bluuebunny/crossref_metadata_2025 huggingface dataset:
AP‐1 is involved in ICOS gene expression downstream of TCR/CD28 and cytokine receptor signaling
Authors: Watanabe Masashi , Nakajima Shinsuke , Ohnuki Kazunobu , Ogawa Shuhei , Yamashita Masakatsu , Nakayama Toshinori , Murakami Yasufumi , Tanabe Kazunari , Abe Ryo
Published: May 2012

It has been proposed that sustained ICOS expression in chronic inflammatory immune conditions, such as autoimmunity and allergy, contributes to symptom exacerbation. Therefore modulation of ICOS gene expression could be a potential therapeutic strategy for such immune diseases. However, the precise molecular mechanisms controlling ICOS gene expression remain poorly understood. In this study, we explored transcription factors involving in ICOS gene expression and examined their roles in a physiological situation. Microarray analysis revealed that one AP‐1 molecule, Fos‐related antigen‐2 (Fra2), was highly correlated with ICOS expression. Ectopic expression of Fra2 and other AP‐1 molecules upregulated ICOS expression on T cells. We identified an AP‐1‐responsive site (AP1‐RE) within the ICOS promoter region and demonstrated AP‐1 actually binds to AP1‐RE upon TCR/CD28 stimulation. Meanwhile, we found several cytokines could upregulate ICOS expression on both naïve and effector T cells in a manner independent of TCR/CD28 stimulation. These cytokine stimuli induced AP‐1 binding to AP1‐RE. Together, our results indicate AP‐1 transcription factors are involved in ICOS gene expression downstream of both TCR/CD28 signaling and cytokine receptor signaling, and suggest AP‐1 activation via cytokine receptor signaling may be one of the mechanisms maintaining high level ICOS expression in chronic inflammatory immune responses.
DC‐derived TSLP promotes Th2 polarization in LPS‐primed allergic airway inflammation
Authors: Zhang Yanlu , Zhou Xueping , Zhou Baohua
Published: May 2012

Thymic stromal lymphopoietin (TSLP) plays important roles in the pathogenesis of allergic diseases. Whether and how TSLP is involved in the initial priming of T helper type‐2 (Th2) differentiation against harmless antigen remains unclear. Using an intranasal sensitization protocol with OVA and LPS, we showed that TSLP signaling is required for low‐dose LPS‐induced Th2 inflammation, but not for high‐dose LPS‐induced Th1 immunity. We further demonstrated that low‐dose LPS‐activated bone marrow‐derived dendritic cells expressed relatively high Tslp but low Il12a, and were able to prime naïve DO11.10 T cells to differentiate into Th2 cells in a TSLP‐dependent manner. After transfer into wild‐type recipient mice, the low‐dose LPS‐activated OVA‐loaded dendritic cells (DCs) induced airway eosinophilia, but primed neutrophil‐dominated airway inflammation when TSLP‐deficient DCs were used. These studies demonstrate that TSLP released by DCs in response to a low concentration of LPS plays a role in priming Th2 differentiation and thus may serve as a polarizing third signal, in addition to antigen/MHC class II and co‐stimulatory factors, from antigen‐presenting DCs to direct effector T‐cell differentiation.
Melanoma cells become resistant to NK‐cell‐mediated killing when exposed to NK‐cell numbers compatible with NK‐cell infiltration in the tumor
Authors: Balsamo Mirna , Vermi William , Parodi Monica , Pietra Gabriella , Manzini Claudia , Queirolo Paola , Lonardi Silvia , Augugliaro Raffaella , Moretta Alessandro , Facchetti Fabio , Moretta Lorenzo , Mingari Maria Cristina , Vitale Massimo
Published: May 2012

During the past few years, a number of studies reported that different melanoma cell lines could be extensively lysed in vitro by IL‐2‐activated NK cells at appropriate effector/target ratios. Here, we show, by histological evaluation of different melanoma lesions, that NK/target‐cell ratios compatible with those allowing efficient melanoma cell killing in vitro are hardly reached at the tumor site. We then investigated the outcome of cocultures established at low NK/melanoma cell ratios. After initial NK‐mediated lysis, residual melanoma cells acquired resistance to IL‐2‐activated NK cells. This reflected primarily an increased expression, on melanoma cells, of classical and nonclassical HLA class I molecules, accompanied by a partial downregulation of NKG2D‐ligands, and was dependent on NK‐mediated IFN‐γ release. Consistently, melanoma lesions showed a higher HLA class I expression on tumor cells that were proximal to infiltrating NK cells. In long‐term cocultures, the “protective phenotype” acquired by melanoma cells was lost over time. However, this phenomenon was counteracted by downregulation of relevant activating receptors in cocultured NK cells. Analysis of different NK‐cell‐activating cytokines indicated that IL‐15 can partially overcome this novel tumor escape mechanism suggesting that IL‐15, rather than IL‐2, may be eligible for NK‐cell‐based immunotherapy.
Deletion of the ST2 proximal promoter disrupts fibroblast‐specific expression but does not reduce the amount of soluble ST2 in circulation
Authors: Lipsky Brian P. , Toy Dean Y. , Swart David A. , Smithgall Molly D. , Smith DirkE.
Published: May 2012

IL‐33 signals through ST2, which is expressed either as a full‐length signaling receptor or a truncated soluble receptor that can suppress IL‐33 activity. Previous data suggest that soluble ST2 mRNA in fibroblasts is coupled to a serum‐inducible proximal promoter, while full‐length ST2 expression in immune cells is directed from a distal promoter. In order to better understand the function of the alternative promoters and how they ultimately affect the regulation of IL‐33, we generated a mouse in which the ST2 proximal promoter is deleted. Promoter deletion had no impact on ST2 expression in mast cells or their ability to respond to IL‐33. In contrast, it resulted in a complete loss of both soluble and full‐length ST2 mRNA in fibroblasts, which corresponded with both an inability to secrete soluble ST2 and a defect in IL‐33 responsiveness. Importantly, in spite of the fibroblast defect, soluble ST2 concentrations were not reduced in the serum of naïve or allergen‐exposed knockout mice. In summary, we found that ST2 promoter usage is largely cell‐type dependent but does not dictate splicing. Moreover, the proximal promoter is not a major driver of circulating soluble ST2 under the conditions tested.
Thymocyte death by neglect: Contribution of engulfing macrophages
Authors: Szondy Zsuzsa , Garabuczi Éva , Tóth Katalin , Kiss Bea , Köröskényi Krisztina
Published: May 2012

The thymus provides the microenvironment in which thymocytes develop into mature T cells, and interactions with thymic stromal cells are thought to provide the necessary signals for thymocyte maturation. Recognition of self‐MHC by T cells is a basic requirement for mature T‐cell functions, and those thymocytes that do not recognize the peptide‐loaded self‐MHC molecules found in the thymus, and therefore lack a TCR signal, undergo a default death pathway named “death by neglect” in the thymic cortex. In the absence of this TCR signaling, it has been suggested that binding of glucocorticoids to — or the ligation of certain cell surface molecules, such as CD8, CD24, CD45, or CD99 on — these neglected thymocytes will induce them to enter the apoptotic program. Apoptotic thymocytes are cleared by the surrounding macrophages and, as a consequence, these macrophages are known to release various molecules, such as adenosine, retinoids, TGF‐β, ATP, and carbon monoxide. Interestingly, all these molecules have been described to induce or promote apoptosis in thymocytes in the absence of TCR signaling. Here, we propose that thymic macrophages, because they continually engulf apoptotic cells, might constantly provide these cell death‐inducing signals, and thus contribute to the formation of a thymic milieu that ensures the effective induction of “death by neglect”.
CD4+T‐cell immunity after pandemic influenza vaccination cross‐reacts with seasonal antigens and functionally differs from active influenza infection
Authors: Schmidt Tina , Dirks Jan , Enders Martin , Gärtner Barbara C. , Uhlmann‐Schiffler Heike , Sester Urban , Sester Martina
Published: May 2012

Antigen‐specific antibodies are well characterized after vaccination with pandemic H1N1 or seasonal influenza vaccines. However, knowledge on cellular immunity toward pandemic H1N1 after vaccination and infection and cross‐reactivities toward seasonal antigens is limited. Nineteen individuals were vaccinated with the pandemic H1N1 vaccine. Among those, ten had been prevaccinated against seasonal influenza. CD4+ T cells specific for pandemic H1N1 and for seasonal vaccine, and antibodies were monitored using flow cytometry and ELISA/neutralization assays, respectively. In addition, seven patients with acute pandemic influenza infection were analyzed. Pandemic H1N1 vaccination induced a strong 4.63‐fold (IQR 4.16) increase in antigen‐specific CD4+ T cells that was more pronounced in individuals not prevaccinated with seasonal influenza (p = 0.01). T‐cell levels toward seasonal vaccine concomitantly rose by 2.71‐fold (IQR 2.26). Likewise, prevaccination with seasonal influenza induced a less pronounced increase in specific antibodies. Influenza‐specific T cells in vaccinees had a Th1 phenotype mainly coexpressing IFN‐γ and IL‐2, whereas patients with active pandemic influenza showed a shift toward cells predominantly expressing IFN‐γ. In conclusion, T cells toward seasonal influenza antigens cross‐react with pandemic H1N1 antigens and affect induction of specific T cells after pandemic influenza vaccination. In addition, the cytokine patterns of specific T cells during acute H1N1 infection and after vaccination differ, and the predominantly dual‐positive cytokine profile of vaccine‐induced T cells suggests sufficient functionality to confer successful virus control.
Dynamic manipulation of modes in an optical waveguide using dielectrophoresis
Authors: Kayani Aminuddin A. , Khoshmanesh Khashayar , Nguyen Thach G. , Kostovski Gorgi , Chrimes Adam F. , Nasabi Mahyar , Heller Daniel A. , Mitchell Arnan , Kalantar‐zadeh Kourosh
Published: May 2012

The emergence of optofluidics has brought a high degree of tuneability and reconfigurability to optical devices. These possibilities are provided by characteristics of fluids including mobility, wide range of index modulation, and abrupt interfaces that can be easily reshaped. In this work, we created a new class of optofluidic waveguides, in which suspended mesoparticles were employed to greatly enhance the flexibility of the system. We demonstrated tuneable quasi single mode waveguides using spatially controllable mesoparticles in optofluidics. The coupling of waveguiding modes into the assembly of mesoparticles produces strong interactions and resonant conditions, which promote the transitions of the waveguiding modes. The modal response of the system depends on the distribution of packed particles above the polymeric rib waveguide which can be readily controlled under the appropriate combination of dielectrophoresis and hydrodynamic forces.
Critical evaluation and further development of methods for testing ecotoxicity at multiple pH using Daphnia magna and Pseudokirchneriella subcapitata
Authors: Rendal Cecilie , Trapp Stefan , Kusk Kresten Ole
Published: May 2012

To meet the requirements of risk assessment legislature regarding the ecotoxicity of ionizing compounds, the present study attempts to establish easy, robust methods for testing ecotoxicity at various pH levels. An overview is given of the buffering methods found in the literature. This is supplemented by a series of experiments where toxicity and ability to stabilize pH of seven common buffering compounds was tested on Daphnia magna and Pseudokirchneriella subcapitata. We consider a buffer applicable at a given concentration if the pH drift is below 0.2 pH units, and if there are no toxic effects. Twenty-four- and 48-h acute toxicity tests with D. magna were carried on a series of organic buffers with pH monitoring. Based on the experimental results it is possible to give recommendations for buffer concentrations for use in toxicity testing with D. magna at pH levels in the range of pH 6.0–7.8 for 48 h exposure, and pH 6.0–9.5 for 24 h exposure. Forty-eight- and 72-h growth inhibition tests with P. subcapitata were carried out, and recommendations for buffer concentrations at pH 7.5 and 8.0 are made for both 48 and 72 h of exposure. Environ. Toxicol. Chem. 2012; 31: 1843–1852. © 2012 SETAC
Musk strawberries: the flavour of a formerly famous fruit reassessed
Authors: Pet'ka Ján , Leitner Erich , Parameswaran Baskaran
Published: May 2012

Fruits of wildly growing and naturally ripened musk strawberries (Fragaria moschata) from two highland sites were studied during two seasons. The fruits were characterized on site by two flavourists and a full profile of the volatile compounds was extracted immediately after collection using solid phase extraction and analysed by gas chromatography–mass spectrometry (GC‐MS) and gas chromatography–olfactometry (GC‐O). The aroma of whole fruits was characterized with dynamic headspace coupled to GC‐MS. Single fruits evaluated on site were characterized by green, spicy, seedy and sweet exotic notes, while the fruit bunches showed a complex tropical smell. More than 100 distinctive volatile compounds were detected by GC‐MS. Some of them are reported for the first time in the Fragaria species, most notably the abundant coniferyl alcohol. GC‐O revealed that mesifuran, eugenol, methyl butyrate, furaneol and 3‐mercaptohexyl acetate were the key components of the highland musk strawberry flavour. A comparison of the volatile pattern with the much more known woodland strawberry (F. vesca) from a nearby location contrasted the differences in the volatile composition of the musk strawberry. Repeated analysis of fruits harvested 2 years after the first vintage confirmed these data. Copyright © 2012 John Wiley & Sons, Ltd.
Biallelic inactivation of NF1 in a sporadic plexiform neurofibroma
Authors: Beert Eline , Brems Hilde , Renard Marleen , Ferreiro Julio Finalet , Melotte Cindy , Thoelen Reinhilde , De Wever Ivo , Sciot Raf , Legius Eric , Debiec‐Rychter Maria
Published: May 2012

Plexiform neurofibromas are a major cause of morbidity in individuals with neurofibromatosis type 1 (NF1). Sporadically, these tumors appear as an isolated feature without other signs of NF1. A role for the NF1 gene in solitary plexiform neurofibromas has never been described. In this study, we report a 13‐year‐old boy who was diagnosed with a plexiform neurofibroma, without other NF1 diagnostic criteria. The tumor was partially resected and analyzed using different techniques: karyotyping, fluorescence in situ hybridization (FISH), and microarray comparative genomic hybridization (aCGH). Tumor Schwann cell culture and subsequent karyotyping showed a rearrangement involving chromosomes 1 and 17, namely an insertion of chromosomal bands 1p36‐35 at 17q11.2. FISH demonstrated that the insertion interrupted the NF1 gene. In addition, a deletion was detected affecting the other NF1 allele. Whole‐genome aCGH analysis of the resected tumor confirmed the presence of an 8.28 Mb deletion including the NF1 gene locus in ∼15–20% of tumor cells. We conclude that biallelic NF1 inactivation was at the origin of the isolated plexiform neurofibroma in this patient. The insertion is most likely the “first hit” and the large deletion the “second hit.” © 2012 Wiley Periodicals, Inc.
Exploring Virtual Worlds in a Computerised Hamster Wheel
Authors: Gantenberg Julia , Schill Kerstin , Zetzsche Christoph
Published: May 2012

A new experimental platform called “VirtuSphere” is helping scientists to understand how we perceive and process spatial information. Their findings will help scientists to develop the next generation of assistive devices and robotic systems.
Evaluation and critique of Healthcare Failure Mode and Effect Analysis applied in a radiotherapy case study
Authors: Chadwick Liam , Fallon Enda F.
Published: May 2012

Healthcare Failure Mode and Effect Analysis (HFMEA®) has been developed to support the proactive analysis of health care treatment processes. Its use has been reported in several studies with both positive and negative comments regarding the quality of the results and the efficacy of the method. This paper examines the HFMEA® methodology and discusses the challenges that were encountered during its application to a proactive analysis of radiotherapy patient record systems in a large public hospital. Several weaknesses in the HFMEA® method and particularly in the HFMEA Decision TreeTM are also discussed. Recommendations to support future HFMEA® studies and also for the re‐design of the HFMEA Decision TreeTM to overcome the identified weaknesses are included. Based on these recommendations, a new decision tree is presented to guide future analyses. © 2012 Wiley Periodicals, Inc.
Adoption of ISO 9000 Practices in Manufacturing Companies: The Perspective of the Shop‐Floor Workers
Authors: Heras‐Saizarbitoria Iñaki , Cilleruelo Ernesto , Zamanillo Ibon
Published: May 2012

This article analyzes the adoption of ISO 9000 practices in manufacturing organizations from the perspective of the shop‐floor workers. For that purpose, extensive exploratory fieldwork based on both in‐depth interviews of employees, as well as on participant observation, was carried out in four Basque manufacturing organizations that have adopted the standard. The results show that the employees have a complex, critical, and heterogeneous perspective on the adoption of ISO 9000. In a set of organizations, most of the employees give an account of interesting evidence of how the ceremonial adoption of ISO 9000 occurs, whereas in other organizations the majority describe how ISO 9000 is adopted substantively. The results and the conclusions of the article have implications for academic theory, managers, and public policy. © 2012 Wiley Periodicals, Inc.
Flow and heat transfer of couple‐stress permeable fluid sandwiched between viscous fluid layers
Authors: Umavathi J.C.
Published: May 2012

The fully developed flow and heat transfer in a horizontal channel consisting of couple‐stress permeable fluid sandwiched between viscous fluid layers is investigated analytically. The channel walls are maintained at two different constant temperatures. The transport properties of the fluids in all regions are assumed to be constant. The governing equations are linear ordinary differential equations and hence closed form solutions are obtained. Effects of physical parameters such as viscosity ratio, thermal conductivity ratio, Eckert number, and Prandtl number on the flow are reported. An interesting and new approach is incorporated to analyze the flow for strong, weak, and comparable porosity with couple‐stress parameter. The variation of rate of heat transfer for different values of couple stress parameter and porosity is also discussed. © 2012 Wiley Periodicals, Inc. Heat Trans Asian Res; Published online in Wiley Online Library (wileyonlinelibrary.com/journal/htj). DOI 10.1002/htj.21007
Analysis of precipitation extremes with the assessment of regional climate models over the Willamette River Basin, USA
Authors: Halmstad Andrew , Najafi Mohammad Reza , Moradkhani Hamid
Published: May 2012

An appropriate, rapid and effective response to extreme precipitation and any potential flood disaster is essential. Providing an accurate estimate of future changes to such extreme events due to climate change are crucial for responsible decision making in flood risk management given the predictive uncertainties. The objective of this article is to provide a comparison of dynamically downscaled climate models simulations from multiple model including 12 different combinations of General Circulation Model (GCM)–regional climate model (RCM), which offers an abundance of additional data sets. The three major aspects of this study include the bias correction of RCM scenarios, the application of a newly developed performance metric and the extreme value analysis of future precipitation. The dynamically downscaled data sets reveal a positive overall bias that is removed through quantile mapping bias correction method. The added value index was calculated to evaluate the models' simulations. Results from this metric reveal that not all of the RCMs outperform their host GCMs in terms of correlation skill. Extreme value theory was applied to both historic, 1980–1998, and future, 2038–2069, daily data sets to provide estimates of changes to 2‐ and 25‐year return level precipitation events. The generalized Pareto distribution was used for this purpose. The Willamette River basin was selected as the study region for analysis because of its topographical variability and tendency for significant precipitation. The extreme value analysis results showed significant differences between model runs for both historical and future periods with considerable spatial variability in precipitation extremes. Copyright © 2012 John Wiley & Sons, Ltd.
Targeted therapy for triple‐negative breast cancer: Where are we?
Authors: Duffy Michael J. , McGowan Patricia M. , Crown John
Published: May 2012

Breast cancers that are negative for estrogen receptor (ER), progesterone receptors (PR) and HER2, using standard clinical assays, have been dubbed triple‐negative (TN). Unlike other molecular subtypes of invasive breast cancer, validated targeted therapies are currently unavailable for patients with TN breast cancer. Preclinical studies however, have identified several potential targets such as epidermal growth factor receptor (EGFR), SRC, MET and poly ADP ribose polymerase 1/2 (PARP1/2). Because of tumor heterogeneity, it is unlikely that any single targeted therapy will be efficacious in all patients with TN breast cancer. The rational way forward for treating these patients is likely to be biomarker‐driven, combination targeted therapies or combination of targeted therapy with cytotoxic chemotherapy.
AKT as locus of cancer angiogenic robustness and fragility
Authors: Radisavljevic Ziv
Published: May 2012

Angiogenesis get full robustness in metastatic cancer, relapsed leukemia or lymphoma when complex positive feedback loop signaling systems become integrative. A cancer hypoxic microenvironment generates positive loops inducing formation of the vascular functional shunts. AKT is an upstream angiogenic locus of integrative robustness and fragility activated by the positive loops. AKT controls two downstream nodes the mTOR and NOS in nodal organization of the signaling genes. AKT phosphorylation is regulated by a balance of an oxidant/antioxidant. Targeting AKT locus represents new principle to control integrative angiogenic robustness by the locus chemotherapy. J. Cell. Physiol. 228: 21–24, 2013. © 2012 Wiley Periodicals, Inc.
Nanog promotes osteogenic differentiation of the mouse mesenchymal cell line C3H10T1/2 by modulating bone morphogenetic protein (BMP) signaling
Authors: Ogasawara Toru , Ohba Shinsuke , Yano Fumiko , Kawaguchi Hiroshi , Chung Ung‐il , Saito Tadahito , Yonehara Yoshiyuki , Nakatsuka Takashi , Mori Yoshiyuki , Takato Tsuyoshi , Hoshi Kazuto
Published: May 2012

How the pluripotency of stem cells is maintained and the role of transcription factors in this maintenance remain major questions. In the present study, in order to clarify the mechanism underlying the pluripotency of stem cells for the advancement of regenerative medicine, we examined the effect of forced Nanog expression in mesenchymal cells, with a particular focus on osteogenic differentiation. The human mesenchymal stromal cells (hMSCs) or mouse mesenchymal cell line C3H10T1/2 cells were transduced with the Nanog gene or control green fluorescent protein (GFP) gene by using retrovirus vectors. Short‐term, forced Nanog gene expression had few effects on the terminal osteogenic differentiation of either hMSCs or C3H10T1/2 cells. To determine its long‐term effects, we established C3H10T1/2 cells expressing Nanog constitutively. Constitutive Nanog expression strongly induced osteogenic differentiation of C3H10T1/2 cells. In regard to cell proliferation, constitutive Nanog expression only repressed the proliferation of the cells treated with rhBMP‐2. Moreover, Nanog also had the potential to promote the proliferation of C3H10T1/2 cells in the absence of rhBMP‐2. Constitutive Nanog expression enhanced phosphorylation of Smad1/5/8 and suppressed Cdk4 and cyclinD1. The promoter activities of both the osteocalcin and Id‐1 genes were activated in cells expressing Nanog constitutively. To identify downstream molecules of Nanog involved in the promotion of osteogenic differentiation, we performed a DNA microarray analysis and discovered that NFATc1 was one of the downstream effectors of Nanog. These results indicate that Nanog functions as a modulator of BMP signaling in C3H10T1/2 cells probably through a genome reprogramming process. J. Cell. Physiol. 228: 163–171, 2013. © 2012 Wiley Periodicals, Inc.
Prostaglandin F2α: A bone remodeling mediator
Authors: Agas Dimitrios , Marchetti Luigi , Hurley Marja M. , Sabbieti Maria Giovanna
Published: May 2012

Prostaglandin F2α (PGF2α) plays multiple roles on bone metabolism by regulating a wide range of signaling pathways. PGF2α, via activation of PKC, stimulates Na‐dependent inorganic phosphate (Pi) transport system in osteoblasts; up‐regulates interleukin (IL)‐6 synthesis; increases vascular endothelial growth factor (VEGF). In addition, PGF2α acts as a strong mitogenic and survival agent on osteoblasts, and these effects are, at least in part, mediated by the binding of fibroblast growth factor‐2 (FGF‐2) to the specific receptor FGFR1. The understanding of PGF2α intracellular network, albeit complex to clarify, provides molecular bases useful to identify the players of osteoblast proliferation, apoptosis, and the associated angiogenic processes. Indeed, the molecular mechanism that underline PGF2α‐regulated bone metabolism may be a promising platform for the development of novel targeted therapies in the treatment of bone disorders and disease. J. Cell. Physiol. 228: 25–29, 2013. © 2012 Wiley Periodicals, Inc.
Autophagy and ionizing radiation in tumors: The “survive or not survive” dilemma
Authors: Palumbo Silvia , Comincini Sergio
Published: May 2012

Autophagy is a so‐called “self‐eating” system responsible for degrading long‐lived proteins and cytoplasmic organelles, whose products are recycled to maintain cellular homeostasis. This ability makes autophagy a good candidate for a survival mechanism in response to several stresses, including the tumor cell transformation. In particular, recent studies suggested that autophagy functions as a pro‐death mechanism within different tumor contexts. It is, however, widely reported that autophagy represents both a survival mechanism or contributes directly to cell death fate. This interplay of the autophagy functions has been observed in many types of cancers and, in some cases, autophagy has been demonstrated to both promote and inhibit antitumor drug resistance. From a therapeutical point of view, the effects of the modulation of the tumor cell autophagic status, in response to ionizing radiations, are presently of particular relevance in oncology. Accordingly, this review also provides a perspective view on future works for exploring the modulation of autophagic indices in tumor cells as a novel molecular‐based adjuvant strategy, in order to improve radiotherapy and chemotherapy effects in cancer patients. J. Cell. Physiol. 228: 1–8, 2013. © 2012 Wiley Periodicals, Inc.
Acrosome Reaction in the Epididymal Sperm of Chinchilla lanigera. Effect of Progesterone, A23187 and Stimulation of a GABAA‐Like Receptor
Authors: GRAMAJO‐BÜHLER MARÍA CECILIA , ZELARAYÁN LILIANA , LÓPEZ LUIS ALBERTO , SÁNCHEZ‐TORANZO GRACIELA
Published: May 2012

Little is known about the reproductive biology of Chinchilla lanigera in captivity. Currently there are no parameters to evaluate the quality of the semen of males that are sold as breeding stock, so they are selected according to their phenotypic characteristics, which cannot guarantee their reproductive efficiency. We worked with sexually mature males; the sperm were collected from the cauda epididymis by puncture. Capacitated sperm were incubated at 37 °C with different concentrations of progesterone. The results showed a biphasic behavior, where the increase in the percentage of the acrosome reaction (AR) was significant between 2.5–5 μM, and a maximum response with a dose of 20 μM. Also the suspension of sperm incubated with different concentrations of ionophore A23187 inducing response is obtained with a concentration of 10 μM. Stimulation of the Gamma‐aminobutyric acid (GABA)A‐like receptor by treating the sperm with different concentrations of muscimol, a specific receptor agonist, also induce acrosome reaction. The dose‐response curve showed a biphasic effect with a maximum response at a concentration of 2.5 μM. The pretreatment of sperm suspension with a specific receptor antagonist, picrotoxin, a decrease was observed in the percentage of reacted perm. These studies represent a first step towards the establishment of standard parameters to determine the functionality and fertilizing capacity of Chinchilla sperm. In addition, our results indicate that the Chinchilla spermatozoa can undergo the AR when treated with A23187 or progesterone at high percentages. It also shows a clear involvement of GABAA‐like receptors in acrosome exocytosis in sperm obtained by epididymal cauda. J. Exp. Zool.317:259–265, 2012. © 2012 Wiley Periodicals, Inc.
David and Goliath: A Mitochondrial Coupling Problem?
Authors: SALIN KARINE , ROUSSEL DAMIEN , REY BENJAMIN , VOITURON YANN
Published: May 2012

An organism's size, known to affect biological structures and processes from cellular metabolism to population dynamics, depends upon the duration and rate of growth. However, it is still poorly understood how mitochondrial function affects the energetic basis of growth, especially in ectotherms, which represent a huge majority of animal biodiversity. Here, we present an intraspecies comparison of neighboring populations of frogs (Rana temporaria) that have large differences in body mass even at the same age. By investigating liver mitochondrial bioenergetics, we find that frogs with high growth rates and large body sizes exhibit higher ATP synthesis rates and more efficient oxidative phosphorylation compared to the smaller frogs with low growth rates. This higher energy transduction efficiency is not associated with significant increased oxidative capacity or membrane potential values, but instead may rely on a higher mitochondrial phosphorylation system activity in combination with a lower inner membrane proton leakage. Overall, the present study introduces the mitochondrial energy transduction system as an important mechanism for balancing physiological and ecological trade‐offs associated with body size. Whether phenotype differences in mitochondrial function result from local ecological constraints or reflect a natural genetic variability within wild populations of common frogs remains an open question. However, our findings highlight the need for closer consideration of all aspects of mitochondrial metabolism for a better understanding of the physiological basis of the link between size, metabolism, and energy production in wild‐dwelling organisms. J. Exp. Zool.317:283–293, 2012. © 2012 Wiley Periodicals, Inc
Involvement of p38MAPKin the Induction ofHsp70 During Acute Thermal Stress in Red Blood Cells of the Gilthead Sea Bream,Sparus aurata
Authors: FEIDANTSIS KONSTANTINOS , PÖRTNER HANS O. , MARKOU THOMAIS , LAZOU ANTIGONE , MICHAELIDIS BASILE
Published: May 2012

The present study aimed to examine the expression and activation ofMAPKs (p38MAPK,ERK1/2, andJNKs) in red blood cells (RBCs) of the gilthead sea bream,Sparus aurata,during thermal stress and investigate their involvement in the expression of heat shock proteins. The data showed that only p38MAPKis detected inRBCs ofSparus aurataand it is phosphorylated and activated during exposure to increased temperature. Induction ofHsp70 in thermally stressedRBCs was abolished in the presence of the p38MAPKinhibitor,SB203580, suggesting the involvement of the kinase in this response. This mechanism might play a cytoprotective role in theRBCs of the gilthead sea bream.J. Exp. Zool.317:303–310, 2012. © 2012 Wiley Periodicals, Inc.
Supraphysiological Cortisol Elevation Alters the Response of Wild Bluegill Sunfish to Subsequent Stressors
Authors: McCONNACHIE SARAH H. , O'CONNOR CONSTANCE M. , GILMOUR KATHLEEN M. , IWAMA GEORGE K. , COOKE STEVEN J.
Published: May 2012

Wild fish are frequently exposed to multiple stressors, but the influence of previous or ongoing stress on an animal's subsequent response is poorly understood. Using wild‐caught bluegill sunfish (Lepomis macrochirus) as a model, we used exogenous hormone implants to experimentally raise circulating cortisol in a group of fish for ∼10 days. We also maintained sham‐treated and control groups of fish. We subjected all animals to a secondary stressor in the form of either a heat challenge or fasting challenge. We compared survival, body condition, and plasma‐borne indicators of physiological status among cortisol‐treated, sham‐treated, and control groups following the secondary stressor. In order to compare short‐ and long‐term effects of cortisol treatment, we initiated the secondary stressor either 4 or 30 days following initial cortisol treatment. Cortisol‐treated fish succumbed to the fasting challenge sooner than sham‐treated and control fish at both 4 and 30 days. Interestingly, cortisol‐treated fish lost equilibrium sooner than sham‐treated and control fish during the heat challenge when conducted at 30 days, but not at 4 days. These results demonstrate that multiple simultaneous stressors have cumulative effects on bluegill sunfish. Furthermore, these results demonstrate that supraphysiological cortisol doses alter the long‐term responses of bluegill sunfish to additional challenges, even after apparent recovery. Such cumulative and long‐term effects may be an important factor in mediating the response of wild animals to natural and anthropogenic stressors, and should be considered in ecological studies. J. Exp. Zool.317:321–332, 2012. © 2012 Wiley Periodicals, Inc
The viral etiology of an influenza‐like illness during the 2009 pandemic
Authors: Thiberville S.D. , Ninove L. , Hai V. Vu. , Botelho‐Nevers E. , Gazin C. , Thirion L. , Salez N. , de Lamballerie X. , Charrel R. , Brouqui P.
Published: May 2012

Many viruses are known to cause influenza‐like illness (ILI); however, in nearly 50% of patients, the etiologic agent remains unknown. The distribution of viruses in patients with ILI was investigated during the 2009 A/H1N1 influenza pandemic (A/H1N1p). From June 2009 to January 2010, 660 patients with suspected influenza were questioned and examined, and nasal swabs were collected. All patient samples were tested for influenza virus, and 286 negative nasal swabs were tested further for 18 other respiratory viruses using real‐time RT‐PCR. Two waves of ILI were observed in the epidemic curve (weeks 35–42 and 42–49). At least eight viruses co‐circulated during this period: human rhinovirus (HRV) (58), parainfluenza 1–4 viruses (PIV) (9), human Coronavirus (hCoV) OC43 (9), enterovirus (5), adenovirus (AdV) (4), and human metapneumovirus (hMPV) (2); however, 204 samples remained negative for all viruses tested. ILI symptoms, according to the Centers for Disease Control and Prevention criteria for ILI definition, were reported in 75% of cases. These patients had positive swabs for A/H1N1p, HRV, hCoV‐OC43, PIV, AdV, and hMPV without significant difference with non‐ILI patients. This study found that many respiratory viruses circulated during this period and that the A/H1N1p did not impact on the kinetics of other respiratory viruses. The proportion of non‐documented cases remains high. ILI could not distinguish A/H1N1p infection from that due to other respiratory viruses. However, in multivariate anlaysis, cough, chills, hyperemia, and dyspnea were associated significantly with influenza virus versus other respiratory viruses. J. Med. Virol. 84: 1071–1079, 2012. © 2012 Wiley Periodicals, Inc.
Characterization of HIV‐1 subtypes and drug resistance mutations among individuals infected with HIV in Georgia
Authors: Dvali Natia , Parker Monica M. , Chkhartishvili Nikoloz , Sharvadze Lali , Gochitashvili Nino , Abutidze Akaki , Karchava Marine , DeHovitz Jack A. , Tsertsvadze Tengiz
Published: May 2012

In order to describe HIV‐1 subtypes and drug resistance mutations in Georgia, blood samples from 153 patients infected with HIV‐1 collected from 2006 to 2008 were genotyped. Of these, 126 samples were from newly diagnosed, antiretroviral (ARV)‐naïve patients and 27 from ARV‐treated patients. Partial pol region sequences were used to identify drug resistance mutations and to conduct phylogenetic analysis for subtype determination. The results indicated that 138 (90.2%) patients harbored subtype A viruses, 11 (7.2%) carried subtype B virus, two subtype G (1.3%), one (0.6%) subtype F and one (0.6%) 03_AB recombinant. All subtype A strains clustered with the Former Soviet Union A (A FSU) subtype. Among patients with no prior exposure to ARVs, mutations associated with resistance were detected in five patients: three (2.4%) patients had reverse transcriptase (RT) inhibitor mutations and two other patients had the protease (PI) inhibitor associated mutation M46I. PI mutation V77I was found in 42 of subtype A isolates. Of 27 ARV‐treated patients, 22 (81.5%) harbored at least one nucleoside reverse transcriptase inhibitors (NRTI), a non‐NRTI (NNRTI) and/or a PI mutation. The most common NRTI resistance mutation was M184V/I (74.1%). Frequency of thymidine analog mutations was relatively low (25.9%). With regard to NNRTI mutations, G190S/A was the most frequent mutation, which might be a preferred mutations for subtype A. Georgia's HIV epidemic continues to be dominated by Subtype A FSU. The prevalence of transmitted drug resistance is low, but has the potential to increase with increasing use of ARVs. J. Med. Virol. 84: 1002–1008, 2012. © 2012 Wiley Periodicals, Inc.
ITPA gene polymorphisms significantly affect hemoglobin decline and treatment outcomes in patients coinfected with HIV and HCV
Authors: Osinusi Anu , Naggie Susanna , Poonia Seerat , Trippler Martin , Hu Zonghui , Funk Emily , Schlaak Joerg , Fishbein Dawn , Masur Henry , Polis Michael , Kottilil Shyam
Published: May 2012

Published studies have described a strong association with a single‐nucleotide polymorphism (SNP) in the inosine triphosphate pyrophosphatase (ITPA) gene and ribavirin (RBV)‐induced hemolytic anemia in HCV‐infected patients receiving pegylated interferon (pegIFN) and RBV. This study sought to evaluate the effect of these polymorphisms on anemia, hemoglobin reduction, HCV kinetics, and treatment outcomes. Sixty‐three patients coinfected with HIV and HCV and 58 patients infected with HCV only were treated with pegIFN/RBV were genotyped using the ABI TaqMan allelic discrimination kit for the 2 ITPA SNP variants rs1127354 and rs7270101. A composite variable of ITPA deficiency using both SNPs was created as previously reported. Statistical analysis was performed using Mann‐Whitney test or Chi square/Fishers exact test for categorical data and mixed model analysis for multiple variables. Thirty‐five patients (30%) were predicted to have reduced ITPA activity. ITPA deficiency was found to be protective against the development of hemoglobin reduction >3 g/dl over the course of treatment. The rates of hemoglobin reduction >3 g/dl decreased in correlation with the severity of ITPA deficiency. ITPA deficiency was associated with slower hemoglobin decline early in treatment (week 4, P = 0.020) and rapid virologic response (RVR) at week 4 (P = 0.017) in patients coinfected with HIV and HCV. ITPA polymorphisms are associated with hemoglobin decline and in patients coinfected with HIV and HCV it is also associated with early virologic outcomes. Determination of ITPA polymorphisms may allow prediction of RBV‐induced anemia and earlier initiation of supportive care to ensure optimal therapeutic outcomes. J. Med. Virol. 84: 1106–1114, 2012. Published 2012. This is a US Government work and as such is in the public domain in the United States of America.
Type 1 diabetes associated and tissue transglutaminase autoantibodies in patients without type 1 diabetes and coeliac disease with confirmed viral infections
Authors: Sarmiento Luis , Galvan José A , Cabrera‐Rode Eduardo , Aira Lazaro , Correa Consuelo , Sariego Susel , Fonseca Magile , Cubas‐Dueñas Ileana , Hung Lai Heng , Resik Sonia , Cilio Corrado M.
Published: May 2012

Coeliac disease and type 1 diabetes are autoimmune diseases that may share the same initiating environmental factors. In this study, the occurrence of type 1 diabetes associated autoantibodies (GADA and IA‐2A) and tissue transglutaminase autoantibodies (TGA) was determined in patients with confirmed viral infections and no signs of type 1 diabetes or coeliac disease. Serum samples from 82 Cuban patients tested positive for PCR and IgG specific to enterovirus (HEV, serotype echovirus 16, 20 samples), Epstein–Barr virus (EBV, 20 samples), cytomegalovirus (CMV, 21 samples), and hepatitis C virus (HCV, 21 samples); and sera from 164 controls negative serologically to EBV, CMV, HCV, and echovirus 16 were enrolled in the study. All subjects were screened for GADA, IA‐2A, and TGA. The prevalence of TGA in patients infected with HEV, EBV, CMV, or HCV was 55% (11/20), 25% (5/20), 9.5% (2/21), and 9.5% (2/21), respectively. GADA and IA‐2A were found in 15% (3/20) and 25% (5/20) of patients infected with HEV. None of the patients infected by EBV, CMV, and HCV had GADA or IA‐2A. All children infected with HEV who were positive for type 1 diabetes‐associated autoantibodies were also TGA‐positive. None of the sera from uninfected subjects were positive for GADA, IA‐2A or TGA. In conclusion, TGA can develop during infection with HEV, EBV, CMV, or HCV, while the emergence of islet cell related autoantibodies is restricted to HEV infections. The findings suggest that HEV may be a shared environmental factor for the development of islet and gut‐related autoimmunity. J. Med. Virol. 84: 1049–1053, 2012. © 2012 Wiley Periodicals, Inc.
Human T‐lymphotropic virus type I (HTLV‐I)‐specific CD8+ cells accumulate in the lungs of patients infected with HTLV‐I with pulmonary involvement
Authors: Kawabata Takashi , Higashimoto Ikkou , Takashima Hiroshi , Izumo Shuji , Kubota Ryuji
Published: May 2012

Pulmonary involvement has been identified in human T‐lymphotropic virus type I (HTLV‐I) carriers and patients with HTLV‐I‐associated myelopathy/tropical spastic paraparesis (HAM/TSP). However, the relationship between HTLV‐I infection and lung disease is poorly understood. The occurrence of HTLV‐I‐specific immune responses in the lungs of patients infected with HTLV‐I with pulmonary involvement was investigated. The frequency of HTLV‐I‐specific CD8+ cells and the amount of HTLV‐I proviral DNA were determined in bronchoalveolar lavage fluid cells and peripheral blood mononuclear cells (PBMCs) from five patients with HAM/TSP and one HTLV‐I carrier who had pulmonary involvement. HTLV‐I‐specific CD8+ cells were detected by flow cytometry using human leukocyte antigen/antigen complex multimers. The analysis of bronchoalveolar lavage fluid revealed lymphocytosis in five of six patients. HTLV‐I provirus was detected in the bronchoalveolar lavage fluid cells of all patients, and the proviral load in these cells was comparable to that in PBMCs. The frequency of HTLV‐I‐specific CD8+ cells in the bronchoalveolar lavage fluid cells was 5.1 times higher than that in PBMCs. Immunohistochemically, clusters formed by HTLV‐I‐specific CD8+ cells were detected in lung tissue by in situ tetramer staining. No samples were available from patients infected with HTLV‐I without lung disorders. Whether accumulation of CD8+ cells is specific to patients with pulmonary involvement remains unclear. These results indicate that HTLV‐I‐specific CD8+ cells accumulate and HTLV‐I‐infected cells exist in the lungs of patients infected with HTLV‐I with pulmonary involvement. J. Med. Virol. 84: 1120–1127, 2012. © 2012 Wiley Periodicals, Inc.
Fe protein over‐expression can enhance the nitrogenase activity of Azotobacter vinelandii
Authors: Nag Papri , Pal Subrata
Published: May 2012

The effects of over‐expression of NifH (Fe protein) on nitrogenase activity in Azotobacter vinelandii UW cells were studied by expressing an extra nifH gene under the control of the inducible meta‐toluic acid pathway promoter Pm. The total amount of protein in UW/pJB654‐N reacting to anti‐NifH antibody was 2–3 fold of that in control UW when both the strains were grown to exponential phase in the presence of 4 μM m‐toluic acid. As a consequence UW/pJB654‐N showed two‐fold higher acetylene reduction activity and released 70% higher amounts of ammonium into the growth medium than the control. Concomitant changes were observed also in the cellular levels of siderophores and iron superoxide dismutase (FeSOD). Thus, our results indicating that increased level of Fe protein in the cell can enhance nitrogen fixation activity of A. vinelandii may have biotechnological significance.
Biodegradation of keratinous waste by Chryseobacterium sp. RBT isolated from soil contaminated with poultry waste
Authors: Gurav Ranjit G. , Jadhav Jyoti P.
Published: May 2012

In the present study, a feather degrading bacterial strain was isolated from poultry waste disposal site, Kolhapur, India. The bacterium was identified as Chryseobacterium sp. RBT using 16S rRNA gene sequence analysis. Chryseobacterium sp. RBT showed rapid hydrolysis of native feathers within 30 h and produced the highest level of keratinase activity (98.3 U/ml). Keratin containing wastes viz. silk, human hair, wool and chicken feathers were tested for keratin degrading ability of the bacterium. Amongst the tested substrates, the Chryseobacterium sp. RBT showed more specificity towards chicken feathers (98.6% degradation) with maximum keratinase activity (98.3 U/ml) and solubilized protein concentration (3.84 mg/ml). Effect of various physico‐chemical parameters (temperature, pH, carbon and nitrogen sources) on keratinase production was monitored. The maximum keratinase activity was observed at pH (8.6) and temperature (50 °C). Molasses (1.0% w/v) acted as an inducer and enhanced the keratinolytic activity by two fold, while starch worked as an inhibitor. The goat skin when treated with crude keratinase enzyme (2% v/v), showed complete dehairing within 12 h. Hence, Chryseobacterium sp. RBT shows potential as a candidate for treating the keratinous waste in an ecofriendly manner.
Differential catalytic action of Brevibacillus laterosporus on two dissimilar azo dyes Remazol red and Rubine GFL
Authors: Kurade Mayur B. , Waghmode Tatoba R. , Tamboli Dhawal P. , Govindwar Sanjay P.
Published: May 2012

This comparative study disclosed the diverse catalytic activities of Brevibacillus laterosporus on two different azo dyes. It decolorized 100% of Remazol red and 95% of Rubine GFL within 30 and 48 h respectively, under static condition at 50 mg l−1 dye concentration. Significant increase was observed in azo reductase, NADH‐DCIP reductase, veratryl alcohol oxidase and tyrosinase in cells obtained after decolorization of Remazol red; whereas these values were much different with complete inhibition of azo reductase during decolorization of Rubine GFL. The plausible pathway of dye degradation obtained from Gas chromatography‐Mass spectroscopy (GC‐MS) data confirmed the different metabolic fate of these structurally unidentical dyes. FTIR and HPTLC analysis of extracted metabolites confirmed the biodegradation, while phytotoxicity study assured the detoxification of both the dyes studied. The results obtained in this study suggests, i) sulpho and hydroxyl group present at ortho position to azo group stimulated reduction of azo bond by azo reductase in Remazol red, ii) the same reduction was totally hampered due to presence of ethyl‐amino propanenitrile group at para position to azo group in Rubine GFL.