bluuebunny/crossref_metadata_2025 huggingface dataset:
Structure and properties of nanocomposites prepared from ball milled 6061aluminium alloy with ZrO2 nanoparticles
Authors: Dutkiewicz J. , Lityńska L. , Maziarz W. , Haberko K. , Pyda W. , Kanciruk A.
Published: September 2009

Ball milling of 6061 aluminum alloy and nanocrystalline ZrO2 powder allowed to obtain homogeneous mixture of nanocrystalline aluminum solid solution of grain size near 50 nm and ZrO2 particles. The ZrO2 particles are partially transformed after milling from tetragonal to the monoclinic crystal structure. After hot pressing in vacuum at temperature of 380 °C bulk nanocomposites were obtained of porosity below 1% consisting of aluminum solid solution of the average grain size near 80 nm and nano‐size ZrO2 particles. Partial recrystallization occurred after hot pressing particularly near particles boundaries, where elongated recrystallized grains free from ZrO2 nanoparticles of thickness near 1 μm and length of a few μm were identified. Estimated fraction of recrystallized grains was below 10%. The compression strength approaching 1000 MPa was obtained in composites containing 20% ZrO2 nanoparticles, what is more than reported for ultra and nano‐grain 6XXX aluminum alloys. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Growth and optical homogeneity investigations of Fe and Fe:Mn codoped LiNbO3 crystals
Authors: Kar S. , Verma Sunil , Khan M. S. , Bartwal K. S.
Published: September 2009

Lithium niobate (LiNbO3) crystals doped with Fe and Fe:Mn were grown by Czochralski technique. The doping concentrations of Fe and Mn were optimized. Transmission studies reveal broad absorption band centered at 488 nm. The UV cutoff observed for Fe doped LiNbO3 is 358 nm whereas for Fe:Mn codoped LiNbO3 is 352 nm. This decrease in UV cutoff for Fe and Mn codoped LiNbO3 compared to only Fe doped LiNbO3 is due to the increase in Li/Nb ratio. Optical homogeneity was assessed using conoscopy and birefringence interferometry. Dark and photo conductivity measurements prove that LiNbO3 is a negative photo conducting material. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Inflammatory events as detected in cervical smears and squamous intraepithelial lesions
Authors: Roeters Anne M. E. , Boon Mathilde E. , van Haaften Maarten , Vernooij F. , Bontekoe Tj. Romke , Heintz A. Peter M.
Published: September 2009

The Dutch cytological coding system, KOPAC, enables to code for eight inflammatory events, that is koilocytosis (related to human papillomavirus (HPV)), Trichomonas, dysbacteriosis [related to bacterial vaginosis (BV)], Candida, Gardnerella, Actinomyces, Chlamydia, and non‐specific inflammation (leucocytosis). This study presents an analysis of 1,008,879 smears. Of each smear, the age of the woman and the reason for smear taking (screening or indication) was available. The cytoscores (per mille) for these codes were calculated. For the screening smears, the cytoscores were for koilocytosis (HPV) 2.6, for Trichomonas vaginalis 1.9, for dysbacteriosis 31.4, for Candida albicans 9.8, for Gardnerella vaginalis 0.7, for Actinomyces 6.9, for Chlamydia 0.8, and for non‐specific inflammatory changes 66.4. For the calculation of the Odds Ratio (OR), normal smears were used as a reference. The cytoscores for Chlamydia and Gardnerella covaried with high grade SIL (HSIL), with an OR of 7 and 12, respectively. In addition, the OR for Trichomonas vaginalis, for dysbacteriosis, and for leucocytosis proved to be significantly high in the indication smears. This study provides an oversight of HSIL and the full range of cervical infections as detected by cytology, proving that this infectious byproduct of screening can be very valuable. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.
Fine‐needle aspiration of primary osseous lesions: A cost effectiveness study
Authors: Layfield Lester J. , Dodd Leslie G. , Hirschowitz Sharon , Crabtree Susie Newman
Published: September 2009

Fine‐needle aspiration (FNA) is not widely used in the work‐up of osseous lesions because of concerns regarding its high incidence of nondiagnostic specimens. Although several studies have shown that FNA is less expensive than surgical biopsy, the authors are aware of only one prior study evaluating the cost effectiveness of FNA, which includes the cost of incisional or core needle biopsies necessary to establish a diagnosis when the initial FNA was noncontributory. A computerized search of the pathology records of three medical centers was performed to obtain all FNAs of primary osseous lesions. For each FNA case, all subsequent core needle, incisional or excisional biopsies were recorded as was the result of the definitive operative procedure. The cost of obtaining the definitive diagnosis was calculated for each case including the cost of FNA, imaging guidance utilized, and cost of subsequent surgical biopsy when necessary. The cost of an alternate approach using only surgical biopsy was calculated. The average per patient costs of these two protocols were compared.

A total of 165 primary bone tumors underwent FNA. One hundred six of these yielded a definitive cytologic diagnosis. In 59 cases, FNA yielded a result insufficient for definitive therapy necessitating surgical biopsy. FNA investigation of the 165 bone lesions cost 575,932 (average of 3,490 per patient). Surgical biopsy alone would have cost 5,760 per patient. FNA resulted in a cost savings of 2,215 per patient. Diagn. Cytopathol. 2010 © 2009 Wiley‐Liss, Inc.
Nipple aspirate fluid and ductoscopy to detect breast cancer
Authors: Sauter Edward R. , Klein‐Szanto Andres , MacGibbon Brenda , Ehya, Hormoz
Published: September 2009

We prospectively performed cytologic assessment and image analysis (IA) on matched nipple aspirate fluid (NAF) and mammary ductoscopy (MD) specimens to determine (1) the accuracy of these methods in cancer detection and (2) whether the two collection methods provide complementary information.

NAF and MD specimens were collected from 84 breasts from 75 women (nine bilateral samples) who underwent breast surgery. Cytologic evaluation was performed on all samples. IA was performed on slides with sufficient epithelial cells.

Cytologic evaluation proved more accurate in patients without pathologic spontaneous nipple discharge (PND) than those with PND, mainly because of the potential false positive diagnosis in the latter. While the sensitivity of NAF and MD cytology was low (10% and 14%, respectively), both were 100% specific in cancer detection in the non‐PND cohort. Combining NAF and MD cytology information improved sensitivity (24%) without sacrificing specificity. Similar to cytology, IA was more accurate in patients without PND having high specificity (100% for aneuploid IA), but relatively low sensitivity (36%). Combining NAF and MD cytology with aneuploid IA improved the sensitivity (45%) while maintaining high specificity (100%). The best predictive model was positive NAF cytology and/or MD cytology combined with IA aneuploidy, which resulted in 55% sensitivity and 100% specificity in breast cancer detection.

Cytologic evaluation and IA of NAF and MD specimens are complementary. The presence of atypical cells arising from an intraductal papilloma in ductoscopic specimens is a potential source of false positive diagnosis in patients with nipple discharge. Diagn. Cytopathol. 2010 © 2009 Wiley‐Liss, Inc.
Intraoperative imprint cytology of central neurocytoma: The great oligodendroglioma mimicker
Authors: Klysik Michal , Gavito Jose , Boman Darius , Miranda Roberto N. , Hanbali Fadi , De Las Casas Luis E.
Published: September 2009

Intraoperative cytologic evaluation of brain tumors has been used either to render a preliminary interpretation or more often as a complement to frozen section examination. Central neurocytoma is a intraventricular neoplasm, typically located in the region of the foramen of Monro, affecting mostly young to middle age adults. Histologically, central neurocytomas are characterized by monotonous cells with round nuclei and neuronal differentiation within a rich capillary network. Their distinction during intraoperative consultations from oligodendroglioma, ependymoma (mainly clear cell ependymoma), and non‐Hodgkin lymphoma can be a diagnostic challenge. We report a case of a 19‐year‐old female with an intraventricular tumor where imprint cytology preparations were crucial for the intraoperative diagnosis of central neurocytoma. Imprint cytology preparations show a round cell neoplasm associated with neuropil clumps and short straight capillaries admixed with tumor cell clusters. To the best of our knowledge, only a few cases describing the cytologic findings of central neurocytomas have been reported in the medical literature. The differential diagnosis, tissue correlation, clinical‐radiologic features, and ancillary studies are discussed. Diagn. Cytopathol. 2010. © 2009 Wiley‐Liss, Inc.
Identification of starmaker‐like in medaka as a putative target gene of Pax2 in the otic vesicle
Authors: Bajoghli Baubak , Ramialison Mirana , Aghaallaei Narges , Czerny Thomas , Wittbrodt Joachim
Published: September 2009

Otoliths in bony fishes are involved in the function of the ear in the senses of balance and hearing. In a large‐scale random in situ hybridization screen of genes expressed in the medaka developing ear, we identified starmaker‐like (stm‐l) gene, a novel homologue of zebrafish starmaker and human dentine sialo‐phosphoprotein (dspp) gene. Despite the absence of sequence similarity between these genes, here we describe their similar genomic structure and expression patterns hinting for a conserved function. In medaka fry, stm‐l is expressed in various organs such as otoliths, teeth, gills, and kidney. Additionally, our results provide evidence that stm‐l is a putative downstream target gene of Pax2 transcription factor and Pax2 itself has a promoting function in otolith formation. Developmental Dynamics 238:2860–2866, 2009. © 2009 Wiley‐Liss, Inc.
Analysis of gene expression in PTHrP−/− mammary buds supports a role for BMP signaling and MMP2 in the initiation of ductal morphogenesis
Authors: Hens Julie , Dann Pamela , Hiremath Minoti , Pan Tien‐Chi , Chodosh Lewis , Wysolmerski John
Published: September 2009

Parathyroid hormone–related protein (PTHrP) acts on the mammary mesenchyme and is required for proper embryonic mammary development. In order to understand PTHrP's effects on mesenchymal cells, we profiled gene expression in WT and PTHrP−/− mammary buds, and in WT and K14‐PTHrP ventral skin at E15.5. By cross‐referencing the differences in gene expression between these groups, we identified 35 genes potentially regulated by PTHrP in the mammary mesenchyme, including 6 genes known to be involved in BMP signaling. One of these genes was MMP2. We demonstrated that PTHrP and BMP4 regulate MMP2 gene expression and MMP2 activity in mesenchymal cells. Using mammary bud cultures, we demonstrated that MMP2 acts downstream of PTHrP to stimulate ductal outgrowth. Future studies on the functional role of other genes on this list should expand our knowledge of how PTHrP signaling triggers the onset of ductal outgrowth from the embryonic mammary buds. Developmental Dynamics 238:2713–2724, 2009. © 2009 Wiley‐Liss, Inc.
Expression of the oscillating gene her1 is directly regulated by hairy/enhancer of split, T‐box, and suppressor of hairless proteins in the zebrafish segmentation clock
Authors: Brend Tim , Holley Scott A.
Published: September 2009

Somites are segmental units of the mesoderm in vertebrate embryos that give rise to the axial skeleton, muscle, and dermis. Somitogenesis occurs in a periodic manner and is governed by a segmentation clock that causes cells to undergo repeated oscillations of gene expression. Here, we present a detailed analysis of cis‐regulatory elements that control oscillating expression of the zebrafish her1 gene in the anterior presomitic mesoderm. We identify binding sites for Her proteins and demonstrate that they are necessary for transcriptional repression. This result confirms that direct negative autoregulation of her gene expression constitutes part of the oscillator mechanism. We also characterize binding sites for fused somites/Tbx24 and Suppressor of Hairless proteins and show that they are required for activation of her1 expression. These data provide the foundation for a precise description of the regulatory grammar that defines oscillating gene expression in the zebrafish segmentation clock. Developmental Dynamics 238:2745–2759, 2009. © 2009 Wiley‐Liss, Inc.
Haptoglobin deficiency facilitates the development of autoimmune inflammation
Authors: Galicia Georgina , Maes Wim , Verbinnen Bert , Kasran Ahmad , Bullens Dominique , Arredouani Mohamed , Ceuppens Jan L.
Published: September 2009

Haptoglobin (HP) is an acute phase protein synthesized by liver cells in response to IL‐6. HP has been demonstrated to modulate the immune response and to have anti‐inflammatory activities. To analyze HP's effect on autoimmune inflammation, we here studied the course of EAE induced by immunization of Hp knockout (Hp−/−) and syngeneic WT mice with myelin oligodendrocyte glycoprotein peptide (MOG35–55). Hp−/−mice suffered from a more severe disease that was associated with increased expression of IL‐17A, IL‐6, and IFN‐γ mRNA in the CNS and with a denser cellular infiltrate in the spinal cord. During the recovery phase, a significantly higher number of myeloid DC, CD8+ cells, IL‐17+ CD4+ and IFN‐γ+ CD4+ cells persisted in the CNS of Hp−/− mice. Absence of HP affected the priming and differentiation of T cells after MOG35–55 immunization, as levels of Th2 cytokines produced in response to MOG stimulation by Hp−/− T cells were reduced. These results suggest that HP plays a modulatory and protective role on autoimmune inflammation of the CNS.
Combination of host susceptibility and Mycobacterium tuberculosis virulence define gene expression profile in the host
Authors: Beisiegel Martin , Mollenkopf Hans‐Joachim , Hahnke Karin , Koch Markus , Dietrich Isabell , T. Reece Stephen , Kaufmann Stefan H. E.
Published: September 2009

Progression and outcome of tuberculosis is governed by extensive crosstalk between pathogen and host. Analyses of global changes in gene expression during immune response to infection with Mycobacterium tuberculosis (M.tb) can help identify molecular markers of disease state and progression. Global distribution of M.tb strains with different degrees of virulence and drug resistance, especially for the immunocompromised host, make closer analyses of host responses more pressing than ever. Here, we describe global transcriptional responses of inducible nitric oxide synthase‐deficient (iNOS–/–) and WT mice infected with two related M.tb strains of markedly different virulence, namely the M.tb laboratory strains H37Rv and H37Ra. Both hosts exhibited highly similar resistance to infection with H37Ra. In contrast, iNOS–/– mice rapidly succumbed to H37Rv, whereas WT mice developed chronic course of disease. By differential analyses, virulence‐specific changes in global host gene expression were analyzed to identify molecular markers characteristic for chronic versus acute infection. We identified several markers unique for different stages of disease progression and not previously associated with virulence‐specific host responses in tuberculosis.
Synthesis and Physical Properties of Two Chiral Terpyridyl Europium(III) Complexes with Distinct Crystal Polarity
Authors: Li Dong‐Ping , Li Cheng‐Hui , Wang Jun , Kang Ling‐Cheng , Wu Tao , Li Yi‐Zhi , You Xiao‐Zeng
Published: September 2009

By utilizing the chiral terpyridyl‐type ligand L(–) [L =“dipineno”‐(4,5:4″,5″)‐fused 2,2′:6′,2″‐terpyridine or 2,6‐bis(7,7‐dimethyl‐5,6,7,8‐tetrahydro‐6,8‐methanoisoquinolin‐3‐yl)pyridine], two novel EuIII complexes with the formula [Eu(tta)3L](C3H6O)·0.5(H2O) (1) and [Eu(bta)3(L)](C3H6O)(2) (tta = 2‐thenoyltrifluoroacetonate, bta = benzoyltrifluoroacetonate) were synthesized and characterized by X‐ray crystallography and spectroscopic methods. The EuIII atoms in 1 and 2 are nine‐coordinate in a geometrical environment of a distorted, monocapped square antiprism. Complex 1 crystallizes in the polar space group P21, while complex 2 crystallizes in the nonpolar space group P212121. The chirality of both complexes is evidenced by the circular dichroism (CD) spectra. Both complexes show the characteristic photoluminescence of the EuIII ion. In contrast to complex 2, complex 1 exhibits brilliant triboluminescence, an intense NLO property and ferroelectric behaviour at room temperature. The distinct chiral optical properties of complexes 1 and 2 originate from the differing crystal polarities.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)
Nutritional value of rapeseed oil and its high oleic/low linolenic variety – A call for differentiation
Authors: Barth Christian A.
Published: September 2009

To offer the best choice of healthy and acceptable food to the consumer a coordination of plant breeding, food processing and nutrition science is required. Here the nutritional aspects of the high oleic/low linolenic (HOLLi) varieties of rapeseed with a low α‐linolenic acid content of about 3% are reviewed. The content of α‐linolenic acid amounting to around 9% is the hallmark of the positive nutritional value of the original (erucic acid free) 00 varieties of rapeseed oil (“canola” quality in North America). n‐3 fatty acids are endowed with the property to protect the cardiovascular system from chronic disease and the consumption of food containing n‐3 fatty acids is explicitly recommended by national and international nutritional and medical authorities. Although the use of HOLLi with a low n‐3 fatty acid content can be unavoidable for specific purposes, because of technological and health considerations the continuous future consumption of the original rapeseed oil with around 9% of α‐linolenic acid by the consumer should have high priority from the standpoint of public health. To pursue this aim confusion of the consumer must be avoided by creating a new name and a new brand for HOLLi varieties.
Physical‐chemical characteristics and oxidative stability of oil obtained from lyophilized raspberry seed
Authors: Šućurović Aleksandra , Vukelić Nikola , Ignjatović Ljubiša , Brčeski Ilija , Jovanović Dušan
Published: September 2009

Fresh raspberry (Rubus idaeus), cultivar Willamette, was freeze‐dried (lyophilization). A byproduct of lyophilization is “fine dust” of raspberry consisting of finely ground raspberry fruit body and seed. The seeds were separated. The seed oil was isolated and its physical and chemical characteristics were determined. Parameters that characterize the seed and quality of the oil were examined, including fatty acid composition, oxidative stability under different storage conditions, and radical‐scavenging activity. The fatty acid composition was determined by GC/FID and the contents of the dominant fatty acids were found as: oleic 16.92%, linoleic 54.95%, and α‐linolenic acid 23.97%. The oxidative stability of the oil was poor. The induction period by Rancimat test at 100 °C was 5.2 h. The radical‐scavenging activity is similar to that of resveratrol [1,3‐benzenediol 5‐(1E‐2‐4‐hydroxy‐phenyl‐ethyl)]. Although this product is used in the candy industry, it would be far more useful if raspberry oil of satisfactory quality could be extracted. This paper demonstrates that sifted lyophilized seeds can be used for the extraction of oils. This process allows for maximal usage of the byproducts, reduces losses and it increases the development of new products.
Insights into the physicochemical properties of coffee oil
Authors: Calligaris Sonia , Munari Marina , Arrighetti Gianmichele , Barba Luisa
Published: September 2009

The lipid fraction of roasted coffee is an interesting ingredient that could be used in a large number of food formulations. Coffee oil has peculiar flavouring as well as nutraceutical characteristics. The feasibility of the use of coffee oil as ingredient greatly depends not only on its chemical characteristics but also on its physical properties. The crystallisation and melting properties of the coffee oil extracted from Arabica roasted coffee powder were determined by using synchrotron X‐ray diffraction coupled with differential scanning calorimetry. The fatty acid composition and the flavour profile were also assessed by using GC and GC‐MS analyses, respectively. The main fatty acids found in coffee oil are linoleic and palmitic acid. Significant amounts of stearic and oleic acid are also present. These chemical characteristics are linked to the phase transition behaviour. The crystallisation of coffee oil occurs at 6.5 ± 0.3 °C, independently of the cooling rate applied (from 0.5 to 10 °C/min). A unique crystalline structure was identified: a double chain length (2L) β' structure (55.29 Å). The sole formation of the β' form indicates that this metastable crystal is the only one that one should expect in foods containing coffee oil stored below 7 °C.
Temperature‐dependent solubility of wax compounds in ethanol
Authors: Holser Ronald Alan
Published: September 2009

The ability of ethanol to dissolve wax compounds, as an alternative to traditional lipid solvents, was investigated for the recovery of cuticular lipids from biomass. The solubilities of fatty esters with carbon chain lengths from 40 to 54 were measured in ethanol over a temperature range of 30–80°C. The greatest increase in solubility was observed between 40° and 60°C for the long chain waxes that are characteristic of flax cuticle lipids. The solubility of a 52‐carbon wax increased by a factor of four over this temperature range. The Van't Hoff equation was used to estimate enthalpy of solution values. Ethanol was an effective lipid solvent at these modestly elevated temperatures and offers an economical method to recover lipid co‐products from biomass prior to conversion to bioethanol.
Catalytic cleavage of methyl oleate or oleic acid
Authors: Köckritz Angela , Blumenstein Michael , Martin Andreas
Published: September 2009

Different reaction pathways are discussed for the Os‐catalyzed oxidation of methyl oleate and oleic acid using O2/aldehyde as oxidation system. Monomethyl azelate and pelargonic acid were the main products obtained in yields of approximately 50–70% starting from methyl oleate. Besides, varying amounts of methyl 9,10‐epoxystearate and methyl 9,10‐dihydroxystearate were found as by‐products. Azelaic acid and pelargonic acid were obtained exclusively from oleic acid used as reactant. Some mechanistic considerations led to the conclusion that the observed products are formed in parallel reaction paths. The effective oxidant for the scission of the C=C double bond seems to be very likely an in situ formed peracid generated by Os‐catalysis from O2/aldehyde. Additional investigations concerning the cleavage of oleic acid and methyl oleate with in situ formed performic acid from H2O2/formic acid corroborate this assumption.
Numerical algorithms for modelling electrodeposition: Tracking the deposition front under forced convection from megasonic agitation
Authors: Hughes Michael , Strussevitch Nadia , Bailey Christopher , McManus Kevin , Kaufmann Jens , Flynn David , Desmulliez Marc P. Y.
Published: September 2009

Electrodeposition is a widely used technique for the fabrication of high aspect ratio microstructures. In recent years, much research has been focused within this area aiming to understand the physics behind the filling of high aspect ratio vias and trenches on substrates and in particular how they can be made without the formation of voids in the deposited material. This paper reports on the fundamental work towards the advancement of numerical algorithms that can predict the electrodeposition process in micron scaled features. Two different numerical approaches have been developed, which capture the motion of the deposition interface and 2‐D simulations are presented for both methods under two deposition regimes: those where surface kinetics is governed by Ohm's law and the Butler–Volmer equation, respectively.

In the last part of this paper the modelling of acoustic forces and their subsequent impact on the deposition profile through convection is examined. Copyright © 2009 John Wiley & Sons, Ltd.
Astrocytic poly(ADP‐ribose) polymerase‐1 activation leads to bioenergetic depletion and inhibition of glutamate uptake capacity
Authors: Tang Kim San , Suh Sang Won , Alano Conrad C. , Shao Zongjun , Hunt Waylon T. , Swanson Raymond A. , Anderson Christopher M.
Published: September 2009

Poly(ADP‐ribose) polymerase‐1 (PARP‐1) is a ubiquitous nuclear enzyme involved in genomic stability. Excessive oxidative DNA strand breaks lead to PARP‐1‐induced depletion of cellular NAD+, glycolytic rate, ATP levels, and eventual cell death. Glutamate neurotransmission is tightly controlled by ATP‐dependent astrocytic glutamate transporters, and thus we hypothesized that astrocytic PARP‐1 activation by DNA damage leads to bioenergetic depletion and compromised glutamate uptake. PARP‐1 activation by the DNA alkylating agent, N‐methyl‐N′‐nitro‐N‐nitrosoguanidine (MNNG), caused a significant reduction of cultured cortical astrocyte survival (EC50 = 78.2 ± 2.7 μM). HPLC revealed MNNG‐induced time‐dependent reductions in NAD+ (98%, 4 h), ATP (71%, 4 h), ADP (63%, 4 h), and AMP (66%, 4 h). The maximal [3H]glutamate uptake rate (Vmax) also declined in a manner that corresponded temporally with ATP depletion, falling from 19.3 ± 2.8 in control cells to 2.1 ± 0.8 nmol/min/mg protein 4 h post‐MNNG. Both bioenergetic depletion and loss of glutamate uptake capacity were attenuated by genetic deletion of PARP‐1, directly indicating PARP‐1 involvement, and by adding exogenous NAD+ (10 mM). In mixed neurons/astrocyte cultures, MNNG neurotoxicity was partially mediated by extracellular glutamate and was reduced by co‐culture with PARP‐1−/− astrocytes, suggesting that impairment of astrocytic glutamate uptake by PARP‐1 can raise glutamate levels sufficiently to have receptor‐mediated effects at neighboring neurons. Taken together, these experiments showed that PARP‐1 activation leads to depletion of the total adenine nucleotide pool in astrocytes and severe reduction in neuroprotective glutamate uptake capacity. © 2009 Wiley‐Liss, Inc.
Electrical coupling of astrocytes in rat hippocampal slices under physiological and simulated ischemic conditions
Authors: Xu Guangjin , Wang Wei , Kimelberg Harold K. , Zhou Min
Published: September 2009

Mammalian protoplasmic astrocytes are extensively coupled through gap junction channels but the biophysical properties of these channels under physiological and ischemic conditions in situ are not well defined. Using confocal morphometric analysis of biocytin‐filled astrocytic syncytia in rat hippocampal CA1 stratum radiatum we found that each astrocyte directly couples, on average, to 11 other astrocytes with a mean interastrocytic distance of 45 μm. Voltage‐independent and bidirectional transjunctional currents were always measured between directly coupled astrocyte pairs in dual voltage‐clamp recordings, but never from astrocyte–NG2 glia or astrocyte–interneuron pairs. The electrical coupling ratio varied considerably among astrocytes in developing postnatal day 14 rats (P14, 0.5–12.4%, mean = 3.6%), but became more constant in young adult P21 rats (0.18–3.9%, mean = 1.6%), and the coupling ratio declined exponentially with increasing pair distance. Electrical coupling was not affected by short‐term oxygen–glucose deprivation (OGD) treatment, but showed delayed inhibition in an acidic extracellular pH of 6.4. Combination of acidic pH (6.4) and OGD, a condition that better represents cerebral ischemia in vivo, accelerated the inhibition of electrical coupling. Our results show that, under physiological conditions, 20.7–24.2% of K+ induced currents can travel from any astrocytic soma in CA1 stratum radiatum to the gap junctions of the nearest neighbor astrocytes, but this should be severely inhibited as a consequence of the OGD and acidosis seen in the ischemic brain. © 2009 Wiley‐Liss, Inc.
Cooperative expression of junctional adhesion molecule‐C and ‐B supports growth and invasion of glioma
Authors: Tenan Mirna , Aurrand‐Lions Michel , Widmer Valerie , Alimenti Alessandro , Burkhardt Karim , Lazeyras François , Belkouch Marie‐Claude , Hammel Philippe , Walker Paul R. , Duchosal Michel A. , Imhof Beat A. , Dietrich Pierre‐Yves
Published: September 2009

Brain invasion is a biological hallmark of glioma that contributes to its aggressiveness and limits the potential of surgery and irradiation. Deregulated expression of adhesion molecules on glioma cells is thought to contribute to this process. Junctional adhesion molecules (JAMs) include several IgSF members involved in leukocyte trafficking, angiogenesis, and cell polarity. They are expressed mainly by endothelial cells, white blood cells, and platelets. Here, we report JAM‐C expression by human gliomas, but not by their normal cellular counterpart. This expression correlates with the expression of genes involved in cytoskeleton remodeling and cell migration. These genes, identified by a transcriptomic approach, include poliovirus receptor and cystein‐rich 61, both known to promote glioma invasion, as well as actin filament associated protein, a c‐Src binding partner. Gliomas also aberrantly express JAM‐B, a high affinity JAM‐C ligand. Their interaction activates the c‐Src proto‐oncogene, a central upstream molecule in the pathways regulating cell migration and invasion. In the tumor microenvironment, this co‐expression may thus promote glioma invasion through paracrine stimuli from both tumor cells and endothelial cells. Accordingly, JAM‐C/B blocking antibodies impair in vivo glioma growth and invasion, highlighting the potential of JAM‐C and JAM‐B as new targets for the treatment of human gliomas. © 2009 Wiley‐Liss, Inc.
Proteomic analysis of astroglial connexin43 silencing uncovers a cytoskeletal platform involved in process formation and migration
Authors: Olk Stephan , Turchinovich Andrey , Grzendowski Michael , Stühler Kai , Meyer Helmut E. , Zoidl Georg , Dermietzel Rolf
Published: September 2009

Connexin43 (Cx43) is the most abundant gap junction protein of the brain, where it is predominantly expressed in astrocytes. Recent studies imply a role of Cx43 in the regulation of important cellular processes, including migration, proliferation, and shape formation. These processes are assumed to be reflected by the proteome of the Cx43 expressing cells. To analyze the influence of Cx43 on the astrocytic proteome, we used RNA interference to downregulate the expression of this connexin in cultures of mouse astrocytes. We applied difference gel electrophoresis (DIGE) to compare silenced astrocytes with control cells. The differential proteome analysis revealed 15 significantly regulated proteins (between 1.2‐ and 1.6‐fold), of which six are known to belong to a group of cytoskeletal proteins involved in cortical platform formation. Astrocytes treated with Cx43 small interfering (si)RNA showed an increased expression of the cytoskeletal proteins: actin, tropomyosin, microtubule‐associated protein RP/EB1, transgelin, and GFAP, and a decreased expression of cofilin‐1. Quantitative immunocytochemistry and Western blotting revealed similar results showing an upregulation of actin, tubulin, tropomyosin, EB1, transgelin and GFAP, and a downregulation of Ser‐3‐phosphorylated cofilin. Furthermore, Cx43 silencing led to phenotypical changes in cell morphology, migratory activity, and cell adhesion. Our results provide mechanistic clues for an understanding of Cx43 interaction with cellular motor activities such as migration and process formation in astrocytes. © 2009 Wiley‐Liss, Inc.
Injection of fresh river water into a saline floodplain aquifer in an attempt to improve the condition of river red gum (Eucalyptus camaldulensis Dehnh.)
Authors: Berens Volmer , White Melissa G. , Souter Nicholas J.
Published: September 2009

The delivery of environmental flows to riverine floodplains is often constrained by water scarcity and infrastructure. This means that areas of floodplain are unable to receive environmental water through mechanisms like dam release, weir pool surcharge and artificial flooding of low lying areas. This study trials aquifer injection as a method of providing environmental flows to areas of stressed river red gum (Eucalyptus camaldulensis) on the floodplain of the lower River Murray. The objective was to inject fresh river water at the top of the saline alluvial aquifer to reduce root zone salinity and improve the condition of E. camaldulensis. Localized radial freshening of ∼10 m was observed at each of the five injection wells when 4·9 Ml (megalitres) of river water was injected into the aquifer over 45 days. Injection was abandoned after 45 days due to aquifer and well clogging, and increases in groundwater head, which caused four of the five wells to breach the confining clay layer. The trial resulted in localized and short‐lived freshening of the groundwater, which reduced salinity in the associated capillary fringe. This did not result in an improvement in the tree condition as they lay beyond the lateral extent of the freshening and were unable to access the water. Solutions to technical issues such as aquifer clogging and well breaching, as well as a need to increase the lateral extent of freshening, are required before shallow aquifer injection can be recommended as a method of delivering environmental flows. Copyright © 2009 John Wiley & Sons, Ltd.
Altered molecular pathways in melanocytic lesions
Authors: Scatolini Maria , Grand Maurizia Mello , Grosso Enrico , Venesio Tiziana , Pisacane Alberto , Balsamo Antonella , Sirovich Roberta , Risio Mauro , Chiorino Giovanna
Published: September 2009

To identify gene expression changes in melanocytic lesions, biopsies from 18 common nevi (CMN), 11 dysplastic nevi (DN), 8 radial and 15 vertical growth phase melanomas (RGPM, VGPM), and 5 melanoma metastases (MTS) were analyzed using whole genome microarrays. The comparison between CMN and RGPM showed an enrichment of Gene Ontology terms related to inter and intracellular junctions, whereas the transition from RGPM to VGPM underlined the alteration of apoptosis. Upregulation of genes involved in dsDNA break repair and downregulation of cellular adhesion genes were observed in MTS with respect to VGPM. DN exhibited rather heterogeneous molecular profiles, with some proliferation genes expressed at higher levels than in CMN, altered regulation of transcription compared to RGPM and a subset of processes, such as mismatch repair, equally expressed as in VGPM. Furthermore, the expression profile of genes involved into cellular detoxification and antigen presentation split them into two classes, with different proliferation potential. Finally, molecular profiling of individual lesions identified altered biological processes, such as regulation of apoptosis, regulation of transcription and T‐cell activation, not associated with specific histological classes but rather with subgroups of samples without apparent relationship. This holds true for dysplastic nevi in particular. Our data indicate that generally the intersection between stage specific and sample specific molecular alterations may lead to a more precise determination of the individual progression risk of melanocytic lesions.
Beneficial and detrimental effects of human endogenous retroviruses
Authors: Kurth Reinhard , Bannert Norbert
Published: September 2009

In this mini review, we aim to evaluate the structure and function of Human Endogenous Retroviruses (HERVs) with respect to the benefit they may have for humans or the damage they may cause. Emphasis is laid on their putative roles, if any, in pregnancy, in gene regulation and in cancer. As a basis for this discussion it will first be necessary to briefly describe the structure and function of retroelements, including HERVs, before addressing their positive or negative effects at the cellular and organismal level. Finally, we will give an outlook in which we will attempt to define priorities for future research.
Sunlight, hormone replacement status and colorectal cancer risk in postmenopausal women
Authors: Freedman D. Michal , Rajaraman Preetha , Fuhrman Barbara , Hoffbeck Richard , Alexander Bruce H.
Published: September 2009

A reanalysis of the Women's Health Initiative (WHI) randomized clinical trial found a significant interaction between supplementation with vitamin D/calcium and estrogen therapy and the risk of colorectal cancer risk, with reduced risks from supplementation limited to the placebo arms of the estrogen trials. To explore whether the vitamin D effects are modified by estrogen therapy, we report a largely cross‐sectional, analysis of the association between sun exposure, which is an important vitamin D source, and colorectal cancer risk among postmenopausal women in the U.S. Radiologic Technologists study. Among 21,695 participants, there were a total of 108 cases. Sun exposure was based on time outdoors and on ambient ultraviolet radiation (UV) exposure based on residence linked to erythemal exposures derived from the Total Ozone Mapping Spectrometer database. Although there was no relationship between outdoor time or ambient UV measure and colorectal cancer risk in current hormone replacement therapy (HRT) users, in never/past HRT users, there was an inverse association with higher ambient UV exposure, RR for highest vs. lowest tertile = 0.40; 95% CI 017, 0.93; p for trend = 0.04. Non‐significant lower risks were also associated with higher levels of outdoor time (≥3.5 hr/week) in never/past HRT users. The interaction between both indicators of sun exposure and HRT and CRC risk was not significant. These data, although exploratory, are consistent with evidence from the WHI suggesting a decrease in colorectal cancer risk may be associated with vitamin D exposure among postmenopausal women who are not taking HRT, but not among current HRT users.
Kallikrein‐related peptidase‐4 initiates tumor–stroma interactions in prostate cancer through protease‐activated receptor‐1
Authors: Wang Wenbin , Mize Gregory J. , Zhang Xiaotun , Takayama Thomas K.
Published: September 2009

In prostate cancer, the mechanism by which the stromal cells surrounding the cancer epithelium become reactive and overproduce growth factors is unclear. Furthermore, the precise process of how these stromal cells stimulate the cancer epithelium is not fully understood. We recently found that protease‐activated receptor‐1 (PAR‐1) in these reactive stromal cells is upregulated. To investigate the role of PAR‐1 in the stromal–epithelial interaction, WPMY‐1 stromal myofibroblasts were stimulated with PAR‐1 agonists including thrombin and PAR‐1 activating peptide. We show that WPMY‐1 cells have functional PAR‐1 by signaling through ERK1/2. Conditioned media (CM) from PAR‐1 agonists‐treated WPMY‐1 cells stimulate the epithelial LNCaP cells leading to ERK1/2 activation and cell proliferation. Cytokine array analysis of the CM demonstrates that PAR‐1 induces stromal cells to release numerous cytokines, of which interleukin 6 (IL‐6) is the major factor responsible for mitogenic signaling in LNCaP cells. CM further induces expression of prostate‐specific kallikrein‐related peptidase‐3 (KLK3/PSA) and KLK4 in LNCaP cells via the IL‐6 pathway. Moreover, KLK4 functions as a potent agonist of PAR‐1 by cleaving the receptor at the proper site on cell surface. KLK4 triggers transmembrane signaling and upregulates IL‐6 in WPMY‐1 cells through PAR‐1. Immunohistochemical analysis indicates that PAR‐1 is predominantly expressed in peritumoral stroma while KLK4 is produced exclusively by the epithelial cancer cells. These data provide evidence for a novel double‐paracrine mechanism whereby cancer epithelium produces KLK4 to activate PAR‐1 in the surrounding stroma, which in‐turn releases cytokines (IL‐6) that stimulate cancer cells to proliferate and increase production of KLKs.
Increased local dopamine secretion has growth‐promoting effects in cholangiocarcinoma
Authors: Coufal Monique , Invernizzi Pietro , Gaudio Eugenio , Bernuzzi Francesca , Frampton Gabriel A. , Onori Paolo , Franchitto Antonio , Carpino Guido , Ramirez Jonathan C. , Alvaro Domenico , Marzioni Marco , Battisti Guido , Benedetti Antonio , DeMorrow Sharon
Published: September 2009

Cholangiocarcinoma is a devastating cancer of biliary origin with limited treatment options. Symptoms are usually evident after blockage of the bile duct by the tumor, and at this late stage, they are relatively resistant to chemotherapy and radiation therapy. Therefore, it is imperative that alternative treatment options are explored. We have previously shown that serotonin metabolism is dysregulated in cholangiocarcinoma leading to an increased secretion of serotonin, which has growth‐promoting effects. Because serotonin and dopamine share the degradation machinery, we evaluated the secretion of dopamine from cholangiocarcinoma and its effects on cell proliferation. Using 4 cholangiocarcinoma cell lines and human biopsy samples, we demonstrated that there was an increase in mRNA and protein expression of the dopamine synthesis enzymes tyrosine hydroxylase and dopa decarboxylase in cholangiocarcinoma. There was increased dopamine secretion from cholangiocarcinoma cell lines compared to H69 and HIBEC cholangiocytes and increased dopamine immunoreactivity in human biopsy samples. Furthermore, administration of dopamine to all cholangiocarcinoma cell lines studied increased proliferation by up to 30%, which could be blocked by the pretreatment of the D2 and D4 dopamine receptor antagonists, whereas blocking dopamine production by α‐methyldopa administration suppressed growth by up to 25%. Administration of α‐methyldopa to nude mice also suppressed cholangiocarcinoma tumor growth. The data presented here represent the first evidence that dopamine metabolism is dysregulated in cholangiocarcinoma and that modulation of dopamine synthesis may represent an alternative target for the development of therapeutic strategies.
Leucinostatin A inhibits prostate cancer growth through reduction of insulin‐like growth factor‐I expression in prostate stromal cells
Authors: Kawada Manabu , Inoue Hiroyuki , Ohba Shun‐Ichi , Masuda Tohru , Momose Isao , Ikeda Daishiro
Published: September 2009

Targeting stroma in tumor tissues is an attractive new strategy for cancer treatment. We developed in vitro coculture system, in which the growth of human prostate cancer DU‐145 cells is stimulated by prostate stromal cells (PrSC) through insulin‐like growth factor I (IGF‐I). Using this system, we have been searching for small molecules that inhibit tumor growth through modulation of tumor‐stromal cell interactions. As a result, we have found that leucinostatins and atpenins, natural antifungal antibiotics, inhibit the growth of DU‐145 cells cocultured with PrSC more strongly than that of DU‐145 cells alone. In this study we examined the antitumor effects of these small molecules in vitro and in vivo. When DU‐145 cells were coinoculated with PrSC subcutaneously in nude mice, leucinostatin A was found to significantly suppress the tumor growth more than atpenin B. The antitumor effect of leucinostatin A in vivo was not obtained against the tumors of DU‐145 cells alone. RT‐PCR experiments revealed that leucinostatin A specifically inhibited IGF‐I expression in PrSC without effect on expressions of other IGF axis molecules. Leucinostatins and atpenins are known to abrogate mitochondrial functions. However, when we used mitochondrial DNA‐depleted, pseudo‐ρ0 cells, we found that one of leucinostain A actions certainly depended on mitochondrial function, but it actually inhibited the growth of DU‐145 cells more strongly in coculture with pseudo‐ρ0 PrSC and reduced IGF‐I expression in pseudo‐ρ0 PrSC. Taken together, our results suggested that leucinostatin A inhibited prostate cancer cell growth through reduction of IGF‐I expression in PrSC.
Genetic disparity between primary tumours, disseminated tumour cells, and manifest metastasis
Authors: Stoecklein Nikolas H. , Klein Christoph A.
Published: September 2009

Recent genetic analyses of paired samples from primary tumours and disseminated tumour cells have uncovered a bewildering genetic disparity. It was therefore proposed that ectopically residing tumour cells disseminate early and develop independently into metastases parallel to the primary tumour. Alternatively, these cells may represent an irrelevant cell population unable to spawn metastases whereas only cells that disseminated late in primary tumour development (which therefore are similar to the primary tumour) will form manifest metastasis. Here, we review comparative analyses of paired samples from primary tumours and disseminated tumour cells or primary tumours and metastases. The data demonstrate a striking disparity, questioning the use of primary tumours as surrogate for the genetics of systemic cancer. In the era of molecular therapies that build upon genetic defects of tumour cells, these data call for a direct diagnostic pathology of systemic cancer.
BMP‐2 modulates β‐catenin signaling through stimulation of Lrp5 expression and inhibition of β‐TrCP expression in osteoblasts
Authors: Zhang Ming , Yan Ying , Lim Yong‐bin , Tang Dezhi , Xie Rong , Chen Ann , Tai Peter , Harris Stephen E. , Xing Lianping , Qin Yi‐Xian , Chen Di
Published: September 2009

Canonical BMP and Wnt signaling pathways play critical roles in regulation of osteoblast function and bone formation. Recent studies demonstrate that BMP‐2 acts synergistically with β‐catenin to promote osteoblast differentiation. To determine the molecular mechanisms of the signaling cross‐talk between canonical BMP and Wnt signaling pathways, we have used primary osteoblasts and osteoblast precursor cell lines 2T3 and MC3T3‐E1 cells to investigate the effect of BMP‐2 on β‐catenin signaling. We found that BMP‐2 stimulates Lrp5 expression and inhibits the expression of β‐TrCP, the F‐box E3 ligase responsible for β‐catenin degradation and subsequently increases β‐catenin protein levels in osteoblasts. In vitro deletion of the β‐catenin gene inhibits osteoblast proliferation and alters osteoblast differentiation and reduces the responsiveness of osteoblasts to the BMP‐2 treatment. These findings suggest that BMP‐2 may regulate osteoblast function in part through modulation of the β‐catenin signaling. J. Cell. Biochem. 108: 896–905, 2009. © 2009 Wiley‐Liss, Inc.