bluuebunny/crossref_metadata_2025 huggingface dataset:
Novel Mechanism of Antibodies to Hepatitis B Virus in Blocking Viral Particle Release from Cells†
Authors: Neumann Avidan U. , Phillips Sandra , Levine Idit , Ijaz Samreen , Dahari Harel , Eren Rachel , Dagan Shlomo , Naoumov Nikolai V.
Published: May 2010

Antibodies are thought to exert antiviral activities by blocking viral entry into cells and/or accelerating viral clearance from circulation. In particular, antibodies to hepatitis B virus (HBV) surface antigen (HBsAg) confer protection, by binding circulating virus. Here, we used mathematical modeling to gain information about viral dynamics during and after single or multiple infusions of a combination of two human monoclonal anti-HBs (HepeX-B) antibodies in patients with chronic hepatitis B. The antibody HBV-17 recognizes a conformational epitope, whereas antibody HBV-19 recognizes a linear epitope on the HBsAg. The kinetic profiles of the decline of serum HBV DNA and HBsAg revealed partial blocking of virion release from infected cells as a new antiviral mechanism, in addition to acceleration of HBV clearance from the circulation. We then replicated this approach

in vitro

, using cells secreting HBsAg, and compared the prediction of the mathematical modeling obtained from the

in vivo

kinetics.

In vitro

, HepeX-B treatment of HBsAg-producing cells showed cellular uptake of antibodies, resulting in intracellular accumulation of viral particles. Blocking of HBsAg secretion also continued after HepeX-B was removed from the cell culture supernatants.

Conclusion:

These results identify a novel antiviral mechanism of antibodies to HBsAg (anti-HBs) involving prolonged blocking of the HBV and HBsAg subviral particles release from infected cells. This may have implications in designing new therapies for patients with chronic HBV infection and may also be relevant in other viral infections.
Beneficial Paracrine Effects of Cannabinoid Receptor 2 on Liver Injury and Regeneration†
Authors: Teixeira-Clerc Fatima , Belot Marie-Pierre , Manin Sylvie , Deveaux Vanessa , Cadoudal Thomas , Chobert Marie-Noele , Louvet Alexandre , Zimmer Andreas , Tordjmann Thierry , Mallat Ariane , Lotersztajn Sophie
Published: May 2010

The cannabinoid receptor 2 (CB2) plays a pleiotropic role in innate immunity and is a crucial mediator of liver disease. In this study, we investigated the impact of CB2 receptors on the regenerative process associated with liver injury. Following acute hepatitis induced by carbon tetrachloride (CCl4), CB2 was induced in the nonparenchymal cell fraction and remained undetectable in hepatocytes. Administration of CCl4 to CB2−/− mice accelerated liver injury, as shown by increased alanine/aspartate aminotransferase levels and hepatocyte apoptosis, and delayed liver regeneration, as reflected by a retarded induction of hepatocyte proliferating cell nuclear antigen expression; proliferating cell nuclear antigen induction was also delayed in CB2−/− mice undergoing partial hepatectomy. Conversely, following treatment with the CB2 agonist JWH-133, CCl4-treated WT mice displayed reduced liver injury and accelerated liver regeneration. The CCl4-treated CB2−/− mice showed a decrease in inducible nitric oxide synthase and tumor necrosis factor-α expression, and administration of the nitric oxide donor moldomine (SIN-1) to these animals reduced hepatocyte apoptosis, without affecting liver regeneration. Impaired liver regeneration was consecutive to an interleukin-6 (IL-6)-mediated decrease in matrix metalloproteinase 2 (MMP-2) activity. Indeed, CCl4-treated CB2−/− mice displayed lower levels of hepatic IL-6 messenger RNA and increased MMP-2 activity. Administration of IL-6 to these mice decreased MMP-2 activity and improved liver regeneration, without affecting hepatocyte apoptosis. Accordingly, administration of the MMP inhibitor CTTHWGFTLC to CCl4-treated CB2−/− mice improved liver regeneration. Finally,

in vitro

studies demonstrated that incubation of hepatic myofibroblasts with JWH-133 increased tumor necrosis factor-α and IL-6 and decreased MMP-2 expressions.

Conclusion:

CB2 receptors reduce liver injury and promote liver regeneration following acute insult, via distinct paracrine mechanisms involving hepatic myofibroblasts. These results suggest that CB2 agonists display potent hepatoprotective properties, in addition to their antifibrogenic effects.
HB‐EGF orchestrates the complex signals involved in triple‐negative and trastuzumab‐resistant breast cancer
Authors: Yotsumoto Fusanori , Oki Eiji , Tokunaga Eriko , Maehara Yoshihiko , Kuroki Masahide , Miyamoto Shingo
Published: May 2010

A number of therapeutic strategies targeting epidermal growth factor receptor (EGFR) have not always led to success in the present state of breast cancer therapy. Notably, there is currently no way to treat trastuzumab‐resistant and triple‐negative breast cancer (TNBC). Here, we found that heparin‐binding epidermal growth factor‐like growth factor (HB‐EGF), a member of the EGFR ligands, was predominantly expressed in breast cancer and that treatment with crossreacting material 197 (CRM197), a specific inhibitor of HB‐EGF, blocked ERK as well as AKT activation via complexes of EGFR and unknown growth factor receptors in TNBC or through complexes of EGFR and human epidermal growth factor receptor‐2 in trastuzumab‐resistant breast cancer, caused significant cell apoptosis and inhibited tumor growth. Accordingly, we can provide a novel concept that a certain EGFR ligand is recognized as a rational target against breast cancer. In addition, it is plausible that CRM197 could be an effective anticancer agent for molecularly targeted therapies.
Immortalization of oral keratinocytes by functional inactivation of the p53 and pRb pathways
Authors: Smeets Serge J. , van der Plas Marlon , Schaaij‐Visser Tieneke B.M. , van Veen Eva A.M. , van Meerloo Johan , Braakhuis Boudewijn J.M. , Steenbergen Renske D.M. , Brakenhoff Ruud H.
Published: May 2010

A subgroup of head and neck squamous cell carcinomas (HNSCCs) contains high‐risk human papillomavirus‐type 16 (HPV16). The viral E6 and E7 oncoproteins inactivate the p53 and pRb proteins, respectively. We examined the causative effect of HPV16 E6 and E7 expression on the immortalization of normal oral keratinocytes (OKCs) and compared the resulting phenotype with alternative ways of p53‐ and pRb‐pathway abrogation frequently found in HNSCCs without HPV. Primary OKCs were conditionally immortalized with temperature‐sensitive SV40 large T‐antigen and human telomerase, allowing these cells to return to their senescent primary state after temperature shift. HPV16 E6 and E7 were introduced to overcome senescence, determined with population doubling (PD) as read‐out. For comparison, we downregulated p53 and p16 by short hairpin RNA genes and expressed mutant p53R(175)H and cyclinD1. Expression of HPV16 E6 caused an extended life span similar to expression of mutant p53R(175)H or p53 knockdown. Expression of mutant p53R(175)H seemed to cause additional activation of the hypoxia and WNT signaling pathways. HPV16 E7 expression had no direct effect on lifespan, similar to p16 knockdown or cyclinD1 expression. In combination with HPV16 E6 or other functional inactivations of p53, abrogation of the pRb‐pathway by either HPV16 E7 or other manipulations caused an immortal phenotype. Our data show the causative role of HPV16 E6/E7 in early squamous carcinogenesis. Activity of each gene could be mimicked by other genetic events frequently found in HNSCC without HPV. This data provides the experimental proof of causal association of HPV in HNSCC carcinogenesis and further support the crucial role of the p53‐ and pRb‐pathways.
Does a helicase activity help mismatch repair in eukaryotes?
Authors: Song Limin , Yuan Fenghua , Zhang Yanbin
Published: May 2010

Mismatch repair (MMR) corrects innate DNA replication infidelities. Many components of eukaryotic MMR have been identified, the molecular mechanism of MMR has been largely demonstrated, and furthermore the nick‐directed MMR reactions have been reconstituted with purified human proteins in vitro. However, some fundamental questions still remain to be answered. One such question is whether a DNA helicase activity is required for MMR in eukaryotes. This short review presents an overview of the interactions between eukaryotic DNA helicases and MMR factors, and suggests a possible mechanism for how DNA helicases may be involved in repair of DNA mismatches. © 2010 IUBMB IUBMB Life 62(7): 548–553, 2010
Validation of the intact rat weanling uterotrophic assay with notes on the formulation and analysis of the positive control chemical in vehicle
Authors: Tyl Rochelle W. , Marr Melissa C. , Brown Sherri S. , Dolbow Emily A. , Myers Christina B.
Published: May 2010

The intact female weanling version in the Organization for Economic Cooperation and Development (OECD) uterotrophic assay Test Guideline (TG) 440 is proposed as an alternative to the adult ovariectomized female version, because it does not involve surgical intervention (vs the ovariectomized version) and detects direct/indirect‐acting estrogenic/anti‐estrogenic substances (vs the ovariectomized version which detects only direct‐acting estrogenic/anti‐estrogenic substances binding to the estrogen receptor). This validation study followed OECD TG 440, with six female weanling rats (postnatal day 21) per dose group and six treatment groups. Females were weighed and dosed once daily by oral gavage for three consecutive days, with one of six doses of 17α‐ethinyl estradiol in corn oil at 5 ml kg−1at 0 and 0.1–10 µg kg−1per day. On postnatal day 24, the juvenile females were euthanized by CO2asphyxiation, weighed, livers weighed and uteri weighed wet and blotted. The presence or absence of vaginal patency was recorded. Absolute and relative (to terminal body weight) uterine wet and blotted weights and uterine luminal fluid weights were significantly increased at 3.0 and 10.0 (bothP< 0.01) µg kg−1per day, and increased to ∼140% of control values at 1.0 µg kg−1per day (not statistically significantly).In vivobody weights, weight changes, feed consumption, liver weights and terminal body weights were unaffected. Vaginal patency was not acquired in any female at any dose, although vaginal puckering was observed in one female at 10.0 µg kg−1per day. Therefore, this intact weanling uterotrophic assay is validated in our laboratory for use under US and European endocrine toxicity testing programs/legislation. Copyright © 2010 John Wiley & Sons, Ltd.
The use of FTIR spectroscopy to assess quantitative changes in the biochemical composition of microalgae
Authors: Wagner Heiko , Liu Zhixin , Langner Uwe , Stehfest Katja , Wilhelm Christian
Published: May 2010

A mid‐infrared spectroscopic method was developed for the simultaneous and quantitative determination of total protein, carbohydrate and lipid contents of microalgal cells. Based on a chemometric approach, measured FTIR (Fourier transform infrared) spectra from algal cells were reconstructed by a partial least square algorithm, using the spectra of the reference substances to determine their relative contribution to the overall cell spectrum. From this specific absorption, absolute macromolecular cell composition [pg cell–1] can be calculated using calibration curves, which have been validated by independent biochemical methods. The future potential of this method for photosynthesis research is shown by its application to follow time‐resolved changes in the cellular composition of microalgae during an illumination period of several hours. We show how the macromolecular composition can be investigated by FTIR spectroscopy methods. This can substantially increase the efficiency of screening processes like bioreactor monitoring and may be beneficial in metabolic engineering of algal cells. (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Chloride intracellular channel 5 modulates adipocyte accumulation in skeletal muscle by inhibiting preadipocyte differentiation
Authors: Li Feng‐Na , Yin Jing‐Dong , Ni Jian‐Jun , Liu Ling , Zhang Hai‐Yan , Du Min
Published: May 2010

Intramuscular fat, the total lipid deposited within skeletal muscle, has been regarded as a potential factor responsible for meat quality in animal production and insulin resistance in humans. The objective of present study was to identify candidate genes which control intramuscular fat accumulation through using animal models. PIC pigs (lean‐type) and Rongchang pigs (obese‐type) were used. By scanning the mRNA samples of longissimus dorsi muscle with Affymetrix Gene‐Chip microarray technology, sus scrofa chloride intracellular channel 5 (CLIC5) was isolated, and its mRNA abundance and protein expression level were reversely related with the intramuscular fat content of pigs. Furthermore, over‐expression of CLIC5 dramatically increased the proliferation of 3T3‐L1 preadipocytes, while inhibited adipocytic differentiation accompanied by the down‐regulation of c/EBPα, LPL, and PPARγ protein. Our results suggest that CLIC5 might be a crucial regulator of adipose accumulation in skeletal muscle of pigs. J. Cell. Biochem. 110: 1013–1021, 2010. © 2010 Wiley‐Liss, Inc.
Regulation of AP‐1 Activity in Glucose‐Stimulated Insulinoma Cells
Authors: Müller Isabelle , Endo Takeshi , Thiel Gerald
Published: May 2010

The transcriptional activity of AP‐1 has been analyzed in glucose‐stimulated INS‐1 insulinoma cells using a chromosomally embedded AP‐1‐responsive reporter gene. We show that AP‐1 activity was significantly elevated in glucose‐treated INS‐1 cells. Preincubation of the cells with nifedipine or expression of the Ca2+ binding protein parvalbumin in the cytoplasm of INS‐1 cells reduced AP‐1 activity. Thus, activation of L‐type Ca2+ channels and an elevated cytoplasmic Ca2+ concentration are crucial to connecting glucose stimulation with enhanced AP‐1 activity. Expression of dominant negative forms of A‐Raf, MKK4 or MKK6 and pharmacological inhibition of MEK and p38 revealed that extracellular signal‐regulated protein kinase, p38 and c‐Jun NH2‐terminal protein kinase participate in the upregulation of AP‐1 activity. Expression of dominant‐negative mutants of c‐Jun and Elk‐1 reduced AP‐1 transcriptional activity in INS‐1 cells indicating that c‐Jun and ternary complex factors are involved in the regulation of AP‐1 activity in glucose‐stimulated insulinoma cells. J. Cell. Biochem. 110: 1481–1494, 2010. © 2010 Wiley‐Liss, Inc.
Sodium 5,6‐benzylidene‐L‐ascorbate induces oxidative stress, autophagy, and growth arrest in human colon cancer HT‐29 cells
Authors: Cheung F.W.K. , Che C.T. , Sakagami H. , Kochi M. , Liu W.K.
Published: May 2010

Our previous studies have demonstrated the oxidative stress properties of sodium ascorbate (SAA) and its benzaldehyde derivative (SBA) on cancer cell lines, but the molecular mechanisms mediating their cytotoxicity remain unclear. In this study, we treated human colon cancer HT‐29 cells with SAA and SBA, and found a significant exposure time‐dependent increase of cytotoxicity in both treatments, with a higher cytotoxicity for 24 h with SAA (IC50 = 5 mM) than SBA (IC50 = 10 mM). A short‐term treatment of cells with 10 mM SAA for 2 h revealed a destabilization of the lysosomes and subsequent induction of cell death, whereas 10 mM SBA triggered a remarkable production of reactive oxidative species, phosphorylation of survival kinase AKT, expression of cyclin kinase‐dependent inhibitor p21, and induction of transient growth arrest. The crucial role of p21 mediating this cytotoxicity was confirmed by isogenic derivatives of the human colon carcinoma HCT116 cell lines (p21+/+ and p21−/−), and immunoprecipitation studies with p21 antibody. The SAA cytotoxicity was blocked by co‐incubation with catalase, whereas the SBA cytotoxicity and its subsequent growth arrest were abolished by N‐acetyl‐L‐cysteine (NAC), but was not affected by PI3K phosphorylation inhibitor LY294002, or catalase, suggesting two separated oxidative stress pathways were mediated by these two ascorbates. In addition, neither active caspase 3 nor apoptotic bodies but autophagic vacuoles associated with increased LC3‐II were found in SBA‐treated HT‐29 cells; implicating that SBA induced AKT phosphorylation‐autophagy and p21‐growth arrest in colon cancer HT‐29 cells through an NAC‐inhibitable oxidative stress pathway. J. Cell. Biochem. 111: 412–424, 2010. © 2010 Wiley‐Liss, Inc.
First principles studies on the interaction of O2 with X@Al12 (X = Al−, P+, C, Si) clusters
Authors: Lu QI Liang , Chen Li Li , Wan Jian Guo , Wang Guang Hou
Published: May 2010

The interaction of O2 with the doped icosahedral X@Al12 (X = Al−, P+, C, Si) clusters with 40 valence electrons were investigated using density functional theory methods. A different behavior exhibited between Al13− and X@Al12 (X = P+, C, Si) when they interact with O2. The dissociation of O2 on Al13− is strongly dependent on spin state of oxygen molecule. But X@Al12 (X = P+, C, and Si) is not the case. The transform of spin moment from O2 to Al13− is much faster. Small molecularly binding energy and relatively high energy barrier show that these clusters are all reluctant reacts with the ground state O2. © 2010 Wiley Periodicals, Inc. J Comput Chem 2010
High expression of IMPACT protein promotes resistance to indoleamine 2,3‐dioxygenase‐induced cell death
Authors: Habibi Darya , Jalili Reza B. , Forouzandeh Farshad , Ong Christopher J. , Ghahary Aziz
Published: May 2010

Indoleamine 2,3‐dioxygenase (IDO), a tryptophan degrading enzyme, is a potent immunomodulatory factor. IDO expression in fibroblasts selectively induces apoptosis in immune cells but not in primary skin cells. However, the mechanism(s) of this selective effect of IDO‐induced low tryptophan environment is not elucidated. The aim of present study was to investigate whether the activity of general control non‐derepressible‐2(GCN2) kinase stress‐responsive pathway and its known inhibitor, protein IMPACT homolog, in immune and skin cells are differentially regulated in response to IDO‐induced low tryptophan environment. IDO‐expressing human fibroblasts were co‐cultured with Jurkat cells, human T cells, fibroblasts, or keratinocytes. Activation of GCN2 pathway was significantly higher in immune cells exposed to IDO‐expressing environment relative to that of skin cells. In contrast, IMPACT was highly and constitutively expressed in skin cells while its expression was very low in stimulated T cells and undetectable in Jurkat cells. A significant IDO‐induced suppressive as well as apoptotic effect was demonstrated in IMPACT knocked down fibroblasts co‐cultured with IDO‐expressing fibroblasts. Proliferation of Jurkat cells, stably transduced with IMPACT‐expressing vector, was rescued significantly in tryptophan‐deficient but not IDO‐expressing environment. This may be due to the ability of IMPACT to recover the effects of IDO‐mediated tryptophan depletion (GCN2 dependent) but not the effects of IDO‐generated cytotoxic metabolites. These findings collectively suggest for the first time that high expression of protein IMPACT homolog in non‐immune cells such as skin cells acts as a protective mechanism against IDO‐induced GCN2 activation, therefore, makes them resistant to the amino acid‐deprived environment caused by IDO. J. Cell. Physiol. 225: 196–205, 2010. © 2010 Wiley‐Liss, Inc.
Modulation of vascular endothelial cell senescence by integrin β4
Authors: Sun ChunHui , Liu Xia , Qi Lei , Xu JiPing , Zhao Jing , Zhang Yun , Zhang ShangLi , Miao JunYing
Published: May 2010

Increasing evidence has demonstrated that the senescence of vascular endothelial cells (VECs) has critical roles in the pathogenesis of vascular dysfunction. Finding important factors that regulate VEC senescence will help provide novel therapeutic strategies for vascular disorders. Previously, we found that integrin β4 was involved in VEC senescence. However, the mechanism underlying VEC senescence mediated by integrin β4 remains poorly understand. In this study, we used a mouse in vivo model and showed that the level of integrin β4 in the endothelium of mouse thoracic aorta was increased during natural aging and atherosclerosis. Furthermore, we found that H‐ras, caveolin‐1, and AP‐1 were implicated in the senescent signal pathway mediated by integrin β4 in human umbilical vein ECs (HUVECs). Knockdown of integrin β4 could attenuate HUVEC senescent features, including increased interleukin‐8 (IL‐8) release and decreased endothelial nitric oxide synthase (eNOS) and NO levels and mitochondrial membrane potential in vitro. Our findings provide new clues illustrating the mechanism of VEC senescence. Integrin β4 might be a potential target for therapy in cardiovascular diseases. J. Cell. Physiol. 225: 673–681, 2010. © 2010 Wiley‐Liss, Inc.
Biosorption of Zn(II) by orange waste in batch and packed‐bed systems
Authors: Pérez Marín Ana Belén , Aguilar María Isabel , Ortuño Juan Francisco , Meseguer Víctor Francisco , Sáez José , Lloréns Mercedes
Published: May 2010

BACKGROUND: This research provides new insights into the biosorption of zinc on a waste product from the orange juice industry. Optimal operating conditions maximizing percentage zinc removal were determined in batch and fixed‐bed systems. Biomass was characterized by FTIR spectroscopy and by major cation content in order to better understand the biosorpion mechanism. Zn‐loaded orange waste was proposed to be used as an alternative fuel in cement kilns.

RESULTS: Sorption capacity was strongly affected by biosorbent dose and solution pH, and was not strongly sensitive to particle size under the experimental conditions studied. Equilibrium data were successfully described by a Langmuir model and sorption kinetic data were adequately modelled with the pseudo‐second‐order and Elovich rate equation. The biomass was found to possess high sorption capacity (qmax = 0.664 mmol g−1) and biosorption equilibrium was established in less than 3 h. Experimental breakthrough curves were adequately fitted to the Thomas model and the dose–response model, obtaining sorption capacities in continuous assays higher than those found in batch mode. Characterization of the biomass suggested the possible contribution of carboxyl and hydroxyl groups of biomass in Zn2+ biosorption and it also highlighted the important role of light metal ions in a possible ion‐exchange mechanism.

CONCLUSIONS: Orange waste could be used as an effective and low‐cost alternative biosorbent material for zinc removal from aqueous solution. Copyright © 2010 Society of Chemical Industry
Bilateral spontaneous internal carotid artery dissection with both early and very late recanalization: A case report
Authors: Vicenzini Edoardo , Ricciardi Maria Chiara , Sirimarco Gaia , Di Piero Vittorio , Lenzi Gian Luigi
Published: May 2010

Spontaneous bilateral internal carotid artery dissection has frequently been described in the literature as a cause of stroke. In more than half of the patients with internal carotid artery dissection, recanalization occurs early after the event and is unusual later than 6 months after onset of the dissection. We describe a patient with ischemic stroke due to left internal carotid artery occlusion in the extracranial segment. The patient was treated with anticoagulants and early vessel recanalization did not occur. Ten months later, he developed contralateral internal carotid occlusion in the intracranial tract, which was followed by early complete recanalization. Anticoagulation therapy was continued and, 16 months after the initial event, the left internal carotid artery unexpectedly also reopened. © 2010 Wiley Periodicals, Inc. J Clin Ultrasound, 2011
Convenient synthesis of substituted 2‐(2‐iminocoumarin‐3‐yl)thieno[2,3‐d]pyrimidin‐4‐ones
Authors: Shynkarenko Pavlo E. , Vlasov Sergiy V. , Kovalenko Sergiy M. , Shishkina Svitlana V. , Shishkin Oleg V. , Chernykh Valentin P.
Published: May 2010

magnified image

The interaction of both 2‐iminocoumarin‐3‐carbonitriles and 2‐iminocoumarin‐3‐carbothioamides with 2‐aminothiophen‐3‐carboxamides lead to formation of 3‐(4‐oxo‐3,4‐dihydrothieno[2,3‐d]pyrimidin‐2‐yl)‐2‐iminocoumarins in two steps. The simplicity of the procedure, as well as the high yields of the target products make this method to be a good alternative of Knoevenagel condensation for the synthesis of 3‐(4‐oxo‐3,4‐dihydrothieno[2,3‐d]pyrimidin‐2‐yl)‐2‐iminocoumarins. J. Heterocyclic Chem., (2010).
Comparative morphology of the foot structure of four genera of Loxosomatidae (Entoprocta): Implications for foot functions and taxonomy
Authors: Iseto Tohru , Hirose Euichi
Published: May 2010

Entoprocta is a group of mostly cryptic, benthic invertebrates with a sedentary lifestyle. Here, we investigate the morphology of the entoproct foot, which is an important structure in attachment and locomotion. We describe the foot structure of four solitary entoprocts, Loxosoma monilis, Loxosomella stomatophora, Loxocorone allax, and Loxomitra mizugamaensis, by means of light and transmission electron microscopy. Gland cells containing secretory granules were found in the foot of all the four species. In L. monilis, the gland cells densely paved the underside of the disc‐shaped foot, but no duct or groove was found. In L. stomatophora and L. allax, a foot gland was present at the frontal end of a foot groove. The foot gland was a solid cell mass in the former species but a sac‐like structure in the latter. Two types of groove accessory cells were recognized in both species; groove bulge cells (GBCs) showed large cytoplasmic bulges extending into the groove lumen, while groove microvillus cells have microvillus mats in the lateral wall of the groove. The bulges of GBCs in L. stomatophora are slender and attached to one another with desmosomes, forming appendages that extend down to the substratum, hinting at their contribution to attachment and locomotion. The bulges in L. allax form large swellings that fill the groove lumen and are connected to the surrounding cells with hemidesmosomes. In the liberated buds of L. mizugamaensis, tripartite gland cell masses were found at the basal end of the stalk, but no groove was found. A small invagination, which may be the opening of the gland, was found at the center of the foot tip, where the liberated buds attach themselves to the substratum and then metamorphose into adults. No openings were found at the lateral terminal wings, which support locomotion in Loxomitra species. J. Morphol. 271:1185–1196, 2010. © 2010 Wiley‐Liss, Inc.
Different responses of E:Z‐isomers of pesticides in liquid chromatography–electrospray ionization–mass spectrometry
Authors: Mayer‐Helm Bernhard , Hofbauer Ludwig , Pani Jutta
Published: May 2010

The mass spectrometric behavior of four pairs of stereoisomers was investigated by liquid chromatography–electrospray ionization–mass spectrometry (LC–ESI–MS). The E‐ and Z‐isomers of the pesticides chlorfenvinphos, dimethomorph, mevinphos and phosphamidon—each with one double bond—were chosen for this study. The MS response of the individual isomers was investigated by infusing the isomers individually into the MS or after the separation of isomer mixtures via high‐performance liquid chromatography(HPLC). In the case of dimethomorph, the same MS response was found for the two isomers. In contrast to that, the individual isomers of chlorfenvinphos, mevinphos and phosphamidon showed different MS response both in the single ion monitoring (SIM) mode in single quadrupole MS and multiple reaction monitoring (MRM) mode in tandem MS. The MS response of the isomers partly depends on (1) the declustering potential of the precursor ion in the SIM mode, (2) the selected transition and (3) the collision energy in the MRM mode. Consequently, quantification by summation of the peak areas of the isomers is inaccurate due to over‐ or underestimating of one of the stereoisomers. Accurate quantitative results can only be achieved when the compound‐specific MS parameters are separately determined for each isomer. This can be done by using pure isomers or by the determination of the MS parameters after HPLC separation and the measurement of the actual isomer ratio with an independent technique. Copyright © 2010 John Wiley & Sons, Ltd.
BDNF‐Akt‐Bcl2 antiapoptotic signaling pathway is compromised in the brain of autistic subjects
Authors: Sheikh Ashfaq M. , Malik Mazhar , Wen Guang , Chauhan Abha , Chauhan Ved , Gong Cheng‐Xin , Liu Fei , Brown William T. , Li Xiaohong
Published: May 2010

Although the pathogenesis of autism is not understood, emerging evidence points to apoptotic mechanisms being involved in this disorder. However, it is not known whether apoptosis signaling is deregulated in the brain of autistic subjects. This study investigates how the apoptosis‐related proteins are regulated in the autistic brain. Our studies show that Bcl2 is significantly decreased, whereas the expression of p53 is increased, in the brain of autistic subjects in comparison with age‐matched controls. We also found that the expression and phosphorylation/activation of Akt kinase that regulates Bcl2 are significantly decreased in the autistic brain. The down‐regulation of Akt may result from a decreased concentration of brain‐derived neurotrophic factor (BDNF), the growth factor that modulates Akt activities. These results suggest that down‐regulation of the BDNF‐Akt‐Bcl2 antiapoptotic signaling pathway in the autistic brain could be one of the underlying mechanisms responsible for the pathogenesis of autism. © 2010 Wiley‐Liss, Inc.
Short‐term low‐frequency electrical stimulation enhanced remyelination of injured peripheral nerves by inducing the promyelination effect of brain‐derived neurotrophic factor on Schwann cell polarization
Authors: Wan Lidan , Zhang Song , Xia Rong , Ding Wenlong
Published: May 2010

Electrical stimulation (ES) has been found to aid repair of nerve injuries and have been shown to increase and direct neurite outgrowth during stimulation. However, the effect of ES on peripheral remyelination after nerve damage has been investigated less well, and the mechanism underlying its action remains unclear. In the present study, the crush‐injured sciatic nerves in rats were subjected to 1 hr of continuous ES (20 Hz, 100 μsec, 3 V). Electron microscopy and nerve morphometry were performed to investigate the extent of regenerated nerve myelination. The expression profiles of P0, Par‐3, and brain‐derived neurotrophic factor (BDNF) in the injuried sciatic nerves and in the dorsal root ganglion neuron/Schwann cell cocultures were examined by Western blotting. Par‐3 localization in the sciatic nerves was determined by immunohistochemistry to demonstrate Schwann cell polarization during myelination. We reported that 20‐Hz ES increased the number of myelinated fibers and the thickness myelin sheath at 4 and 8 weeks postinjury. P0 level in the ES‐treated groups, both in vitro and in vivo, was enhanced compared with the controls. The earlier peak of Par‐3 in the ES‐treated groups indicated an earlier initiation of Schwann cell myelination. Additionally, ES significantly elevated BDNF expression in nerve tissues and in cocultures. ES on the site of nerve injury potentiates axonal regrowth and myelin maturation during peripheral nerve regeneration. Furthermore, the therapeutic actions of ES on myelination are mediated via enhanced BDNF signals, which drive the promyelination effect on Schwann cells at the onset of myelination. © 2010 Wiley‐Liss, Inc.
Nuclear factor‐κB/p65 responds to changes in the Notch signaling pathway in murine BV‐2 cells and in amoeboid microglia in postnatal rats treated with the γ‐secretase complex blocker DAPT
Authors: Cao Q. , Li P. , Lu J. , Dheen S.T. , Kaur C. , Ling E.‐A.
Published: May 2010

Microglial cells constitutively express Notch‐1 and nuclear factor‐κB/p65 (NF‐κB/p65), and both pathways modulate production of inflammatory mediators. This study sought to determine whether a functional relationship exists between them and, if so, to investigate whether they synergistically regulate common microglial cell functions. By immunofluorescence labeling, real‐time polymerase chain reaction (RT‐PCR), flow cytometry, and Western blot, BV‐2 cells exhibited Notch‐1 and NF‐κB/p65 expression, which was significantly up‐regulated in cells challenged with lipopolysaccharide (LPS). This was coupled with an increase in expression of Hes‐1, tumor necrosis factor‐α (TNF‐α), and interleukin‐1β (IL‐1β). In BV‐2 cells pretreated with N‐[N‐(3,5‐difluorophenacetyl)‐1‐alany1]‐S‐phenyglycine t‐butyl ester (DAPT), a γ‐secretase inhibitor, followed by LPS stimulation, Notch‐1 expression level was enhanced but that of all other markers was suppressed. Additionally, Hes‐1 expression and NF‐κB nuclear translocation decreased as shown by flow cytometry. Notch‐1's modulation of NF‐κB/p65 was also evidenced in amoeboid microglial cells (AMC) in vivo. In 5‐day‐old rats given intraperitoneal injections of LPS, Notch‐1, NF‐κB/p65, TNF‐α, and IL‐1β immunofluorescence in AMC was markedly enhanced. However, in rats given an intraperitoneal injection of DAPT prior to LPS, Notch‐1 labeling was augmented, but that of TNF‐α and IL‐1β was reduced. The results suggest that blocking of Notch‐1 activation with DAPT would reduce the level of its downstream end product Hes‐1 along with suppression of NF‐κB/p65 translocation, resulting in suppressed production of proinflammatory cytokines. It is concluded that Notch‐1 signaling can trans‐activate NF‐κB/p65 by amplifying NF‐κB/p65‐dependent proinflammatory functions in activated microglia. © 2010 Wiley‐Liss, Inc.
Inhibiting nerve growth factor or its receptors downregulates calcitonin gene‐related peptide expression in rat lumbar dorsal root ganglia innervating injured intervertebral discs
Authors: Orita Sumihisa , Ohtori Seiji , Nagata Maiko , Horii Manato , Yamashita Masaomi , Yamauchi Kazuyo , Inoue Gen , Suzuki Munetaka , Eguchi Yawara , Kamoda Hiroto , Arai Gen , Ishikawa Tetsuhiro , Miyagi Masayuki , Ochiai Nobuyasu , Kishida Shunji , Takaso Masashi , Aoki Yasuchika , Takahashi Kazuhisa
Published: May 2010

Nerve growth factor (NGF) and its dual structurally unrelated receptors, tropomyosin‐related kinase A (TrkA) or p75 neurotrophin receptor (p75NTR), cause the pathogenesis of discogenic pain. To investigate the sensory innervation of injured rat lumbar intervertebral disc (IVD), we examined the expression of neuropeptides such as calcitonin gene‐related peptide (CGRP) at dorsal root ganglia (DRG) by inhibiting NGF or its dual receptors. Sprague–Dawley rats with multiply punctured L5–L6 IVD were used. Six experimental groups were prepared: naïve, sham control, and four agent‐treated groups with punctured IVD (vehicle, anti‐NGF antibody, anti‐TrkA antibody, and anti‐p75NTR antibody). Retrograde neurotracer Fluoro‐Gold (FG) was applied together except for the naïve group. Their lumbar DRG were harvested and immunolabeled for CGRP. FG‐labeled DRG neurons were most prevalent at L1 and L2 DRG, and the proportion of FG‐labeled CGRP‐immunoreactive DRG neurons in the vehicle group was significantly elevated (p < 0.05) compared with the sham group, while those of antibody‐treated groups, especially in the anti‐p75NTR group, significantly decreased compared with the vehicle group (p < 0.05). Direct intradiscal application of antibody to NGF or its receptors suppressed CGRP expression, and p75NTR antagonism induced the most profound suppression. © 2010 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:1614–1620, 2010
Revised age estimate of the Mjáuvøtn tephra A on the Faroe Islands based on Bayesian modelling of 14C dates from two lake sequences
Authors: Olsen Jesper , Gudmundsdóttir Esther Ruth , Björck Svante , Odgaard Bent V. , Heinemeier Jan
Published: May 2010

Tephra horizons are potentially perfect time markers for dating and cross‐correlation among diverse Holocene palaeoenvironmental records such as ice cores and marine and terrestrial sequences, but we need to trust their age. Here we present a new age estimate of the Holocene Mjáuvøtn tephra A using accelerator mass spectrometry 14C dates from two lakes on the Faroe Islands. With Bayesian age modelling it is dated to 6668–6533 cal. a BP (68.2% confidence interval) – significantly older and better constrained than the previous age. Copyright © 2010 John Wiley & Sons, Ltd.
Pigments used in Roman wall paintings in the Vesuvian area
Authors: Aliatis Irene , Bersani Danilo , Campani Elisa , Casoli Antonella , Lottici Pier Paolo , Mantovan Silvia , Marino Iari‐Gabriel
Published: May 2010

Powdered pigments found in bowls from the Pompeii archaeological site and some wall‐painting fragments from the Vesuvian area (conserved in the National Archaeological Museum of Naples) were investigated by microscopic Raman and FTIR spectroscopies, X‐ray diffraction and scanning electron microscopy‐energy dispersive X‐ray. Brown, red and yellow pigments are common ochres based on goethite and haematite. The blue pigment is Egyptian blue: the presence of tridymite and cristobalite indicates firing temperatures in the 1000–1100 °C range. Pink pigments were prepared both with purely inorganic materials, by mixing haematite and Egyptian blue (violet hue), or presumably by adding an organic dye to an aluminium‐silica matrix. A white powder found in a bowl is composed mainly of the unusual pigment huntite (CaMg3(CO3)4). Celadonite is found in the green samples from the wall paintings, together with Egyptian blue and basic lead carbonate, while the heterogeneous green pigment in a bowl shows malachite mixed with goethite, Egyptian blue, haematite, carbon, cerussite and quartz. Copyright © 2010 John Wiley & Sons, Ltd.
Effect of obesity on laparoscopy‐assisted distal gastrectomy compared with open distal gastrectomy for gastric cancer
Authors: Makino Hirochika , Kunisaki Chikara , Izumisawa Yusuke , Tokuhisa Motohiko , Oshima Takashi , Nagano Yasuhiko , Fujii Shoichi , Kimura Jun , Takagawa Ryo , Kosaka Takashi , Ono Hidetaka A. , Akiyama Hirotoshi , Endo Itaru
Published: May 2010

Background and Objectives

This study compared surgical outcomes between patients undergoing laparoscopy‐assisted distal gastrectomy (LADG) and those undergoing open distal gastrectomy (ODG) from the viewpoint of obesity.

Methods

Between June 2002 and May 2008, 146 patients with preoperatively diagnosed early gastric cancer who underwent LADG (n = 90) or ODG (n = 56) were enrolled in this study and compared in terms of clinicopathological findings and operative outcome. The visceral fat area (VFA) and subcutaneous fat area (SFA) were assessed as identifiers of obesity using FatScan software. The relationship between obesity and operative outcomes after LADG and ODG was evaluated.

Results

There were no significant correlations between intraoperative blood loss (IBL) and any obesity‐related factors, or between operation time (OT) and any obesity‐related factors in the LADG group. There was a significant correlation between IBL and BMI (r = 0.486, P = 0.0001), IBL and VFA (r = 0.456, P = 0.0003), IBL and SFA (r = 0.311, P = 0.0193), OT and BMI (r = 0.406, P = 0.0017), OT and VFA (r = 0.314, P = 0.0178), and between OT and SFA (r = 0.382, P = 0.0034) in the ODG group.

Conclusions

LADG may be a useful operative manipulation that is not influenced by obesity, whereas ODG may be influenced by obesity even after reaching the surgical plateau. J. Surg. Oncol. 2010;102:141–147. © 2010 Wiley‐Liss, Inc.
Indications and approach to surgical resection of lung metastases
Authors: Kaifi Jussuf T. , Gusani Niraj J. , Deshaies Isabelle , Kimchi Eric T. , Reed Michael F. , Mahraj Rickhesvar P. , Staveley‐O'Carroll Kevin F.
Published: May 2010

Pulmonary metastasectomy is a curative option for selected patients with cancer spread to the lungs. Complete surgical removal of pulmonary metastases can improve survival and is recommended under certain criteria. Specific issues that require consideration in a multidisciplinary setting when planning pulmonary metastasectomy include: adherence to established indications for resection, the surgical strategy including the use of minimally invasive techniques, pulmonary parenchyma preservation, and the role of lymphadenectomy. J. Surg. Oncol. 2010;102:187–195. © 2010 Wiley‐Liss, Inc.
Does serum carcinoembryonic antigen elevation in patients with postoperative stage II colorectal cancer indicate recurrence? Comparison with stage III
Authors: Hara Masayasu , Sato Mikinori , Takahashi Hiroki , Takayama Satoru , Takeyama Hiromitsu
Published: May 2010

Background and Objectives

The aim of this study was to determine the accuracy of postoperative monitoring of serum carcinoembryonic antigen (CEA) to detect or rule out recurrence in patients with stage II colorectal cancer (CRC) by comparing results with stage III.

Methods

A total of 303 patients with CRC who underwent curative surgery were enrolled. Serum CEA was assayed, and radiological examination was performed routinely for 5 years after surgery. Yearly recurrence rates, sensitivities, specificities, likelihood ratios, and posttest probabilities were calculated.

Results

Sensitivity and specificity of CEA monitoring in stage II patients are almost same as those in stage III. Whereas recurrences occurred early in stage III, they occurred almost as frequently in both early and late stage II. The obtained posttest probability of recurrence in stage II patients with CEA elevation was significantly lower (only 30% or less) than those in stage III (approximately 80%).

Conclusion

Elevation of CEA in patients with stage II CRC does not represent recurrence with high probability. One of the reasons for the unreliability of CEA monitoring was its high false‐positive rate. Another tumor marker with a lower false‐positive rate is necessary to follow‐up stage II CRC patients. J. Surg. Oncol. 2010;102:154–157. © 2010 Wiley‐Liss, Inc.
Outcome of selected breast cancer patients with micrometastasis or isolated tumor cells in sentinel node biopsy and no completion axillary lymph node dissection
Authors: Meretoja Tuomo J. , Vironen Jaana H. , Heikkilä Päivi S. , Leidenius Marjut H.
Published: May 2010

Background and Objectives

Axillary lymph node dissection (ALND) is the standard of care in patients with tumor‐positive sentinel nodes (SN). However, approximately half of these patients do not have additional metastases in their axilla and therefore do not benefit from completion ALND. Our aim was to examine the outcome of highly selected breast cancer patients with tumor‐positive SN without completion ALND.

Methods

Altogether 48 patients with tumor‐positive SN without ALND were included in this study. Twenty‐two patients had micrometastasis and 26 had isolated tumor cells (ITC) in their sentinel node biopsy. The median follow‐up time was 37 months (range 9–78).

Results

No axillary recurrences occurred during the follow‐up. One patient had a local recurrence. Distant metastases as the first event were observed in two patients. One of them died in breast cancer. Nine patients died from intercurrent causes.

Conclusions

Omitting ALND seems safe in selected breast cancer patients with SN micrometastasis or ITC. J. Surg. Oncol. 2010;102:215–219. © 2010 Wiley‐Liss, Inc.
Evaluation of selective composite cryogel for bromate removal from drinking water
Authors: Hajizadeh Solmaz , Kirsebom Harald , Galaev Igor Y. , Mattiasson Bo
Published: May 2010

Bromate, which is a potential carcinogen, should be removed from drinking water to levels of less than 10 μg/L. A chitosan‐based molecularly imprinted polymer (MIP) and a sol–gel ion‐exchange double hydrous oxide (Fe2O3·Al2O3·xH2O) adsorbent (inorganic adsorbent) were prepared for this purpose. The sorption behavior of each adsorbent including sorption kinetics, isotherms, effect of pH and selective sorption were investigated in detail. Sorption experimental results showed that the MIP adsorbents had better selectivity for bromate, even in the presence of high concentrations of nitrate, as compared to the inorganic adsorbent. It was found that pH does not affect the adsorption of bromate when using the inorganic adsorbent. Additionally, both adsorbents were immobilized in a polymeric cryogel inside plastic carriers to make them more practical for using in larger scale. Regeneration of the cryogels either containing MIP or inorganic adsorbents were carried out by 0.1 M NaOH and 0.1 M NaCl, respectively. It was found that the regenerated MIP and inorganic adsorbents could be used at least three and five times, respectively, without any loss in their sorption capacity.
Extraction of lutetium(III) from aqueous solutions by employing a single fibre‐supported liquid membrane
Authors: Trtić‐Petrović Tatjana M. , Kumrić Ksenija R. , Đorđević Jelena S. , Vladisavljević Goran T.
Published: May 2010

Transport behaviour of Lu(III) across a polypropylene hollow fibre‐supported liquid membrane containing di(2‐ethylhexyl)phosphoric acid (DEHPA) in dihexyl ether as a carrier has been studied. The donor phase was LuCl3 in the buffer solution consisting of 0.2 M sodium acetate at pH 2.5–5.0. A miniaturised system with a single hollow fibre has been operated in a batch mode. The concentration of Lu(III) was determined by indirect voltammetric method using Zn–EDTA complex. The effect of pH and volume of the donor phase, DEHPA concentration in the organic (liquid membrane) phase, the time of extraction and the content of the acceptor phase on the Lu(III) extraction and stripping behaviour was investigated. The results were discussed in terms of the pertraction and removal efficiency, the memory effect and the mean flux of Lu(III). The optimal conditions for the removal of 177Lu(III) from labelled 177Lu‐radiopharmaceuticals were discussed and identified. The removal efficiency of Lu(III) greater than 99% was achieved at pH of the donor phase between 3.5 and 5.0 using DEHPA concentration in the organic phase of 0.47 M and the ratio of the donor to the acceptor phase of 182.
Estimation of global structural and transport properties of peptides through the modeling of their CZE mobility data
Authors: Piaggio Maria V. , Peirotti Marta B. , Deiber Julio A.
Published: May 2010

Peptide electrophoretic mobility data are interpreted through a physicochemical CZE model, providing estimates of the equivalent hydrodynamic radius, hydration, effective and total charge numbers, actual ionizing pK, pH‐near molecule and electrical permittivity of peptide domain, among other basic properties. In this study, they are used to estimate some peptide global structural properties proposed, providing thus a distinction among different peptides. Therefore, the solvent drag on the peptide is obtained through a characteristic friction power coefficient of the number of amino acid residues, defined from the global chain conformation in solution. As modeling of the effective electrophoretic mobility of peptides is carried out in terms of particle hydrodynamic size and shape coupled to hydration and effective charge, a packing dimension related to chain conformation within the peptide domain may be defined. In addition, the effective and total charge number fractions of peptides provide some clues on the interpretation of chain conformations within the framework of scaling laws. Furthermore, the model estimates transport properties, such as sedimentation, friction and diffusion coefficients. As the relative numbers of ionizing, polar and non‐polar amino acid residues vary in peptides, their global structural properties defined here change appreciably. Needs for further research are also discussed.