bluuebunny/crossref_metadata_2025 huggingface dataset:
The effects of body mass index on complications and survival outcomes in patients with cervical carcinoma undergoing curative chemoradiation therapy
Authors: Kizer Nora T. , Thaker Premal H. , Gao Feng , Zighelboim Israel , Powell Matthew A. , Rader Janet S. , Mutch David G. , Grigsby Perry W.
Published: October 2010

BACKGROUND:

The effect of body mass index (BMI) on treatment outcomes for patients with locally advanced cervical carcinoma who receive definitive chemoradiation is unclear.

METHODS:

The cohort in this study included all patients with cervical carcinoma (n = 404) who had stage IB1 disease and positive lymph nodes or stage ≥IB2 disease and received treatment at the authors' facility between January 1998 and January 2008. The mean follow‐up was 47.2 months. BMI was calculated using the National Institute of Health online calculator. BMI categories were created according to the World Health Organization classification system. Primary outcomes were overall survival, disease‐free survival, and complication rate. Univariate and multivariate analyses were performed. Kaplan‐Meier survival curves were generated and compared using Cox proportional hazard models.

RESULTS:

On multivariate analysis, compared with normal weight (BMI 18.5‐24.9 kg/m2), a BMI <18.5 kg/m2 was associated with decreased overall survival (hazard ratio, 2.37; 95% confidence interval, 1.28‐4.38; P < .01). The 5‐year overall survival rate was 33%, 60%, and 68% for a of BMI <18.5 kg/m2, a BMI from 18.5 kg/m2 to 24.9 kg/m2, and a BMI >24.9 kg/m2, respectively. A BMI <18.5 kg/m2 was associated with increased risk of grade 3 or 4 complications compared with a BMI >24.9 kg/m2 (radiation enteritis: 16.7% vs 13.6%, respectively; P = .03; fistula: 11.1% vs 8.8%, respectively; P = .05; bowel obstruction: 33.3% vs 4.4%, respectively; P < .001; lymphedema: 5.6% vs 1.2%, respectively; P = .02).

CONCLUSIONS:

Underweight patients (BMI <18.5 kg/m2) with locally advanced cervical cancer had diminished overall survival and more complications than normal weight and obese patients. Cancer 2011. © 2010 American Cancer Society.
Subcutaneous or intravenous administration of romiplostim in thrombocytopenic patients with lower risk myelodysplastic syndromes
Authors: Sekeres Mikkael A. , Kantarjian Hagop , Fenaux Pierre , Becker Pamela , Boruchov Adam , Guerci‐Bresler Agnes , Hu Kuolung , Franklin Janet , Wang Yow‐Ming C. , Berger Dietmar
Published: October 2010

BACKGROUND:

Romiplostim is a peptibody protein that augments thrombopoiesis by activating the thrombopoietin receptor.

METHODS:

In this phase 2, multicenter, open‐label study, 28 thrombocytopenic patients with lower risk myelodysplastic syndromes (MDS) were assigned to receive romiplostim 750 μg administered subcutaneously either weekly or biweekly or administered as biweekly intravenous injections for 8 weeks. Patients also could enter a 1‐year study extension phase.

RESULTS:

At least 1 adverse event was observed in 93% of patients. The most common adverse events were fatigue and headache (18% for both, and 5 events were grade 3 or 4. There was 1 serious treatment‐related adverse event in the biweekly intravenous cohort (hypersensitivity). This hypersensitivity resolved without discontinuation of study treatment. No patients developed neutralizing antibodies or bone marrow fibrosis. Of the patients who completed 8 weeks of treatment, 57% had a complete platelet response, an additional 8% had a major platelet response, and 61% did not require a platelet transfusion during this period. Weekly subcutaneous injections achieved the highest mean trough concentrations.

CONCLUSIONS:

The safety and efficacy profiles of romiplostim in this study suggested that weekly subcutaneous administration of 750 μg romiplostim is an appropriate starting dose for future clinical studies in patients with MDS and thrombocytopenia. Cancer 2011. © 2010 American Cancer Society.
Concordance in histological and biological parameters between first and second primary breast cancers
Authors: Huo Dezheng , Melkonian Stephanie , Rathouz Paul J. , Khramtsov Andrey , Olopade Olufunmilayo I.
Published: October 2010

BACKGROUND:

Women with breast cancer are more likely to have a second breast cancer than women in the general population are to have a primary cancer. However, the biological relationship between primary and second breast cancers is not clear.

METHODS:

A total of 30,617 patients diagnosed with bilateral breast cancers between 1990 and 2007 were identified through 17 cancer registries of the Surveillance, Epidemiology, and End Results program. Logistic regression with odds ratios (ORs) and 95% confidence intervals (CIs) was used to model strength of association in hormone receptor status, grade, and histology between 2 cancers.

RESULTS:

There was a strong association in estrogen receptor status between 2 bilateral tumors (OR, 7.64; 95% CI, 7.00‐8.35). The strength of association in estrogen receptor status depended on the time interval between the first and second tumors and age at diagnosis. The OR was 25.9 for synchronous tumors (within 1 month) and 3.69 for metachronous tumors separated by ≥10 years. The strength of association was stronger in patients whose first cancer was diagnosed before age 50 (OR, 11.7) versus after age 50 (OR, 5.71). A similar pattern was observed for progesterone receptor, grade, and histological type, but with relatively weaker association.

CONCLUSIONS:

The strong concordance in hormone receptor status of primary and second breast cancers suggests that 2 breast cancers arise in a common milieu and that tumor subtypes are predetermined in the early stage of breast carcinogenesis. Cancer 2011. © 2010 American Cancer Society.
Long‐term outcomes in the second‐line treatment of chronic myeloid leukemia
Authors: Jabbour Elias , Cortes Jorge , Kantarjian Hagop
Published: October 2010

Chronic myeloid leukemia (CML) is a progressive and often fatal myeloproliferative neoplasm. The hallmark of CML is an acquired chromosomal translocation known as the Philadelphia chromosome (Ph), which results in the synthesis of the breakpoint cluster region‐Abelson murine leukemia (BCR‐ABL) fusion oncoprotein, a constitutively active tyrosine kinase. The introduction of imatinib, a tyrosine kinase inhibitor (TKI) that is specific for BCR‐ABL, was a major breakthrough in CML therapy. Although most patients respond to first‐line imatinib therapy, some experience loss of response (resistance) or require treatment discontinuation because of toxicity (intolerance). For patients with CML, failure on standard‐dose imatinib therapy (400 mg daily), imatinib dose escalation (600‐800 mg daily) is a second‐line option. However, high‐dose imatinib is not an appropriate approach for patients who experience drug toxicity, and there remain questions over the durability of responses achieved with this strategy. Alternative second‐line options include the TKIs dasatinib and nilotinib. A substantial amount of long‐term data for these agents is available. Although both are potent and specific BCR‐ABL TKIs, dasatinib and nilotinib exhibit unique pharmacologic profiles and response patterns relative to different patient characteristics, such as disease stage and BCR‐ABL mutation status. To optimize therapeutic benefit, clinicians should select treatment based on each patient's historic response, adverse‐event tolerance, and risk factors. Cancer 2010. © 2010 American Cancer Society.
Fine‐needle aspiration cytopathology—An accurate diagnostic modality in mammary carcinoma with osteoclast‐like giant cells
Authors: Jacquet Solène‐Florence , Balleyguier Corinne , Garbay Jean‐Rémi , Bourgier Céline , Mathieu Marie‐Christine , Delaloge Suzette , Vielh Philippe
Published: October 2010

BACKGROUND:

Invasive ductal carcinoma with osteoclast‐like giant cells (OGCs) is a very rare breast tumor the main characteristic of which is the presence of multinucleated cells of histiocytic nature.

METHODS:

The authors report a study of 8 consecutive cases of fine‐needle aspiration cytopathology (FNAC) of breast nodules in which OGCs and malignant epithelial cells were associated and diagnosed as mammary carcinoma with OGCs. These cases were selected over a period of 5 years from more than 6000 patients who were examined during a weekly, single‐day, multidisciplinary breast clinic. The corresponding biopsies and surgical specimens were examined histologically in an immunohistochemical study using a histiocytic marker (cluster of differentiation 68 [CD68]).

RESULTS:

Conventional histologic analysis made it possible to diagnose 5 of the 8 cases as mammary carcinoma with OGCs; whereas, in the other 3 cases, OGCs were not detected without the help of immunohistochemistry.

CONCLUSIONS:

FNAC appeared to be a very efficient way to diagnose breast carcinoma with OGCs, because it detected forms with only a few OGCs that usually are not observed at histologic diagnosis. Consequently, the current results indicated that mammary carcinoma with OGCs may be more frequent than reported previously. Cancer (Cancer Cytopathol) 2010. © 2010 American Cancer Society.
Hydrogen Sulfide Adsorption on MOFs and MOF/Graphite Oxide Composites
Authors: Petit Camille , Mendoza Barbara , Bandosz Teresa J.
Published: October 2010

Composites of a copper‐based metal‐organic framework (MOF) and graphite oxide (GO) were tested for hydrogen sulfide removal at ambient conditions. In order to understand the mechanisms of adsorption, the initial and exhausted samples were analyzed by various techniques including X‐ray diffraction, Fourier transform infrared spectroscopy, thermogravimetric analyses, and sorption of nitrogen. Compared to the parent materials, an enhancement in hydrogen sulfide adsorption was found. It was the result of physical adsorption of water and H2S in the pore space formed at the interface between the MOF units and the graphene layers where the dispersive forces are the strongest. Besides physisorption, reactive adsorption was found as the main mechanism of retention. H2S molecules bind to the copper centers of the MOF. They progressively react with the MOF units resulting in the formation of copper sulfide. This leads to the collapse of the MOF structure. Water enhances adsorption in the composites as it allows the dissolution of hydrogen sulfide.
Parental division of labor, coordination, and the effects of family structure on parenting in monogamous prairie voles (Microtus ochrogaster)
Authors: Ahern Todd H. , Hammock Elizabeth A.D. , Young Larry J.
Published: October 2010

Family relationships help shape species‐typical social and emotional development, but our understanding of how this shaping occurs is still relatively limited. Prairie voles are a socially monogamous and biparental species that is well situated to complement traditional animal models, such as rats and mice, in investigating the effects of family experience. In this series of studies, we aimed to test hypotheses relating to how prairie vole families function under undisturbed, standard laboratory conditions. In the first study, we compared the parental behavior of primiparous biparental (BP) and single‐mother (SM) prairie vole family units for 12 postnatal days and then tested for sex differences, behavioral coordination, and family structure effects. Under BP conditions, nest attendance was coordinated and shared equally by both sexes, while pup‐directed and partner‐directed licking and grooming (LG) were coordinated in a sex and social‐context‐dependent manner. Contrary to our expectations, SMs showed no evidence of strong parental compensation in response to the lack of the father, indicating a minimal effect of family structure on maternal behavior but a large effect on pup care. In the second study, we examined the effects of these BP and SM rearing conditions on family dynamics in the next generation and found that SM‐reared adult parents exhibited lower rates of pup‐directed LG in comparison to BP‐reared counterparts. Situated in the context of human family dynamics and psychology, these results suggest that the study in prairie voles may help improve our understanding of family systems and how perturbations to these systems can affect adults and offspring. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 53:118–131, 2011
Age‐related changes in visual recognition memory during infancy and early childhood
Authors: Morgan Kirstie , Hayne Harlene
Published: October 2010

Age‐related changes in long‐term memory during infancy and early childhood were examined using the Visual Recognition Memory (VRM) procedure. Independent groups of 1‐, 2‐, 3‐, and 4‐year‐olds were familiarized with a visual stimulus and were tested either immediately or after a delay that ranged from 24 hr to 6 months. Although all age groups exhibited a significant novelty preference when tested immediately after familiarization, clear age‐related differences emerged over longer retention intervals. We conclude that age‐related increases in basic retention are a fundamental aspect of mammalian memory development and, in humans, these increases may play a vital role in the offset of childhood amnesia. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 53:157–165, 2011
Rhoh deficiency reduces peripheral T‐cell function and attenuates allogenic transplant rejection
Authors: Porubsky Stefan , Wang Shijun , Kiss Eva , Dehmel Stefan , Bonrouhi Mahnaz , Dorn Tatjana , Luckow Bruno , Brakebusch Cord , Gröne Hermann‐Josef
Published: October 2010

Rhoh is a hematopoietic system‐specific GTPase. Rhoh‐deficient T cells have been shown to have a defect in TCR signaling manifested during their thymic development. Our aims were to investigate the phenotype of peripheral Rhoh‐deficient T cells and to explore in vivo the potential benefit of Rhoh deficiency in a clinically relevant situation, in which T‐cell inhibition is desirable. In murine allogenic kidney transplantation, Rhoh deficiency caused a significant 75% reduction of acute and chronic transplant rejection accompanied by 75% lower alloantigen‐specific antibody levels and significantly better graft function. This effect was independent of the lower T‐cell numbers in Rhoh‐deficient recipients, because injection of equal numbers of Rhoh‐deficient or control T cells into kidney transplanted mice with SCID led again to a significant 60% reduction of rejection. Mixed lymphocyte reaction revealed that the weaker alloreactivity was associated with a 85% lower cytotoxicity and a 50–80% lower cytokine release in Rhoh‐deficient T cells without an influence on the secretion itself. Antigen uptake and presentation in DC were unaffected by Rhoh deficiency. These findings stress the importance of Rhoh for the function of peripheral T cells.
CD4+CD25+ Treg regulate the contribution of CD8+ T‐cell subsets in repopulation of the lymphopenic environment
Authors: Almeida Afonso R. M. , Ciernik Ilja F. , Sallusto Federica , Lanzavecchia Antonio
Published: October 2010

Peripheral T‐cell expansion is of major relevance for immune function after lymphopenia. In order to promote regeneration, the process should result in a peripheral T‐cell pool with a similar subpopulation structure as before lymphopenia. We investigated the repopulation of the CD8+ central‐memory T cells (TCM) and effector‐memory T cells (TEM) pools after adoptive transfer of sorted CD8+ T cells from naïve, TCM and TEM subsets into T‐cell‐deficient hosts. We show that the initial kinetics of expansion are distinct for each subset and that the contribution to the repopulation of the CD8+ T‐cell pool by the progeny of each subset is not a mere function of its initial expansion. We demonstrate that CD4+CD25+ Treg play a major role in the repopulation of the CD8+ T‐cell pool and that CD8+ T‐cell subsets impact on each other. In the absence of CD4+CD25+ Treg, a small fraction of naïve CD8+ T cells strongly proliferates, correlating with further expansion and differentiation of co‐expanding CD8+ T cells. CD4+CD25+ Treg suppress these responses and lead to controlled repopulation, contributing decisively to the maintenance of recovered TCM and TEM fractions, and leading to repopulation of each pool with progeny of its own kind.
Genetic proof for the transient nature of the Th17 phenotype
Authors: Kurschus Florian C. , Croxford Andrew L. , P. Heinen André , Wörtge Simone , Ielo Daniele , Waisman Ari
Published: October 2010

IL‐17‐producing CD4+ T cells (Th17) have been classified as a new T helper cell subset. Using an IL‐17 fate mapping mouse strain, which genetically fixes the memory of IL‐17 expression, we demonstrate that IL‐17A/F‐expressing T helper cells generated either in vitro or in vivo are not a stable T‐cell subset. Upon adoptive transfer of IL‐17F‐reporter‐positive Th17 cells to RAG‐deficient or WT animals, encephalitogenic Th17 cells partially lose IL‐17 expression and upregulate IFN‐γ. Additionally, we show that Th1 cells can convert in vivo to IL‐17A/IFN‐γ‐coexpressing cells in the mesenteric lymph nodes (mLN). Our data classify IL‐17A and IL‐17F as cytokines produced transiently in response to the local microenvironment, thus showing that IL‐17 expression does not define an end‐stage T helper cell subset.
First Examples of α‐(1→4)‐Glycosylation Reactions on Ionic Liquid Supports
Authors: Pépin Matthieu , Hubert‐Roux Marie , Martin Claudette , Guillen Fréderic , Lange Catherine , Gouhier Géraldine
Published: October 2010

We have developed the first α‐(1→4)‐glycosylation reactions on ionic liquid supports. The purification steps involve simple liquid/liquid extractions. The positive influence of the soluble support on the stereoselectivity was proved, and the recyclable character of the ionic liquid was demonstrated.
Manipulating particles in microfluidics by floating electrodes
Authors: Yalcin Sinan E. , Sharma Ashutosh , Qian Shizhi , Joo Sang W. , Baysal Oktay
Published: October 2010

Various particle manipulations including enrichment, movement, trapping, separation, and focusing by floating electrodes attached to the bottom wall of a straight microchannel under an imposed DC electric field have been experimentally demonstrated. In contrast to a dielectric microchannel possessing a nearly uniform surface charge (or ζ potential), the metal strip (floating electrode) is polarized under the imposed electric field, resulting in a nonuniform distribution of the induced surface charge with a zero net surface charge along the floating electrode's surface, and accordingly induced‐charge electroosmotic flow near the metal strip. The induced induced‐charge electroosmotic flow can be regulated by controlling the strength of the imposed electric field and affects both the hydrodynamic field and the particle's motion. By using a single floating electrode, charged particles could be locally concentrated in a section of the channel or in an end‐reservoir and move toward either the anode or the cathode by controlling the strength of the imposed electric field. By using double floating electrodes, negatively charged particles could be concentrated between the floating electrodes, subsequently squeezed to a stream flowing in the center region of the microchannel toward the cathodic reservoir, which can be used to focus particles.
Is bisphenol A a weak carcinogen like the natural estrogens and diethylstilbestrol?
Authors: Cavalieri Ercole L. , Rogan Eleanor G.
Published: October 2010

Bisphenol A (BPA) displays weak estrogenic properties and could be a weak carcinogen by a mechanism similar to that of estrone (E1), estradiol (E2) and the synthetic estrogen diethylstilbestrol, a human carcinogen. A wide variety of scientific evidence supports the hypothesis that certain estrogen metabolites, predominantly catechol estrogen‐3,4‐quinones, react with DNA to cause mutations that can lead to the initiation of cancer. One of the major pathways of estrogen metabolism leads to the 4‐catechol estrogens, 4‐OHE1(E2), which are oxidized to their quinones, E1(E2)‐3,4‐Q. The quinones react with DNA to form predominantly the depurinating adducts 4‐OHE1(E2)‐1‐N3Ade and 4‐OHE1(E2)‐1‐N7Gua. This process constitutes the predominant pathway in the initiation of cancer by estrogens. One pathway of BPA metabolism is hydroxylation of one of its symmetric benzene rings to form its catechol, 3‐OHBPA. Subsequent oxidation to BPA‐3,4‐quinone would lead to reaction with DNA to form predominantly the depurinating adducts 3‐OHBPA‐6‐N3Ade and 3‐OHBPA‐6‐N7Gua. The resulting apurinic sites in the DNA could generate mutations in critical genes that can initiate human cancers. The catechol of BPA may also alter expression of estrogen‐activating and deactivating enzymes, and/or compete with methoxylation of 4‐OHE1(E2) by catechol‐O‐methyltransferase, thereby unbalancing the metabolism of estrogens to increase formation of E1(E2)‐3,4‐Q and the depurinating estrogen‐DNA adducts leading to cancer initiation. Thus, exposure to BPA could increase the risk of developing cancer by direct and/or indirect mechanisms. Knowledge of these mechanisms would allow us to begin to understand how BPA may act as a weak carcinogen and would be useful for regulating its use. © 2010 IUBMB IUBMB Life, 62(10): 746–751, 2010
TIEG1 negatively controls the myoblast pool indispensable for fusion during myogenic differentiation of C2C12 cells
Authors: Miyake Masato , Hayashi Shinichiro , Iwasaki Shunsuke , Uchida Takafumi , Watanabe Kouichi , Ohwada Shyuichi , Aso Hisashi , Yamaguchi Takahiro
Published: October 2010

The transforming growth factor (TGF)‐β inducible early gene (TIEG)‐1 is implicated in the control of cell proliferation, differentiation, and apoptosis in some cell types. Since TIEG1 functioning may be associated with TGF‐β, a suppressor of myogenesis, TIEG1 is also likely to be involved in myogenesis. Therefore, we investigated the function of TIEG1 during myogenic differentiation in vitro using the murine myoblasts cell line, C2C12. TIEG1 expression increased during differentiation of C2C12 cells. Constitutive expression of TIEG1 reduced survival and decreased myotube formation. Conversely, knocking down TIEG1 expression increased the number of viable cells during differentiation, and accelerated myoblast fusion into multinucleated myotubes. However, expression of the myogenic differentiation marker, myogenin, remained unaffected by TIEG1 knockdown. The mechanism underlying these events was investigated by focusing on the regulation of myoblast numbers after induction of differentiation. The knockdown of TIEG1 led to changes in cell cycle status and inhibition of apoptosis during the initial stages of differentiation. Microarray and real‐time PCR analyses showed that the regulators of cell cycle progression were highly expressed in TIEG1 knockdown cells. Therefore, TIEG1 is a negative regulator of the myoblast pool that causes inhibition of myotube formation during myogenic differentiation. J. Cell. Physiol. 226: 1128–1136, 2011. © 2010 Wiley‐Liss, Inc.
The hippo pathway in biological control and cancer development
Authors: Chan Siew Wee , Lim Chun Jye , Chen Liming , Chong Yaan Fun , Huang Caixia , Song Haiwei , Hong Wanjin
Published: October 2010

The Hippo pathway is an evolutionally conserved protein kinase cascade involved in regulating organ size in vivo and cell contact inhibition in vitro by governing cell proliferation and apoptosis. Deregulation of the Hippo pathway is linked to cancer development. Its first core kinase Warts was identified in Drosophila more than 15 years ago, but it gained much attention when other core components of the pathway were identified 8 years later. Major discoveries of the pathway were made during past several years. The core kinase components Hippo, Salvador, Warts, and Mats in the fly and Mst1/2, WW45, Lats1/2, and Mob1 in mammals phosphorylate and inactivate downstream transcriptional co‐activators Yorkie in the fly, Yes‐associated protein (YAP) and transcriptional co‐activator with PDZ‐binding motif (TAZ) in mammals, respectively. Phosphorylated Yorkie, YAP, and TAZ are sequestered in the cytoplasm by interaction with 14‐3‐3 proteins. Here we review recent progresses of this pathway by focusing on how these proteins communicate with each other and how loss of regulation results in cancers. J. Cell. Physiol. 226: 928–939, 2011. © 2010 Wiley‐Liss, Inc.
The transforming growth factor‐beta (TGF‐β) mediates acquisition of a mesenchymal stem cell‐like phenotype in human liver cells
Authors: Caja Laia , Bertran Esther , Campbell Jean , Fausto Nelson , Fabregat Isabel
Published: October 2010

Transforming growth factor‐beta (TGF‐β) mediates several and sometime opposite effects in epithelial cells, inducing growth inhibition, and apoptosis but also promoting an epithelial to mesenchymal transition (EMT) process, which enhances cell migration and invasion. TGF‐β plays relevant roles in different liver pathologies; however, very few is known about its specific signaling and cellular effects in human primary hepatocytes. Here we show that TGF‐β inhibits proliferation and induces pro‐apoptotic genes (such as BMF or BIM) in primary cultures of human fetal hepatocytes (HFH), but also up‐regulates anti‐apoptotic genes, such as BCL‐XL and XIAP. Inhibition of the epidermal growth factor receptor (EGFR), using gefitinib, abrogates the increase in the expression of the anti‐apoptotic genes and significantly enhances cell death. Simultaneously, TGF‐β is able to induce an EMT process in HFH, coincident with Snail up‐regulation and a decrease in E‐cadherin levels, cells showing mesenchymal proteins and reorganization of the actin cytoskeleton in stress fibers. Interestingly, these cells show loss of expression of specific hepatic genes and increased expression of stem cell markers. Chronic treatment with TGF‐β allows selection of a population of mesenchymal cells with a de‐differentiated phenotype, reminiscent of progenitor‐like cells. Process is reversible and the mesenchymal stem‐like cells re‐differentiate to hepatocytes under controlled experimental conditions. In summary, we show for the first time that human hepatocytes may respond to TGF‐β inducing different signals, some of them might contribute to tumor suppression (growth inhibition and apoptosis), but others should mediate liver tumor progression and invasion (EMT and acquisition of a stem‐like phenotype). J. Cell. Physiol. 226: 1214–1223, 2011. © 2010 Wiley‐Liss, Inc.
Fibroblast cytoskeletal remodeling contributes to connective tissue tension
Authors: Langevin Helene M. , Bouffard Nicole A. , Fox James R. , Palmer Bradley M. , Wu Junru , Iatridis James C. , Barnes William D. , Badger Gary J. , Howe Alan K.
Published: October 2010

The visco‐elastic behavior of connective tissue is generally attributed to the material properties of the extracellular matrix rather than cellular activity. We have previously shown that fibroblasts within areolar connective tissue exhibit dynamic cytoskeletal remodeling within minutes in response to tissue stretch ex vivo and in vivo. Here, we tested the hypothesis that fibroblasts, through this cytoskeletal remodeling, actively contribute to the visco‐elastic behavior of the whole tissue. We measured significantly increased tissue tension when cellular function was broadly inhibited by sodium azide and when cytoskeletal dynamics were compromised by disrupting microtubules (with colchicine) or actomyosin contractility (via Rho kinase inhibition). These treatments led to a decrease in cell body cross‐sectional area and cell field perimeter (obtained by joining the end of all of a fibroblast's processes). Suppressing lamellipodia formation by inhibiting Rac‐1 decreased cell body cross‐sectional area but did not affect cell field perimeter or tissue tension. Thus, by changing shape, fibroblasts can dynamically modulate the visco‐elastic behavior of areolar connective tissue through Rho‐dependent cytoskeletal mechanisms. These results have broad implications for our understanding of the dynamic interplay of forces between fibroblasts and their surrounding matrix, as well as for the neural, vascular, and immune cell populations residing within connective tissue. J. Cell. Physiol. 226: 1166–1175, 2011. © 2010 Wiley‐Liss, Inc.
Identification of heat shock protein 5, calnexin and integral membrane protein 2B as Adam7‐interacting membrane proteins in mouse sperm
Authors: Han Cecil , Park Inju , Lee Boyeon , Jin Sora , Choi Heejin , Kwon Jun Tae , Kwon Yong‐il , Kim Do Han , Park Zee Yong , Cho Chunghee
Published: October 2010

In mammals, sperm acquire their motility and ability to fertilize eggs in the epididymis. This maturation process involves the acquisition of particular proteins from the epididymis. One such secretory protein specifically expressed in the epididymis is Adam7 (a disintegrin and metalloprotease 7). Previous studies have shown that Adam7 that resides in an intracellular compartment of epididymal cells is transferred to sperm membranes, where its levels are dependent on the expression of Adam2 and Adam3, which have critical roles in fertilization. Here, using a proteomics approach based on mass spectrometry, we identified proteins that interact with Adam7 in sperm membranes. This analysis revealed that Adam7 forms complexes with calnexin (Canx), heat shock protein 5 (Hspa5), and integral membrane protein 2B (Itm2b). Canx and Hspa5 are molecular chaperones, and Itm2b is a type II integral membrane protein implicated in neurodegeneration. The interaction of Adam7 with these proteins was confirmed by immunoprecipitation‐Western blot analysis. We found that Adam7 and Itm2b are located in detergent‐resistant regions known to be highly correlated with membrane lipid rafts. We further found that the association of Adam7 with Itm2b is remarkably promoted during sperm capacitation owing to a conformational change of Adam7 that occurs in concert with the capacitation process. Thus, our results suggest that Adam7 functions in fertilization through the formation of a chaperone complex and enhanced association with Itm2b during capacitation in sperm. J. Cell. Physiol. 226: 1186–1195, 2011. © 2010 Wiley‐Liss, Inc.
CXCL5/ENA78 increased cell migration and epithelial‐to‐mesenchymal transition of hormone‐independent prostate cancer by early growth response‐1/snail signaling pathway
Authors: Kuo Po‐Lin , Chen Yen‐Hsu , Chen Tun‐Chieh , Shen Kun‐Hung , Hsu Ya‐Ling
Published: October 2010

Prostate cancers that are resistant to hormone therapy are more invasive and have greater ability to spread to other organs than androgen‐dependent prostate cancers. Furthermore, this type of prostate cancer is also highly resistant to current forms of chemotherapy. This study analyzed CXCL5/ENA78, which is highly expressed in androgen‐independent prostate cancers, and is responsible for cell migration and epithelial‐to‐mesenchymal transition in two androgen‐independent prostate cancer cell lines. Inducement of PC‐3 and DU145 cancer progression by CXCL5/ENA78 is associated with increased Raf/MEK/ERK activation, and the upregulation of early growth response‐1 (Egr‐1) and Snail. Blockade of Egr‐1 decreased Snail upregulation and cell migration, indicating that Egr‐1 is required in CXCL5/ENA78‐mediated Snail enhancement and cell migration. In addition, Egr‐1 siRNA also decreased the effect of CXCL5/ENA78 on p27 inhibition, Cdk4 induction and cell proliferation, suggesting Egr‐1 is also involved in CXCL5/ENA78‐mediated cell growth. Moreover, blocking ERK1/2 by siRNA suppressed CXCL5/ENA78‐induced Egr‐1 enhancement, cell migration, and proliferation. Our study suggests that inhibition of CXCL5/ENA78‐mediated ERK/Egr‐1/Snail signaling is an attractive therapeutic target for androgen‐independent prostate cancer. J. Cell. Physiol. 226: 1224–1231, 2011. © 2010 Wiley‐Liss, Inc.
Differential role of Rho GTPases in intestinal epithelial barrier regulation in vitro
Authors: Schlegel Nicolas , Meir Michael , Spindler Volker , Germer Christoph‐Thomas , Waschke Jens
Published: October 2010

Maintenance of intestinal epithelial barrier functions is crucial to prevent systemic contamination by microbes that penetrate from the gut lumen. GTPases of the Rho‐family such as RhoA, Rac1, and Cdc42 are known to be critically involved in the regulation of intestinal epithelial barrier functions. However, it is still unclear whether inactivation or activation of these GTPases exerts barrier protection or not. We tested the effects of Rho GTPase activities on intestinal epithelial barrier functions by using the bacterial toxins cytotoxic necrotizing factor 1 (CNF‐1), toxin B, C3 transferase (C3 TF), and lethal toxin (LT) in an in vitro model of the intestinal epithelial barrier. Incubation of cell monolayers with CNF‐1 for 3 h induced exclusive activation of RhoA whereas Rac1 and Cdc42 activities were unchanged. As revealed by FITC‐dextran flux and measurements of transepithelial electrical resistance (TER) intestinal epithelial permeability was significantly increased under these conditions. Inhibition of Rho kinase via Y27632 blocked barrier destabilization of CNF‐1 after 3 h. In contrast, after 24 h of incubation with CNF‐1 only Rac1 and Cdc42 but not RhoA were activated which resulted in intestinal epithelial barrier stabilization. Toxin B to inactivate RhoA, Rac1, and Cdc42 as well as Rac1 inhibitor LT increased intestinal epithelial permeability. Similar effects were observed after inhibition of RhoA/Rho kinase signaling by C3 TF or Y27632. Taken together, these data demonstrate that both activation and inactivation of RhoA signaling increased paracellular permeability whereas activation of Rac1 and Cdc42 correlated with stabilized barrier functions. J. Cell. Physiol. 226: 1196–1203, 2011. © 2010 Wiley‐Liss, Inc.
Substance P stimulates the recovery of bone marrow after the irradiation
Authors: An You Sun , Lee EunAh , Kang Mi‐Hyun , Hong Hyun Sook , Kim Mi‐Ra , Jang Won Seok , Son Youngsook , Yi Jae Youn
Published: October 2010

The therapeutic use of ionizing radiation (e.g., X‐rays and γ‐rays) needs to inflict minimal damage on non‐target tissue. Recent studies have shown that substance P (SP) mediates multiple activities in various cell types, including cell proliferation, anti‐apoptotic responses, and inflammatory processes. The present study investigated the effects of SP on γ‐irradiated bone marrow stem cells (BMSCs). In mouse bone marrow extracts, SP prolonged activation of Erk1/2 and enhanced Bcl‐2 expression, but attenuated the activation of apoptotic molecules (e.g., p38 and cleaved caspase‐3) and down‐regulated Bax. We also observed that SP‐decreased apoptotic cell death and stimulated cell proliferation in γ‐irradiated mouse bone marrow tissues through TUNEL assay and PCNA analysis. To determine how SP affects bone marrow stem cell populations, mouse bone marrow cells were isolated and colony‐forming unit (CFU) of mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs) was estimated. SP‐pretreated ones showed higher CFUs of MSC and HSC than untreated ones. Furthermore, when SP was pretreated in cultured human MSC, it significantly decreased apoptotic cells at 48 and 72 h after γ‐irradiation. Compared with untreated cells, SP‐treated human MSCs showed reduced cleavage of apoptotic molecules such as caspase‐8, ‐9, ‐3, and poly ADP‐ribose polymerase (PARP). Thus, our results suggest that SP alleviates γ‐radiation‐induced damage to mouse BMSCs and human MSCs via regulation of the apoptotic pathway. J. Cell. Physiol. 226: 1204–1213, 2011. © 2010 Wiley‐Liss, Inc.
Role of Smad2/3 and p38 MAP kinase in TGF‐β1‐induced epithelial–mesenchymal transition of pulmonary epithelial cells
Authors: Kolosova Irina , Nethery David , Kern Jeffrey A.
Published: October 2010

Idiopathic pulmonary fibrosis is characterized by myofibroblast accumulation, extracellular matrix (ECM) remodeling, and excessive collagen deposition. ECM‐producing myofibroblasts may originate from epithelial cells through epithelial to mesenchymal transition (EMT). TGF‐β1 is an inducer of EMT in pulmonary epithelial cells in vitro and in vivo, though the mechanisms are unclear. We hypothesized that TGF‐β1 induced EMT through Smad‐dependent and ‐independent processes. To test this hypothesis, we studied the roles and mechanisms of TGF‐β1‐induced Smad and p38 mitogen‐activated protein kinase (MAPK) signaling in EMT‐related changes in pulmonary epithelial cells. Exposure of pulmonary epithelial 1HAEo− cells to TGF‐β1 resulted in morphological and molecular changes of EMT over a 96‐h period; loss of cell–cell contact, cell elongation, down‐regulation of E‐cadherin, up‐regulation of fibronectin, and up‐regulation of collagen I. Both Smad2/3 and p38 MAPK signaling pathways were activated by TGF‐β1. However, neither Smad2/3 nor p38 MAPK were required for the down‐regulation of E‐cadherin, yet p38 MAPK was associated with fibronectin up‐regulation. Both Smad2/3 and p38 MAPK had a role in regulation of TGF‐β1‐induced collagen expression. Furthermore, these data demonstrate that Smads and p38 MAPK differentially regulate EMT‐related changes in pulmonary epithelial cells. J. Cell. Physiol. 226: 1248–1254, 2011. © 2010 Wiley‐Liss, Inc.
The inhibitory effect of ginsan on TGF‐β mediated fibrotic process
Authors: Ahn Ji‐Yeon , Kim Mi‐Hyoung , Lim Min‐Jin , Park Sarah , Lee Sae‐lo‐oom , Yun Yeon‐Sook , Song Jie‐Young
Published: October 2010

Transforming growth factor‐beta (TGF‐β) plays a central role in the development of fibrosis by stimulating extracellular matrix accumulation, and signals either directly or indirectly through types I, II, and III (TβRI, II, and III) TGF‐β receptor complexes. Ginsan, a polysaccharide extracted from Panax ginseng, has multiple immunomodulatory effects. Here, we examine whether ginsan regulates the fibrogenic process by interfering with TGF‐β signaling pathways. TGF‐β treatment of murine or human normal lung fibroblasts enhanced the levels of several fibrotic markers, including smooth muscle alpha actin (α‐SMA), collagen‐1, and fibronectin. Interestingly, ginsan treatment either before or after TGF‐β administration led to significant reductions in all of α‐SMA, collagen‐1, and fibronectin expression levels. Ginsan not only inhibited phosphorylation of Smad2 and Smad3, but also attenuated pERK and pAKT signaling induced by TGF‐β. Moreover, ginsan restored TβRIII protein expression, which was significantly downregulated by TGF‐β, but reduced TβRI and TβRII protein levels. In a murine model of bleomycin (BLM)‐induced pulmonary fibrosis, ginsan significantly suppressed accumulation of collagen, α‐SMA, and TGF‐β. These data collectively suggest that ginsan acts as an effective anti‐fibrotic agent in the treatment of pulmonary fibrosis by blocking multiple TGF‐β signaling pathways. J. Cell. Physiol. 226: 1241–1247, 2011. © 2010 Wiley‐Liss, Inc.
Leptin increases motility and integrin up‐regulation in human prostate cancer cells
Authors: Huang Chih‐Yang , Yu Hsin‐Shan , Lai Tung‐Yuan , Yeh Yu‐Lan , Su Cheng‐Chuan , Hsu His‐Hsien , Tsai Fuu‐Jen , Tsai Chang‐Hai , Wu Hsi‐Chin , Tang Chih‐Hsin
Published: October 2010

Prostate cancer is the most commonly diagnosed malignancy in men and shows a predilection for metastasis to distant organs. Leptin, an adipocyte‐derived cytokine that is closely associated with obesity, has recently been shown to be involved in carcinogenesis and cancer progression. The aim of this study was to investigate whether leptin is associated with the motility of prostate cancer cells. We found that leptin increased the migration of human prostate cancer cells and expression of αvβ3 integrin on these cells. Leptin‐mediated migration and increased integrin expression were attenuated by OBRl receptor antisense oligonucleotide (ODN). Activation of insulin receptor substrate (IRS‐1), phosphatidylinositol 3‐kinase (PI3K), Akt, and NF‐κB pathways after leptin treatment was demonstrated. Furthermore, leptin‐induced integrin expression and migration activity were inhibited by specific inhibitors; small interfering RNAs (siRNAs); and mutants of the IRS‐1, PI3K, Akt, and NF‐κB cascades. Therefore, this study shows that leptin stimulates the migration of human prostate cancer cells, one of the mechanisms underlying leptin‐directed migration was transcriptional up‐regulation of αvβ3 integrin expression through the OBR1/IRS‐1/PI3K/Akt/NF‐κB signal transduction pathway. J. Cell. Physiol. 226: 1274–1282, 2011. © 2010 Wiley‐Liss, Inc.
Neutrophil differentiation from human‐induced pluripotent stem cells
Authors: Morishima Tatsuya , Watanabe Ken‐ichiro , Niwa Akira , Fujino Hisanori , Matsubara Hiroshi , Adachi Souichi , Suemori Hirofumi , Nakahata Tatsutoshi , Heike Toshio
Published: October 2010

Induced pluripotent stem (iPS) cells are of potential value not only for regenerative medicine, but also for disease investigation. The present study describes the development of a neutrophil differentiation system from human iPS cells (hiPSCs) and the analysis of neutrophil function and differentiation. The culture system used consisted of the transfer of hiPSCs onto OP9 cells and their culture with vascular endothelial growth factor (VEGF). After 10 days, TRA 1‐85+CD34+VEGF receptor‐2 (VEGFR‐2)high cells were sorted and co‐cultured with OP9 cells in the presence of hematopoietic cytokines for 30 days. Floating cells were collected and subjected to morphological and functional analysis. These hiPSC‐derived neutrophils were similar to peripheral blood mature neutrophils in morphology, contained functional neutrophil specific granules, and were equipped with the basic functions such as phagocytosis, superoxide production, and chemotaxis. In the process of differentiation, myeloid cells appeared sequentially from immature myeloblasts to mature segmented neutrophils. Expression patterns of surface antigen, transcription factors, and granule proteins during differentiation were also similar to those of granulopoiesis in normal bone marrow. In conclusion, differentiation of mature neutrophils from hiPSCs was successfully induced in a similar process to normal granulopoiesis using an OP9 co‐culture system. This system may be applied to elucidate the pathogenesis of various hematological diseases that affect neutrophils. J. Cell. Physiol. 226: 1283–1291, 2011. © 2010 Wiley‐Liss, Inc.
Retinoic acid receptor‐mediated signaling protects cardiomyocytes from hyperglycemia induced apoptosis: Role of the renin‐angiotensin system
Authors: Guleria Rakeshwar S. , Choudhary Rashmi , Tanaka Takemi , Baker Kenneth M. , Pan Jing
Published: October 2010

Diabetes mellitus (DM) is a primary risk factor for cardiovascular diseases and heart failure. Activation of the retinoic acid receptor (RAR) and retinoid X receptor (RXR) has an anti‐diabetic effect; but, a role in diabetic cardiomyopathy remains unclear. Using neonatal and adult cardiomyocytes, we determined the role of RAR and RXR in hyperglycemia‐induced apoptosis and expression of renin‐angiotensin system (RAS) components. Decreased nuclear expression of RARα and RXRα, activation of apoptotic signaling and cell apoptosis was observed in high glucose (HG) treated neonatal and adult cardiomyocytes and diabetic hearts in Zucker diabetic fatty (ZDF) rats. HG‐induced apoptosis and reactive oxygen species (ROS) generation was prevented by both RAR and RXR agonists. Silencing expression of RARα and RXRα, by small interference RNA, promoted apoptosis under normal conditions and significantly enhanced HG‐induced apoptosis, indicating that RARα and RXRα are required in regulating cell apoptotic signaling. Blocking angiotensin type 1 receptor (AT1R); but, not AT2R, attenuated HG‐induced apoptosis and ROS generation. Moreover, HG induced gene expression of angiotensinogen, renin, AT1R, and angiotensin II (Ang II) synthesis were inhibited by RARα agonists and promoted by silencing RARα. Activation of RXRα, downregulated the expression of AT1R; and RXRα silencing accelerated HG induced expression of angiotensinogen and Ang II synthesis, whereas there was no significant effect on renin gene expression. These results indicate that reduction in the expression of RARα and RXRα has an important role in hyperglycemia mediated apoptosis and expression of RAS components. Activation of RAR/RXR signaling protects cardiomyocytes from hyperglycemia, by reducing oxidative stress and inhibition of the RAS. J. Cell. Physiol. 226: 1292–1307, 2011. © 2010 Wiley‐Liss, Inc.
Gene profiling reveals hydrogen sulphide recruits death signaling via the N‐methyl‐D‐aspartate receptor identifying commonalities with excitotoxicity
Authors: Chen Minghui Jessica , Peng Zhao Feng , Manikandan Jayapal , Melendez Alirio J. , Tan Gek San , Chung Ching Ming , Li Qiu‐Tian , Tan Theresa M. , Deng Lih Wen , Whiteman Matthew , Beart Philip M. , Moore Phillip K. , Cheung Nam Sang
Published: October 2010

Recently the role of hydrogen sulphide (H2S) as a gasotransmitter stimulated wide interest owing to its involvement in Alzheimer's disease and ischemic stroke. Previously we demonstrated the importance of functional ionotropic glutamate receptors (GluRs) by neurons is critical for H2S‐mediated dose‐ and time‐dependent injury. Moreover N‐methyl‐D‐aspartate receptor (NMDAR) antagonists abolished the consequences of H2S‐induced neuronal death. This study focuses on deciphering the downstream effects activation of NMDAR on H2S‐mediated neuronal injury by analyzing the time‐course of global gene profiling (5, 15, and 24 h) to provide a comprehensive description of the recruitment of NMDAR‐mediated signaling. Microarray analyses were performed on RNA from cultured mouse primary cortical neurons treated with 200 µM sodium hydrosulphide (NaHS) or NMDA over a time‐course of 5–24 h. Data were validated via real‐time PCR, western blotting, and global proteomic analysis. A substantial overlap of 1649 genes, accounting for over 80% of NMDA global gene profile present in that of H2S and over 50% vice versa, was observed. Within these commonly occurring genes, the percentage of transcriptional consistency at each time‐point ranged from 81 to 97%. Gene families involved included those related to cell death, endoplasmic reticulum stress, calcium homeostasis, cell cycle, heat shock proteins, and chaperones. Examination of genes exclusive to H2S‐mediated injury (43%) revealed extensive dysfunction of the ubiquitin‐proteasome system. These data form a foundation for the development of screening platforms and define targets for intervention in H2S neuropathologies where NMDAR‐activated signaling cascades played a substantial role. J. Cell. Physiol. 226: 1308–1322, 2011. © 2010 Wiley‐Liss, Inc.
G‐protein coupled receptor kinase 5 mediates lipopolysaccharide‐induced NFκB activation in primary macrophages and modulates inflammation in vivo in mice
Authors: Patial Sonika , Shahi Shipra , Saini Yogesh , Lee Taehyung , Packiriswamy Nandakumar , Appledorn Daniel M. , LaPres John J. , Amalfitano Andrea , Parameswaran Narayanan
Published: October 2010

G‐protein coupled receptor kinase‐5 (GRK5) is a serine/threonine kinase discovered for its role in the regulation of G‐protein coupled receptor signaling. Recent studies have shown that GRK5 is also an important regulator of signaling pathways stimulated by non‐GPCRs. This study was undertaken to determine the physiological role of GRK5 in Toll‐like receptor‐4‐induced inflammatory signaling pathways in vivo and in vitro. Using mice genetically deficient in GRK5 (GRK5−/−) we demonstrate here that GRK5 is an important positive regulator of lipopolysaccharide (LPS, a TLR4 agonist)‐induced inflammatory cytokine and chemokine production in vivo. Consistent with this role, LPS‐induced neutrophil infiltration in the lungs (assessed by myeloperoxidase activity) was markedly attenuated in the GRK5−/− mice compared to the GRK5+/+ mice. Similar to the in vivo studies, primary macrophages from GRK5−/− mice showed attenuated cytokine production in response to LPS. Our results also identify TLR4‐induced NFκB pathway in macrophages to be selectively regulated by GRK5. LPS‐induced IκBα phosphorylation, NFκB p65 nuclear translocation, and NFκB binding were markedly attenuated in GRK5−/− macrophages. Together, our findings demonstrate that GRK5 is a positive regulator of TLR4‐induced IκBα–NFκB pathway as well as a key modulator of LPS‐induced inflammatory response. J. Cell. Physiol. 226: 1323–1333, 2011. © 2010 Wiley‐Liss, Inc.