bluuebunny/crossref_metadata_2025 huggingface dataset:
Viscoelastic properties of injectable bone cements for orthopaedic applications: State‐of‐the‐art review
Authors: Lewis Gladius
Published: April 2011

Injectable bone cements (IBCs) are used for a variety of orthopaedic applications, examples being poly (methyl methacrylate) (PMMA) bone cements used for anchoring total joint replacements (TJRs) (high load‐bearing application), PMMA bone cements used in the vertebral body augmentation procedures of vertebroplasty (VP) and balloon kyphoplasty (BKP) (medium load‐bearing application), and calcium phosphate‐based and calcium sulfate‐based cements used as bone void fillers/bone graft substitutes (low load‐bearing application). For each of these applications, the viscoelastic properties of the cement are very important. For example, (1) creep of the cement has an influence on the longevity of a cemented TJR (for example, creep allows the cement to remodel, thereby maximizing the contact area of the cement‐bone interface and, hence, minimizing stress concentration at that interface); and (2) in VP and BKP, the likelihood of cement extravasation is directly related to the profile of the viscosity‐versus‐time elapsed from commencement of mixing of the cement. There are a few reviews of the literature on a number of viscoelastic properties of some IBCs but a comprehensive review of the literature on all viscoelastic properties of all IBCs is lacking. The objective of this contribution is to present such a review. In addition, a number of ideas for future study in the field of viscoelastic properties of IBCs are described. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2011.
SHIP2 multiple functions: A balance between a negative control of PtdIns(3,4,5)P3 level, a positive control of PtdIns(3,4)P2 production, and intrinsic docking properties
Authors: Erneux Christophe , Edimo William's Elong , Deneubourg Laurence , Pirson Isabelle
Published: April 2011

The SH2 domain containing inositol 5‐phosphatase 2 (SHIP2) belongs to the family of the mammalian inositol polyphosphate 5‐phosphatases. The two closely related isoenzymes SHIP1 (or SHIP) and SHIP2 contain a N‐terminal SH2 domain, a catalytic domain, potential PTB domain‐binding sites (NPXY), and C‐terminal proline‐rich regions with consensus sites for SH3 domain interactions. In addition, SHIP2 contains a unique sterile alpha motif (SAM) domain that could be involved in SAM–SAM domain interactions with other proteins or receptors. SHIP2 also shows the presence of an ubiquitin interacting motif at the C‐terminal end. SHIP2 is essentially a PI(3,4,5)P3 5‐phosphatase that negatively controls PI(3,4,5)P3 levels in intact cells and produce PI(3,4)P2. Depending on the cells and stimuli, PI(3,4)P2 could accumulate at important levels and be a “second messenger” by its own. It could interact with a very large number of target proteins such as PKB or TAPP1 and 2 that control insulin sensitivity. In addition to its catalytic activity, SHIP2 is also a docking protein for a large number of proteins: Cytoskeletal, focal adhesion proteins, scaffold proteins, adaptors, protein phosphatases, and tyrosine kinase associated receptors. These interactions could play a role in the control of cell adhesion, migration, or endocytosis of some receptors. SHIP2 could be acting independently of its phosphatase activity being part of a protein network of some receptors, e.g., the EGF receptor or BCR/ABL. These non‐catalytic properties associated to a PI phosphatase have also been reported for other enzymes of the metabolism of myo‐inositol such as Ins(1,4,5)P3 3‐kinases, inositol phosphate multikinase (IPMK), or PTEN. J. Cell. Biochem. 112: 2203–2209, 2011. © 2011 Wiley‐Liss, Inc.
Comprehensive patterns in microRNA regulation of transcription factors during tumor metastasis
Authors: Reshmi G. , Sona Charles , Pillai M. Radhakrishna
Published: April 2011

In spite of a large body of information about the upstream regulators of metastasis, a process that often plays a limiting factor in therapeutic outcome of cancer patients, the impact of regulatory microRNA patterns remains obscure. This review describes computational analysis of coordinated regulation of genes by di‐directional regulation of microRNA and transcription factors that specifically regulate the process of metastasis. We discovered several unexpected modes of regulatory patterns between microRNAs and transcription factors. For example, we found a double positive feedback loop regulated by the hub transcription factor ZEB1 and miR‐200 during epithelial–mesenchymal transition. This review further explains flow of information and how such components coordinate various adaptable controls of microRNAs and thus, contribute to regulation of transcription factors in context of cancer metastasis. Information described here provides a regulatory framework for future experimental analyses and discoveries of new insights into post‐transcriptional gene regulation at the microRNA level in cancer metastasis. J. Cell. Biochem. 112: 2210–2217, 2011. © 2011 Wiley‐Liss, Inc.
Over‐expression of FoxM1 leads to epithelial–mesenchymal transition and cancer stem cell phenotype in pancreatic cancer cells
Authors: Bao Bin , Wang Zhiwei , Ali Shadan , Kong Dejuan , Banerjee Sanjeev , Ahmad Aamir , Li Yiwei , Azmi Asfar S. , Miele Lucio , Sarkar Fazlul H.
Published: April 2011

FoxM1 is known to play important role in the development and progression of many malignancies including pancreatic cancer. Studies have shown that the acquisition of epithelial‐to‐mesenchymal transition (EMT) phenotype and induction of cancer stem cell (CSC) or cancer stem‐like cell phenotypes are highly inter‐related, and contributes to drug resistance, tumor recurrence, and metastasis. The molecular mechanism(s) by which FoxM1 contributes to the acquisition of EMT phenotype and induction of CSC self‐renewal capacity is poorly understood. Therefore, we established FoxM1 over‐expressing pancreatic cancer (AsPC‐1) cells, which showed increased cell growth, clonogenicity, and cell migration. Moreover, over‐expression of FoxM1 led to the acquisition of EMT phenotype by activation of mesenchymal cell markers, ZEB1, ZEB2, Snail2, E‐cadherin, and vimentin, which is consistent with increased sphere‐forming (pancreatospheres) capacity and expression of CSC surface markers (CD44 and EpCAM). We also found that over‐expression of FoxM1 led to decreased expression of miRNAs (let‐7a, let‐7b, let‐7c, miR‐200b, and miR‐200c); however, re‐expression of miR‐200b inhibited the expression of ZEB1, ZEB2, vimentin as well as FoxM1, and induced the expression of E‐cadherin, leading to the reversal of EMT phenotype. Finally, we found that genistein, a natural chemo‐preventive agent, inhibited cell growth, clonogenicity, cell migration and invasion, EMT phenotype, and formation of pancreatospheres consistent with reduced expression of CD44 and EpCAM. These results suggest, for the first time, that FoxM1 over‐expression is responsible for the acquisition of EMT and CSC phenotype, which is in part mediated through the regulation of miR‐200b and these processes, could be easily attenuated by genistein. J. Cell. Biochem. 112: 2296–2306, 2011. © 2011 Wiley‐Liss, Inc.
Cigarette smoke‐induced skeletal muscle atrophy is associated with up‐regulation of USP‐19 via p38 and ERK MAPKs
Authors: Liu Qian , Xu Wei‐Guo , Luo Yong , Han Feng‐Feng , Yao Xiao‐Hong , Yang Tian‐Yun , Zhang Yue , Pi Wei‐Feng , Guo Xue‐Jun
Published: April 2011

Ubiquitin‐specific proteases (USPs) deubiquitinate ubiquitin–protein conjugates in the ubiquitin–proteasome system. Previous research shows that ubiquitin‐specific protease‐19 (USP‐19) is up‐regulated in mammalian skeletal muscle in some degradative conditions, such as including fasting, diabetes, dexamethasone treatment, and cancer, and its function is associated with muscle atrophy. However, it is still unclear whether USP‐19 is involved in muscle atrophy induced by chronic obstructive pulmonary disease. Rats exposed to chronic cigarette smoke and L6 myotubes incubated with cigarette smoke extract (CSE) were studied here. Using western blot analysis and quantitative real‐time polymerase chain reaction (qPCR), we observed over‐expression of USP‐19 and down‐regulation of myosin heavy chain (MHC) in both models. Moreover, CSE exposure inhibited myogenic differentiation and myotube formation in L6 myotubes. To explore the mechanism underlying these effects, we investigated the levels of phosphorylated mitogen‐activated protein kinases (MAPKs) and total MAPKs. Exposing myotubes to CSE resulted in the general activation of MAPKs such as p38, JNK, and ERK1/2. The ERK inhibitor PD98059 and the p38 inhibitor SB203580 significantly blocked the increase in USP‐19 gene expression induced by CSE. Our findings suggest that USP‐19 is associated with muscle atrophy in response to cigarette smoke and is a potential therapeutic target. CSE promotes myotube wasting in culture partly by inhibiting myogenic differentiation and acts via p38 and ERK MAPK to stimulate expression of USP‐19 in vitro. J. Cell. Biochem. 112: 2307–2316, 2011. © 2011 Wiley‐Liss, Inc.
TCERG1 inhibits C/EBPα through a mechanism that does not involve sequestration of C/EBPα at pericentromeric heterochromatin
Authors: Moazed Banafsheh , Banman Shanna L. , Wilkinson Grey A. , Roesler William J.
Published: April 2011

Transcriptional elongation regulator 1 (TCERG1) is a nuclear protein that participates in multiple events that include regulating the elongation of RNA polymerase II and coordinating transcription and pre‐mRNA processing. More recently, we showed that TCERG1 is also a specific inhibitor of the transcription factor CCAAT enhancer binding protein α (C/EBPα). Interestingly, the inhibition of C/EBPα by TCERG1 is associated with the relocalization of TCERG1 from the nuclear speckle compartment to the pericentromeric regions where C/EBPα resides. In the present study, we examined additional aspects of C/EBPα‐induced redistribution of TCERG1. Using several mutants of C/EBPα, we showed that C/EBPα does not need to be transcriptionally competent or have anti‐proliferative activity to induce TCERG1 relocalization. Moreover, our results show that C/EBPα does not need to be localized to the pericentromeric region in order to relocalize TCERG1. This conclusion was illustrated through the use of a V296A mutant of C/EBPα, which is incapable of binding to the pericentromeric regions of heterochromatin and thus takes on a dispersed appearance in the nucleus. This mutant retained the ability to redistribute TCERG1, however in this case the redistribution was from the nuclear speckle pattern to the dispersed phenotype of C/EBPα V296A. Moreover, we showed that TCERG1 was still able to inhibit the activity of the V296A mutant. While we previously hypothesized that TCERG1 might inhibit C/EBPα by keeping it sequestered at the pericentromeric regions, our new findings indicate that TCERG1 can inhibit C/EBPα activity regardless of the latter's location in the nucleus. J. Cell. Biochem. 112: 2317–2326, 2011. © 2011 Wiley‐Liss, Inc.
Minimum MD simulation length required to achieve reliable results in free energy perturbation calculations: Case study of relative binding free energies of fructose‐1,6‐bisphosphatase inhibitors
Authors: Rathore R.S. , Aparoy P. , Reddanna P. , Kondapi A. K. , Rami Reddy M.
Published: April 2011

In an attempt to establish the criteria for the length of simulation to achieve the desired convergence of free energy calculations, two studies were carried out on chosen complexes of FBPase‐AMP mimics. Calculations were performed for varied length of simulations and for different starting configurations using both conventional‐ and QM/MM‐FEP methods. The results demonstrate that for small perturbations, 1248 ps simulation time could be regarded a reasonable yardstick to achieve convergence of the results. As the simulation time is extended, the errors associated with free energy calculations also gradually tapers off. Moreover, when starting the simulation from different initial configurations of the systems, the results are not changed significantly, when performed for 1248 ps. This study carried on FBPase‐AMP mimics corroborates well with our previous successful demonstration of requirement of simulation time for solvation studies, both by conventional and ab initio FEP. The establishment of aforementioned criteria of simulation length serves a useful benchmark in drug design efforts using FEP methodologies, to draw a meaningful and unequivocal conclusion. © 2011 Wiley Periodicals, Inc. J Comput Chem, 2011
Detecting coached feigning using the test of Memory Malingering (TOMM) and the structured inventory of Malingered Symptomatology (SIMS)
Authors: Jelicic Marko , Ceunen Erik , Peters Maarten J.V. , Merckelbach Harald
Published: April 2011

Undergraduate students were administered the Test of Memory Malingering (TOMM) and the Structured Inventory of the Malingered Symptomatology (SIMS) and asked to respond honestly, or instructed to feign cognitive dysfunction due to head injury. Before both instruments were administered, symptom‐coached feigners were provided with some information about brain injury, while feigners who received a mix of symptom‐coaching and test‐coaching were given the same information plus advice on how to defeat symptom validity tests. Results show that, although the accuracy of both instruments appears to be somewhat reduced by a mix of symptom coaching and test coaching, the TOMM and SIMS are relatively resistant to different kinds of coaching. © 2011 Wiley Periodicals, Inc. J Clin Psychol 67:1–6, 2011.
Targeting mitochondria as a therapeutic target in cancer
Authors: Wenner Charles E.
Published: April 2011

Knowledge of re‐programming in cancer cells with metabolic differences from their normal counterparts has resulted in new examination of therapeutic approaches. Several studies of the role of tumor mitochondria in cancer have led to the development of non‐genotoxic therapies which target mitochondrial proteins, function. The now well‐established functions of mitochondria in apoptosis provide novel targets for tumor cell suicide. Mitochondria serve as a central hub for responses to cellular stress as well as injury. The alterations in cancer cells which result in protection from apoptosis can be targeted to inhibit proliferation. Because of the reprogramming of cancer cell metabolism involving increased glycolysis, it appears that blocking InsP3R Ca2+ release or adaptive pathways in response to hypoxia by targeting HIF‐1 or metabolic enzymes encoded by the HIF‐1 gene represents a feasible therapeutic approach to cancer. A very early in vitro event found in tumor cells following resveratrol addition is an increase in intracellular Ca2+, measurable within seconds. Ca2+ release is also observed with non‐toxic flavonoids and a goal to identify the sentinel targets of resveratrol as a model compound involved in calcium activation seems worthwhile. New findings of the relationship between autophagy and apoptosis are discussed. The contribution of reactive oxygen species (ROS) generated by mitochondria is also considered. New data as to how cyclophilins and VDAC are involved in mitochondrial hexokinase protection of factors that induce apoptosis are reviewed. In addition, chemotherapeutic approaches based on Akt‐activated mTORC1 are described, and their relationship to the role of aerobic glycolysis in this protection. J. Cell. Physiol. © 2011 Wiley Periodicals, Inc.
High expression of ubiquitin conjugates and NF‐κB in unstable human intracranial atherosclerotic plaques
Authors: Sun Ruihong , Xiao Ling , Duan Shurong
Published: April 2011

Arteries from the first segment of the middle cerebral and adjacent normal blood vessel specimens were collected from 19 patients who had died from cerebral infarction. According to morphologic examination the atherosclerotic plaque present was classified as stable or unstable. The expression of ubiquitin conjugates and nuclear factor kappa B (NF‐κB) was assessed and found significantly higher in plaques than normal tissue and higher in unstable plaques than in stable plaques. Ubiquitin conjugates and NF‐κB were positively correlated with each other. It was concluded that the ubiquitin proteasome system and NF‐κB possibly play a role in the development of atherosclerotic plaques in intracranial arteries. J. Cell. Physiol. 227: 784–788, 2012. © 2011 Wiley Periodicals, Inc.
Tumorigenic polyploid cells contain elevated ROS and ARE selectively targeted by antioxidant treatment
Authors: Roh Meejeon , van der Meer Riet , Abdulkadir Sarki A.
Published: April 2011

Polyploidy has been linked to tumorigenicity mainly due to the chromosomal aberrations. Elevated reactive oxygen species (ROS) generation, on the other hand, has also been associated with oncogenic transformation in most cancer cells. However, a possible link between ploidy and ROS is largely unexplored. Here we have examined the role of ROS in the tumorigenicity of polyploid cells. We show that polyploid prostate and mammary epithelial cells contain higher levels of ROS due to their higher mitochondrial contents. ROS levels and mitochondrial mass are also higher in dihydrocytochalasin B (DCB)‐induced polyploid cells, suggesting that higher levels of ROS observed in polyploid cell can occur due to cytokinesis failure. Interestingly, polyploid cells were more sensitive to the inhibitory effect of the antioxidant, N‐Acetyl‐L‐cysteine (NAC), than control diploid cells. Treatment of polyploid/diploid cells with NAC led to the selective elimination of polyploid cells over time and abrogated the tumorigenicity of polyploid cells. This effect was partially mediated via the Akt signaling pathway. We next explored a possible role for ROS in promoting chromosomal instability by analyzing the effects of ROS on the mitotic stage of the cell cycle. Enhancing ROS levels by treating cells with hydrogen peroxide delayed not only entry into and but also exit from mitosis. Furthermore, increasing ROS levels significantly increased taxol resistance. Our results indicated that increased ROS in polyploid cells can contribute to tumorigenicity and highlight the therapeutic potential of antioxidants by selectively targeting the tumorigenic polyploid cells and by reversing taxol resistance. J. Cell. Physiol. 227: 801–812, 2012. © 2011 Wiley Periodicals, Inc.
PML as a potential predictive factor of oxaliplatin/fluoropyrimidine‐based first line chemotherapy efficacy in colorectal cancer patients
Authors: Vincenzi Bruno , Santini Daniele , Perrone Giuseppe , Graziano Francesco , Loupakis Fotios , Schiavon Gaia , Frezza Anna Maria , Ruzzo Anna Maria , Rizzo Sergio , Crucitti Pierfilippo , Galluzzo Sara , Zoccoli Alice , Rabitti Carla , Muda Andrea Onetti , Russo Antonio , Falcone Alfredo , Tonini Giuseppe
Published: April 2011

PML regulates a wide range of pathways involved in tumorigenesis, such as apoptosis, which is also one of the main mechanisms through which oxaliplatin and fluoropyrimidine exert their antineoplastic activity. The present study aims to investigate PML expression as a predictive factor of oxaliplatin/fluoropyrimidine therapy efficacy. Seventy‐four metastatic colorectal cancer patients who received oxaliplatin/floropyrimidine‐based first line therapy have been included in this retrospective study. PML expression was assessed by immunohistochemistry. PML down‐regulation was detected in 39 (52.7%) patients (14 complete and 25 partial PML loss). RR was significantly lower (25.6%) in patients with PML down‐regulation than in patients with preserved PML expression (60%) (P = 0.006). Median TTP was 5.5 months when PML was down‐regulated versus 11.9 months in case of preserved PML expression (P < 0.0001). A statistical significant difference was also detected in OS (15.6 and 24.5 months, respectively, P = 0.003). The impact of PML down‐regulation on TTP and OS was statistically significant also in a multivariate model. This study represents the first evidence of a possible correlation between PML protein expression and outcome of metastatic colorectal cancer patients treated with oxaliplatin/fluoropyrimidine‐based first line therapy. J. Cell. Physiol. 227: 927–933, 2012. © 2011 Wiley Periodicals, Inc.
Functional redundancy of type II BMP receptor and type IIB activin receptor in BMP2‐induced osteoblast differentiation
Authors: Liu Hongbin , Zhang Rongrong , Chen Di , Oyajobi Babatunde O. , Zhao Ming
Published: April 2011

Signaling pathways for bone morphogenetic proteins (BMPs) are important in osteoblast differentiation. Although the precise function of type I BMP receptors in mediating BMP signaling for osteoblast differentiation and bone formation has been characterized previously, the role of type II BMP receptors in osteoblasts is to be well clarified. In this study, we investigated the role of type II BMP receptor (BMPR‐II) and type IIB activin receptor (ActR‐IIB) in BMP2‐induced osteoblast differentiation. While osteoblastic 2T3 cells expressed BMPR‐II and ActR‐IIB, loss‐of‐function studies, using dominant negative receptors and siRNAs, showed that BMPR‐II and ActR‐IIB compensated each other functionally in mediating BMP2 signaling and BMP2‐induced osteoblast differentiation. This was evidenced by two findings. First, unless there was loss of function of both type II receptors, isolated disruption of either BMPR‐II or ActR‐IIB did not remove BMP2 activity. Second, in cells with loss of function of both receptors, restoration of function of either BMPR‐II or ActR‐IIB by transfection of the wild‐type forms, restored BMP2 activity. These findings suggest a functional redundancy between BMPR‐II and ActR‐IIB in osteoblast differentiation. Results from experiments to test the effects of transforming growth factor β (TGF‐β), activin, and fibroblast growth factor (FGF) on osteoblast proliferation and differentiation suggest that inhibition of receptor signaling by double‐blockage of BMPR‐II and ActR‐IIB is BMP‐signaling specific. The observed functional redundancy of type II BMP receptors in osteoblasts is novel information about the BMP signaling pathway essential for initiating osteoblast differentiation. J. Cell. Physiol. 227: 952–963, 2012. © 2011 Wiley Periodicals, Inc.
Allodynic skin in post‐herpetic neuralgia: Histological correlates
Authors: Buonocore Michelangelo , Gatti Anna Maria , Amato Georgia , Aloisi Anna Maria , Bonezzi Cesare
Published: April 2011

Most post‐herpetic neuralgia (PHN) patients suffer from tactile allodynia (pain evoked by lightly touching the skin) and it is frequently the dominant clinical manifestation. The pathophysiology of tactile allodynia in PHN patients is poorly understood and this is one of the major limits to the development of appropriate therapies. Epidermal nerve fibres (ENFs) are free nerve endings of small‐diameter A‐delta and C primary afferents, which can easily be assessed by neurodiagnostic skin biopsy (NSB). The aim of this study was to establish the correlation between the residual epidermal innervation of the allodynic skin and the intensity of tactile allodynia in that area. Twenty‐five patients (13 males and 12 females) with PHN were enrolled. Eighteen patients had PHN in the thoracic dermatome, four in the cervical, two in the trigeminal and one in the lumbar. The severity of allodynia evoked by a paintbrush was graded according to an eleven‐point numerical scale. A skin biopsy was obtained from the maximal allodynia area and from the contralateral skin. Nerve fibres were labelled with indirect immunofluorescence. Results showed that epidermal innervation was lower in the allodynic skin than in the contralateral skin, although there was great variability among patients. There was no correlation between severity of allodynia and epidermal innervation of the PHN skin. In conclusion, the present study further indicates peripheral nervous system involvement in PHN but does not support a direct correlation between epidermal innervation changes and tactile allodynia. J. Cell. Physiol. 227: 934–938, 2012. © 2011 Wiley Periodicals, Inc.
Distinct cellular responses induced by saporin and a transferrin–saporin conjugate in two different human glioblastoma cell lines
Authors: Cimini A. , Mei S. , Benedetti E. , Laurenti G. , Koutris I. , Cinque B. , Cifone M.G. , Galzio R. , Pitari G. , Di Leandro L. , Giansanti F. , Lombardi A. , Fabbrini M.S. , Ippoliti R.
Published: April 2011

Glioblastoma multiforme (GBM) is the most common primary brain tumour in adults, with a median survival of ∼12–18 months post‐diagnosis. GBM usually recurs within 12 months post‐resection, with poor prognosis. Thus, novel therapeutic strategies to target and kill GBM cells are urgently needed. The marked difference of tumour cells with respect to normal brain cells renders glioblastoma a good candidate for selective targeted therapies. Recent experimental strategies focus on over expressed cell surface receptors. Targeted toxins represent a new class of selective molecules composed by a potent protein toxin and a carrier ligand. Targeted toxins approaches against glioblastoma were under investigation in phase I and II clinical trials with several immunotoxins (IT)/ligand toxins such as IL4‐Pseudomonas aeruginosa exotoxin A (IL4‐PE, NBI‐3001), tumour growth factor fused to PE38, a shorter PE variant, (TGF)alpha‐TP‐38, IL13‐PE38, and a transferrin‐C diphtheriae toxin mutant (Tf‐CRM107). In this work, we studied the effects of the plant ribosome‐inactivating saporin and of its chimera transferrin–saporin against two different GBM cell lines. The data obtained here indicate that cell proliferation is affected by the toxin treatments but that different mechanisms are used, directly linked to the presence of an active or inactive p53. A model is proposed for these alternative intracellular pathways. J. Cell. Physiol. 227: 939–951, 2012. © 2011 Wiley Periodicals, Inc.
Resveratrol suppresses tumorigenicity and enhances radiosensitivity in primary glioblastoma tumor initiating cells by inhibiting the STAT3 axis
Authors: Yang Yi‐Ping , Chang Yuh‐Lih , Huang Pin‐I , Chiou Guang‐Yuh , Tseng Ling‐Ming , Chiou Shih‐Hwa , Chen Ming‐Hsiung , Chen Ming‐Teh , Shih Yang‐Hsin , Chang Chin‐Hong , Hsu Chuan‐Chih , Ma Hsin‐I , Wang Chin‐Tien , Tsai Lo‐Lin , Yu Cheng‐Chia , Chang Charn‐Jung
Published: April 2011

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor. Patients diagnosed with GBM have a poor prognosis, and it has been reported that tumor malignancy and GBM recurrence are promoted by STAT3 signaling. As resveratrol (RV), a polyphenol in grapes, is reported to be a potent and non‐toxic cancer‐preventive compound, the aim of this study was to investigate the therapeutic effect and molecular mechanisms of RV on GBM‐derived radioresistant tumor initiating cells (TIC). Firstly, our results showed that primary GBM‐CD133+ TIC presented high tumorigenic and radiochemoresistant properties as well as increased protein levels of phosphorylated STAT3. We consistently observed that treatment with shRNA‐STAT3 (sh‐STAT3) or AG490, a STAT3 inhibitor, significantly inhibited the cancer stem‐like cell properties and radioresistance of GBM‐CD133+ in vitro and in vivo. Furthermore, treatment of GBM‐CD133+ with 100 µM RV induced apoptosis and enhanced radiosensitivity by suppressing STAT3 signaling. Microarray results suggested that RV or AG490 inhibited the stemness gene signatures of GBM‐CD133+ and facilitated the differentiation of GBM‐CD133+ into GBM‐CD133− or astrocytoma cells. Finally, xenotransplant experiments indicated that RV or sh‐STAT3 therapy could significantly improve the survival rate and synergistically enhance the radiosensitivity of radiation‐treated GBM‐TIC. In summary, RV can reduce in vivo tumorigenicity and enhance the sensitivity of GBM‐TIC to radiotherapies through the STAT3 pathway. J. Cell. Physiol. 227: 976–993, 2012. © 2011 Wiley Periodicals, Inc.
Recent advances in computational analysis of mass spectrometry for proteomic profiling
Authors: Sun Clement S. , Markey Mia K.
Published: April 2011

The proteome, defined as an organism's proteins and their actions, is a highly complex end‐effector of molecular and cellular events. Differing amounts of proteins in a sample can be indicators of an individual's health status; thus, it is valuable to identify key proteins that serve as ‘biomarkers’ for diseases. Since the proteome cannot be simply inferred from the genome due to pre‐ and posttranslational modifications, a direct approach toward mapping the proteome must be taken. The difficulty in evaluating a large number of individual proteins has been eased with the development of high‐throughput methods based on mass spectrometry (MS) of peptide or protein mixtures, bypassing the time‐consuming, laborious process of protein purification. However, proteomic profiling by MS requires extensive computational analysis. This article describes key issues and recent advances in computational analysis of mass spectra for biomarker identification. Copyright © 2011 John Wiley & Sons, Ltd.
Formation of metal complex ions from amino acid in the presence of Li+, Na+ and K+ by electrospray ionization: metal replacement of hydrogen in the ligands
Authors: Jang Soonmin , Ju Song Min , Kim Hyunsik , Choi Sung‐Seen
Published: April 2011

Alkali metal cations easily form complexes with proteins in biological systems; understanding amino acid clusters with these cations can provide useful insight into their behaviors at the molecular level including diagnosis and therapy of related diseases. For the purpose of characterization of basic interaction between amino acids and alkali metal, each of the 20 naturally occurring amino acids were ionized in the presence of lithium, sodium and potassium cations by electrospray ionization, and the resulting product ions were analyzed. We focus our attention on the gas phase alkali metal ion‐proton exchanged complexes in current study, specifically complexes with serine, threonine, asparagine and glutamine, which share characteristic pattern unlike other amino acids. All amino acids generated [M + H]+ and [M + Na]+ ions, where M stands for the neutral amino acid. Serine, threonine, asparagine and glutamine generated cluster ions of [nM − nH + (n + 1)Na]+ and [nM − (n − 1)H + (n − 1)Na + K]+, where n = 1–7. While the (M − H + Li) and (M − H + K) species were not observed, the neutral (M − H + Na) species formed by proton–sodium cation exchange had a highly stable cyclic structure with ketone and amine ligand sites, suggesting that (M − H + Na) serves as a building block in cluster ion formation. Cluster ion intensity distributions of [nM − nH + (n + 1)Na]+ and [nM − (n − 1)H + (n − 1)Na + K]+ showed a magic number at n = 3 and 4, respectively. Extensive B3LYP‐DFT quantum mechanical calculations were carried out to elucidate the geometry and energy of the cluster ions, and they provided a reasonable explanation for the stability and structure of the cluster ions. Copyright © 2011 John Wiley & Sons, Ltd.
T‐cell response to human papillomavirus type 52 L1, E6, and E7 peptides in women with transient infection, cervical intraepithelial neoplasia, and invasive cancer
Authors: Chan Paul K.S. , Liu Shih‐Jen , Cheung Jo L.K. , Cheung T.H. , Yeo Winnie , Chong Pele , Man Stephen
Published: April 2011

The E6 and E7 proteins encoded by human papillomaviruses (HPV) are prime targets for therapeutic vaccine development. Ninety‐five women with HPV 52 infection (33 transient infections, 17 cervical intraepithelial neoplasia grade II, 15 cervical intraepithelial neoplasia grade III, and 30 invasive cervical cancers) were examined for T‐cell responses using interferon‐γ enzyme‐linked immunospot (IFN‐γ ELISPOT) assay. Of the 29 peptides (13 L1, 10 E6, and 6 E7) screened positive by an in vitro peptide‐binding assay, 14 were positive by the IFN‐γ ELISPOT assay. Positive epitopes for HLA A11 were located at amino acid positions 103–111, 332–340, 342–350, and 373–381 of the L1 protein; and at 27–35 and 86–94 of the E6 protein; and at 1–9 and 27–35 of the E7 protein. A24‐specific epitopes included 60–68 and 98–106 of the L1 protein, 42–50 and 59–67 of the E6 protein, and 24–32 of the E7 protein. Only one epitope (99–107) of the E6 protein showed positive responses for HLA A2 subjects. Overall, T‐cell responses against L1 were observed mainly in subjects who had cleared infection; whereas responses against E6 and E7 were confined mainly to subjects who had developed cervical neoplasia. The proportion of subjects showing detectable T‐cell responses was low across all grades of cervical neoplasia suggesting that immune evasion mechanisms had set on early in the course of disease progression. This study provides the first set of T‐cell epitopes mapped for HPV 52, which can be considered for further evaluation as targets for immunotherapy. J. Med. Virol. 83:1023–1030, 2011. © 2011 Wiley‐Liss, Inc.
Prevalence, risk factors and virological profile of chronic hepatitis b virus infection in pregnant women in India
Authors: Pande Chandana , Sarin Shiv Kumar , Patra Sharda , Bhutia Kazila , Mishra Siddhartha Kumar , Pahuja Sangeeta , Jain Manjula , Srivastava Shikha , Dar Sadaf Bashir , Trivedi Shubha Sagar , Mukhopadhyay Chinmay K. , Kumar Ashish
Published: April 2011

A large program was conducted by the Government of India to study the prevalence and profile of chronic hepatitis B virus (HBV) infection and its risk factors in pregnant women attending a tertiary care hospital in India. From September 2004 to December 2008 consecutive pregnant women attending the antenatal clinic were screened and those found positive for HBsAg were enrolled. Healthy non‐pregnant women of child‐bearing age, who presented for blood donation during the same period, served as controls. Women with symptoms of liver disease or those aware of their HBsAg status were excluded. Of the 20,104 pregnant women screened, 224 (1.1%) and of the 658 controls, 8 (1.2%) were HBsAg positive (P = ns). Previous blood transfusions and surgery were significant risk factors for infection with HBV. Of the women who were HBsAg positive, the ALT levels were normal in 54% of the women and HBV DNA levels were above 2,000 IU/ml in 71% of women. The median HBV DNA levels were higher in women who were HBeAg positive compared to the HBeAg negative group. The most common HBV genotype was D (84%) followed by A + D and A (8% each). In conclusion, the prevalence of HBsAg positivity among asymptomatic pregnant women in North India is 1.1% with 71% having high HBV DNA levels. These women may have a high risk of transmitting infection to their newborns. J. Med. Virol. 83:962–967, 2011. © 2011 Wiley‐Liss, Inc.
Occult hepatitis C virus infection in Iranian patients with cryptogenic liver disease
Authors: Bokharaei‐Salim Farah , Keyvani Hossein , Monavari Seyed Hamid Reza , Alavian Seyed Moayed , Madjd Zahra , Toosi Mohssen Nassiri , Alizadeh Amir Houshang Mohammad
Published: April 2011

The diagnosis of cryptogenic liver disease is made when after extensive evaluations, recognizable etiologies of chronic liver disease are excluded. In this study, the presence of hepatitis C virus (HCV) RNA was tested in peripheral blood mononuclear cells (PBMCs) taken from Iranian patients who although were found negative for plasma HCV RNA and anti‐HCV antibodies, suffered from chronic liver disease of unknown etiology. From September 2007 to March 2010, 69 patients from Tehran with chronic liver disease of unknown etiology who were referred to our center were enrolled in the present study. PBMCs were isolated from 10 mL peripheral blood specimens. HCV‐RNA status was tested in plasma and PBMCs samples by reverse‐transcription polymerase chain reaction (RT‐PCR). HCV‐RNA was detected in HCV‐positive PBMCs specimens by RT‐PCR method. HCV genotypes were subsequently analyzed in HCV‐positive samples using restriction fragment length polymorphism (RFLP) assay; then HCV genotypes were confirmed by sequencing of 5′ non‐coding fragments after cloning PCR products into pJET1.2/blunt cloning vector. HCV‐RNA was detected in PBMCs specimens belonging to 7 (10%) out of 69 patients. Genotyping of the HCV‐RNA isolated from PBMCs showed that 3 (43%) patients with occult HCV infection had genotype 1b, 2 (29%) had genotype 1a, and another 2 (29%) had genotype 3a. The results of this study suggest that patients with chronic liver disease of unknown etiology may have occult HCV infection in the absence of anti‐HCV antibodies and plasma HCV‐RNA. It has been suggested that in the absence of liver biopsy specimens, analysis of PBMC sample for HCV‐RNA would be informative. J. Med. Virol. 83:989–995, 2011. © 2011 Wiley‐Liss, Inc.
T1846 and A/G1913 are associated with acute on chronic liver failure in patients infected with hepatitis B virus genotypes B and C
Authors: Yan Tao , Li Ke , Li Fan , Su Haibin , Mu Jinsong , Tong Shuping , Patel Mitul , Xia Jie , Wands Jack R. , Wang Huifen
Published: April 2011

The aim of this study was to determine whether mutations in the hepatitis B virus (HBV) genome are associated with the onset of acute on chronic liver failure (ACLF). For the longitudinal study, full‐length HBV genomes were cloned and sequenced from four ACLF patients and compared with sequences from matching samples collected before ACLF. For the cross‐sectional study, 166 serum samples were obtained, including 49 samples from patients with ACLF. The results of longitudinal study showed that C53T, A1846T, and G1896A were the most common mutations in association with ACLF. In the cross‐sectional study 61.2% patients with ACLF presented with T1846, which was higher than patients with chronic hepatitis B (CHB) (11.1%), liver cirrhosis (LC) (31.1%), and hepatocellular carcinoma (HCC) (33.3%). Prevalence of A/G1913 was 42.9% in patients with ACLF, also higher than patients with CHB (2.2%), LC (17.8%), and HCC (11.1%). There were no differences in HBV genotype and patients' HBeAg status among patients with ACLF, LC, and HCC. However, prevalence of T1846 was much higher in patients infected with genotype B (57.1%) than genotype C (30.4%). A/G1913 was higher in HBeAg negative patients (28%) than HBeAg positive patients (13.2%). Results of a multivariable analysis showed that T1846 and A/G1913 were independent factors for ACLF (OR = 3.373 and 4.244, respectively). Interestingly, T1846 destroys an ATG codon of a small open reading frame in the preC region, which may increase core protein expression. We conclude that T1846 and A/G1913 in the preC/C gene are closely associated with ACLF. J. Med. Virol. 83:996–1004, 2011. © 2011 Wiley‐Liss, Inc.
Molecular characterization of hepatitis C virus genotype 4 sequences in HIV‐coinfected patients from Argentina
Authors: Bolcic F. , Jones L.R. , Laufer N. , Quarleri J.
Published: April 2011

The prevalence of hepatitis C virus genotype 4 (HCV‐4) is increasing in different parts of the World but in Latin America the data are still scarce. We aimed to characterize HCV‐4 isolates from 383 HIV‐coinfected patients in Argentina. Sequence analyses were based on the non‐structural 5B region of HCV. Results from 18 patients indicated a genetic heterogeneity that involved three genotype 4 subtypes. Sequences were ascribed to subtype 4d (67%), 4a (22%), and 4m (11%). In spite of different sources of transmission were defined among patients, no statistical association was found with the genotype 4 subtype. The scenario is also compatible with multiple importation of the epidemic and there is no evidence for transmission‐specific clusters or network‐like transmission of HCV‐4. This HCV‐4 does not represent a recent introduction in Argentina, it circulates in all transmission groups and its presence is increasing among HIV‐infected patients. J. Med. Virol. 83:935–940, 2011. © 2011 Wiley‐Liss, Inc.
Rotavirus A genotype P[4]G2: Genetic diversity and reassortment events among strains circulating in Brazil between 2005 and 2009
Authors: Gómez Mariela Martínez , de Mendonça Marcos César Lima , Volotão Eduardo de Mello , Tort Luis Fernando López , da Silva Marcelle Figueira Marques , Cristina Juan , Leite Jose Paulo Gagliardi
Published: April 2011

Group A rotaviruses (RV‐A) are the leading cause of severe gastroenteritis in infants and young children worldwide. Due to the epidemiologic complexity of RV‐A, especially in developing countries, it is important to determine which genotypes are circulating, principally after the introduction in March 2006 of the monovalent (P[8]G1) Rotarix® vaccine in Brazil by the National Immunization Program. In Phase III trials with Rotarix®, the prevalence of genotype P[4]G2 was extremely low, and therefore, evaluation of heterotypic immunization against this genotype was performed by meta‐analysis statistics tests. Different studies have shown the re‐emergence of genotype P[4]G2 in Brazil, since 2005, as well as in other countries, suggesting that it could be a continental phenomenon related to the temporal variability in the genotype's naturally occurring distribution. It is important to note that genotype P[4]G2 does not share VP4 or VP7 antigens with the vaccine strain. Therefore, we performed a phylogenetic analysis based on VP4 (VP8*), VP7, VP6, and NSP4 genes of RV‐A genotype P[4]G2 samples isolated from the five regions of Brazil between 2005 and 2009. This study revealed that different genetic variants of RV‐A genotype P[4]G2 circulated in Brazil between 2005 and 2009, and that this variability is determined mainly by: occurrence of point mutations; reassortment events; and widespread global gene flow. The results obtained in this study are important to our understanding of the epidemiology and evolution of RV‐A genotype P[4]G2 and demonstrate the importance of continuous monitoring and molecular characterization of RV‐A strains circulating in human and animal populations. J. Med. Virol. 83:1093–1106, 2011. © 2011 Wiley‐Liss, Inc.
Effects of hepatitis C virus on suppressor of cytokine signaling mRNA levels: Comparison between different genotypes and core protein sequence analysis
Authors: Pascarella Stéphanie , Clément Sophie , Guilloux Kévin , Conzelmann Stéphanie , Penin François , Negro Francesco
Published: April 2011

Glucose metabolism disturbances, including insulin resistance and type 2 diabetes, are frequent and important cofactors of hepatitis C. Increasing epidemiological and experimental data suggest that all major genotypes of hepatitis C virus (HCV), albeit to a different extent, cause insulin resistance. The HCV core protein has been shown to be sufficient to impair insulin signaling in vitro through several post‐receptorial mechanisms, mostly via the activation of suppressor of cytokine signaling (SOCS) family members and the consequent decrease of insulin receptor substrate‐1 (IRS‐1). The levels of IRS‐1 and SOCS were investigated upon expression of the core protein of HCV genotypes 1–4. Furthermore, the core protein sequences were analyzed to identify the amino acid residues responsible for IRS‐1 decrease, with particular regard to SOCS mRNA deregulation. The results suggest that the activation of SOCS family members is a general mechanism associated with the common HCV genotypes. A rare genotype 1b variant, however, failed to activate any of the SOCS tested: this allowed to analyze in detail the distinct amino acid sequences responsible for SOCS deregulation. By combining approaches using intergenotypic chimeras and site‐directed mutagenesis, genetic evidence was provided in favor of a role of amino acids 49 and 131 of the HCV core‐encoding sequence in mediating SOCS transactivation. J. Med. Virol. 83:1005–1015, 2011. © 2011 Wiley‐Liss, Inc.
Uteroglobulin‐related protein 1 and severity of respiratory syncytial virus infection in children admitted to hospital
Authors: Hashimoto Koichi , Katayose Masahiko , Sakuma Hiroko , Kawasaki Yukihiko , Sumikoshi Makoto , Sakata Hiroshi , Sato Masatoki , Ohara Shinichiro , Abe Yusaku , Watanabe Masahiro , Sato Toshiko , Ishibashi Kei , Suzutani Tatsuo , Munakata Mitsuru , Hosoya Mitsuaki
Published: April 2011

There are several reports suggesting that genetic factors contribute to the severity of infection with the respiratory syncytial virus (RSV). Infants hospitalized with lower respiratory tract infection (LRTI) due to RSV are at a significantly increased risk for both recurrent wheezing and childhood asthma. Uteroglobin‐related protein 1 (UGRP1) is a secretory protein expressed in the airways, and speculated to have anti‐inflammatory activity. The presence of the −112G/A polymorphism in the UGRP1 promoter was found to have a significant correlation with asthma phenotype. Also plasma UGRP1 levels were shown to be associated both with this polymorphism and the severity of asthma. The study population consisted of 62 previously healthy infants, ≤12 months of age, who were hospitalized with RSV LRTI, and a control group of 99 healthy adults. Genotyping was performed by restriction fragment length polymorphism. UGRP1 serum levels were determined using ELISA. There were no significant differences in the overall distribution of UGRP1 −112G/A polymorphism genotypes or alleles between the hospitalized infants and healthy adults. A comparison of serum UGRP1 concentration measured at the time of admission and discharge between patients with and without the −112A allele revealed that there was no relation between the presence of the −112A allele and serum UGRP1 in hospitalized infants with RSV infection. Furthermore, there was no relationship between severity of RSV infection and genotype or serum UGRP1 concentration. These results suggest that UGRP1 does not have a major role in the development of severe RSV infection. J. Med. Virol. 83:1086–1092, 2011. © 2011 Wiley‐Liss, Inc.
Clinical and Molecular epidemiologic analyses of norovirus‐associated sporadic gastroenteritis in adults from Beijing, China
Authors: Gao Yan , Jin Miao , Cong Xv , Duan Zhaojun , Li Hui‐ying , Guo Xiaolin , Zuo Ying , Zhang Yanmin , Zhang Yi , Wei Lai
Published: April 2011

Noroviruses (NoVs) are a major cause of nonbacterial acute gastroenteritis. The molecular epidemiology of NoV infections in China has not been well characterized. To study the incidence of NoV infections and the nature of the circulating NoV genotypes, 403 specimens were collected from adult outpatients with acute gastroenteritis in Beijing, China, between October 2007 and September 2008. Samples were examined for NoV by reverse transcriptase‐polymerase chain reaction (RT‐PCR) and sequences corresponding to both the NoV polymerase and capsid regions were characterized. Among the 403 specimens, 48 (11.9%) were positive for NoV and 1/3 NoV‐associated gastroenteritis cases occurred during the colder months (November and December). Based on polymerase region sequences, 6 NoV genotypes (GII‐4, GII‐b, GII‐6, GI‐2, GI‐3, and GI‐4) were identified with GII‐4 2006b being the most predominant genotype (37/48, 77.1%). Eleven distinct genotype sequences in polymerase and capsid regions were identified, indicating a genetic diversity among NoV isolates. This study suggested NoVs were an important pathogen responsible for sporadic acute gastroenteritis in adults in Beijing, China. J. Med. Virol. 83:1078–1085, 2011. © 2011 Wiley‐Liss, Inc.
Latent and lytic Epstein‐Barr virus gene expression in the peripheral blood of astronauts
Authors: Stowe Raymond P. , Kozlova Elena V. , Sams Clarence F. , Pierson Duane L. , Walling Dennis M.
Published: April 2011

Epstein‐Barr virus (EBV) latent and replicative gene transcription was analyzed in peripheral blood B‐lymphocytes from astronauts who flew on short‐duration (∼11 days) Shuttle missions and long‐duration (∼180 days) International Space Station (ISS) missions. Latent, immediate‐early, and early gene replicative viral transcripts were detected in samples from six astronauts who flew on short‐duration Shuttle missions, whereas viral gene transcription was mostly absent in samples from 24 healthy donors. Samples from six astronauts who flew on long‐duration ISS missions were characterized by expanded expression of latent, immediate‐early, and early gene transcripts and new onset expression of late replicative transcription upon return to Earth. These data indicate that EBV‐infected cells are no longer expressing the restricted set of viral genes that characterize latency but are expressing latent and lytic gene transcripts. These data also suggest the possibility of EBV‐related complications in future long‐duration missions, in particular interplanetary travel. J. Med. Virol. 83:1071–1077, 2011. © 2011 Wiley‐Liss, Inc.