authors: Gates Margaret A. , Wolpin Brian M. , Cramer Daniel W. , Hankinson Susan E. , Tworoger Shelley S.
Published: March 2010

Previous studies have observed an association between ABO blood group and risk of certain malignancies, including ovarian cancer; however, no prospective studies of the association with ovarian cancer risk are available. Using data from 49,153 women in the Nurses' Health Study, we examined the association between ABO blood group and incidence of epithelial ovarian cancer. Study participants reported their blood type and Rh factor in 1996, and 234 women were diagnosed with incident ovarian cancer during 10 years of follow‐up. We used Cox proportional hazards regression to model the incidence rate ratios (RR) and 95% confidence intervals (CI) of ovarian cancer for each blood group category. Compared to women with blood group O, women with blood group AB or B had a nonsignificant 38% increase in ovarian cancer incidence (95% CI = 0.88–2.16 for blood group AB and 0.96–1.99 for blood group B), whereas blood group A was not associated with risk (RR = 0.95, 95% CI = 0.70–1.30). Combining blood groups AB and B, we observed a statistically significant positive association with presence versus absence of the B antigen overall (RR = 1.41, 95% CI = 1.06–1.88) and for the serous invasive subtype (RR = 1.53, 95% CI = 1.08–2.17). In this large, prospective cohort of women, presence of the B antigen was positively associated with ovarian cancer incidence, whereas blood group A was not associated with risk. Additional studies are needed to confirm this association and to explore the mechanisms through which blood group may influence ovarian cancer risk.
Lysophosphatidic acid‐induced expression of periostin in stromal cells: Prognoistic relevance of periostin expression in epithelial ovarian cancer
Authors: Choi Kyung Un , Yun Jeong Sup , Lee Il Hwan , Heo Soon Chul , Shin Sang Hun , Jeon Eun Su , Choi Yoon Ji , Suh Dong‐Soo , Yoon Man‐Soo , Kim Jae Ho
Published: March 2010

Lysophosphatidic acid (LPA) is a bioactive lipid crucial for the initiation and progression of ovarian cancer. Identification of LPA‐induced biomarkers is necessary for predicting prognosis of ovarian cancer patients. Here we report periostin, an extracellular matrix protein, as an LPA‐induced protein in stromal cells and as a prognostic marker in patients with epithelial ovarian cancer (EOC). In human EOC tissues, periostin was mainly expressed in cancer‐associated stromal fibroblasts, but not in cancer cells. The expression levels of periostin highly correlated with poor survival and tumor recurrence of ovarian cancer patients. Treatment of human adipose tissue‐derived stromal cells with LPA or conditioned media from human ovarian adenocarcinoma cell lines, such as SK‐OV‐3 and OVCAR‐3, induced expression of periostin. The periostin expression induced by cancer‐conditioned media was abrogated by silencing of the LPA receptor 1 expression using small hairpin RNA lentivirus. Recombinant periostin stimulated adhesion and invasion of SK‐OV‐3 human ovarian adenocarcinoma cells and induced expression of matrix metalloprotease‐2 in the cancer cells. These results suggest that LPA is associated with the expression of periostin in cancer‐associated fibroblasts of EOC.
A potentially functional polymorphism in the promoter region of miR‐34b/c is associated with an increased risk for primary hepatocellular carcinoma
Authors: Xu Yan , Liu Li , Liu Jibin , Zhang Yixin , Zhu Jian , Chen Jianguo , Liu Sheng , Liu Zheng , Shi Haibin , Shen Hongbing , Hu Zhibin
Published: March 2010

The miR‐34 family members are direct transcriptional targets of tumor suppressor p53, and loss of miR‐34 function can impair p53‐mediated cell cycle arrest and apoptosis. A potentially functional SNP rs4938723 (T > C) was found in the promoter region of pri‐miR‐34b/c (423 bp from the transcription start site), located in the CpG island and might affect transcription factor GATA binding and therefore pri‐miR‐34b/c expression. In our study, we hypothesized that SNPs miR‐34b/c rs4938723 and TP53 Arg72Pro may independently or jointly contribute to primary hepatocellular carcinoma (HCC) susceptibility. We then genotyped the 2 SNPs in a case–control study of 501 patients with primary HCC and 548 cancer‐free controls in a Chinese population. We observed that the variant genotypes of miR‐34b/c rs4938723 were associated with significantly increased HCC risks compared with the wild‐type TT genotype (adjusted OR = 1.37, 95% CI =1.06–1.78 for TC; OR = 1.53, 95% CI = 1.02–2.31 for CC and OR = 1.40, 95% CI = 1.10–1.80 for TC/CC). Furthermore, we found a significant interaction between alcohol drinking and SNP rs4938723 on HCC risk (p = 0.05 for multiplicative and p = 0.01 for additive interaction). However, we did not find any main effect of TP53 Arg72Pro on HCC risk in this population. These findings indicate that the potentially functional SNP rs4938723 in the promoter region of pri‐miR‐34b/c may contribute to the susceptibility of HCC in this Chinese population.
p53 transactivation is involved in the antiproliferative activity of the putative tumor suppressor RBM5
Authors: Kobayashi Takahiko , Ishida Junich , Musashi Manabu , Ota Shuichi , Yoshida Takeshi , Shimizu Yuichi , Chuma Makoto , Kawakami Hiroshi , Asaka Masahiro , Tanaka Junji , Imamura Masahiro , Kobayashi Masanobu , Itoh Hiroshi , Edamatsu Hironori , Sutherland Leslie C. , Brachmann Rainer K.
Published: March 2010

RBM5 (RNA‐binding motif protein 5) is a nuclear RNA binding protein containing 2 RNA recognition motifs. The RBM5 gene is located at the tumor suppressor locus 3p21.3. Deletion of this locus is the most frequent genetic alteration in lung cancer, but is also found in other human cancers. RBM5 is known to induce apoptosis and cell cycle arrest but the molecular mechanisms of RBM5 function are poorly understood. Here, we show that RBM5 is important for the activity of the tumor suppressor protein p53. Overexpression of RBM5 enhanced p53‐mediated inhibition of cell growth and colony formation. Expression of RBM5 augmented p53 transcriptional activity in reporter gene assays and resulted in increased mRNA and protein levels for endogenous p53 target genes. In contrast, shRNA‐mediated knockdown of endogenous RBM5 led to decreased p53 transcriptional activity and reduced levels of mRNA and protein for endogenous p53 target genes. RBM5 affected protein, but not mRNA, levels of endogenous p53 after DNA damage suggest that RBM5 contributes to p53 activity through post‐transcriptional mechanisms. Our results show that RBM5 contributes to p53 transcriptional activity after DNA damage and that growth suppression and apoptosis mediated by RBM5 are linked to activity of the tumor suppressor protein p53.
Lipid peroxidation in the kidney of rats treated with V and/or Mg in drinking water
Authors: Ścibior Agnieszka , Zaporowska Halina , Niedźwiecka Irmina
Published: March 2010

Spontaneous and stimulated lipid peroxidation (LPO) after vanadate and magnesium treatment was studied in kidney supernatants obtained from outbred 5‐month‐old, albino male Wistar rats. The 2‐month‐old animals daily received: group I (control), deionized water to drink; group II, water solution of sodium metavanadate, NaVO3 (SMV, 0.125 mg V ml−1); group III, water solution of magnesium sulfate, MgSO4 (MS, 0.06 mg Mg ml−1); and group IV, water solution of SMV‐MS at the same concentrations as in groups II and III for V and Mg, respectively, over a 12‐week period. FeSO4, NaVO3 and MgSO4 were selected as agents that may modify LPO process in in vitro conditions. Spontaneous malondialdehyde (MDA) levels in kidney supernatants increased significantly in the rats in groups II and IV, compared with groups I and III; and they were also significantly higher in all the groups of rats compared with the liver supernatants. The total antioxidant status (TAS) in groups II and IV tended to be higher too. Vanadium concentration in the kidney of the rats in groups II and IV increased, whereas the kidney Mg content in groups II, III and IV decreased, compared with levels in the liver. As the two‐way ANOVA indicated, the changes in the basal MDA level, TAS and Mg concentration in the liver of rats at combined V and Mg application only resulted from independent action of V. As far as the in vitro results are concerned, in the supernatants obtained from the rats in groups II and IV, a significant increase in MDA level was demonstrated in the presence of 30 µm of exogenous FeSO4 as well as 30, 100, 200 and 400 µm NaVO3 and 100, 200, 400, 600, 800 and 1000 µm MgSO4, compared with groups I and III. The 600, 800 and 1000 µm of exogenous MgSO4 also significantly elevated MDA production in the supernatants obtained from the rats in group III, compared with spontaneously formed MDA in the same supernatants. The three‐way ANOVA showed that the changes in LPO induced by in vitro treatment of kidney supernatants with exogenous Fe or V or Mg (600, 800 and 1000 µm) were a consequence of independent action of those metals and they also resulted from the interactions between exogenous Fe (Feexog) and endogenous V (Vend) and between Vend and exogenous V (Vexog). In conclusion, V (as NaVO3) consumed by the rats with drinking water at a dose of 12.9 mg V kg−1 b.w. per 24 h for 12 weeks increased the basal LPO and markedly enhanced TAS in the renal tissue. Its pro‐oxidant potential was also found in in vitro conditions. The Mg dose (6 mg Mg kg−1 b.w. per 24 h) ingested by the rats together with V (12.7 mg V kg−1 b.w. per 24 h) neither reduced nor intensified the spontaneous LPO, compared with V‐only intoxicated animals; however, the stimulating effect of Mg on LPO was revealed in in vitro conditions.
Response to retreatment of malignant hypercalcemia with the bisphosphonate AHPrBP (APD): Respective role of kidney and bone
Authors: Dr. Thiebaud D. , Jaeger P. , Burckhardt P.
Published: March 2010

Malignant hypercalcemia is caused by both increased bone resorption and enhanced tubular reabsorption of calcium.

First, the response to an infusion of APD was compared in two groups of patients: 23 with breast cancer versus 20 with squamous cell cancer. The decrease in plasma calcium was smaller in the latter group (p < 0.05 at day 14), due to increased tubular reabsorption of calcium (TmCa/GFR 2.20 ± 0.05 versus 2.58 ± 0.06 mmol/liter; p < 0.001), whereas the degree of bone resorption reflected by urinary hydroxyproline was identical. Therefore, at a given initial plasma calcium level, the type of tumor (on which TmCA/GFR depends) seems to be a determinant for the effectiveness of the treatment.

Second, the response to the initial treatment was compared with that to a second treatment with the same dose in 12 patients whose malignant hypercalcemia relapsed. Within 9 days, plasma calcium decreased from 3.46 ± 0.10 to 2.50 ± 0.10 mmol/liter after the first course, but only from 3.37 ± 0.08 to 2.79 ± 0.09 mmol/liter after the second course (p < 0.01). TmCa/GFR was similar before the first and the second treatment and did not vary during the days following the infusion of APD. Initial urinary hydroxyproline was slightly but not significantly higher before the second treatment. It dropped following both APD courses, but to a lesser extent after the second treatment, reflecting higher bone resorption or possible resistance to bisphosphonate.

In conclusion, both the degree of tubular reabsorption of calcium (which depends on tumor type) and that of bone resorption modify the response of malignant hypercalcemia to APD, bone resorption playing the key role for the loss of efficacy in case of retreatment.
A new method for automatic recognition of the radiographic trabecular pattern
Authors: Geraets Wil G. M. , van der Stelt Paul F. , Netelenbos Coen J. , Elders Petra J. M.
Published: March 2010

This study reports a method to describe and analyze the structure of the trabecular pattern seen on radiographs of the distal radius. The structure is measured and related to the bone mineral density (BMD) of the lumbar spine measured by dual-photon absorptiometry.

Radiographs of hand and wrist combined with additional information of the bone mineral density of the vertebrae serve as testing material. With a computer-aided imaging system, a part of the depicted radius is scanned. The image is filtered and segmented into a bilevel picture consisting of a light network with dark meshes. Seven features of the bilevel picture are measured and analyzed.

It is shown that six features correlate significantly with the bone mineral density measured at the lumbar spine, although the correlations between the trabecular pattern and the BMD are too weak to allow precise predictions of BMD values for individuals. Nevertheless, the correlations confirm the existence of a relationship between the radiographic trabecular pattern and the bone mineral density of the lumbar spine. The method is worth being further developed for use on individual patients. It provides a noninvasive tool to make an objective and quantitative assessment of the trabecular pattern.
Serum osteocalcin (BGP) Levels in normal men: A longitudinal evaluation reveals an age-associated increase
Authors: Orwoll Eric S. , Deftos Leonard J.
Published: March 2010

Serum levels of bone gla protein (BGP) have been reported to increase with aging and hence to reflect an age-related increase in bone remodeling activity. To evaluate the relationship between aging and serum BGP levels in a study of longitudinal design, we measured BGP concentrations in 77 normal men at 6 month intervals over a 3 year period. Mean BGP levels at the onset (4.95 ± 1.5 ng/ml) increased significantly during the study (p = 0.004), and the mean of individual BGP slopes was positive (0.38 ± 0.6 ng/ml per year, p = 0.0001). The rate of change in BGP was not related to serum creatinine levels or dietary calcium intake.
Acidosis inhibits 1,25-(OH)2D3 but not cAMP production in response to parathyroid hormone in the rat
Authors: Ro Heung-Kyu , Tembe Vrishali , Krug Thomas , Yang Po-Yao J. , Bushinsky David A. , Dr. Favus Murray J.
Published: March 2010

Parathyroid hormone (PTH) is a major activator of renal proximal tubule 25-hydroxyvitamin D3-1-hydroxylase (1-OHase). Chronic metabolic acidosis (CMA) inhibits 1-OHase and reduces circulating 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] levels in rats fed a low-Ca diet (LCD, 0.002% Ca). To examine the cellular mechanism whereby CMA inhibits 1-OHase, PTH-dependent renal 1-OHase activity and cAMP were measured in proximal tubules isolated from rats fed LCD for 14 days and made acidotic by the addition of 1.5% ammonium chloride to the drinking water. Serum 1,25-(OH)2D3 and proximal tubule 1-OHase activity and cAMP content were lower in acidotic rats. hPTH-(1–34) (10−7 M) in vitro increased cAMP content to equivalent concentrations in tubules from rats with CMA and from nonacidotic controls; however, PTH increased 1-OHase activity only in tubules from nonacidotic animals. Although forskolin increased tubule cAMP content to equivalent levels in tubules from acidotic and nonacidotic rats, 1-OHase activity declined in tubules from nonacidotic rats and remained suppressed in acidotic tubules.

The results suggest that chronic metabolic acidosis inhibits the PTH activation of 1-OHase through alteration of one or more steps in a cAMP-independent messenger system. PTH and forskolin can increase cAMP production by acidotic and nonacidotic proximal tubules; however, 1-OHase activity is not restored to normal in acidotic tubules and nonacidotic tubule 1-OHase may be inhibited.
Aminohydroxybutane bisphosphonate inhibits bone loss due to immobilization in rats
Authors: Thompson David D. , Seedor J. Gregory , Weinreb Miron , Rodan Gideon A. , Rosini Sergio
Published: March 2010

The purpose of this study was to document the effects of aminobutane bisphosphonate (AHBuP) on bone remodeling during immobilization in rats.

Male Sprague-Dawley rats underwent unilateral sciatic neurectomy after receiving two daily subcutaneous injections of 0, 0.01, 0.10, or 1.0 mg P per kg AHBuP. Rats were sacrificed at 24 h or 10 or 20 days postimmobilization. Femora were ashed and tibiae were prepared for histomorphometric analysis.

AHBuP was effective in inhibiting bone loss due to immobilization in a dose-dependent manner. The percentage loss of femoral ash weight due to immobilization decreased in a dose-dependent manner. In vehicle-treated rats, there was a significant decrease in trabecular bone volume (TBV) in the immobilized tibiae compared to the normal tibiae; in AHBuP-treated rats there was a dose-dependent increase in TBV both in the immobilized and control tibiae. The osteoid surface extent was decreased in AHBuP-treated rats in a dose-dependent manner. The mineral apposition rate was altered only in the intact leg of rats treated with 0.1 and 1.0 mg P AHBuP per kg compared to vehicle treated. Osteoclast number per mm was reduced by AHBuP treatment.

In conclusion, aminohydroxybutane bisphosphonate effectively prevented the bone loss due to immobilization in this system.