bluuebunny/crossref_metadata_2025 huggingface dataset:
Modulating α‐actinin‐4 dynamics in podocytes
Authors: Michaud Jean‐Louis R. , Hosseini‐Abardeh Mona , Farah Kevin , Kennedy Chris R. J.
Published: February 2009

Podocytes are epithelial cells that line the outer aspect of renal blood vessels and provide a platform for the kidney's filtering apparatus, the slit diaphragm. Mutations in α‐actinin‐4, an actin bundling protein highly expressed in podocytes, result in increased affinity for actin and cause a familial form of focal segmental glomerulosclerosis. We hypothesized that such gain‐of‐affinity mutations would override α‐actinin‐4's sensitivity to regulatory factors such as calcium (acting via two EF‐hand motifs), and phosphoinositides. We generated calcium‐ (mutEF) and phosphoinositide‐ (mutPI) insensitive variants of α‐actinin‐4, comparing their properties to a disease‐associated mutant (K256E) and to the wildtype (wt) protein. α‐Actinin‐4mutPI displayed increased affinity for actin, while the affinity of α‐actinin‐4mutEF was unchanged. Addition of calcium to actin sedimentation assays caused a decrease in the association of α‐actinin‐4wt with filamentous actin, while phosphoinositides generally increased this association. Similar to α‐actinin‐4K256E, α‐actinin‐4mutPI was mislocalized in cultured podocytes, being preferentially associated with filamentous actin and focal adhesions. Fluorescence recovery after photobleaching experiments revealed a rapid turnover of α‐actinin‐4wt and α‐actinin‐4mutEF along stress fibers and focal adhesions, while the turnover of α‐actinin‐4K256E and α‐actinin‐4mutPI was dramatically reduced at these subcellular locales. Equibiaxial mechanical stimulation of podocytes, a mimic of intraglomerular forces, reduced podocyte surface area by 50%; this decrease was more severe (70%) in the presence of high‐affinity mutants of α‐actinin‐4. These data suggest that dynamic regulation of α‐actinin‐4/actin interactions may be necessary for maintaining podocyte structure in response to glomerular hydrostatic forces. Cell Motil. Cytoskeleton, 2009. © 2009 Wiley‐Liss, Inc.
α‐Skeletal muscle actin mutants causing different congenital myopathies induce similar cytoskeletal defects in cell line cultures
Authors: Vandamme Drieke , Rommelaere Heidi , Lambert Ellen , Waterschoot Davy , Vandekerckhove Joël , Constantin Bruno , Ampe Christophe
Published: February 2009

Central core disease (CCD), congenital fibre type disproportion (CFTD), and nemaline myopathy (NM) are earlyonset clinically heterogeneous congenital myopathies, characterized by generalized muscle weakness and hypotonia. All three diseases are associated with α‐skeletal muscle actin mutations. We biochemically characterized the CCD and CFTD causing actin mutants and show that all mutants fold correctly and are stable. Expression studies in fibroblasts, myoblasts, and myotubes show that these mutants incorporate in filamentous structures. However they do not intercalate between the nascent z‐lines in differentiating muscle cell cultures. We also show that the distribution of mitochondria and of the ryanodine receptors, and calcium release properties from ryanodine receptors, are unchanged in myotubes expressing the CCD causing mutants. CFTD causing mutants induce partly similar phenotypes as NM associated ones, such as rods and thickened actin fibers in cell culture. Our results suggest that molecular mechanisms behind CFTD and NM may be partly related. Cell Motil. Cytoskeleton, 2009. © 2009 Wiley‐Liss, Inc.
The Wave complex is intrinsically inactive
Authors: Derivery Emmanuel , Lombard Bérangère , Loew Damarys , Gautreau Alexis
Published: February 2009

The Wave proteins activate the Arp2/3 complex at the leading edge of migrating cells. The resulting actin polymerization powers the projection of the plasma membrane in lamellipodia and membrane ruffles. The Wave proteins are always found associated with partner proteins. The canonical Wave complex is a stable complex containing five subunits. Even though it is well admitted that this complex plays an essential regulatory role on Wave function, the mechanisms by which Wave proteins are regulated within the complex are still elusive. Even the constitutive activity or inactivity of the complex is controversial. The major difficulty of these assays resides in the long and difficult purification of the Wave complex by a combination of several chromatography steps, which gives an overall low yield and increases the chance of Wave complex denaturation. Here we report a greatly simplified approach to purify the human Wave complex using a stable cell line expressing a tagged subunit and affinity chromatography. This protocol provided us with sufficient amount of pure Wave complex for functional assays. These assays unambiguously established that the Wave complex in its native conformation is intrinsically inactive, indicating that, like WASP proteins, Wave proteins have a masked C‐terminal Arp2/3 binding site at resting state. As a consequence, the Wave complex has to be recruited and activated at the plasma membrane to project migration structures. Importantly, the approach we describe here for multiprotein complex purification is likely applicable to a wide range of human multiprotein complexes. Cell Motil. Cytoskeleton 2009. © 2009 Wiley‐Liss, Inc.
Survival after Hodgkin lymphoma
Authors: Favier Olav , Heutte Natacha , Stamatoullas‐Bastard Aspasia , Carde Patrice , van't Veer Mars B. , Aleman Berthe M. P. , Noordijk Evert M. , Thomas José , Fermé Christophe , Henry‐Amar Michel ,   for the European Organization for Research and Treatment of Cancer (EORTC) Lymphoma Group and the Groupe d'Études des Lymphomes de l'Adulte (GELA)
Published: February 2009

BACKGROUND:

The objective of this study was to analyze cause‐specific excess mortality in adult patients with Hodgkin lymphoma (HL) with respect to treatment modality.

METHODS:

The study population consisted of 4401 Belgian, Dutch, and French patients aged 15 to 69, in all stages of disease, who were treated between 1964 and 2000. Excess mortality was expressed by using a standardized mortality ratio (SMR) and calculating the absolute excess risk (AER). Relative survival was calculated and analyzed using a previously described regression model.

RESULTS:

At a median follow‐up of 7.8 years, 725 of 4401 patients (16.5%) had died, 51% of HL, 10% of treatment‐related toxicity, 18% of second cancer, 5% of cardiovascular diseases, 2% of infections, 8% of other causes, and 6% of an unspecified cause. Overall, the SMR was 7.4 (95% confidence limits [CL], 6.9‐8.0), and the AER was 182.8 (95% CL, 167.7‐198.8). These indicators were 3.8 (95% CL, 3.2‐4.5) and 27.9, respectively, for deaths from a second cancer and 4.0 (95% CL, 2.3‐6.7) and 3.3, respectively for deaths from infection. After 15 years, the observed survival rate was 75%, and the relative survival rate was 80%. In patients with early‐stage disease, the overall excess mortality was associated with age ≥40 years (P = .007), men (P < .001), unfavorable prognosis features (P < .001), and 2 treatments: combined nonstandard nonalkylating chemotherapy plus involved‐field radiotherapy (P = .002) and mantle‐field irradiation alone (P = .003). With follow‐up censored at the first recurrence, no treatment modalities were associated with excess mortality.

CONCLUSIONS:

Progressive disease remained the primary cause of death in patients with HL in the first decades after treatment. Excess mortality in patients with early‐stage disease was linked significantly to treatment modalities that were associated with poor treatment failure‐free survival. Cancer 2009. © 2009 American Cancer Society.
A simplified mesh‐free method with embedded discontinuities
Authors: Wang H. , Wang S.
Published: February 2009

A simplified mesh‐free method with embedded discontinuities is presented. In this method, the crack can be arbitrarily oriented, but its growth is represented discretely by activation of crack surfaces at individual particles, so no representation of the crack surface is needed. The crack is modeled by a local enrichment of the test and trial functions with a sign function, so that the discontinuities are along the direction of the crack. The discontinuity consists of cohesive crack segments that pass through the entire domain of influence at the node. A set of cracking rules is developed to avoid spurious cracking. We show by numerical experiments that complicated crack patterns with branching cracks can be modeled independent of the mesh. Copyright © 2009 John Wiley & Sons, Ltd.
Simulation of sheet metal forming process using reproducing kernel particle method
Authors: Liu H. S. , Xing Z. W. , Yang Y. Y.
Published: February 2009

Numerical simulation of sheet metal forming process using the meshless method called reproducing kernel particle method (RKPM) is studied in this paper. Owing to limitations of meshless approximation under parametric coordinate system, a constrained reproducing kernel (RK) approximation under Cartesian coordinate system is employed to describe the sheet metal deformation. Several technologies are used to solve the crucial problems involved in the numerical procedure. RKPM shape function is modified to impose essential boundary conditions; stabilized conforming nodal integration method is adopted to implement numerical integration of weak formulation; and an algorithm based on moment method is used to treat contact detection during sheet metal forming process. A program based on constrained RK approximation is developed to simulate two sheet metal forming examples: square‐cup deep drawing and hemispherical part drawing. Numerical results are presented to demonstrate the effectiveness of the RKPM in numerical simulation of sheet metal forming. Copyright © 2009 John Wiley & Sons, Ltd.
Pax6 regulation of Math5 during mouse retinal neurogenesis
Authors: Riesenberg Amy N. , Le Tien T. , Willardsen Minde I. , Blackburn David C. , Vetter Monica L. , Brown Nadean L.
Published: February 2009

Activation of the bHLH factor Math5 (Atoh7) is an initiating event for mammalian retinal neurogenesis, as it is critically required for retinal ganglion cell formation. However, the cis‐regulatory elements and trans‐acting factors that control Math5 expression are largely unknown. Using a combination of transgenic mice and bioinformatics, we identified a phylogenetically conserved regulatory element that is required to activate Math5 transcription during early retinal neurogenesis. This element drives retinal expression in vivo, in a cross‐species transgenic assay. Previously, Pax6 was shown to be necessary for the initiation of Math5 mRNA expression. We extend this finding by showing that the Math5 retinal enhancer also requires Pax6 for its activation, via Pax6 binding to a highly conserved binding site. In addition, our data reveal that other retinal factors are required for accurate regulation of Math5 by Pax6. genesis 47:175–187, 2009. © 2009 Wiley‐Liss, Inc.
Dropout from inpatient treatment for anorexia nervosa: Critical review of the literature
Authors: Wallier Jenny , Vibert Sarah , Berthoz Sylvie , Huas Caroline , Hubert Tamara , Godart Nathalie
Published: February 2009

Objective:

High dropout rates from inpatient treatment for Anorexia Nervosa (AN) pose a serious obstacle to successful treatment. Because dropping out of inpatient treatment may have a negative impact on outcome, it is important to understand why dropout occurs so that treatment can be targeted toward keeping patients in care. We therefore conducted a critical literature review of studies on dropout from inpatient treatment for AN.

Method:

Searches of Medline and PsycINFO revealed nine articles on this subject. Two were excluded because they did not differentiate AN from other eating disorders in analyses.

Results:

Results were scarce and conflicting, with methodological issues complicating comparisons. Weight on admission, AN subtype, eating disorder symptoms, greater psychiatric difficulty in general, and the absence of depression were related to dropout in multivariate analyses.

Discussion:

Authors should use a common definition of dropout and continue research on the identified predictors as well as potential predictors such as impulsivity and family factors. © 2009 by Wiley Periodicals, Inc. Int J Eat Disord 2009
Inter‐ and Intrachain Spin‐Transition Processes in One‐Dimensional Polynuclear Iron(III) Complexes of N,N′‐Ethylenebis(acetylacetonylideneimine) Bridged by 1,3‐Bis(4‐pyridyl)propane and 1,4‐Bis(imidazolyl)butane (Eur. J. Inorg. Chem. 6/2009)
Authors: Imatomi Shinya , Hashimoto Shingo , Matsumoto Naohide
Published: February 2009

The cover picture shows two types of 1D polynuclear spin crossover (SCO) iron(III) complexes, in which planar iron(III) species are bridged axially by a linear bridging ligand to give a 1D structure. In the intermediate spin state between the HS and LS states, the one complex consists of alternately arrayed HS and LS chains, whereas the other consists of alternately arrayed HS and LS units within a chain. The results demonstrate two different SCO transmission mechanisms, that is, SCO occurs in a chain‐by‐chain manner for the first complex and between adjacent units within a chain for the second one. Details are discussed in the article by N. Matsumoto et al. on p. 721 ff.
Interrater and intermethod reliability of default mode network selection
Authors: Franco Alexandre R. , Pritchard Aaron , Calhoun Vince D. , Mayer Andrew R.
Published: February 2009

There has been a growing interest in the neuroimaging community regarding resting state data (i.e., passive mental activity) and the subsequent activation of the so‐called default mode network (DMN). Although this network was originally characterized by a pattern of deactivation during active cognitive states, more recent applications of data‐driven techniques such as independent component analysis (ICA) have permitted the analysis of brain activation during extended periods of truly passive mental activity. However, ICA requires the resultant components to be evaluated for “goodness of fit” via either human raters or more automated techniques. To our knowledge, an investigation on the reliability of either technique in determining the component that best corresponds to default‐mode activity has not been performed. Moreover, it is not clear how automated techniques, which are necessarily dependent upon a template mask, are affected by the structures used to compose the mask. The current study investigated both interrater (human‐human) reliability and intermethod (human‐machine) reliability for determining DMN activation in 42 healthy controls. Results indicated that near perfect interrater reliability was achieved, whereas intermethod reliability was only within the moderate range. The latter was significantly improved via a weighted combination of the anterior and posterior cingulate nodes of the DMN. Implications for fully automating the component selection process are discussed. Hum Brain Mapp, 2009. © 2009 Wiley‐Liss, Inc.
Record rewards: the effects of targeted quality incentives on the recording of risk factors by primary care providers
Authors: Sutton Matt , Elder Ross , Guthrie Bruce , Watt Graham
Published: February 2009

Financial incentives may increase performance on targeted activities and have unintended consequences for untargeted activities. An innovative pay‐for‐performance scheme was introduced for UK general practices in 2004. It incentivised particular quality indicators for targeted groups of patients. We estimate the intended and unintended consequences of this Quality and Outcomes Framework (QOF) using dynamic panel probit models estimated on individual patient records from 315 general practices over the period 2000/1–2005/6. We focus on annual rates of recording of blood pressure, smoking status, cholesterol, body mass index and alcohol consumption. The recording of each risk factor is designated as incentivised or unincentivised for each individual based on whether they have one of the diseases targeted by the QOF. The effect on incentivised factors was substantially larger on the targeted patient groups (+19.9 percentage points) than on the untargeted groups (+5.3 percentage points). There was no obvious evidence of effort diversion but there was evidence of substantial positive spillovers (+10.9 percentage points) onto unincentivised factors for the targeted groups. Moreover, provider responses were larger on those indicators for which more stringent standards were set and greater rewards offered. We conclude that the incentives induced providers to improve targeted quality and make investments in quality that extended beyond the scheme. We estimate that the average provider was paid £20 500 for recording 410 additional items of information on the risk factors targeted by the financial incentives. Allowance for the positive spillovers reduces the estimated average reward from £50 to £25 per additional record. Copyright © 2009 John Wiley & Sons, Ltd.
Richter syndrome: molecular insights and clinical perspectives
Authors: Rossi Davide , Gaidano Gianluca
Published: February 2009

Richter syndrome (RS) represents the clinico‐pathologic transformation of chronic lymphocytic leukaemia (CLL) to an aggressive lymphoma, most commonly diffuse large B‐cell lymphoma (DLBCL). The clinical definition of RS is heterogeneous, and encompasses at least two biologically different conditions: (i) CLL transformation to a clonally related DLBCL, that accounts for the majority of cases; (ii) development of a DLBCL unrelated to the CLL clone. In clonally related RS, the pathogenetic link between the CLL and the DLBCL phases is substantiated by the acquisition of novel molecular lesions at the time of clinico‐pathologic transformation. RS is not a rare event in the natural history of CLL, since the cumulative incidence of RS at 10 years exceeds 10%. Prompt recognition of RS is known to be clinically useful, and may be favoured by close monitoring of CLL patients harbouring clinical and/or biological risk factors of RS development. Conventional risk factors that are independent predictors of RS development at the time of CLL diagnosis include: (i) expression of CD38; (ii) absence of del13q14 and (iii) lymph node size ≥3 cm. Other risk factors of RS development include CD38 genotype and usage of specific immunoglobulin variable genes. The molecular pathogenesis of RS has been elucidated to a certain extent. Acquisition ofTP53mutations and/or 17p13 deletion is a frequent molecular event in RS, as it is in other types of transformation from indolent to aggressive B‐cell malignancies. Additional molecular alterations are being revealed by genome wide studies. Once that transformation has occurred, RS prognosis may be predicted by the RS score, based on performance status, LDH, platelet count, tumour size and number of prior therapies. Depending on patient's age and RS score, the therapeutic options for RS may range from conventional immunochemotherapy to allogeneic bone marrow transplantation. Copyright © 2009 John Wiley & Sons, Ltd.
Autologous stem cell transplantation for elderly patients with acute myeloid leukaemia conditioned with continuous infusion idarubicin and busulphan
Authors: Ferrara Felicetto , Palmieri Salvatore , Pedata Mariangela , Viola Assunta , Izzo Tiziana , Criscuolo Clelia , Mele Giuseppina
Published: February 2009

Different studies have suggested the potential utility of autologous stem cell transplantation (ASCT) in acute myeloid leukaemia (AML) of the elderly with encouraging results in selected patients. However, while the introduction of peripheral blood stem cells (PBSC) has consistently reduced morbidity and mortality of the procedure, relapse still represents the major cause of ASCT failure. One possibility to ameliorate therapeutic results could rely on the adoption of conditioning regimens specifically designed for AML. We report therapeutic results from a series of 40 AML patients older than 60 years (median age 67 years) autografted in first complete remission (CR), after conditioning with continuous infusion (c.i.) high dose idarubicin and busulphan. Fourty patients (median age: 67 years) received 2 days c.i. idarubicin at 20 mg/m2/day, followed by 3 days oral or intravenous busulphan (4 mg/kg/day) as conditioning. No case of transplant‐related mortality occurred. Cardiac toxicity was absent, while 31 patients (77%) had grade 3–4 mucositis. After a median follow‐up of 25 months, median disease free and overall survival (OS) for the whole patient population were 13 and 22 months, respectively. Three patients died while in CR from causes unrelated to AML. Better results were achieved in patients with intermediate karyotype as opposed to those with adverse cytogenetics. Our data confirm the feasibility of a conditioning regimen based on high‐dose idarubicin plus busulphan in older selected AML patients and suggest clinical improvement in patients with normal cytogenetics. Copyright © 2009 John Wiley & Sons, Ltd.
Assessing the credibility of instrumental variables inference with imperfect instruments via sensitivity analysis
Authors: Ashley Richard
Published: February 2009

Consistent instrumental variables (IV) estimation requires instruments uncorrelated with model errors, but this assumption is usually both suspect and untestable. Here the asymptotic sampling distribution of the IV parameter estimator is derived for any specified instrument‐error covariance vector. This result makes it possible to quantify the sensitivity of any particular IV inference result to instrument‐error correlations, allowing one to assess the robustness of such inferential conclusions to uncertainty in the validity of the instruments. An application illustrating the value of this sensitivity analysis is given to a study by Acemoglu et al. (2001). Copyright © 2009 John Wiley & Sons, Ltd.
Increased expression of the testicular estrogen receptor alpha in adult mice exposed to low doses of methiocarb
Authors: Han Jiyou , Park Mira , Kim Jae‐Hong , Kim Ahyoung , Won Miae , Lee Dong Ryul , Ko Jeong‐Jae , Yoon Ho , Sim Se‐Hoon , Lee Kangseok , Bae Jeehyeon
Published: February 2009

Methiocarb is a widely used carbamate pesticide and a suspected endocrine disrupter. The objective of this study was to examine the in vivo effects of methiocarb at low doses on testicular expression of steroid receptors, spermatogenesis and sperm quality in adult mice. Eighteen‐week‐old DBA/2 males were treated with daily intraperitoneal injection of methiocarb (0, 0.03, 0.3, 1.0 or 3.0 µg kg−1 of body weight) for 20 days. Kidney and liver weights were significantly increased in the 1.0 or 3.0 µg kg−1 treatment groups (P < 0.05). The testicular expression of estrogen receptor α (ERα) was significantly increased in mice treated with methiocarb as confirmed by Western blot analysis. The sperm production and sperm quality of the methiocarb‐exposed mice were not significantly altered as determined using a computer‐assisted sperm analysis system. Therefore, these results demonstrate, that although the exposure to methiocarb at low doses alters testicular ERα expression in adult mice, both sperm production and quality remain unaffected. Copyright © 2009 John Wiley & Sons, Ltd.
Mechanical force inhibits osteoclastogenic potential of human periodontal ligament fibroblasts through OPG production and ERK‐mediated signaling
Authors: Kook Sung‐Ho , Son Young‐Ok , Hwang Jung‐Min , Kim Eun‐Mi , Lee Choon‐Bong , Jeon Young‐Mi , Kim Jong‐Ghee , Lee Jeong‐Chae
Published: February 2009

Periodontal ligament and gingival fibroblasts play important roles in bone remodeling. Periodontal ligament fibroblasts stimulate bone remodeling while gingival fibroblasts protect abnormal bone resorption. However, few studies had examined the differences in stimulation of osteoclast formation between the two fibroblast populations. The precise effect of mechanical forces on osteoclastogenesis of these populations is also unknown. This study revealed that more osteoclast‐like cells were induced in the co‐cultures of bone marrow cells with periodontal ligament than gingival fibroblasts, and this was considerably increased when anti‐osteoprotegerin (OPG) antibody was added to the co‐cultures. mRNA levels of receptor activator of nuclear factor‐kappaB ligand (RANKL) were increased in both populations when they were cultured with dexamethasone and vitamin D3. Centrifugal forces inhibited osteoclastogenesis of both populations, and this was likely related to the force‐induced OPG up‐regulation. Inhibition of extracellular signal‐regulated kinase (ERK) signaling by a pharmacological inhibitor (10 µM PD98059) or by siERK transfection suppressed the force‐induced OPG up‐regulation along with the augmentation of osteoclast‐like cells that were decreased by the force. These results suggest that periodontal ligament fibroblasts are naturally better at osteoclast induction than gingival fibroblasts, and that centrifugal force inhibited osteoclastogenesis of the periodontal fibroblasts through OPG production and ERK activation. J. Cell. Biochem. 106: 1010–1019, 2009. © 2009 Wiley‐Liss, Inc.
Phosphoregulation of K+‐Cl−cotransporter 4 during changes in intracellular Cl−and cell volume
Authors: Bergeron Marc J. , Frenette‐Cotton Rachelle , Carpentier Gabriel A. , Simard Michael G. , Caron Luc , Isenring Paul
Published: February 2009

It has long been stated that the K+‐Cl−cotransporters (KCCs) are activated during cell swelling through dephosphorylation of their cytoplasmic domains by a protein phosphatase (PP) but that other enzymes are involved by targeting this PP or the KCCs directly. To date, however, the role of signaling intermediates in KCC regulation has been deduced from indirect evidence rather than in vitro phosphorylation studies, and examined after simulation of ion transport through cell swelling orN‐ethylmaleimide treatment. In this study, the oocyte expression system was used to examine the effects of changes in cell volume (CVOL) and intracellular [Cl−] ([Cl−]i) on the activity and phosphorylation levels (PLEV) of KCC4, and determine whether these effects are mediated by PP1 or phorbol myristate acetate (PMA)‐sensitive effectors. We found that (1) low [Cl−]ior low CVOLleads to decreased activity but increased PLEV, (2) high CVOLleads to increased activity but no decrease in PLEVand (3) calyculin A (Cal A) or PMA treatment leads to decreased activity but no increase in PLEV. Thus, we have shown for the first time that one of the KCCs can be regulated through direct phosphorylation, that changes in [Cl−]ior CVOLmodify the activity of signaling enzymes at carrier sites, and that the effectors directly involved do not include a Cal A‐sensitive PP in contrast to the widely held view. J. Cell. Physiol. 219: 787–796, 2009. © 2009 Wiley‐Liss, Inc.
A novel role for the TRPV1 channel in UV‐induced matrix metalloproteinase (MMP)‐1 expression in HaCaT cells
Authors: Lee Young Mee , Kim Yeon Kyung , Kim Kyu Han , Park Su Jung , Kim Sung Joon , Chung Jin Ho
Published: February 2009

Transient receptor potential vanilloid type 1 (TRPV1) is a molecular sensor for detecting adverse stimuli, such as capsaicin, heat, and acid. TRPV1 has been localized in keratinocytes and is suggested to be a mediator of heat‐induced matrix metalloproteinase‐1 (MMP‐1). With regard to the multimodal activation of TRPV1, we hypothesize that TRPV1 might also mediate UV‐induced MMP‐1 in keratinocytes. In HaCaT, a human keratinocyte cell line, we initially confirmed capsaicin‐induced membrane current and Ca2+ influx. UV irradiation induced slow and persistent calcium influx and increased membrane current, which was inhibited by TRPV1 inhibitors (capsazepine and ruthenium red). The UV‐induced MMP‐1 expression in HaCaT was also decreased by TRPV1 inhibitors and was facilitated by capsaicin. Knock‐down of TRPV1 using siRNA transfection also decreased MMP‐1 expression, as well as UV‐induced Ca2+ influx in HaCaT. UV failed to induce MMP‐1 expression in HaCaT cells cultured in Ca2+‐free media. Both the UV‐induced increase in [Ca2+]i and MMP‐1 were suppressed by Gö6976 (a calcium‐dependent PKC inhibitor), but not by rottlerin (a calcium‐independent PKC inhibitor). In addition to a plausible role of TRPV1 in UV‐induced MMP‐1 expression, we showed that UV increased TRPV1 expression in both HaCaT cells and human skin in vivo. From these results, we suggest that UV‐induced MMP‐1 expression might be mediated in part by PKC‐dependent activation of TRPV1 and subsequent Ca2+‐influx in human keratinocytes. J. Cell. Physiol. 219: 766–775, 2009. © 2009 Wiley‐Liss, Inc.
Imaging of mouse experimental melanoma in vivo and ex vivo by combination of confocal and nonlinear microscopy
Authors: Chernyavskiy Oleksandr , Vannucci Luca , Bianchini Paolo , Difato Francesco , Saieh Mustafa , Kubínová Lucie
Published: February 2009

We investigated possibilities of the combination of the one‐ and two‐photon excitation microscopy for examination of the experimental melanoma tissue in vivo, in mice under general anesthesia, and ex vivo on freshly harvested specimens. Our aim was to obtain sufficiently informative images of unstained tumor tissues and their modifications after hyperthermia treatment. The mouse experimental melanoma structure was studied and compared with normal tissue from the same animal by using confocal and nonlinear microscopy techniques based on (i) one‐photon excitation (1PE) fluorescence, (ii) 1PE reflectance, (iii) second harmonic generation imaging, and (iv) two‐photon excitation autofluorescence. We checked different spectral conditions and other settings of image acquisition, as well as combinations of the above imaging modalities, to fully exploit the potential of these techniques in the evaluation of treated and untreated cancer tissue morphology. Our approach enabled to reveal the collagen fiber network in relation with the other tissues, and to identify invasive tumor cells. It also proved to be useful for the examination of interrelationships between functional and morphological aspects based on optical properties of the tissues, especially in studies of changes between the tumor and control tissue, as well as changes induced by physical treatments, e.g., delivery of microwave hyperthermia treatment. These differences were also evaluated quantitatively, when we found out that the maximum Euler–Poincaré characteristic reflects well the melanoma morphological structure. The results showed that the proposed investigative approach could be suitable also for a direct evaluation of tissue modifications induced by clinical interventions. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.
A study of hydrogenated carbon fibers by scanning electron microscopy and confocal laser scanning microscopy
Authors: De La Cal Antonio Madroñero , Aguado‐Serrano Juan , Rojas‐Cervantes Maria Luisa , Adame Elena V. Rosa , Marron Belen Sarmiento , Rosende Africa Castro , Nevshupa Roman
Published: February 2009

The hydrogen absorption process is studied in carbonaceous fibers produced from a mixture of methane and hydrogen. The absorption of the hydrogen was examined in two types of fibers, in “as‐grown” state and after a process of desorption during an annealing to 1.473 K under vacuum. Later to its production process, the fibers withstand an oxidation in air to 973 K. The fibers were examined by means of scanning electron microscopy (SEM) and confocal microscopy by reflection. Differences in the behavior during the oxidation were observed between the fibers in as‐grown state and those subjected to a further annealing. It could be verified that the fibers were really constituted by two different phases. In one of the phases, the storage of the hydrogen absorbed took place, whereas in the other phase there was no alteration. The process of annealing prior to the absorption of the hydrogen has an appreciable effect on the desorption rate of the hydrogen. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.
Localization and function of the anion exchanger Ae2 in developing teeth and orofacial bone in rodents
Authors: Bronckers Antonius L.J.J. , Lyaruu Donacian M. , Jansen Ineke D. C. , Medina Juan F. , Kellokumpu Sakari , Hoeben Kees A. , Gawenis Lara R. , Oude‐Elferink Ronald P.J. , Everts Vincent
Published: February 2009

To explore the functions of the anion exchanger 2 (Ae2) in the development of bones and teeth we examined the distribution of Ae2 in cells involved in the formation of teeth and surrounding bone in young hamsters, mice and rats. In all three species strongest immunostaining for Ae2 was obtained in basolateral membranes of maturation ameloblasts and in osteoclasts resorbing bone. In hamsters a weaker staining was also seen in the Golgi apparatus of secretory ameloblasts, young osteoblasts and osteocytes, odontoblasts and fibroblasts of the forming periodontal ligament. In adult Ae2 mice, in which Ae2‐targeted disruption precluded the expression of Ae2a, Ae2b1 and Ae2b2 isoforms, the immunostaining for Ae2 in ameloblasts and osteoclasts was totally abolished. The enamel formation was abnormal but teeth erupted, osteoclasts in jaw bone were functional and structure of dentin and bone was normal. In another mouse model, Ae2−/− mice in which the expression of all five Ae2 isoforms was disrupted, teeth failed to erupt and the alveolar bone proved poorly formed with giant but apparently functional osteoclasts. Our data indicate that basolaterally located Ae2a, Ae2b1 or Ae2b2 (or a combination of these) is present in maturation ameloblasts critical for the cells' normal functioning. Although isoforms of Ae2 were also present in basolateral membranes of osteoclasts, they proved to be not critical to osteoclast resorption of orofacial bone. Poorly formed bone and the failure of teeth to erupt seen in the Ae2−/− mice with gene disruption affecting all isoforms may result from secondary (systemic) changes that are different from Ae2 mice. J. Exp. Zool. (Mol. Dev. Evol.) 312B:375–387, 2009. © 2009 Wiley‐Liss, Inc.
Seasonal variation of the oviduct of the American alligator, Alligator mississippiensis (Reptilia: Crocodylia)
Authors: Bagwill April , Sever David M. , Elsey Ruth M.
Published: February 2009

The annual oviductal cycle of the American alligator, Alligator mississippiensis, is described using light and electron microscopy. Previous work done by Palmer and Guillette ([1992] Biol Reprod 46:39–47) shed some light on the reproductive morphology of the female alligator oviduct; however, their study was limited and did not report details relating to variation across the reproductive season. We recognize six variable regions of the oviduct: infundibulum, tube, isthmus, anterior uterus, posterior uterus, and vagina. Each area shows variation, to some degree, in the histochemistry and ultrastructure of oviductal secretions. Peak secretory activity occurs during the months of May and June, with the greatest variation occurring in the tube and anterior uterus. During the month of May, high densities of neutral carbohydrates and proteins are found within the tubal and anterior uterine glands. The epithelium of the entire oviduct secretes neutral carbohydrates throughout the year, but many regions lack protein secretions, and the posterior uterine glands show little secretory activity of any type throughout the year. After oviposition, secretory activity decreases drastically, andthe oviduct resembles that of the premating season. This study also provides evidence to support the homology between alligator and bird oviducts. Sperm were observed in glands at the tubal‐isthmus and utero‐vaginal junctions in preovulatory, postovulatory and postovipository females. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.
The theropod furcula
Authors: Nesbitt Sterling J. , Turner Alan H. , Spaulding Michelle , Conrad Jack L. , Norell Mark A.
Published: February 2009

The furcula is a structure formed by the midline fusion of the clavicles. This is the element which is unique to theropods and is important for understanding the link between birds and other theropods. New specimens from basal theropods suggest that the furcula appeared very early in theropod history. We review furcula development, function, and morphology, as well as the anatomical terminology applied to it. Furcular morphology is highly variable in crown‐group avians but is rather conserved among nonavian theropods. Here we review, or describe for the first time, the furculae in many nonavian theropods. Furculae occur in nearly all major clades of theropods, as shown by new theropod specimens from the Early Cretaceous of China and a close inspection of previously collected specimens. Informative phylogenetic characters pertaining to the furcula occur throughout Theropoda, though care should betake to consider taphonomic effects when describing furcular morphology. J. Morphol., 2009. © 2009 Wiley‐Liss, Inc.
An automated GCxGC‐TOF‐MS protocol for batch‐wise extraction and alignment of mass isotopomer matrixes from differential 13C‐labelling experiments: a case study for photoautotrophic‐mixotrophic grown Chlamydomonas reinhardtii cells
Authors: Kempa Stefan , Hummel Jan , Schwemmer Thorsten , Pietzke Matthias , Strehmel Nadine , Wienkoop Stefanie , Kopka Joachim , Weckwerth Wolfram
Published: February 2009

Two dimensional gas chromatography coupled to time‐of‐flight mass spectrometry (GCxGC‐TOF‐MS) is a promising technique to overcome limits of complex metabolome analysis using one dimensional GC‐TOF‐MS. Especially at the stage of data export and data mining, however, convenient procedures to cope with the complexity of GCxGC‐TOF‐MS data are still in development. Here, we present a high sample throughput protocol exploiting first and second retention index for spectral library search and subsequent construction of a high dimensional data matrix useful for statistical analysis. The method was applied to the analysis of 13C‐labelling experiments in the unicellular green alga Chlamydomonas reinhardtii. We developed a rapid sampling and extraction procedure for Chlamydomonas reinhardtii laboratory strain (CC503), a cell wall deficient mutant. By testing all published quenching protocols we observed dramatic metabolite leakage rates for certain metabolites. To circumvent metabolite leakage, samples were directly quenched and analyzed without separation of the medium. The growth medium was adapted to this rapid sampling protocol to avoid interference with GCxGC‐TOF‐MS analysis. To analyse batches of samples a new software tool, MetMax, was implemented which extracts the isotopomer matrix from stable isotope labelling experiments together with the first and second retention index (RI1 and RI2). To exploit RI1 and RI2 for metabolite identification we used the Golm metabolome database (GMD [1] with RI1/RI2‐reference spectra and new search algorithms. Using those techniques we analysed the dynamics of 13CO2 and 13C‐acetate uptake in Chlamydomonas reinhardtii cells in two different steady states namely photoautotroph and mixotroph growth conditions. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Effect of integrated application of chitosan coating and modified atmosphere packaging on overall quality retention in litchi cultivars
Authors: De Reuck Karen , Sivakumar Dharini , Korsten Lise
Published: February 2009

BACKGROUND: The storage life of litchi is limited due to pericarp browning and decay. Modified atmosphere packaging (MAP) showed promising results for ensuring quality retention. However, to improve the efficiency of MAP the integrated treatment of a chitosan coating and MAP was investigated.

RESULTS: The effect of chitosan (1.0 g L−1) + MAP was compared with MAP (control), and was effective in preventing decay, browning and retaining the pericarp colour in the cultivar McLean's Red. Chitosan (1.0 g L−1) + MAP significantly reduced polyphenol oxidase (PPO) and peroxidase (POD) activity, retained membrane integrity, anthocyanin content and prevented the decline of pericarp colour values during storage. The POD activity was greater than the PPO activity in the cultivars McLean's Red and Mauritius. The two cultivars differed in anthocyanin content and the activity of oxidation enzymes. The gas compositions within the packages were compared between chitosan at 1.0 g L−1 and 20.0 g L−1 concentration for both cultivars. Chitosan (20.0 g L−1) + MAP lowered the respiration during storage in both cultivars compared to 1.0 g L−1 + MAP.

CONCLUSION: The McLean's Red cultivar is better suited for chitosan (1.0 g L−1) + MAP integrated treatment than is Mauritius in retaining overall quality. Copyright © 2009 Society of Chemical Industry
Relation between chemical composition of oak wood used in cooperage and sensory perception of model extracts
Authors: Prida Andrei , Heymann Hildegarde , Balanuta Anatol , Puech Jean‐Louis
Published: February 2009

BACKGROUND: Oak extractive substances modify the aroma and taste of ‘oaked’ wines, with an increased intensity of sensory descriptors such as ‘oaky’, ‘spicy’, ‘vanilla’, ‘smoky’, etc. arising during barrel maturation. In this study the relationship between the chemical composition of untoasted oak wood and the sensory assessment of oak extract was investigated. Oak wood samples (140) were analysed both chemically (using gas chromatography/mass spectrometry after extraction with dichloromethane) and using sensory descriptive analysis of oak‐extracted model wine solutions.

RESULTS: Oak samples from the USA, France and Eastern Europe had different chemical compositions in terms of principal odour‐active extractives, and this was reflected by perceived sensory differences.

CONCLUSION: The major sensory differences were associated with cis‐whisky‐lactone concentrations of the wood extracts and were related to sensory descriptors such as coconut, caramel and milk. Other compounds studied (trans‐whisky‐lactone, eugenol and vanillin) were found to have a minor impact on sensory differences between samples in spite of their odour activity. This fact could be explained by the natural differences between samples not being sufficiently large to be perceived. The effects of aroma amplification and/or masking linked to the presence of other odorous chemicals could also explain these observations. Copyright © 2009 Society of Chemical Industry
Evaluating NIR instruments for quantitative and qualitative assessment of intact apple quality
Authors: Paz Patricia , Sánchez María‐Teresa , Pérez‐Marín Dolores , Guerrero José‐Emilio , Garrido‐Varo Ana
Published: February 2009

BACKGROUND: In recent years, near‐infrared reflectance (NIR) instruments have undergone radical changes, becoming much more versatile, more portable, cheaper and better adapted to hostile working areas. In this study, three commercially available spectrophotometers were evaluated for the determination of quality (soluble solid content, firmness and shelf‐life) in intact apples. The three instruments used, which differ primarily in terms of measurement principle and wavelength range, were a scanning monochromator (SM) with a range of 400–2500 nm, a combination of diode array and scanning monochromator (DASM) with a range of 350–2500 nm and a diode array (DA) with a range of 900–1700 nm.

RESULTS: A total of 334 apples (Malus domestica Borkh.), cvs Fuji and Golden Delicious, were used to build calibration models in different spectral regions and using various spectral signal pretreatments. The three NIR instruments evaluated provided good precision for soluble solid content, with r2 values between 0.90 and 0.94 and standard error of cross‐validation (SECV) values ranging from 0.51 to 0.68°Brix; however, firmness measurements were less precise in all three cases (r2 = 0.52–0.57, SECV = 8.28–8.83 N). The performance of the three instruments in classifying apples by shelf‐storage duration (0, 8 and 14 days) was studied using partial least squares discriminant analysis to develop classification models. A total of 86.1% of samples from the mixed‐cultivar group were correctly assigned, compared with 86.6% of samples from single‐cultivar groups.

CONCLUSION: The results obtained suggest that, in general, the three NIR instruments tested provided a similar level of accuracy for the measurement of soluble solid content, firmness and shelf‐life, being slightly better the prediction models developed with the DASM spectrophotometer. Copyright © 2009 Society of Chemical Industry
Electrochemical Polymerization of Bis(3,4‐ethylenedioxythiophene)‐Substituted 1,4‐Diketo‐3,6‐diphenyl‐pyrrolo[3,4‐c]pyrrole (DPP) Derivative
Authors: Zhu Yu , Zhang Kai , Tieke Bernd
Published: February 2009

Synthesis and electropolymerization of the new bisthienyl‐substituted diketopyrrolopyrrole monomer 1,4‐diketo‐2,5‐di‐hexyl‐3,6‐bis{[2‐(3',4'‐ethylenedi‐oxy)thienyl]phenyl}pyrrolo[3,4‐c]pyrrole (EDOT‐DPP‐EDOT) is reported. EDOT‐DPP‐EDOT exhibits a low oxidation potential of 0.67 V versus ferrocene and can be electropolymerized to form a stable, well adherent polymer thin film at the ITO anode. The polymer film is insoluble in organic solvents, non‐fluorescent, but exhibits reversible electrochromic properties with color changes from deep blue in the neutral state via transparent gray to purple red in the oxidized state. The reversibility in air is excellent.

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Well‐Defined Crystalline TiO2 Nanoparticles Generated and Immobilized on a Colloidal Nanoreactor
Authors: Lu Yan , Hoffmann Martin , Yelamanchili Ram Sai , Terrenoire Alexandre , Schrinner Marc , Drechsler Markus , Möller Michael W. , Breu Josef , Ballauff Matthias
Published: February 2009

We synthesized well‐defined, crystalline TiO2 nanoparticles at room temperature by using spherical polyelectrolyte brush particles as a template. The template particles consist of a polystyrene core from which long chains of poly(styrene sodium sulfonate) are grafted. Tetraethylorthotitanate is hydrolyzed in the presence of brush particles leading to the formation of well‐dispersed TiO2 nanoparticles (d = 4∼12 nm). Wide‐angle X‐ray scattering demonstrates that anatase nanoparticles with high crystallinity have been generated at room temperature. The as‐prepared TiO2 nanocomposites present high photocatalytic activity for the degradation of Rhodamine B under UV irradiation. Finally, mesoporous TiO2 structures with defined pore size are formed after calcination.

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Macromol. Chem. Phys. 3–4/2009
Authors: Retsch Markus , Zhou Zuocheng , Rivera Sergio , Kappl Michael , Zhao Xiu Song , Jonas Ulrich , Li Qin
Published: February 2009

Cover: Highly ordered and robust monolayers of colloidal particles can be prepared by a simple and versatile floating method at the air–water interface. The floating monolayer can be efficiently transferred to corrugated and curved surfaces, allowing the stacking of complex layer architectures. Further details can be found in the article by M. Retsch, Z. Zhou, S. Rivera, M. Kappl, X. S. Zhao, U. Jonas, Q. Li on page 230.
Double‐porosity homogenization for perimeter functionals
Authors: Solci Margherita
Published: February 2009

In this paper we are concerned with the overall description of interfacial energies in a composite environment where the local properties of the media are characterized by a large variation of the values of surface tension (double‐porosity homogenization).

We prove some asymptotic results in terms of Γ‐convergence for a sequence of functionals modelling these energies. We show the connection of the Γ‐limit with the geometry of the periodic ‘holes’ of the perforated domain, in particular investigating the relation with the Cheeger constant and the Cheeger set of a convex subset of ℝn.